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Biotage Flash Chromatography

1. Rf CV 0 90 1 10 0 85 1 17 0 80 1 25 0 75 1 33 CV vs Rf Isocratic 0 70 1 40 0 65 1 54 0 60 1 65 10 0 55 1 81 i 0 50 2 00 0 45 2 22 2 0 40 2 50 0 35 2 86 4 0 30 3 33 0 25 4 00 0 20 5 00 S 0 15 6 67 i Qal 92 03 04 06 07 08 09 1 0 10 10 00 Figure 1 Rf to CV correlation isocratic elution 82 When scouting TLC solvent systems it is important to realize a low Rf 0 15 0 35 is preferred because a lower Rf means a greater CV Large CV s indicate increased compound silica contact time improving the chances of component resolution Since CV is a measure of compound retention then ACV is the measure of compound resolution see figure 2 In FLASH purification ACV dictates the sample load range possible for any given cartridge size see table 1 For two adjacent components a large ACV is desirable B i A B A R 0 50 0 40 CV 2 0 2 5 B II A cv 3 0 5 0 B III A CV 5 0 10 Figure 2 The Rf CV relationship is illustrated in this graphic I Although compounds A and B are well resolved on TLC with Rf of 0 5 and 0 4 respectively FLASH purification with the same solvent conditions provides low retention and low resolution ACV 0 5 for A and B respectively II Lowering the Rf A 0 33 B 0 20 provides increased retention and resolution ACV 2 III Extremely high resolution ACV 5 is obtained by further reducing
2. 2 60 2 40 2 20 2 00 1 80 1 1 60 1 40 1 20 1 00 0 80 0 60 0 40 0 20 0 00 L Absorbance c BS ad ERES OUO NAN 21 26 31 Fraction Number 36 41 46 51 56 1 0 0 9 0 8 0 7 0 6 0 5 0 4 0 3 0 2 0 1 0 0 Absorbance 8 Compound 1 0 Compound 3 Compound 5 Compound 7 0 0000000 0 Compound 2 Compound 4 Compound 6 0 0000000 0000 0 000000000000000008 0 i 0 D 9 n e 11 13 15 17 19 21 23 25 27 2931 33 35 37 39 4143 45 47 C Fraction Number Figure 3 The FLASH 40 HPFC cartridge design is more efficient greater resolution R more retentive and delivers higher purity fractions than other cartridge designs Figure 4 FLASH 12 M chromatogram showing purification of 90 mg of crude macrolide fermentation product Four compounds are distinctly visible Highlighted boxes correspond to fractions tested by TLC Figure 5 TLC analysis of collected fractions Fractions 11 15 are pure compound 1 18 21 pure compound 2 24 29 pure compound 3 and 34 47 pure compound 4 Sent to waste below threshold UV Abs mV Average a o o o 8 17 01 8 17 01 3 45 3 50 PM PM 8 17 01 4 00 PM Silica Cartridge Specifications surface area 500 m g porosity 60 particle size 40 63 um C18 Cartridge Specifications surface area 450 m g poros
3. pil m 1 i Cartridge Biotage FLASH 25 M i 25 x 150 mm ker Load 450 mg BE Detection 254 nm au cip Solvent A DCM me 1 i Solvent B MeOH rit Gradient 0 10 B in 500 mL nin Flow rate 32 mL min Figure 2 Gradient FLASH separation quickly isolates desired product from the reaction mixture after each Fmoc de blocking step The Horizon system was programmed with the following conditions Results This multistep solution synthesis yielded 765 mg 83 3 of highly pure greater than 90 Fmoc Tyr t Bu Gly Gly Phe Leu NDMCP in less than eight hours Synthesis of the same compound by traditional solution phase techniques using aqueous extractions was complicated because of the formation of emulsion during the aqueous wash required at each step and the final yield was less than 40 This synthesis required two to three days to complete this multistep synthesis Conclusion Substituting aqueous work up with FLASH chromatography increases the yield and purity of the final product Because no aqueous washes are required there is much less sample handling and no product loss due to emulsions drying with MgSO4 and filtration Also using FLASH chromatography ensures up to 95 purity of isolated intermediate product at each synthetic step with very little product lost The ability to combine Flash purification and synthesis into a single process increased the yield of the overall reaction by 100 and pr
4. Phenol Alkyl Bromide o Br jo o ot Q OH o ioc So B oa o ue Bo IE cane o or S iba oa o e E QUE SONO 72 Table 2 Twelve component a phenoxyacetophenone array prepared using Syntage KP TBD Samplet cartridge AU 0 zo CN G AGN No RE 80 C Product Cartridge FLASH 25 M KP Sil Solvent A Hexane B EtOAc Gradient Step 1 0 40 B in 350 mL Program Step 2 40 100 B in 300 mL Flow Rate 25 mL min TELELE iii CO ag peii pi ea Tiia rs Collected 25 mL min Fraction J HPLC Conditions 20 Crude 60 1 Column H an 120 1 Flash 93 69 IE n Mobile Phase A H20 ACN 95 5 100 0 1 TFA B ACN H20 95 5 li 0 1 TFA 50 Z LE il Gradient B 5 100 in 8 200 400 6 00 i 8 00 10 00 12 00 14 00 min hold 2 min Minutes Flow Rate 1 5 mL min Detection 254nm Conclusion Syntage KP TBD Samplet cartridges easily and successfully integrate synthesis and purification Traditional work up steps are eliminated when the Syntage Samplet cartridges are directly inserted into FLASH purification cartridges This allows fewer solution transfers and sample manipulations and results in a higher yield of final product With each collected product the purity was greater than 93 and the individual yield varied between 53 and 84 Since these on column reactions o
5. Syringe Injection for ZIF SIM for Liquid Samples Low Solubility Optional Air Regulator FLASH Module Pivoting Arm il Stopcock Valve Solvent Reservoir Collect Fractions gelbe Figure 1 FLASH flow diagram FLASH Compression Modules Incorporate ZIF and Samplet Technologies Customize Your FLASH System FLASH compression modules use Biotage patented FLASH cartridges packed with a range of normal and reversed phase media see page 88 for more detailed packing media information The FLASH compression modules are designed with patented ZIF Zero Insertion Force head technology that improves purification of synthetic and natural products by axially compressing the FLASH cartridge Axial compression of the cartridge bed minimizes void space which improves separation efficiency and loading capacity For simplified sample handling the FLASH compression modules accept Samplet sample loading cartridges For low solubility samples that cannot be dissolved in an appropriate volume for Samplet loading a ZIF SIM adds additional purification capabilities ZIF head assembly designed to distribute sample and solvent flow evenly through each cartridge 114 The FLASH 12 Compression Module and 12 mm ID Prepacked Cartridges are Designed for Milligram scale FLASH Separations j FLASH 12 M System with ZIF SIM10 ZIF SIM35 and test tube rac
6. AUTOFLASH 75 systems can utilize our patented sample injection modules the SIM 100 and SIM 500 Components and Accessories Autoflash 75 Ordering Information Add on Components and Accessories Item Description Part Number AUTOFLASH 75 Compression Modules AUTOFLASH 75S 75S compression module AF 005 19160 pressure gauge 4 way injection valve tubing start up kit and manual AUTOFLASH 75M 75M compression module AF 005 19040 pressure gauge 4 way injection valve tubing start up kit and manual AUTOFLASH 75L 75L compression module AF 005 19070 pressure gauge 4 way injection valve tubing start up kit and manual AUTOFLASH 75 Accessories AUTOFLASH 75S Powder coated aluminum barrel with 03711 Barrel pressure gauge and mounting bars uses FLASH 75S cartridges AUTOFLASH 75M Powder coated aluminum barrel with 03710 Barrel pressure gauge and mounting bars uses FLASH 75M cartridges AUTOFLASH 75L Powder coated aluminum barrel with 03706 Barrel pressure gauge and mounting bars uses FLASH 75L cartridges AUTOFLASH Stainless steel sample injection module SIM 1007 ideal for loading low solubility samples SIM 5102 or liquid volumes up to 100 mL for use with AUTOFLASH systems AUTOFLASH Stainless steel sample injection module SIM 5502 SIM 5007 ideal for loading low solubility samples or liquid volumes up to 500 mL For use with AUTOFLASH systems AUTOFLASH Contains all required tubing nuts and AFSU 1000
7. One step three component synthesis of Imidazo 1 2 a annulated Pyridine o HN Br CN HCIO Br SN 4 a N a S EtOH K gt paz NO NH l 170 C 5min N Safety Use of Perchloric acid as catalyst un Optimized on Emrys Liberator 70 yield 2 Directly scaled up 30 times e 68 yield with 5 4 g of product e 1HNMR showed 100 purity e Total time from discovery to 5 4 g isolated 3 h Triphenylphosphine free one pot Wittig olefination HN SN PR Br ANN NO EtOH WN CN 170 C 5min Solid phase supported reagent Heterogeneous solution H Developed amp optimized on Emrvs Liberator 38 yield N Scaled up 30 times in Emrys Advancer 92 yield 1HNMR measured purity of 95 40 System Specifications Reaction vessel Operating temperature Pressure range Temperature increase Power range Agitation Cooling System Control amp Data Management Temperature Humidity Power Air supply Water supply used to cool magnetron Weight Dimensions Three extra vessel entry ports 350 mL Teflon 60 250 C 140 482 F 1 20 bar 2 MPa 290 psi 0 5 4 C sec solvent dependent 0 1200 W Overhead Stirrer Gas cooling instant adiabatic flash cooling Windows XP operating system PIV 2 GHz processor 20 GB hard disc and 256 MB primary memory Includes 17 TFT flat screen keyboard mouse and Biotage Advancer 1 4 software Prints via the LAN
8. CATALOG G 206 Biotage Biotage 2005 Product Catalog BLENDING TECHNOLOGY 3 CUSTOMER SUPPORT AND SERVICE 6 SYNTHESIS SOLUTIONS AND OPTIMIZATION 14 INITIATOR MICROWAVE SYNTHESIS SYSTEMS 30 Initiator Initiator 8 Initiator 60 ADVANCER 38 BIOTAGE MICROWAVE VIALS 44 BIOTAGE PATHFINDER 48 APPLICATIONS CORNER 52 FLASH PURIFICATION SOLUTIONS AND OPTIMIZATION 80 FLASH SvNTAGE AND SAMPLET CARTRIDGES 90 DISCOVERY SCALE FLASH SYSTEMS AND MODULES 112 FLASH 1247 FLASH 2547 FLASH 40 47 FLASH 65i ZIF SIM SP FLASH PURIFICATION SYSTEMS 126 SP1 SP4 QUAD PARALLEL PURIFICATION SYSTEMS 134 Quad UV System Quad3 System DEVELOPMENT SCALE FLASH CHROMATOGRAPHY SYSTEMS 142 FLASH 75i FLASH 150i PARALLEX FLEX HPLC SYSTEMS 154 FLASH AC ACTIVATED CARBON CARTRIDGES 164 PRODUCTION SCALE FLASH SYSTEMS 172 KILOPREP PREPARATIVE HPLC SYSTEMS 180 Kiloprep Cartridges Preparative HPLC Scale up Guidelines SPARE PARTS 190 TERMS CONDITIONS AND WARRANTY 202 Working together today pharmaceutical chemists throughout the world rely on Biotage products as part of their daily workflow Data shows that chemists have performed more than 900 000 microwave syntheses and 1 7 million flash purifications using Biotage products As the innovators of microwave synthesis technology and cartridge ba
9. Meter 50 000 40 000 30 000 20 000 10 000 Cartridge Number Reduced Plate Height 2 Figure 3 This chart clearly demonstrates the consistent performance and high efficiencv reported bv plates per meter and reduced plate height Kiloprep Radial Compression Modules Biotage Kiloprep Radial Compression Modules hold our prepacked cartridges and ensure that the cartridges provide the highest performance Dynamic radial compression reduces the potential for void and channel formation increasing cartridge lifetime Compression modules are designed to hold cartridges of one diameter and two optional lengths see chart below Chemists can scale up directly by selecting a larger compression module and increasing the cartridge diameter Kiloprep Compression Cartridge Cartridge Typical Flow Typical Sample Module Diameter mm Lengths cm Rate per min Load g KPCM100 75 15 or 30 125 250 mL 0 1 50 KPCM250 100 30 or 60 250 500 mL 1 125 KPCM500 150 30 or 60 0 5 1 0 L 2 250 KPCM1000 200 30 or 60 1 2L 5 500 KPCM3000 300 30 or 60 2 4 L 10 1 000 Preparative HPLC Scale Up Guidelines When scaling up a current method a chemist can continue to use the original column and make many injections or select a larger column that will complete the project in many fewer runs If you have already optimized a method on an analytical or semi prep column Biotage can help you select a Kiloprep cartridge that will dramatically
10. Dimensions Interfaces Touch screen Ethernet LAN Archiving and back up Upgrade Path Initiator Eight Rack capacity Dimensions Weight Initiator Sixty Rack Capacity Dimensions Weight 60 250 C 140 482 F 2 5 C sec 36 41 F sec 0 20 bar 2 MPa 290 PSI 0 300 W at 2 45 GHz 2 or 4 vial sizes mL 0 5 2 2 5 0 2 0 5 and 10 20 with EXP function 0 5 5 mL 0 2 20 mL with EXP function Magnetic stirrer 18 32 C 64 90 F 20 95 RH EU 220 240 V 50 Hz 5 A US 120 V 60 Hz 10 amp JP 100V 50 60 Hz 10 A 1100 VA Pressurized air supply gt 60 L min 2 1 cubic feet min 2 5 4 bar 0 25 0 4 MPa 36 58 PSI 48 9 Ib 22 2 kg 14 4 x 15 9 x 16 3 WxDxH 365 x 405 x 415 mm 6 4 complies with IEEE 802 3 ANSI 8802 3 MII Via the LAN The Initiator system can be automated with Robot 8 or Robot 60 4 vials in rack for 0 2 5 mL vials 2 vials in rack for 10 20 mL vials 15 7 x 19 7 x 22 8 WxDxH 400 x 500 x 580 mm 64 Ib 29 kg 30 vials in rack for 0 2 0 5 0 5 2 and 2 5 mL vials 12 vials in rack for 10 20 mL vials 24 6 x 15 9 x 18 5 WxDxH 625 x 405 x 470 mm 77 b 34 9 kg Initiator Ordering Information Ordering Information Part Number System Country Systems 355230 Initiator EU 355286 Initiator US 355287 Initiator JPN 355301 Initiator EXP EU 355302 Initiator EXP US 355303 Initiator EXP JPN 355522 Initiator Eight EU 355524 Initiator Eight US 355526
11. Each cartridge is constructed of rugged medium density polyethylene to resist cracking or splitting There s no pressurized glass and all of the silica is self contained eliminating any exposure to contaminated silica dust FLASH 150 Specifications Cartridge Dimensions Sample Size Flow Rate SIM Volume Reservoir Volume mm ID xcm L g mL min mL L FLASH 150M 150 x 30 3 160 500 1000 1000 37 FLASH 150L 150 x 60 6 320 500 1000 2000 60 Cartridge Diameter Bed Length Packing Column Vol Easy Typical Difficult mm cm Wt kg nom L ACV 6 ACV 2 ACV 1 FLASH 150M 150 30 2 5 4 3 80 160 16 80 3 16 FLASH 150L 150 60 5 8 6 160 320 32 160 6 32 Table 3 Biotage FLASH 150 Sample Sizes Based on TLC Evaluation ACV the difference in column volumes at which two compounds elute calculated from the thin layer chromatography Rf values as 1 Rf 1 Rf gt Ordering Information Add on Components and Accessories Item FLASH 1501 Systems FLASH 150M System FLASH 150L System FLASH 150M Compression Module FLASH 150L Compression Module FLASH 150 Accessories FLASH 150M Barrel FLASH 150L Barrel SIM 1000 Sample Injection Module SIM 2000 Sample Injection Module SIM Top Frits SIM Bottom Frits Grounding Kit 3 way Injection Valve 37 L Solvent Reservoir 60 L Solvent Reservoir Air Manifold SIM 1000 Start up Kit SIM 2000 Start up Kit FLASH 150 Start up Kit Descripti
12. FLASH 150i Packages FLASH 150i Packages FLASH 150i packages include a radial compression module a fully integrated air manifold a solvent reservoir a sample injection module start up kit with all necessary tubing a grounding kit and a user s manual Please order your FLASH 150M Medium or FLASH 150L Long prepacked cartridges separately Flash 150 compression modules are mounted onto stable and robust portable bases which are fitted with casters for easy mobility High Flow rate Operation The FLASH 150 cartridges 150 mm ID are available in two lengths 30 cm 2 5 kg FLASH 150M and 60 cm 5 kg FLASH 150L They are built to withstand operating pressures up to 100 psig With flow rates as high as 1 0 L min these systems allow you to complete runs and scale up faster saving hours even days of separation time Cartridges are Available with a Variety of Media e KP Sil 40 63 um 60 silica e KP C18 HS 40 63 um 90 C18 bonded silica e Mitsubishi Diaion HP20 amp HP20SS SDVB resins see page 95 for more information on cartridges FLASH 150M and 150L compression modules and cartridges 149 150 Patented Radial Compression Biotage s patented radial compression technology eliminates voids and channels that often occur in glass columns resulting in a highly dense packed bed Compounds can then be collected in narrower bands for optimum separation efficiency Easier and Safer to Use
13. Frame and tubing Chemraz O ring 1 Retaining ring 1 Instruction sheet For fraction collection 1 8 tubing FEP 3 Tube with Luer Lok adapter Threaded barrel adapter connects head assembly to 40S or 40M barrel Disposable 10 mL polypropylene 1 4 to 1 16 1 4 and 1 8 nuts and ferrules 3 sets of each SIM Receiver Head Assembly 1 4 x 1 8 reducing union 4 required per set SIM 1040 Spare Parts Item O ring Collar Head Assembly Fitting Kit Tubing Kit Cap Kit Tool Adapter SIM Kit FLASH 65i Spare Item O ring E ring Barrel Retainer Collection Tubing Kit Reducing Union Description SIM Top Head O Ring SIM Barrel Locking Collar SIM Receiver Head Assembly SIM Luer Lok Kit SIM Tubing Kit SIM Plastic Cap Kit 1 SIM cap adapter and 1 retaining ring 5 8 Frit insertion tool Male Luer to 1 8 NPT 35 cc barrels and frits 24 Parts Description Chemraz O ring 1 Retainer clip needed with O ring Barrel Adapter Female Luer 1 16 Barb 1 8 FEP Tubing 24 1 8 Male Luertight fitting 1 8 Knurled nut 1 8 ferrule set SS 1 4 x 1 8 reducing union 4 required per set Part Number FC 022 17024 01800 01818 06981 01985 SU 242 0000 03216 01888 01790 03277 02979 NF 15075 06090 04144 Part Number 01963 01969 06090 01984 01903 03251 01965 01959 FTF 0040 Part Number 08395 03024 0152 08403 04144 FLASH 75i System Spare
14. KP Sil TLC Plates Glass KP Sil TLC Plates Glass Scaling Columns Cartridge KP Sil Silica Scaling Column Scaling Column KP C18 HS Scaling Column Scaling Column Diaion HP20 Scaling Column Scaling Column Diaion HP20SS Scaling Column Scaling Column Description 2 75 cm x 7 5 cm glass backed 5 cm x 10 cm glass backed 10 cm x 10 cm glass backed Description 4 6 mm ID x 25 cm 10 mm ID x 25 cm 4 6 mm ID x 25 cm 10 mm ID x 25 cm 4 6 mm ID x 25 cm 10 mm ID x 25 cm 4 6 mm ID x 25 cm 10 mm ID x 25 cm Qty Case 100 50 25 Part Number S1K0 1107 93050 S1K0 1107 95050 S1L0 1118 93050 S1L0 1118 95050 SFT6 2030 93050 SFT6 2030 95050 SFT6 2530 93050 SFT6 2530 95050 Order TLC Plates and Scaling Columns Part Number TLC 2575 FI TLC 0510 FI TLC 1010 FI 99 The Syntage line of chemistry cartridges are preformatted sample loading cartridges based on Samplet cartridge technology that contain a variety of functionalized media used for scavenging work up and synthesis Unlike traditional synthesis and work up methods using Syntage Samplet cartridges with FLASH purification cartridges combines work up synthesis and purification into one process Biotage FLASH and Syntage chemistry cartridges feature medium pressure 100psi capability for faster flow rates Syntage cartridges clean up crude reaction mixtures while inside a FLASH purification ca
15. Ordering information is found following each product listing How to Order Europe 46 18 56 57 10 Fax 46 18 56 57 05 E mail ordermailbox eu biotage com US or Canada 1 800 446 4752 8 30 a m and 5 30 p m EST Fax 1 434 979 4743 E mail ordermailbox biotage com Japan 81 422 28 1233 Fax 81 422 28 1236 E mail order biotage co jp When placing your order please have available e Your purchase order number e Biotage part number s e Product descriptions e Shipping address e Billing address e Contact person including telephone number e Product user name and department Orders can also be placed using vour VISA Mastercard or American Express account IN THE US ONLY MasterCard Prices and Specifications AMERICAN VA REN Prices in the accompanying Biotage Price Book are suggested list prices and are current at the time of the book s printing Specifications not listed in this catalog can be obtained by contacting Biotage Customer Support and Service or your local distributor Contact Biotage 1 Point Support www biotage com The Biotage Web site offers our customers easy access to current information on new products applications and events Europe Service and Support Telephone 46 18 56 59 11 E mail 1 PointSupport eu biotage com United States Service and Support Telephone 1 800 446 4752 press 3 at the auto attendant E mail 1 pointsupport biotage com Japan Service and Sup
16. SAM TSCL25 SAM TSCL40 SAM KMNO412 SAM KMNO425 SAM KMNO440 SAM TBD12 SAM TBD25 SAM TBD40 SAM NCO312 SAM NCO325 SAM NCO340 SAM CF12 SAM CF25 SAM CF40 Ordering Information Syntage Heating Blocks Item Description Part Number Syntage 12 Solid aluminum block with Teflon coating 09644 Heating Block holds 24 Syntage 12 Samplet cartridges and Syntage 12 sleeves Syntage 12 Sleeves Straight side polyethylene shell for 09641 use with Syntage 12 Samplet cartridges and heating blocks pack of 24 Syntage 25 Solid aluminum block with Teflon coating 09645 Heating Block holds 8 Syntage 25 Samplet cartridges and Syntage 25 sleeves Syntage 25 Sleeves Straight side polyethylene shell for use 09642 with Syntage 25 Samplet cartridges and heating blocks pack of 24 Syntage 40 Solid aluminum block with Teflon coating 09646 Heating Block holds 4 Syntage 40 Samplet cartridges and Syntage 40 sleeves Syntage 40 Sleeves Straight side polyethylene shell for use 09643 with Syntage 40 Samplet cartridges and heating blocks pack of 24 Discovery Scale FLASH Chromatography Systems and Modules 112 FLASH High Yield High Performance Milligram to Gram Scale Purifications Biotage FLASH chromatography products incorporate all the features of our HPFC product line The tools you need for efficient effective FLASH purifications can be purchased as complete FLASH systems or as individual comp
17. Segment 2 Segment 1 to TLC conditions over 10 CV Segment 3 Hold segment 2 for 1 CV These conditions work when your compound of interest has an Rf of 0 4 For example the TLC conditions are 8 2 hexane ethyl acetate and the Rf is 0 4 Set the gradient as follows Segment 1 5 ethyl acetate for 1 CV Segment 2 5 to 20 ethyl acetate over 10 CV Segment 3 hold 20 ethyl acetate for 1 CV Using these conditions a compound with an Rf of 0 4 is the last compound to elute To elute more strongly retained compounds increase the gradient s slope by decreasing the volume in segment 2 or increasing the strong solvent concentration in segments 2 and 3 Patent pending Reversed phase Flash As a technique used for purification of water soluble compounds reversed phase flash purification method development uses an approach different than normal phase The recommended approach for reversed phase includes developing and optimizing the method using HPLC and a Biotage KP C18 scaling column 4 6 x 250 mm The scaling column is packed with the identical C18 phase as the KP C18 FLASH cartridges Begin by creating a gradient on the HPLC from 10 90 acetonitrile or methanol in water at 3 mL min 1 CV min with this gradient Segment 1 10 ACN or MeOH for 1 min Segment 2 10 90 ACN MeOH over 10 min Segment 3 Hold 90 ACN for 2 CV Continue to modify this until the compound of interest is fully separated from its impurities and has a re
18. o o co H 23 eX so 2 2 o 8 lt 3 e S T T T T T T T T T T T T T T T T T 0 2 4 6 8 10 12 14 16 18 Time min Figure 8 100 mg mL injection of synthetic compound separated on a 2 cm I D x 25 cm long C18 column Despite poor peak shapes a method selection containing specific fractionation decisions resulted in only four fractions being collected with three of the fractions being the target compound for a total of 20 75 mL 159 160 THE PARALLEX FLEX PROBLEM SOLUTION 000060000008 i444 OQOJOOOOUeo mE LLLE Figure 9 Automated parallel synthesis is enabling production of arrays of single compounds to meet the demands of high throughput screening HTS programs In order to obtain the most reliable data the compounds must be purified characterized and quantified Flex V3 is a pivotal purification system facilitating complete integration into a high throughput chemistry environment providing high quality compounds High throughput Application The Flex V3 is a proven instrument for the purification of large numbers of compounds Parallel flow streams allow the simultaneous purification of combinatorial arrays in a more efficient and cost effective way than using multiple single systems A four channel system can purify up to 40 samples in one hour Purification however is only one component of the high throughput chemistry process The Flex V3 system is designed to provide an integrated solution to
19. reducing union 4 required per set Horizon Spare Parts Item Tube Assemb Tube Assemb Spare Parts Ki Tube Assemb Tube Assemb Tube Assemb Valve Cell Valve it Y Description Compression module outlet to fraction collector arm 1 8 Compression module outlet to UV detector inlet 1 8 Kit spare tubing amp fittings horizon 1 8 Flow control outlet to UV detector outlet 1 8 Flow control valve to 3 way collect valve to waste 1 8 Flow control outlet to FC arm 1 8 UV collect valve 0 3 mm flow cell Stop flow control valve Part Number 08192 06785 09427 08857 08746 09261 09260 09341 08091 05461 05654 06026 04030 04795 09312 09311 04009 04794 08173 01801 04144 Part Number 08079 08085 08442 08761 08762 08763 08535 08632 08770 196 Horizon Spare Parts continued Item Lamp Cell Window Tubing Kit Reducing Union Description UV lamp for Horizon Detector 0 1 mm flow cell Replacement windows for flow cells Upgrade tubing for outlet of compression module to UV collect valve 1 4 x 1 8 reducing union 4 required per set ZIF SIM Spare Parts Item E Clip ZIF SIM Tubing Kit Description Retaining ring Luer fitting with 1 8 tubing 1 8 knurled nut connects ZIF SIM outlet to compression module ZIF SIM 10 Spare Parts Item Frit Insertion Tool Upper O ring Lower O ring Description Polypropylene frit i
20. where interrupted samples can be rerun System status is immediately visualized by intuitive color coding samples darken on completion and fraction vessels become black as they are filled giving an instant view of available collection capacity 155 156 Reliable Unattended Operation A closed loop feedback has been incorporated into the software to compensate for occasional power loss This feature allows the system to resume operation from the point of interruption and continue purifying active samples Or if necessary samples may be rerun in the future at a more convenient time Solvent levels and fraction vessel capacities are monitored causing the system to pause when a preset warning level is reached requesting additional solvent or collector tubes be added After the solvent is replenished or new fraction collector tubes are added the system will resume to complete the current and queued samples A user friendly manual control enables simple maintenance and troubleshooting A system replica allows users to select a pump loader or collect control screens for diagnostic review Figures 3a and 3b Figure 3 Interactive manual control screen a and pump control screen b allow easy column maintenance and sample recovery where necessary a b Maximum Yield and Purity UV triggered fractionation is simple and reliable giving greater than 90 yields of standard compounds Table 1
21. 1 4 ferrule 1 back 1 4 ferrule 1 4 OD polyethylene tubing 6 03275 1 4 OD 3 8 Male NPT brass bushing 03072 to 1 4 insta tite fitting 1 4 PFA front ferrules 6 03062 1 4 PFA back ferrules 6 Finger tight nuts 2 03061 1 4 PFA front ferrules 2 1 4 PFA back ferrules 2 1 4 x 1 8 reducing union 04144 4 required per set Parts Description Part Number Chemraz O ring 1 03046 Retaining ring 1 Instruction sheet For fraction collection 03071 1 16 OD tubing FEP 12 tubing Union 1 16 to 1 16 SS 1 16 OD tubing SS 3 5 1 8 to 1 16 reducing union Disposable 2 mL polypropylene 5 03276 1 4 to 1 16 tube to tube 02979 For solvent reservoir 01889 1 4 and 1 8 nuts and ferrules NF 15075 3 sets of each For use with FLASH 12 Cartridges FC 022 15024 Frame and tubing SU 242 0000 Short Barrel 02954 Medium Barrel 02942 Upper flow distribution head 02843 Lower flow distribution head 02946 1 4 x 1 8 reducing union 04144 4 required per set 192 FLASH 40i Spare Item FLASH 40i Compression Module FLASH 40S Barrel FLASH 40M Barrel FLASH 40L Barrel FLASH 40i Spare Parts Kit FLASH 12i 40i Start Up Kit O ring Kit Collection Tube Kit Barrel Adapter Syringe Reducing Union Nut and Ferrule Kit Head Assembly Reducing Union Parts Description For use with FLASH 40 Cartridges Short Barrel Medium Barrel Long Barrel Tubing O ring gasket V band clamp
22. 1 5 400 36 min Spare Parts 190 Spare Parts Microwave Synthesis Accessories Item Crimper Decapper Description Hand operated used to crimp 20 mm caps on vials Hand operated used to decap 20 mm caps on vials Microwave Synthesis Miscellaneous Spare Parts Item Tubing kit Syringe Piston Syringe Piston Rack tray Emrys grip vial tongs GSG Booklet GSG Poster Connection Set Vial Rack Cavity Air Guide Cavity Lid Seal Vent Screw Replacement Cavity sleeve Fuse kit Process Needles Solvent bottles Reservoir bottle Vial Adapters Vial Adapters O rings Description Interconnect tubing for syringe pump to needle 500 ul gas tight syringe 500 ul Replacement piston for syringe 10 mL 10 mL Tray for Emrys Synthesizer to hold racks Replacements for gripper Getting Started Guide Booklet Getting Started Guide Poster Air adapters to connecting MAOS to facilities Holds 12 10 20 mL vials Improves cooling of vials O ring blue for EXP MAOS for venting on vials Enables vial venting when required Process chamber cavity sleeve Set of 4 10 A fuses Includes 3 needles 50 mL 6 pack 500 mL bottle and tubing Designed to make 0 2 0 5 mL vials fit into the microwave cavity Kit includes 10 pieces Designed to make 10 20 mL vials fit into the microwave cavity Kit includes 12 pieces Replacement O rings for 10 20 mL adapters Quantity 12 Initiator Spare Parts Item Initiator
23. 2500 1000 2 15 L min FLASH AC 400L 400 x 60 2500 10000 2 15 L min 166 Production Scale FLASH AC FLASH AC Cartridges Provide Predictable Scale up Testing has demonstrated an excellent correlation from a lab scale FLASH AC purification and the actual full scale performance Convert existing batch and deep bed processes to FLASH AC cartridges with a minimal amount of process development and validation Contact Biotage s 1 POINT SUPPORT team for assistance with scale up matters US 1 800 446 4752 EU 46 18 56 59 11 JP 81 422 28 1233 Production Scale FLASH AC FLASH AC 150 mm and 400 mm cartridges are currently in use at major pharmaceutical companies around the world Biotage products provide standard and custom approaches to activated carbon purifications 167 168 FLASH AC Ordering Information Item Description Acid Activated Carbon Cartridges FLASH WAC 12S FLASH WAC 12M FLASH WAC 75S FLASH WAC 75M FLASH WAC 75L FLASH WAC 150M FLASH WAC 150L FLASH WAC 400M FLASH WAC 400L 12 mm x 7 5 cm 12 mm x 15 cm 75 mm x 9 cm 75 mm x 15 cm 75 mm x 30 cm 150 mm x 30 cm 150 mm x 60 cm 400 mm x 30 cm 400 mm x 60 cm Qty Case 20 20 Part Number C1YR 4021 15026 C1YR 4021 15046 C1YR 4021 19163 C1YR 4021 19043 C1YR 4021 19073 C1YR 4021 25075 C1YR 4021 25155 C1YR 4021 50075 C1YR 4021 50155 The FLASH purification systems sold in this catalog readily accept FLASH WAC acid act
24. 4 2 5 CV and the Rf of the impurity A is 0 55 1 8 CV providing a ACV of 0 7 With a ACV this low only a small sample amount can be FLASH purified before overload resolution loss low purity fractions occurs By weakening the solvent strength to 60 hexane and 40 ethyl acetate solvent strength 0 24 Figure 4 middle the Rf of compound B falls to 0 2 5 CV and impurity A s Rf is lowered to 0 3 3 3 CV with a resulting ACV of 1 7 enabling a potential five fold increase in sample load on a FLASH cartridge Table 1 If you find adequate component retention with a particular solvent mixture you can prepare other solvent mixtures of similar strength but different selectivity for comparison Figure 4 bottom Table 3 A solvent mixture s strength is calculated using Solvent Strength g volume proportions and the individual solvent s strength Methanol 0 95 In the example above diluting a solvent mixture with a less l polar solvent hexane from 5090 to 6090 reduces solvent Ethanol 0 88 strength increasing compound retention and resolution ACV Also solvent combinations of similar strength Isopropanol 0 82 but different selectivity can also be compared Both Acetonitrile 0 65 hexane ethyl acetate 50 50 and hexane dichloromethane SEC N 30 70 have solvent strength of 0 3 but ethvl acetate and Ethyl acetate 0 58 dichloromethane provide different selectivitv Tetrahvdrofuran 0 57 Acetone 0
25. 70 hexane dichloromethane both 0 30 solvent strength alters both selectivity and resolution ACV 1 1 Once a solvent system has been selected Rf values measured and ACV values calculated use Table 1 on page 80 to select the correct cartridge for your sample size and ACV The data generated from your TLC method development efforts are applicable to any sized Biotage cartridge Normal phase Gradient Flash Gradient elution enables chemists to speed purification improve recovery and yield and even increase fraction purity In a gradient the stronger eluting solvent concentration is increased over time increasing the solubility of more highly retained components and causing them to elute sooner and in tighter bands compared to isocratic elution Optimize your Purification 85 86 Because solvent strength is increasing during the purification the isocratic CV 1 Rf relationship does not hold In a gradient compounds elute with fewer column volumes than predicted by the isocratic relationship The exact number of elution CV depends on the gradient slope Biotage chemists have developed an applicable algorithm to help chemists transfer TLC Rf to gradient CV This algorithm is incorporated into the SP1 and SP4 graphic user interface software Touch Logic Control If you are using a Biotage Horizon or original SP4 a generally applicable algorithm to use is Segment 1 1 4 the TLC polar solvent concentration for 1 CV
26. 95 253 3 9 99 95 311 3 X b 98 63 223 1 98 93 282 3 CL SQV Ag 98 74 295 2 D 99 95 353 2 Ay Or oe 2 99 95 279 2 OO iet 98 92 298 2 C 99 93 250 1 a 98 85 269 2 ACC 3 c 99 89 321 1 ur 99 88 340 2 T Table 2 Summary of purity and yield achieved for small alkylation array combining the Syntage KP Pr SH Samplet cartridge Conclusion Combining prepacked Syntage KP PrSH Samplet cartridges with Flash chromatography represents a new technique for on column rapid cleanup of excess reagents The work presented in this application note illustrates that the combination of Syntage KP PrSH Samplet cartridges plus Flash chromatography is an efficient and easy technique amenable to parallel solution phase synthesis of tertiary amines This technique provides quick product recovery time by eliminating all the post reaction filtration and washing steps This procedure decreases the number of steps in the synthesis and purification processes since no filtration of the solid support reagent is required Combining scavenging and flash purification in one step provides compounds sufficiently pure for NMR IR and MS confirmation All application notes can be downloaded in Adobe Acrobat format from our Web site at www biotage com BUT Flash Purification Solutions and Optimization 80 Integrated Flash Purification Solutions From synthesis to purification Biotage deli
27. All remaining parameters e g concentration solvent reagent etc should be varied when applicable If the reaction is not proceeding at all or not going to completion e Increase the temperature As long as the reactants reagents can withstand the higher temperature the only limit is the pressure build up in the vial and the security limit of 250 C e Extend the reaction time e Increase the concentration s of reagent s e Change the solvent Some solvents e g water will behave differently at high temperatures as they become less polar This makes some reactions that normally would not work in polar solvents perfectly feasible e Change the reagent s Due to the high temperatures that can be reached sometimes a less reactive but more temperature stable reagent can be used If you see decomposition of reactants reagents products e Lower the temperature e Shorten the reaction time It may be that the desired product is actually formed but then decomposes rapidly at elevated temperatures In some cases it is therefore possible to trap the product by using a shorter reaction time Decrease the concentration s of reagent s e Change to a more temperature stable reagent How Microwave Synthesis Impacts Chemistry Research and Development Microwave synthesis can have a significant impact on chemistry research and development if its strengths namely speed and simplicity are capitalized upon Speed Th
28. Ph D Omar Mneimne M S J Robert Bickler M S and Peter Rahn Ph D Product Horizon HPFC Abstract Synthetic organic chemists worldwide monitor reactions by using some type of chromatographic analysis whether it is thin layer chromatography TLC gas chromatography high performance liquid chromatography HPLC or most recently liquid chromatography mass spectrometry LC MS By far the simplest and most widely used analytical technique is TLC TLC is also used to develop conditions for the purification of organic compounds purifications typically implemented using isocratic elution with normal phase silica columns 24 58 Isocratic elution is effective but can be slow requiring many column volumes of solvent to complete Gradient elution a chromatographic elution technique that increases the solvent strength over time is a method of reducing purification duration Synthetic medicinal chemists under pressure to increase novel lead candidate production need turnkey purification systems and protocols to help them achieve productivity goals In this application four oxazoline analogs were synthesized and purified at gram scale using a generic gradient elution and the new Horizon high performance FLASH chromatography HPFC system The successful chromatographic results are shown in Figure 1 CI 7 31 01 4 33 33 Cl 8 03 01 5 03 56 Run 1 5 100 EtOAc Heptane GRADIENT Run 2 20 80 EtOAc Heptane
29. R3CO 0 TEA DMAP E see oC 10 25 mins 100 180 C 5 20 min N R3 sies OH 2 SECO column CO col 7 A i column i N NH RG N X 10 min R e n RI X Br OTf CI 50 98 80 100 C 5 min 3 SPE 47 99 Sauer D R et al Org Lett 2004 6 16 2793 2796 Kappe O C et al Org Lett 2003 5 8 1205 1208 O OH 1 Benzylamine HOBt DMA PS carbodiimide N 110 C 5 min N 2 Si Carbonate MeOH 5 min filter Sauer D R et al Org Lett 2003 5 24 4721 4724 Polymer Supported Reactions O 1 R NH EDC HCI o HOAt DMF O core ET cd 2 170 C 20 min RI 0 80 R1 0 Chassaing C et al Org Lett 2003 5 11 1851 1853 Pd PPh Cl Cul Z3 CH CH NH DMF xX H R R 120 C 15 25 min 89 98 Gogoll A Erdelyi M J Org Chem 2003 68 6431 6434 NHSO Ar NHSO Ar Cu OAc NMP 200 90 10min oC 10 min 65 82 a Pr T o A Dai W M et al Org Lett 2003 5 16 2919 2922 Review Articles Mavandadi F Lidstrom P Current Topics in Med Chem 2004 4 773 792 Kappe C O Curr Opin Chem Biol 2002 6 314 320 Lew A Krutzik P O Hart M E Chamberlain A R J Combi Chem 2002 4 95 105 Larhed M Hallberg A Drug Discoverv Todav 2001 6 406 416 Lidstrom P Tierney J Tetrahedron 2001 57 9225 9283 Kappe C O Am Lab 2001 33 13 19 Gh LH 5 UKM H The applications group at Biotage uses
30. Software CD Initiator Sixty CD Initiator EXP Software CD Initiator Sixty EXP CD Air hose x 2m Air Control Unit Air Supply connection set Power cable EU Power cable GB Power cable US amp JP Getting Started Guide Description Part Number 353671 353913 Part Number 352289 352508 352509 352510 352511 352512 353010 353067 353068 353480 354836 354974 354494 354878 352296 352291 352024 352076 352078 354459 354367 354838 Part Number 355305 355449 355306 355450 351088 352281 353480 351693 3512669 3511277 355422 Spare Parts Purification FLASH i series Spare Parts The i series of FLASH products preceded the current FLASH line Item V band Clamp Gasket Stopcock Valve Hose Barb Adapter Luer Lok Collar Luer Lok Kit Air Tubing Air Fitting Ferrule Kit Nut and Ferrule Kit Reducing Union FLASH 12i Spare Item O ring kit Collection Tube Kit Syringe Reducing Union Dip Tube Nuts and Ferrules FLASH 12i Compression Module FLASH 12i 40i Start Up Kit FLASH 12S Barrel FLASH 12M Barrel Distribution Head Distribution Head Reducing Union Description Part Number 3 V band clamp for 1 L 08526 solvent reservoirs 3 solvent reservoir gasket 01617 polyethylene 2 way stop flow valve polypropylene 03274 Female Luer to 1 8 tube 01986 Male Luer Lok to 1 8 tube w collar 03229 1 female plastimate Luer Lok port 02838 1 finger tight nut 1 front
31. Three standard compounds with a total load of 40 mg were separated collected and recovered using a fractionation trigger set at 0 2 AU 220 nm Flex Fraction Collection Patented Intelligent algorithms fractionate unresolved components into separate vessels resulting in higher purity This is illustrated by a separation of 100 mg crude sample on a 2 1 x 2 5 cm ABZ C18 column using a simple gradient method where components 1 and 2 have unresolved contaminants Figure 4a The expanded chromatogram shows that although components are not completely resolved they have been collected into different tubes Figure 4b Figure 4a Figure 4b Table 1 Average percent recovery for Propranolol Verapamil Terfenadine UV triggered fractionation 0 Average 976 91 2 92 0 Recovery SD n 3 2 53 2 22 2 16 2 kal x N S X o e o e e y e e e 0 10 20 Time min AU 0 2 0 4 0 6 0 8 1 0 1 2 1 4 1 6 1 8 Time min 157 158 Difficult Separations Three compounds were barely separated by analytical HPLC Figure 5 The B same three compounds were recovered with good yield and high purity using E the Flex HPLC system 26 mg of crude compound was separated with a 2 1 cm x 25 cm column The method chosen was based on polarity and UV absorbance Figures 6a and 6b Data courtesy of GSK Harlow UK B E n Figure 5 Registry Code JWGLFR3 6 A1
32. a user alert Features and Benefits Features and Benefits cont Solvent and waste monitoring Automatically calculates the required solvent amount for a purification sequence monitors the solvent volume used and notifies user to replenish reservoirs when needed Accumulated waste is also measured eliminating potential overflows Variable dual wavelength detector optional Collect fractions using one wavelength while simultaneously monitoring the separation at a second wavelength Flow cells available with either a 0 1 or 0 3 mm flow path Fraction arm positioning Precise fraction arm positioning will not dispense fractions if misaligned eliminating sample loss Fraction Finder Locate compounds of interest faster by pressing the fraction finder button and chromatogram The fraction position is highlighted on the chromatogram and the collection display Print reports Run data and methods are automatically stored Print results and transfer methods to another SP system or PC Autocontinue feature Detect UV absorbance in the flow cell and continue pumping solvent and collecting fractions after the run until the baseline stabilizes SP4 4 column sequential purification Load and run up to 4 samples with just a few icon touches Continually add samples to the queue for uninterrupted sequential purification Flowpaths and waste streams are segregated and automatically flushed between runs eliminating sample to sample cross con
33. allows for higher reproducibility and predictability of results as well as optimization of yields which are usually more difficult when using a domestic microwave oven All research scale microwaves from Biotage are equipped with single mode resonators Why Does Microwave Irradiation Speed Up Chemical Reactions Chemical reactions performed using microwave synthesis techniques are rapid mainly because microwaves are able to quickly achieve higher temperatures and pressures All Biotage microwave synthesizers can achieve temperatures of up to 250 C and pressures of up to 20 bars allowing reactions to be carried out much faster than traditional reflux heating In early literature there were many claims of a specific microwave effect responsible for the observed rate accelerations Later experiments show some of these early reports to be artifacts while others are debatable or difficult to explain An attempt to rationalize a possible specific microwave effect has been published by Perreux et a Most of the reports on specific effects however can be rapidly dismissed due to poor temperature control These inaccuracies in temperature measurements often occur when performing the reactions in domestic ovens with microtitre plates or on solid supports where there are inherent difficulties in measuring the temperature accurately Even with today s specialized equipment it is very difficult to capture the true temperature of a
34. and adsorption processing Like our smaller scale FLASH purification systems the FLASH 400 uses prepacked cartridges 400 mm ID and is designed for radial compression of the packing material providing maximum performance and reliability Biotage FLASH 400 systems are faster safer and easier to use than traditional glass and steel columns Built to last and engineered to perform Biotage FLASH 400 systems are rapidly becoming the first choice of pharmaceutical companies around the world for critical adsorption purification FLASH 400 System Specifications Pressure 100 psig Flow lt 7 L min 80 psig Temperature 5 80 C Wetted Surfaces 316 and 316L j Weight 1000 Ib 450 kg Dimensions 44 x 30 x 90 W x L x H FLASH 400 systems are built with industrial grade components that are appropriate for operations under the U S Food and Drug Administration regulations for current Good Manufacturing Practices CGMP and meet National Electrical Code requirements for explosion proofing CLASS I Divisions 1 amp 2 Groups C amp D FLASH 400 systems have interchangable barrels for use with either 400 mm x 30 cm FLASH 400M or 400 mm x 60 mm FLASH 400L cartridges A system includes a compression module of one size and the module of the other size can be ordered as an interchangeable option e 316 SS ASME rated compression module and radial compression tank s 304 SS frame with industrial style casters
35. are available for use in nonhazardous or explosion proof environments as defined by the National Electrical Code Class I Division 1 Groups C amp D These systems are used to produce multigram to kilogram quantities of peptides oligonucleotides chiral enantiomers or natural products Kiloprep 2000 3000 and 4000 System Configurations Component Standard Configuration Inlet Header A B binary gradient and separate C inlet Sample Loading C inlet on main pump Main Pump Three head double diaphragm Instrumentation Dual channel variable wavelength UV monitor Precolumn pressure Transducer Flow Path Pulse dampener Gradient mixer Fraction Collection Waste or Collect Valve Peak Detection Mode Time UV threshold UV slope Boolean combinations of parameters The Biotage Custom Systems Group tailors each system to your specific needs We offer the greatest range of options for flow rate pressure instrumentation and flow paths Please contact Biotage for a Kiloprep Quotation Workbook and our technical team will help you define the optimum configuration US 1 800 446 4752 EU 001 434 979 2319 KP 2000 KP 3000 KP 4000 Options Tertiary gradient A B C Isolated sample pump Flow meter with feedback control pH meter Conductivity meter Densitometer Thermometer Column delta pressure Filter header with selections for e Filter or Bypass e Filter 1 or Filter 2 Column header with se
36. be deemed acceptance by Customer of the terms and conditions at time of invoice 2 PAYMENT TERMS In addition to the prices invoiced Customer shall pay any tax duty custom or other fee of any nature imposed upon the transaction by any federal state or local government authority In the event Biotage is required to prepay any such tax or fee Customer will reimburse Biotage Payment terms shall be net thirty 30 days In addition Biotage reserves the right in its sole discretion to require C O D payment terms from any Customer Biotage may also refuse to sell to any Customer until all prior overdue accounts are paid in full 3 DELIVERIES AND SHIPMENT Unless otherwise noted on the invoice all Products are delivered F O B to Charlottesville Virginia Customer assumes all risk of loss or damage upon shipment Biotage shall use com mercially reasonable efforts to deliver the Products or provide the Services invoiced hereunder in accordance with the estimated delivery date provided that Biotage shall not be liable for any losses or for general special or consequential damages arising out of any delays 4 WARRANTY The Products and Services shall be provided with the applicable system standard warranty copies of which are available upon request and will be furnished with the Products The applicable standard warranty is in lieu of all other warranties with respect to the products and services expressed or implied includ ing but not li
37. bed level of the loose sample coated silica The pre adsorbed sample loading technique is used when the sample solubility is low or if the dissolution solvent is too strong for the purification This technique can be completed in four easy steps Step 1 The sample is adsorbed onto loose silica or another solid support media Biotage recommends a l part sample load to 2 or 3 parts silica Step 2 The loose sample is loaded directly into the Biotage FLASH purification cartridge Step 3 A corresponding cartridge frit is placed on top of the loose sample inside the cartridge Step 4 The cartridge is loaded into the barrel and sealed using the Dry Loading Compression Head Assembly Features and Benefits e Enables easy purification of low solubility compounds with Biotage FLASH purification cartridges e Maximizes sample loading in a given cartridge e Creates a secure liquid tight seal eliminating potential sample loss e Can be adapted to other FLASH systems with our adapter kit Cartridge Size Maximum Silica Load Cartridge g FLASH 12 2 FLASH 25 8 Use this table as a guide when using the FLASH 40 20 Dry Loading Compression Head Assembly Warning Biotage Samplet cartridges are not compatible with the Dry Loading Compression Head Assembly 122 Ordering Information Dry Loading Compression Head Assembly Ordering Information Item Description Part Number Dry Load
38. by the cus tomer redesigning or repair by Customer or by anyone other than Biotage or its authorized agents or through any cause beyond the reasonable control of Biotage 6 No representative of Biotage is authorized to commit Biotage to any warranty other than the warranty con tained herein Limited Warranty of Biotage Software Performance Biotage warrants for a period of twelve months from the date of shipment the Warranty Period that the Biotage Software will when used in the designated operating environment perform materially in accordance with the applicable specifications The foregoing warranty shall apply only to the most current version of the Biotage Software that was supplied to the Customer by Biotage The limited warranty is subject to the conditions set forth below 1 The Customer must give written notice to Biotage during the Warranty Period with an explanation of the circumstances of any claim that the computer programs or software fail to conform to this limited warranty 2 The Customer s sole and exclusive remedy in the event of any such failure is expressly limited to the correction or replacement of the defective computer program or software or the refund of the fees paid for the non conforming computer program or software 3 Biotage does not warrant that the operation of the computer programs or software will be uninterrupted or error free 4 This limited warranty is for the benefit of and applies
39. capacity of the cartridge A Samplet 100 mg B Direct injection 100 mg HPLC Conditions C Column YMC Pro C18 4 6 x 100 mm 1 142 Solvents A Methanol 0 1 NH4COOH aq 10 90 B Methanol 0 190 NH4COOH aq 90 10 Gradient Equilibrate 2 min at 0 B Linear gradient 0 50 B in 20 min Step to 100 B hold for 2 min Flow rate 2 mL min Injection vol 50 UL Detection 254 nm Compounds 1 Promethazine 100 mg mL in methanol 2 Blue degradation product LOL kin LLES RA N i m zi M i g J n ELLEI Fraction LIT sa N Figure 3 Samplet A and direct injection B impact on degraded promethazine purification using FLASH 12 S C18 cartridges and gradient elution Samplet injection of promethazine sample provides complete compound 1 and degradation product 2 retention and purification while a direct liquid sample injection 1 mL inhibits retention and purification Collected FR 1 FR1 fractions were analyzed by HPLC to verify fraction 0 10 20 24 0 10 20 24 composition and compound purity Minutes Minutes FR 17 FR7 Absorbance 254 nm Absorbance 254 nm Samplet use greatly reduces the solvent effects seen with direct injections increasing compound retention purity and yield Additionally co elution or breakthrough in the void volume is reduced elevating recovery of the compound of interest Application 42 Using TLC to Accurately Predict FLASH
40. entire array of Biotage systems The types of coverage range from standard maintenance and spare parts to advanced training applications support and frequent visits by our field engineers The 1 Point Support Standard Plan With every system purchase from Biotage the 1 POINT SUPPORT standard plan is automatically included This provides customers unlimited access to e our dedicated technical support line e all published Biotage material e tips and procedures for all of our products e our 1 POINT SUPPORT Web site www biotage com which gives customers exclusive access to our user manuals spare parts lists linked to system diagrams special application notes technical bulletins The 1 Point Support Instrument Plan The aim of our individual products support plans is to supply different levels of coverage for travel labor and permanent parts required to maintain a system in like new condition and to enable 100 utilization of the system capabilities Our individual Platinum plans also include e all service and maintenance covered in the standard warranty e product software patches for the software written by Biotage e preventative maintenance visit s scheduled at the customer s request during the coverage period Instrument Plans SER 200 SP1 atinum SP1 Part Number Type Instrumen
41. ferrule 2 03061 Luer Lok Kit Female Luer Lok injection kit 02838 194 FLASH 400 Spare Parts Item O ring O ring O ring O ring O ring O ring O ring Valve Diaphragm Ball Seat O ring O ring Diaphragm Description 387 Viton O ring 387 EPDM O ring 460 Viton O ring 460 EPDM O ring 223 Chemraz O ring 222 Chemraz O ring 460 Chemraz O ring Air Lube Free Wilden Teflon Wilden Teflon Wilden SS Wilden 119 Teflon 120 Teflon Neoprene Wilden FLASH Miscellaneous Spare Parts Item Tubing Tubing Air Tubing Air Tubing Air Tubing Air Tubing Description 1 8 OD FEP tubing Sold by the foot 1 4 OD FEP tubing Sold by the foot 1 4 OD RED air tubing Sold by the foot 1 4 OD YELLOW air tubing Sold by the foot 1 4 OD GREEN air tubing Sold by the foot 1 4 OD BLUE air tubing Sold by the foot FLASH Spare Parts Item E Clip V band Clamp Reducing Union Description Retaining ring 3 V band clamp for 1 L solvent reservoirs 1 4 x 1 8 reducing union 4 required per set FLASH 12 Spare Parts Item Lower ZIF O ring Upper ZIF O ring Barrel O ring FLASH 25 Lower ZIF O ring Upper ZIF O ring Barrel O ring FLASH 40 Lower ZIF O ring Description Chemraz 107 O ring Chemraz 108 O ring Chemraz 108 O ring Chemraz 208 O ring Chemraz 209 O ring Chemraz 209 O ring Chemraz 215 O ring Part Number 03020 03019 02939 03011 03010 06875 0
42. in ethyl acetate Gradient Equilibrate 2 CV column volumes Program Step 1 0 0 B for 2 CV Step 2 0 100 B for 5 CV linear Step 3 100 B for 10 CV Flow Rate 12 mL min Figure 9 KP PrSH Representative Scavenging Procedure Compound 1 was prepared on 0 1 mmol scale in the presence of excess 4 methoxyphenacyl bromide 2 0 eq and diisopropylethylamine 3 0 eq in THF The solution was stirred for 30 minutes and 0 5 mL was transferred to a PrSH12 Syntage Samplet cartridge dried on a heating block inserted into a FLASH 12 M silica cartridge and purified using the following method on a Horizon HPFC system Compound 1 was isolated in 93 yield with an HPLC purity of 99 o Il Br AT aoso Compound 1 XY N rities NH Ep 2 KP PrSH YS Lan 2 0 4 0 Crude Reaction Mixture Purification on Horizon System Solvent A EtOAc Hexane 20 80 Solvent B EtOAc Gradient 0 100 B in 500 mL linear Flow Rate 35 mL min fo up m 4 i Pit Ae Li HPLC analysis of purified fraction 720 40 60 80 100 120 4140 Minute Figure 10 KP TsCL Representative Synthesis Procedure Using a Tertiary Amine Synthesis Method compound 1 was synthesized on 0 1 mmol scale using a Syntage TsCI 12 chemistry cartridge isolated using a FLASH 12 M silica cartridge and puri fied using the following method on a Horizon HPFC system resulting in 81 yield with an HPLC purity of 97 3 uc Compoun
43. methylbenzimidazole brucine and promethazine having a similar separation profile even though twice as much sample was loaded and purified using the Samplet The increased pressure capability of the Horizon HPFC system enabled a separation to occur in 12 minutes at a flow rate of 13 mL min A similar separation would take nearly an hour to complete using glass column technology A 300 mg introduced with Samplet B 150 mg direct injection 1 mL nO KA AU m aH MW MR iN iH ba bE di D H m mi m m am ma a Hi Hd CENT RC IEEE UFR 17182 EMT CCPUMD PM COPUND APR 17182 PUMA USERS ki kA bii SN Db de d O HOI M d x ai R Um em m m R R R R m m du ii Figure 2 FLASH Elution Conditions Cartridge FLASH 12 M C18 Solvents A DI H20 with 0 1 NHACOOH B 90 methanol in H20 with 0 1 NH4COOH Gradient Equilibrate 12 mL at 0 B Linear gradient 0 100 B in 90 mL Hold 100 B for 24 mL step to 0 B for 60 mL Flow rate 13 mL min Detection 254 nm Compounds 1 l Methvibenzimidazole 2 Brucine 3 Promethazine 62 Conclusion Reversed phase purifications using KP C18 HS cartridges with the Horizon HPFC system demonstrate high loading capacity and throughput Because of ruggedly constructed cartridges and the Horizon s high flow high pressure pump flow rate limitations are eliminated reducing overall sample run time Use of Samplet technology further increases purification throughput by increasing the loading
44. mm a b Figure 10 a The Flex V3 database is updated with fractions containing the target mass and are confirmed Yes b The mass spectral data for each sample can be reviewed in OpenLynx browser by right clicking on the sample information 161 FLASH AC Activated Carbon Prepacked Cartridges 164 FLASH AC Activated Carbon Cartridges e High performance easy to use cartridges for the purification of pharmaceuticals and fine chemicals e Available in 12 mm 75 mm 150 mm and 400 mm cartridge diameters e Provide a clean efficient and convenient process for carbon adsorption in a cGMP current Good Manufacturing Practices environment e Engineered for optimum adsorption kinetics and fluid hydraulics for simple and effective purifications e Custom packing services of specific carbon media available please inquire Biotage FLASH AC cartridges are well suited for the pretreatment of compounds that will be crystallized for final purification Pretreatment with FLASH AC can effectively remove contaminants that cocrystallize with the product FLASH AC cartridges are recommended for the following applications e Removal of reaction byproducts color and other contaminants e Removal of catalysts s Removal of lipopolysaccharide LPS pyrogens e Clean up of degraded reagents Predictable scale up from research through production How It Works L D Figure 1 Th
45. mounting bars Powder coated aluminum barrel with stainless steel endcaps V band clamps pressure indicator an ASME rated safety relief valve and mounting bars Powder coated aluminum barrel with pressure relief valve and mounting bars accepts FLASH 75S cartridges Powder coated aluminum barrel with pressure relief valve and mounting bars accepts FLASH 75M cartridges Powder coated aluminum barrel with pressure relief valve and mounting bars accepts FLASH 75L cartridges Stainless steel sample injection module ideal for loading low solubility samples or liquid volumes up to 100 mL Stainless steel sample injection module ideal for loading low solubility samples or liquid volumes up to 500 mL 25 x 1 5 frits for the SIM 100 module 25 x 3 frits for the SIM 500 module 25 stainless steel frits and sealing rings replaces old style SIM bottom frits 25 frits and sealing rings for SIM 100 or 500 used with frit holder FSS 0075 Grounding kit includes Teflon coated wires and clips to dissipate static charges 3 way vertical injection valve for direct liquid injections complete with Luer Lok injection port 4 L stainless steel solvent reservoir ASME rated complete with solvent shut off and relief valves 12 L stainless steel solvent reservoir ASME rated complete with solvent shut off and relief valves Air manifold safely regulates the flow of air pressure and solvent s to FLASH 75 systems 100 mL SIM a cartrid
46. only to the Customer 5 This limited warranty does not apply to any third party operating system software included with the person al computer provided to Customer by Biotage as any such computer software will be subject to applicable man ufacturer s warranties 6 The limited warranty shall not apply to any Biotage Software that has been improperly installed or modified by Customer or any third party or which has been the subject of neglect misuse abuse misapplication or alter ation or has been used in violation of the applicable software license agreement 7 No representative of Biotage is authorized to commit Biotage to any warranty other than the warranty con tained herein PURIFICATION CARTRIDGES and MICROWAVE REACTION VIALS CONSUMABLES Biotage warrants that Biotage Consumables will be of good quality and workmanship and will meet the appli cable product specification This warranty applies only to the initial test performed at the Customer s facility upon the initial start up of the consumables and expires at the time the user applies an actual sample to the consumable If the cartridge is packed with media provided by the Customer the Biotage warranty applies only to the plastic tube frits and labor required for packing and testing the cartridge Biotage will not be liable for 204 any damage to media provided by the Customer that may be caused when Biotage packs such media in accor dance with Bi
47. procedure with microwave synthesis is generally of very low complexity Until recently there has been an uncertainty about the likelihood of success using microwave synthesis However during the past couple of years there has been a real break through in this area as indicated by the vast number of reviews published showing the utility of this technique The extreme simplicity the proven likelihood of good results and the wide reaction range that can be easily performed has made microwave synthesis one of the most valuable tools for the chemist during the past few years This simple dogma can have enormous effects on the chemistry development as shown by Chris Sarko and his colleagues at the second Coherent Synthesis conference in San Diego April 2002 The same group showed that the technology enhanced chemistry development by over 1000 in individual optimization cycles and that an overall productivity enhancement of 200 400 was achieved in their library syntheses 26 References 1 Galema S A Chem Soc Rev 1997 26 233 238 2 Gabriel C Gabriel S Grant E H Halstead B S J Mingos D M P Chem Soc Rev 1998 27 213 3 Gedye R et al The use of microwave ovens for rapid organic synthesis Tetrahedron Lett 1986 27 279 282 4 Giguere R J et al Application of commercial microwave ovens to organic synthesis Tetrahedron Lett 1986 27 4945 4948 5 Langa F et al Microwave irradiation more t
48. reduce the time and expense of scaling up To determine the optimum sample size flow rate and gradient duration for scale up use the following steps 1 Sample Size If an initial method was developed on a 4 6 mm ID x 25 cm column with a 6 mg sample size what is the appropriate sample size for a Biotage Kiloprep 100M cartridge 75 mm ID x 15 cm long Mass Prep Mass Scaling x dia2 prep x length prep Preparative sample g 0 006 75 4 6 2 15 25 0 957 g 2 Flow Rate As you scale up a method you will also need to adjust flow rate to keep the linear velocity constant In many cases Biotage cartridges can be run at higher flow rates to further reduce the purification time Therefore this equation is a starting point for development Flow Rate Prep Flow Rate Scaling x dia2 prep In the same example the chemist ran the 4 6 mm ID analytical column at 1 5 mL min what is the flow rate for the KP100M cartridge Preparative flow rate mL min 1 5 75 4 6 2 400 mL min 3 Gradient Duration If your current method uses a linear gradient the gradient duration must also be adjusted for the new larger column Gradient Duration Prep Gradient Duration Scaling x dia2 prep x length prep x flow rate scaling In the same example the current method used a 60 min linear gradient for 10 to 70 what is the appropriate gradient duration for the KP100M cartridge Preparative HPLC gradient duration min 60 75 4 6 2 15 25
49. selectivity gt compared to toluene Elution order 1 fat red 7B 2 methyl yellow 3 Sudan IV 4 Sudan blue Application Note 28 Performance of Parallex Flex HPLC By Jack Liu Ph D Product Parallex Flex Abstract The Parallex Flex Flexible Purification System is a walk up preparative HPLC high performance liquid chromatography system configured with up to four independent channels eliminating purification stage bottlenecks in the drug discovery process Many drug companies choose to purify their compounds prior to biological testing The Parallex Flex parallel purification systems make this method of prepurification efficient and effective The Parallex Flex uses 24 48 and 96 well microtiter plate formats for automated sample injection Each of the four flow paths is independent simultaneously each can run a different gradient with a different sample on a different column Each flow stream is monitored by a preparative dual wavelength UV detector that allows the fraction collector to collect fractions based on slope and or threshold settings at either wavelength This tool minimizes mixed fractions and the total number of fractions collected High purity fractions are collected into 24 48 or 96 well microtiter plates or test tubes Application 28 addresses the issues of cross contamination and sample recovery Cross contamination was determined by conducting an experiment in which four samples each containi
50. time up to 10X s Eliminate loose carbon to reduce reactor and piping cleaning Eliminate the need for recrystallization and improve product yields production rates and solvent consumption s Simplify operation ready to use cartridge reduces set up and clean up times Quality Assurance e Manufactured in an ISO 9002 certified facilitv Completely traceable materials s Plastic components meet 21 CFR 177 1520 requirements for all components Certificate of compliance for activated carbon media Superior Packing Technology Our proprietary packing technique is critical to the high performance of the FLASH AC cartridges The radial compression of the cartridge eliminates early breakthrough and provides predictable scale up A poorly packed column allows solution to pass through only partially treated leading to early contaminant breakthrough and poor carbon utilization Biotage s high performance FLASH AC media provides the optimum balance between adsorption kinetics and hydraulic pressure drop for maximum performance FLASH AC Specifications Cartridge Dimensions Typical Load Flow Rate mm x cm L mL min FLASH AC 12 S 12x 7 5 0 1 5 20 15 FLASH AC 12M 12x15 0 1 5 20 15 FLASH AC 75 S 75x9 50 300 50 600 FLASH AC 75M 75x 15 50 300 50 600 FLASH AC 75L 75 x 30 100 600 50 600 FLASH AC 150 M 150 x 30 300 1500 0 2 2 4 L min FLASH AC 150L 150 x 60 600 3000 0 2 2 4 L min FLASH AC 400 M 400 x 30
51. to evaluate the solvent strength of these mixtures by varying the polar component percentage A total of 12 solvent systems were transferred and simultaneously run on a 12 channel Quad3 Parallel FLASH Purification system to show how TLC separations translate to flash separations TLC results show that the three hydrazine dye analogs fat red 7B methyl yellow and Sudan IV cannot be separated when an oxygen containing solvent is used regardless of concentration only methylene chloride or toluene efficiently separate all four dyes Figures 1 and 2 Without using TLC for method development and optimization successful FLASH purification would not have been possible A B Solvent Front Solvent Front A 6 D B c e c D E E A Figure 1 B TLC separations showing the effect of different solvents on elution profile selectivity A A mixture of hexane group and ethyl acetate group Vla B same oam Origin sample separated with methylene chloride group V Note how the elution order of 8 the five spots has changed Hexane EtOAc 100 CH Cl 56 5 10 15 20 Toluene Solvent Solvent Solvent Solvent Solvent Front Front Front Front Front 1 0 0 8 e 1 0 6 S 5 2 M 3 P 8 U 0 0 a ai im Oriain Solvent Origin Origin Origin Origin g Strength 0 04 0 07 0 10 0 12 0 29 Figure 2 TLC separation of dyes using ethyl acetate heptane mixtures and toluene Note the difference in
52. 0 peptide purification KP C4 WP 300 15 20 peptide purification Cartridge Description Qty Case Part Number Silica FLASH 12 S KP Sil 12 x 75 mm 20 FPK0 1107 15026 FLASH 12 M KP Sil 12 x 150 mm 20 FPK0 1107 15046 FLASH 25 S KP Sil 25 x 75 mm 20 FPK0 1107 16026 FLASH 25 M KP Sil 25 x 150 mm 20 FPK0 1107 16046 FLASH 40 S KP Sil 40 x 75 mm 20 FPK0 1107 17026 FLASH 40 M KP Sil 40 x 150 mm 20 FPK0 1107 17046 FLASH Cartridge Ordering Information Cartridge C18 FLASH 12 S FLASH 124M FLASH 25 S FLASH 254M FLASH 40 S FLASH 404 M NH Amine FLASH 12 S FLASH 124M FLASH 25 S FLASH 254M FLASH 40 S FLASH 404 M C18WP FLASH 40 S C18WP FLASH 40 M C18WP CAWP FLASH 40 S CAWP FLASH 40 M CAWP Description KP C18 HS 12 x 75 mm KP C18 HS 12 x 150 mm KP C18 HS 25 x 75 mm KP C18 HS 25 x 150 mm KP C18 HS 40 x 75 mm KP C18 HS 40 x 150 mm KP NH 12 x 75 mm KP NH 12 x 150 mm KP NH 25 x 75 mm KP NH 25 x 150 mm KP NH 40 x 75 mm KP NH 40 x 150 mm KP C18 WP 40 x 75 mm KP C18 WP 40 x 150 mm KP C4 WP 40 x 75 mm KP C4 WP 40 x 150 mm Qty Case e e NNNN 20 20 10 10 Part Number FPLO 1118 15025 FPLO 1118 15045 FPLO 1118 16025 FPLO 1118 16045 FPLO 1118 17020 FPLO 1118 17040 FPNH 12S FPNH 12M FPNH 25S FPNH 25M FPNH 40S FPNH 40M FPV1 0906 17020 FPV1 0906 17040 FPV1 1104 17020 FPV1 1104 17040 FLASH Samplets Ordering Information Cartridge Silica FLASH
53. 1 Zone 2 Zone 3 Zone 4 Zone 5 Description 1 4 inch Trough Drain Tube Horizon Flash Collector Set 1 4 316 SS 1 4 Tube 316 SS 1 4 OD x 040 Wall FEP Teflon Mercury UV Fixed Wavelength 254nm UV Variable Wavelength 1 4 x 1 8 reducing union 4 required per set Description Per hour Per hour Per day Local travel 1 50 miles or 1 75 km Local travel 51 100 miles or 76 150 km Travel 101 200 miles or 151 300 km Travel 201 500 miles or 301 750 km Travel 501 1000 miles or 751 1500 km Travel 1001 2000 miles or 1501 3000 km Special travel Part Number 08432 01634 01635 00089 09427 09427 08856 08632 04144 Part Number SER 002 0001 SER 002 0002 SER 0001 LBR SER 0ZA TRVL SER 0Z1 TRVL SER 0Z2 TRVL SER 0Z3 TRVL SER 0Z4 TRVL SER 0Z5 TRVL 202 Limited Warranty FLASH QUAD HORIZON PIONEER Sp4 PARALLEX CHROMATOGRAPHY SYSTEMS INITIATOR CREATOR OPTIMIZER amp ADVANCER SYNTHESIS SYSTEMS Limited Instrumentation Warranty Biotage warrants for a period of twelve months from the date of shipment that the instrumentation products included in any FLASH Quad Horizon Pioneer Sp4 Parallex Initiator Emrys Creator Emrys Optimizer Emrys Advancer or other Chromatography or Synthesis Systems the System shall be free from defects in material and workmanship under normal use and service and when used in compliance with the applicable operating instructions The li
54. 12 Silica Samplet FLASH 25 Silica Samplet FLASH 40 Silica Samplet cis FLASH 12 C18 Samplet FLASH 25 C18 Samplet FLASH 40 C18 Samplet NH Amine FLASH 12 NH Samplet FLASH 25 NH Samplet FLASH 40 NH Samplet CAWP FLASH 40 C4WP Samplet C18WP FLASH 40 C18WP Samplet Description KP Sil 12 mm Samplet KP Sil 25 mm Samplet KP Sil 40 mm Samplet KP C18 HS 12 mm Samplet KP C18 HS 25 mm Samplet KP C18 HS 40 mm Samplet KP NH 12 mm Samplet KP NH 25 mm Samplet KP NH 40 mm Samplet KP C4 WP 40 mm Samplet KP C18 WP 40 mm Samplet Qty Case 48 20 20 48 20 20 48 20 20 20 20 Order FLASH Samplets Part Number SAM 1107 1421J SAM 1107 16016 SAM 1107 17016 SAM 1118 1421J SAM 1118 16016 SAM 1118 17016 SAM NH12 SAM NH25 SAM NH40 SAM 1104 17016 SAM 0906 17016 95 FLASH 12i 40i and 65i Cartridge Ordering Information Qty case Cartridge Silica FLASH 12S Silica FLASH 12S Silica FLASH 12M Silica FLASH 12M Silica FLASH 40S Silica FLASH 40S Silica FLASH 40M Silica FLASH 40M Silica FLASH 40L Silica FLASH 65i Silica cis FLASH 12S C18 FLASH 12M C18 FLASH 40S C18 FLASH 40M C18 FLASH 40L C18 FLASH 65i C18 Diaion HP20 FLASH 40S FLASH 40M Diaion HP20SS FLASH 40S FLASH 40M C18 WP FLASH 40S FLASH 40M C4 WP FLASH 40S FLASH 40M Description KP Sil 12 mm x 75 mm KP Sil 12 mm x 75 mm KP Sil 12 mm x 150 mm KP Sil 12 mm x 150 mm KP Sil
55. 2 000 2000 135 75 200 Pilot KP2000 450 4 500 2 000 135 150 300 Production KP2000EF 750 7 500 1 200 80 150 400 Production KP3000 950 9 500 1 500 100 200 600 Production KP3000EF 1 500 15 000 1500 100 200 800 Production KP4000 1 700 17 000 1 200 80 300 1 000 Production Biotage Kiloprep cartridges are available from 75 mm ID to 300 mm ID LAs Lou 70 minutes 50 minutes Column 4 6 x 220mm Column 7 5 x 15cm v Cartridge Packing C18 12pm 100A Packing C18 12pm 100 Flow Rate 1 5mL min Flow Rate 400mL min Sample 6mg of a 34 mer peptide Sample 300mg of a 34 mer peptide Figure 1 High resolution reverse phase separation of a 34 amino acid synthetic peptide SO Ph i Column 7 5 x 15cm Kiloprep Cartridge Pis D F Packing 12pm Shandon Hyperprep silica uo E Mobile Phase 45 50 5 Hexane Methlyene 3 E Chloride Ethvl Acetate Flow Rate 300mL min Sample 500mg po CH Oe L A Bae Z isomer E isomer e wo QT E isomer Z lt g S A ze Z n fa e 0 5 10 15 20 25 30 35 Time Minutes Figure 2 High resolution normal phase separation of fluorosulfone Z amp E isomers 181 Kiloprep 100 The Kiloprep 100 series are compact bench top HPLC systems designed for use in a laboratory environment These systems produce milligram to multigram quantities of peptides oligonucleotides enantiomers or natural products The Kiloprep 100 systems are for use in research or in process dev
56. 254M 4045 40 M 65i 755 75M 75L 1150M 150L 400M 400L TS 1 2 4 9 11 22 78 47 78 1156 625 11250 4444 8889 124M 1 2 4 6 11 39 23 39 78 313 625 2222 4444 2546 1 2 3 5 18 11 18 36 144 288 11024 2048 25 M 1 1 3 9 5 9 18 72 144 512 1024 g 1045 1 2 7 4 7 14 56 113 400 800 40 M 1 4 2 4 7 28 156 1200 400 t 65i 1 1 1 2 8 16 57 114 B 75s 1 2 3 13 27 95 190 V 75M 1 2 16 57 114 75L 1 28 57 150M 1 2 7 14 150L 1 4 400M 400L 1 Table 4 Scale up chart for Biotage FLASH cartridges 87 FLASH Syntage and Samplet Cartridges FLASH HPFC cartridges were developed to meet the high performance requirements of our FLASH HPFC Systems Automated cartridge packing systems efficiently pack Biotage FLASH cartridges to minimize performance variability Food grade polyethylene barrels are packed with the highest quality silica amino and C18 materials available Strict ISO quality controls at Biotage and years of production experience ensure con sistent performance from cartridge to cartridge FLASH cartridges are available in three diameters 12 mm 25 mm 40 mm and two lengths 7 5 cm S and 15 cm M to fulfill a broad range of sample sizes FLASH cartridges are designed to utilize Samplet cartridges for quick and convenient sample introduction directly on top of the cartridge Samplet cartridges simplify and improve sample preparation for mu
57. 40 S 40 mm IDx75mm 40 mg 2 d 30 100 mL min 60 1L FLASH 40 M 40 mm ID x 150 mm 80 mg 5g 30 100 mL min 60 1L FLASH 40 Samplet 38 mm ID x 20 mm 10 mL sues Please refer to page 117 for all FLASH System ordering information Ordering Information FLASH Ordering Information Component 1 Component 2 Component 3 Options A FLASH 12 S Compression Module O No ZIF SIM A Short Barrel 12 B FLASH 12 M Compression Module C FLASH 254 S 1 ZIF SIM10 Compression Module B Short Barrel 25 D FLASH 25 M Compression Module E FLASH 40 S 2 ZIF SIM35 Compression Module FS1 R F FLASH 40 4 M Compression C Short Barrel 40 Any FLASH Module System and 1 Liter G FLASH 12 S and 3 ZIF SIM60 Reservoir 25 S Compression Modules H FLASH 12 M and D Medium Barrel 12 25 M Compression Modules 4 ZIF SIM10 J FLASH 12 S and and 35 40 S Compression Modules K FLASH 12 M and E Medium Barrel 25 40 M Compression Modules 5 ZIF SIM10 and 60 L FLASH 25 S and 40 S Compression Modules M FLASH 25 M and F Medium Barrel 40 40 M Compression Modules 6 ZIF SIM35 and 60 N FLASH 12 25 40 S Compression Modules P FLASH 12 25 40 M 7 ZIF SIM10 35 G Air Regulator with Compression Modules and 60 filter Part Number FS 1 R__ Instructions Simply choose one component from each of the component columns and then as many options
58. 40 mm x 75 mm KP Sil 40 mm x 75 mm KP Sil 40 mm x 150 mm KP Sil 40 mm x 150 mm KP Sil 40 mm x 200 mm KP Sil 65 mm x 200 mm KP C18 HS 12 mm x 75 mm KP C18 HS 12 mm x 150 mm KP C18 HS 40 mm x 75 mm KP C18 HS 40 mm x 150 mm KP C18 HS 40 mm x 200 mm KP C18 HS 65 mm x 200 mm 40 mm x 75 mm 40 mm x 150 mm 40 mm x 75 mm 40 mm x 150 mm 40 mm x 75 mm 40 mm x 150 mm 40 mm x 75 mm 40 mm x 150 mm 20 100 20 100 12 144 12 144 12 6 KA e KA KA MM N 12 12 Part number FKO 1107 15026 FKO 1107 15028 FKO 1107 15046 FKO 1107 15048 FKO 1107 17024 FKO 1107 17028 FKO 1107 17044 FKO 1107 17048 FKO 1107 17174 FKO 1107 1804C FLO 1118 15025 FLO 1118 15045 FLO 1118 17020 FLO 1118 17040 FLO 1107 17170 FLO 1118 18040 FT6 2030 17024 FT6 2030 17044 FT6 2530 17020 FT6 2530 17040 FV1 0906 17020 FV1 0906 17040 FV1 1104 17020 FV1 1104 17040 FLASH 75i and Cartridge Silica FLASH 75S Silica FLASH 75M Silica FLASH 75L Silica FLASH 75S Silica FLASH 75M Silica FLASH 75L Silica FLASH 150M Silica FLASH 150L Silica C18 FLASH 75S FLASH 75M FLASH 75L FLASH 150M Diaion HP20 FLASH 75S FLASH 75M FLASH 75L FLASH 75S FLASH 75M FLASH 75L FLASH 150M FLASH 150L Diaion HP20SS FLASH 75S FLASH 75M FLASH 75L FLASH 150M FLASH 150L Description KP Sil 75 mm x 9 cm KP Sil 75 mm x 15 cm KP Sil 75 mm x 30 cm KP Sil 75 mm x 9 cm KP Sil 75 mm x 15 cm KP Sil 75 mm x 30 cm KP Sil 150 m
59. 56 Formula Solvent A x solvent A strength Solvent B x solvent B strength Dichloromethane 0 42 100 100 Chloroform 0 40 Examples Ether 0 38 Hexane ethyl acetate 50 50 Toluene 0 29 Solvent strength 0 5 x 0 01 0 5 x 0 58 0 30 Hexane 0 01 Heptane 0 01 Hexane ethvl acetate 60 40 Solvent strength 0 6 x 0 01 0 4 x 0 58 0 24 Isooctane 0 01 Hexane dichloromethane 30 70 Solvent strength 0 3 x 0 01 0 7 x 0 42 0 30 84 Optimal R range Hexane EtOAc 50 50 Solvent stre ngth 0 30 A B L ACV 0 7 0 1 2 3 4 5 6 Column Volumes Hexane EtOAc 60 40 Solvent stre ngth 0 24 0 1 2 3 4 5 6 Column Volumes A Hexane DCM 3 7 Solvent stre ngth 0 30 A e ACV 21 1 B 0 1 2 3 4 5 6 Column Vo lumes Figure 4 Examples of solvent strength on compound retention and resolution The top TLC shows two sample components resolved with a 50 50 hexane ethyl acetate solvent system ACV 0 7 Neither the component of interest B nor the impurity A has an Rf value within the optimal 0 15 0 35 range This leads to poor flash purification top chromatogram After adjusting the solvent to 6096 hexane 409 6 ethyl acetate the Rf values for both A and B fall into the optimal zone middle TLC FLASH chromatography with these conditions middle chromatogram shows increased compound retention and greatly improved resolution ACV 1 7 Replacing 50 50 hexane ethyl acetate with 30
60. 62 ID FEP 1 8 Flangeless 1 4 28 Peek Flangeless 1 8 Tefzel Mercury UV Fixed Wavelength 254nm UV Variable Wavelength Replacement cell 0 1 mm pathlength Replacement cell 0 3 mm pathlength Part Number 09624 09702 09472 05516 08192 06785 9566 4795 4030 9468 08399 09664 01 09664 02 09664 03 09664 04 09376 04144 09314 09311 09312 09665 09666 04144 00088 04795 04030 09695 08394 08372 00088 08399 04030 09427 09427 08856 08632 200 SP4 Spare Parts Item Fraction Collector UV collect valve Needle Nut Ferrule Valve Assembly Fuse Adapter Adapter Operator s Manual Software Sp4 Pump Valve Disk Lip seal Ring Valve Valve Tube Assembly Tube Assembly Tube Assembly Tube Assembly Filter Tubing Nut Ferrule Tube Assembly Tube Assembly Union Tube Assembly Nut Tube Assembly Ferrule Tubing Nut Ferrule Tube Assembly Nut Nut Tubing Nut Nut Ferrule Description 1 6 inch OD x 030 Inch ID x 1 83L 1 6inch Flangeless Fer 1 4 28 Peek Flangeless 1 16 Tefzel UV Collect 3 Way 5 X 20MM 2 5A 250V 40mm to 25mm Flash Module Rack 40mm to 12mm Flash Module Rack CD SP4 10 psi Max NC Collar Mount Check 3 16 inch Balls Assembly Quad3 0 500 OD x 312 ID 302 SST Spring Wear 500 OD X 370 ID Quadi 30 psi Max NC Collar Mount 150 psi Max NC Collar Mount Solvent Inlet 5 Color Red Sp4B Sp1 Solvent Inlet 5 Color Blue Sp4B Sp1 Solvent In
61. 8648 04493 04494 04495 04496 04497 04498 04499 Part Number 00088 00089 00546 01354 00547 01487 Part Number 03024 08526 04144 Part Number 04593 02948 02948 06178 06221 06221 08558 FLASH 12 Spare Parts continued Item Upper ZIF O ring Barrel O ring Reducing Union Description Chemraz 218 O ring Chemraz 218 O ring 1 4 x 1 8 reducing union 4 required per set Part Number 08557 08557 04144 Biotage recommends annual replacement of Chemraz O rings to maintain FLASH compression modules V1 SP4 Spa re Parts The Vi SP4 system preceded the current SP4 system Item Lipseal Ring Wear UV lamp Replacement Windows Valve Valve Valve UV Collect Valve Valve Spring Cprsn O Ring O Ring Ferrule Nut Ferrule Nut Ferrule Nut Tubing Inlet Filters Reducing Union Description 0 500 OD x 312 ID 302 SST Spring 500 OD x 370 ID Quadi for Sp4 Detector for flow cells Solenoid 2 Way NO 24VDC 100 PSI 1 4 28 Prots Select 3 Ports Select 4 Ports UV Collect valve for Horizon and Sp4 UV Collect Valve 3 way 0075 Wire 3 8 Long K 22 316 SS 014 Chemraz 007 Chemraz Flangeless 1 8 Tefzel 1 8 inch Flangeless Fer 5 16 24 Peek Flangeless 3 16 Tubing 3 16 Flangeless 5 16 24 Peek Natural Flangeless 1 16 Tefzel 1 6 inch Flangeless Fer 1 4 28 Peek 1 6 inch OD x 030 Inch ID x 1 83 Long Flash Collector Solvent inlet filter disks 3 8 1 4 x 1 8
62. B1A Variable Wavelength 0 1 mm cell US CSA SP1 B1B Variable Wavelength 0 1 mm cell UK CE SP1 B1C Variable Wavelength 0 1 mm cell EU CE SP1 B2A Variable Wavelength 0 3 mm cell US CSA SP1 B2B Variable Wavelength 0 3 mm cell UK CE SP1 B2C Variable Wavelength 0 3 mm cell EU CE Accessories Item Description Part Number Collection Racks Collection Rack 13 x 100 mm Test tube rack 120 fractions 2 pk HRAK 013 Collection Rack 16 x 100 mm Test tube rack 96 fractions 2 pk HRAK 016 Collection Rack 16 x 150 mm Test tube rack 96 fractions 2 pk QRAK 016 Collection Rack 17 x 130 mm Test tube rack 80 fractions 2 pk HRAK 017 Collection Rack 18 x 150 mm Test tube rack 48 fractions 2 pk QRAK 018 Collection Rack 25 x 150 mm Test tube rack 48 fractions 2 pk QRAK 025 Collection Rack 120 mL bottle rack 24 fractions 1 pk HRAK 120 Collection Rack 240 mL bottle rack 12 fractions 1 pk HRAK 240 Collection Rack 13 x 100 mm IsoBlock rack 128 fractions 4 pk BLK 013 Collection Rack 16 x 100 mm IsoBlock rack 128 fractions 4 pk BLK 016 Collection Rack 16 x 150 mm IsoBlock rack 96 fractions 4 pk BLK 116 Collection Rack 18 x 150 mm IsoBlock rack 48 fractions 4 pk BLK 018 Collection Rack 25 x 150 mm IsoBlock rack 48 fractions 4 pk BLK 025 IsoBlock tray required for IsoBlock racks 1 pk 09047 Cartridge Modules Compression module FLASH 12 S compression module 1 pk 07857 Compression module FLASH 12 M compression module 1 pk 07858
63. Biotage FLASH CV FLASH Relative TLC CV FLASH Relative TLC Rf CV Error Rf CV Error 0 84 1 19 1 25 4 76 0 82 1 22 1 25 2 44 0 34 2 94 1 75 3 00 1 96 0 33 3 03 1 81 3 00 1 01 0 30 3 33 0 39 3 25 2 56 0 28 3 57 0 54 3 25 9 89 0 18 5 56 2 23 5 50 1 01 0 25 4 00 0 43 5 50 27 27 0 10 10 00 4 44 9 25 8 11 0 09 11 11 7 11 9 25 20 12 Table 1 Shows improved flash purification predictability when the TLC silica and flash silica are the same Much greater reli ability in calculated elution volumes solvent consumption is possible In this example the yellow dye has an RF of 0 18 5 56 CV on the Biotage FLASH TLC plate but an RF of 0 25 4 00 CV on the competitive plate With more accurate CV calculations better throughput purity yield and solvent cost savings will be realized Conclusion TLC to flash method transfer accuracy is improved when TLC plates and flash cartridges made with identical silica from the same vendor are used In the example cited above the Biotage FLASH TLC plate Rf data correlate to flash CV with greater accuracy than a competitive plate with the same silica specifications For synthetic chemists the benefits of matched TLC plates and flash cartridges are better purification throughput increased compound purity and yield and reduced solvent cost Application Note 43 Eliminating Aqueous Work Up in Multiple Step Solution Phase Synthesis using Flash Chromatography By Shahnaz Ghassemi Ph D Abstract Post reacti
64. Biotage s world class field service organization serves customers on site to provide the highest quality personalized support Upgrade to Initiator 8 or 60 Initiator Eight The 8 position sample bed provides automation to medicinal chemists for rapid optimization of reaction conditions and analog synthesis The ability to use both large and small vials in combination at any time and in any order without manual intervention provides the medicinal chemist flexibility and the ability to rapidly scale up compounds of interest Initiator Sixty The 60 position sample bed supports the production of focused libraries multiuser environments and scale up by scale out Flexible operation enables the use of both large and small vials in combination at any time and in any order without manual intervention Intuitive Touch Logic Control as simple as 1 2 3 Step 1 Experiment Editor Select Rack type Select the number of reactions and vial position s to load Enter conditions for each vial step 2 View experiment select user and run Step 2 Edit Your Method Specify reaction time temperature vial size and select absorption properties of your sample Select to use fixed hold time if needed n cu ie e a iae os cadi T Step 3 WATT me m nnnm HE LE ri sr rE T Status SS A Check run status remaining processing p time and queue status View temperat
65. C18 Samplets and air dried in a fume hood until no solvent was organoleptically detected All three samples were separated on KP C18 HS cartridges using a Horizon automated HPFC system Separation performance and loading capacity were compared using a direct liquid injection through a valve and the Samplet injection technique The promethazine solution was purified using a FLASH 12 S cartridge and the three component mixtures were separated using a FLASH 12 M cartridge In all instances 1 mL of sample was applied Compound retention and peak shape were compared for each injection technique Promethazine fractions were analyzed for peak purity by HPLC Results In each case the Samplet increased compound retention and loading capacity by eliminating solvent effects Drying the Samplets prior to purification increased compound retention Figures 2 and 3 because the dissolution solvent removed by evaporation no longer competed with the compounds interaction with the cartridge media Also compounds eluted with narrower peakwidth Figure 2 higher purity no mixed peak containing both promethazine and its degradant and greater yield Figure 3 when the sample was introduced onto the cartridge using the Samplet Higher loading capacity is also evident when using a Samplet In figure 2 sample A 300 mg mL was inject ed using a Samplet and sample B 150 mg mL was directly injected using a syringe The data shows the three components 1
66. Compression module FLASH 25 S compression module 1 pk FC 022 16024 Compression module FLASH 25 M compression module 1 pk FC 022 16044 Compression module FLASH 40 S compression module 1 pk 07975 Compression module FLASH 40 M compression module 1 pk 07976 Compression module FLASH 65i compression module 1 pk 07868 Compression modules FLASHPac All 12 S thru 40 M compression 08615 module components Module Stand FLASH 65i compression module stand 09185 holds up to 4 compression modules Flow path module SP4 flow path module 09487 Leak detection kit Contains leak collection tray and removable optical sensor 09713 3 way liquid injection valve FLV VLV 1000 Zif Simi0 FZIM 0010 Zif Sim35 FZIM 0035 Zif Sim60 FZIM 0060 Installation and Training See page 6 in the Customer Support and Service Section for installation and training information Quad Parallel Purification Systems 134 QUAD PARALLE The Biotage Quad Parallel Purification Systems Were Designed by Chemists for Chemists Quad systems address the high throughput purification needs of medicinal chemists and are available in two configurations the 4 channel Quad UV system and the 4 to 12 channel Quad3 system You can customize these systems to meet your current purification requirements and upgrade them as productivity needs change Choose from a selection of modules and cartridge sizes depending on the sample load and application req
67. Flash purification step using a combination of DCM MeOH and ammonia The use of solid supported reagents combines the advantages of solution phase synthesis which includes easy monitoring of the reaction progress by LC MS TLC or NMR techniques and solid phase chemistry which employs excess reagent without adding additional purification steps The advantages of using the Syntage silica supported cartridges include the following 1 2 3 4 5 Eliminates aqueous work up Increases rate of reaction Eliminates need to wash and filter solid support Decreases solvent consumption Facilitates easy automation for parallel solution phase synthesis Syntage Solid Bonded Reagent Cartridges for Synthesis The Syntage KP TsOH Samplet cartridge is utilized as a reagent to synthesize benzofurans by starting with 2 3 1 co phenoxyacetophenone in toluene using the following protocol 300 mg of the Q phenoxvacetophenone was dissolved in 800 mL of toluene and transferred onto a Syntage KP TsOH Samplet cartridge Figure 2 Figure 2 Syntage KP TsOH cartridge being loaded with a phenoxyacetophenone The Syntage KP TsOH Samplet cartridges were placed into the heating blocks and heated for four hours at 70 C in the oven The Svntage KP TsOH Samplet cartridge containing the starting material and product are directly placed into the FLASH 12 cartridge and purified using FLASH chromatography Figure 3 Fi
68. ISOCRATIC 1400 1200 1000 800 mV 600 400 ji l I I i I I I 1 120 180 240 300 360 420 480 540 600 660 Elution Volume mL Gradient Conditions FLASH cartridge FLASH 25 M 25 x 150 mm Silica Waste 100 A 48 mL Flow 30 mL min Gradient 0 100 B in 480 mL Mobile phase A Heptane Detection UV 254 nm B Ethyl acetate FC format 16 x 150 mm test tube Figure 1 Comparison of isocratic 2 and gradient 1 elution techniques Gradient elution provides enhanced selectivity and resolution due to gradual solvent strength change over time Application Note 40 Purification of Heterocyclic Amines on a New FLASH Media KP NH Silica MarthaJoy M Spano and J Robert Bickler Product Syntage KP NH Cartridges Abstract Much of pharmaceutical research is centered on nitrogen heterocycle chemistry These basic amines are difficult to purify using traditional silica chromatography because of strong interactions between acidic silica and the molecules basic amine groups These interactions cause band spreading and poor compound recovery Biotage Inc recently developed KP NH an alternative to amine modified solvents and reversed phase chromatography for organic amine purification KP NH has a slightly alkaline nitrogenous surface chemistry that provides an organic amine friendly environment capable of high sample loads improved selectivity and recovery compared to silica This application no
69. Initiator Eight JPN 355521 Initiator Eight EXP EU 355523 Initiator Eight EXP US 355525 Initiator Eight EXP JPN 355435 Initiator Sixty EU 355437 Initiator Sixty US 355439 Initiator Sixty JPN 355434 Initiator Sixty EXP EU 355436 Initiator Sixty EXP US 355438 Initiator Sixty EXP JPN Upgrade Modules Initiator Robot Eight 355380 Initiator Robot Sixty 355381 Initiator EXP Upgrade 355420 Emrys PathFinder 355239 Web Subscription Accessories Initiator Waste Tray Inserts 355366 Biotage Microwave Vials 0 2 0 5 mL Qty 100 355458 Biotage Microwave Vials 0 5 2 mL Qty 100 352016 Biotage Microwave Vials 0 5 2 mL Qty 3x100 354625 Biotage Microwave Vials 2 5 mL Qty 100 351521 Biotage Microwave Vials 2 5 mL Qty 3x100 354624 Biotage Microwave Vials 10 20 mL Qty 50 354833 Microwave Vial Caps 352298 Crimper 353671 Decapper 353913 Vial Adapters 0 2 0 5 mL Qty 10 355459 Vial Adapters 10 20 mL Qty 12 355367 O rings for Vial Adapters 10 20 mL Qty 10 354838 Initiator Cavity Air Guide 354974 Vial Rack Initiator 8 Vial 4 rack 0 2 5 mL 355391 Vial Rack Initiator 8 Vial 2 rack 10 20 mL 355390 Vial Rack Initiator 60 Vial 30 rack 0 2 5 mL 353478 Vial Rack Initiator 60 Vial 12 rack 10 20 mL 354836 Magnetic Stir Bars 10 20 mL Qty 5 353930 Screwdriver Insex 354107 Screwdriver Torx TX20 355415 Venting Screw Replacement 354878 Installation and Training See page 6 in the Customer Support and Service Section for installation and training in
70. LO 1118 50070 FLASH 400 Upgrades Cartridge Seal Adapter With heavy duty thread 1 07635 Cartridge Insertion Tool Pneumatic press 1 08327 FLASH 400 Barrel Option Barrel Use with 400M FB 012 50070 Barrel Use with 400L FB 012 50150 Must be ordered with system SIM Sample Injection Module SIM 10000 Injection Module SIM 10000 SIM 10000 Cartridge Includes two tubes with bottom frit FTF 10000 assemblies and four frits for packing FLASH WAC acid activated carbon classified washed high efficiency adsorption FLASH WAC 400M 13 2 C1YR 4021 50075 FLASH WAC 400L 26 2 C1YR 4021 50155 176 The Biotage MP 8000 is an automatic pumping and control system designed for use with the FLASH 4007 system Biotage tailors the MP 8000 to meet individual requirements for process control and data recording MP 8000 System Benefits e Fully automates the purification process Including e Sample loading e Binary gradient elution e UV detection e Fraction collection e Safe operation e Class 1 Division 2 Groups C amp D and other ratings e Monitors multiple alarm conditions e Explosion proof The MP 8000 provides monitoring and automation with the precision necessary for the purification of valuable compounds The units are built with industrial grade components appropriate for operations under U S Food and Drug Administration regulations for current Good Manufacturing Practices cGMP They also meet the explosion proofi
71. MPLC Start Up Kit ferrules to connect an AUTOFLASH system to a MPLC pump 148 E IEEE Figure 4 Purify kilogram per day quantities of promising drug candidates with the Biotage 150i ote p ru EROS system and our 150 mm ID cm Qu a LG E De r For Solid Injections prepacked FLASH 150 cartridges OF E sm AM 190 d Injection Air Manifold A DA 1 000 mL _ For Liquid or 2 000 mL Injections 1 f Bl House Air e FLASH 150i A Radial Compression Module Solvent Collect Reservoir Fractions 37 L or 60 L Save Weeks of Separation Time ll The power and performance of the FLASH 150i svstem is clear in this application which saved a customer nearly four weeks of time on a 1 kilogram project FLASH 150M Glass Column Solvent Front Solvent Front e e e o oo 8 6e ee ee ee oo ern 6 e 9 6 0 D 8 Qe X oW os Crude 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 Crude 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 Figure 5 FLASH 150M vs glass column Table 2 FLASH 150 vs Traditional Glass Column FLASH 150M Glass Column Column size 150 mm x 30 cm 120 mm x 66 cm 2 5 kg 3 kg Sample load 180 g 450 g Flow rate 500 mL min 70 mL min No of fractions 45 30 Purification time 90 min 430 min Pure compound 70 5 g run 45 g run Percent recovery 87 22 2 Purification Throughput 120 g hr 63 g hr
72. P20SS SDVB resins e KP Alumina 40 um 60 A neutral alumina Ld FLASH 75S 75M and 75L compression modules See Table 1 for sample sizes Cartridge Diameter Bed Length Packing Wt g Column Vol Easy Typical Difficult mm cm nom mL ACV 6 ACV 2 ACV 1 FLASH 75S 75 9 200 320 5 10 1 5 0 2 1 FLASH 75M 75 15 400 535 10 20 2 10 0 4 2 FLASH 75L 75 30 800 1 070 20 40 4 20 0 8 4 Lo Table 1 Biotage FLASH 75 suggested sample sizes based on TLC evaluation Sample Range g Faster than Glass Separations In this comparison between a FLASH 75L prepacked cartridge and a glass column fractions were collected in 2 5 hours using an 11 x 20 cm glass column while a FLASH 75L 75 mm x 30 cm cartridge required just 40 minutes to complete the separation There were fewer mixed fractions and a greater vield of pure product from the FLASH 75L Glass Column 2 5 hr FLASH 75L Cartridge 40 min Solvent Front Solvent Front ee L 00000 0000 688888888 8888888888886866 6 688888888 00000000 C 123456789 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 C1234567 89 10 11 12 13 14 15 16 17 18 19 20 21 Figure 2 Glass Column vs FLASH 75L Cartridge FLASH U The Biotage FLASH UV detector recorder module simplifies manual flash purification and improves collected compound purity and yield The UV detector monitors displays and records the separation in real time Instead of collecting by v
73. Parts Item Description Part Number Extraction Tool FLASH cartridge extraction tool 02131 O ring Viton O ring 00778 V band Clamp 4 V Band clamp 08527 Valve 100 psi relief valve 00790 Valve Pressure relief valve 07072 Gasket 1 5 EPDM 01644 Clamp 1 5 SS sanitary clamp 00440 Frits Top 1 5 frits 25 FTF 0125 Frits Bottom frit and sealing rings 25 FBS 1025 Injection Valve Ferrule 1 4 PFA 6 03062 Nut and Ferrule Kit 1 4 SS nut w PFA ferrule 2 03061 Tube adapter 1 4 to 1 8 tube to tube adapter 03332 Luer Lok Kit Female Luer Lok injection kit 02838 Sold by the foot FLASH 75i SIM 100 Spare Parts Item Description Part Number Insertion Tool Top frit insertion tool 1 5 diameter 01534 Gasket 1 5 polyethylene 01642 Gasket 1 5 Viton 01643 FLASH 150i Compression Module Spare Parts Item Description Part Number O ring Viton O ring 01420 V band Clamp 8 V band clamp for FLASH 150 01419 compression module Extraction Tool FLASH 150 cartridge extraction tool 01428 SIM 500 1000 2000 Spare Parts Item Description Part Number Gasket 3 polyethylene 01617 Gasket 3 Viton 01615 Gasket 3 EPDM 01616 Clamp 3 SS sanitary clamp 00444 Frits Top 3 frits 25 FTF 0225 Frits Bottom frit and sealing rings 25 FBS 1025 for the stainless steel frit holder Insertion Tool Top frit insertion tool 3 diameter 01534 Injection Valve Ferrule 1 4 PFA ferrule 6 03062 Nut and Ferrule Kit 1 4 SS nut w PFA
74. Purification Results J Robert Bickler Abstract Although many TLC and flash grade silicas have the same physical specifications surface area porosity etc differences in their qualities exist These differences manifest themselves as gross errors in the calculated CV variable selectivity and unreliable loading capacity calculations Because of these real method transfer problems it is very important that the TLC silica and the flash silica be identical and from the same vendor Red Orange Black Yellow Blue When TLC and flash silicas are identical the equation 1 Rf CV holds where Rf is the retention factor of a compound separated by TLC and CV is the number of column volumes required to elute a compound in flash Cartridge loading capacity is based on the difference in CV ACV between two adjacent compounds Discussion A test of Biotage and a competitor s TLC plates using a 5 dye test mix with identical elution conditions highlights the performance differences Although Rf values are similar for most compounds red orange black and blue the yellow dye shows a remarkably different Rf value 0 18 5 56 CV on a Biotage TLC plate but an Rf of 0 25 4 00 CV on the competitor s plate Table 1 When this dye mix was purified on a Biotage FLASH 12 M cartridge the yellow dye eluted in 5 50 CV as predicted by the Biotage TLC plate Table 1 TLC Rf to Flash CV Correlation Biotage Calc ACV Biotage Comp Calc ACV
75. Sample Name Sample ID 19110172 UV2 220nm Absorbance Units Time min Figure 6a A 8 6 mg B 3 5 mg C 1 6 mg z ti F 8 B E Figure 6b Parallex Flex Robust Loading Samples may be injected from individual vials or deep well plates To minimize the risk of precipitation in the injection loop the sample is surrounded by an air gap and a sandwich solvent This prevents contact with the aqueous conditions at the start of the gradient Figure 7 The Biotage Flex V3 loop loading feature facilitates the successful injection of several hundred milligrams A 100 mg injection of synthetic sample is shown below Figure 8 Fractionation was minimized by triggering fractionation at 0 5 AU on 254 nm Despite poor peak shape 77 of the target compound was recovered with a purity greater than 97 The number of fractions can be further minimized and concentration increased by using a 0 1 mm path length flow cell A needle and loop wash reduces carry over to less than 0 2 even at 100 mg loads based on studies with methyl blue which represents a worst case Figure 7 Typical sample loading where the ki alba solvent sample is separated from Satie e the solvent representing Solvent initial gradient conditions by air and a sandwich solvent such as DMSO DMSO methanol 1 1 or acetonitrile water 1 1 ____ AIR GAPS e
76. ack A Quad3 Tube Rack 6 x 20 Fractions 2 pk Cartridge Module 6 x 20 load 4 d pe l l pk B 13 mm Test Tube Rack Pump Module Cartridges 4 x 20 Fractions 2 pk 4 Channel B 13 mm B Quad3 0 Test Cartridge Module No Collector Tube Rack C 16 mm Test Tube Rack 8 Channel 4 x 20 6 x 16 Fractions 2 pk 12 mm Fractions 2 Quad3 Cartridges 2 pk Pump Module D 16 mm Test Tube Rack 8 Channel c Quad3 C 16 mm 4 x 16 Fractions 2 pk Cartridge Module Test 12 Channel Tube Rack 12 mm 6x16 E 18 mm Test Tube Rack Cartridges Fractions 4 x 12 Fractions 2 pk 2 pk G Quad 63b 3 Quad3 oP ier D 16 mm F 25 mm Test Tube Rack Pump Module p Test 4 x 12 Fractions 2 pk Basic Compression Quad 12 Channel Modules 4 40 S i bos bd System RCM Quad FLASH Fractions G Scintillation Vial Rack Channels A1 A2 Collector 2 pk 4 x 10 Fractions 2 pk A3 and A4 E 18 mm For the Qu Test Tube Rack ad UV System H 13 mm Microtiter Block D Quad12 25 4x12 6 x 5 Fractions 8 pk 4 Channel Fractions 12 and 25 mm 2 pk Cartridges J Quad UV Module F 25 mm Test Tube Rack K Quad 4 40 rack 4 E Quad12 25 40 M Compression 4 Channel 4x12 Modules 4 40 S Barrels L 12 mm Fractions and Quad Tubing Kit for Cartridges 2 pk channels B1 B2 B3 and B4 4 4 channel Quadi Pump 2 L Continuous Gradient Module F Quadi2 25 TA FLASH g scintillation Module Collector i 4 Channel 4 Channel Vial Rack cannot be sca and Quad 4x 10 M S
77. age KP PrSH SPE 62 Yellow Orange Flash Chromatography 83 85 3 None alone Syntage KP PrSH 99 93 None Samplet cartridge and FLASH Cartridge Table 1 Purity and yield comparison for three different purification techniques Synthesis of Small Array Using Syntage Samplet Cartridges The general alkylation technique shown above was used to produce a small array of cyclic tertiary amines in one day All the reaction mixtures were purified using the Horizon HPFC system with the Syntage KP PrSH Samplet cartridges inserted into the head of the FLASH cartridge With this new technique all the isolated amines were produced with greater than 98 purity requiring no further workup to confirm the amines structures using high field H NMR FT IR and ES MS to calculate the purity and confirm the structures Table 3 Flash Purification Alone EESEESERESEES Minutes Figure 5 Comparison of Flash separation with and without using a Syntage KP PrSH Samplet cartridge The Syntage KP PrSH Samplet cartridge removes the excess alkyl halide starting material providing a higher purity product with higher yield Combining Syntage KP PrSH cartridge with Flash Purification Minutes Flash Purification Conditions Eluent A Hexane EtOAc 8 2 B EtOAc Gradient 0 35 Bin 120 mL 35 100 B in 90 mL Fraction size 9 mL Flow rate 25 mL min Purity Yield ES MS Purity Yield ES MS CO G 100
78. ally KP PrSH will scavenge metal catalysts used in organic synthesis including Cu Pd and Pb aen Figure 1 Structure of Syntage KP PrSH cartridge General Synthesis and Purification procedure for solution phase alkylation reaction In a 5 mL vial 0 6 mmole of amine and 1 2 mmole of alkyl halide were reacted with 1 6 mmole diisopropylamine in THF Figure 2 The reaction mixture was stirred for 30 minutes at room temperature and then transferred onto a Syntage KP PrSH 25 mm cartridge dried in an oven at 80 C for 10 minutes and then the Samplet cartridge was directly inserted into a FLASH 25 M silica cartridge Figure 3 Products were isolated using a Biotage automated FLASH Horizon HPFC system Advantage of coupling KP PrSH cartridges to flash chromatography Figure 2 indicates the step where the Syntage KP PrSH Samplet cartridge was employed in the alkylation reaction s purification step Figure 3 chromatographically shows the TLC and HPLC chromatographic results after reacting 1 2 3 4 tetrahvdroisoquinoine with 2 bromo 4 methoxyacetophenone When the reaction is complete excess starting material and byproducts must be removed prior to further biological screening or use of this intermediate product Q DIEA KP PrSH Br CO IO THF Flash OMe Chromatography Excess 2 N 9 TEL Figure 2 Figure 2 76 Purity Yield Color Reaction Mixture 42 Yellow Orange Synt
79. amic Field Tuning minimizes reflected microwave energy Extra vessel entry ports Allows in situ sampling during run addition of reagents better control of exothermals or the creation of inert atmospheric conditions Adaptable for real time reaction monitoring through e g Raman spectroscopy Best in class safety features Breech lock closure with pressure release valve and auto mated safety interlock ensure maximum operator safety System pressure control rated up to 20 bar Compatible with the IRORI X Kan system Run Array microwave assisted solid phase chemistry in batch format using IRORI X Kans This technique is beneficial for the introduction of problematic diversity elements or for the rapid microwave optimization of solid phase chemistry and validation screening of building blocks for diversity steps The IRORI X Kan system with CleVap combines multiwell dispensing cleavage transfer collection formatting and evaporation This minimizes operator intervention improving high throughput walk away operation by up to 1152 Kans per run at two runs per day By pooling microreactors into common building block groups the process is very efficient for synthesizing one discrete compound in each microreactor Improved Yield and Speed Experience the same advantages achieved using microwave synthesis at discovery scale in the scale up laboratory Two representative reactions illustrate improvements in speed yield and purity
80. and forth in a matrix they generate heat by friction The amount of heat generated by a given reaction mixture is a complex function of its dielectric property volume geometry concentration viscosity and temperature Thus two samples irradiated at the same power level for the same period of time will most likely end up with rather different final temperatures Essentially the ability of a substance to heat in a microwave field is dependent upon two factors 1 the efficiency with which the substance adsorbs the microwave energy normally described by its dielectric properties and 2 the efficiency with which the adsorbed energy can be converted to heat described by the loss factor A convenient way to evaluate the ability of two closely related substances to convert microwave energy into heat is to compare their respective loss tangent values where the loss tangent is defined as the tangent of the ratio of the loss factor and the dielectric properties Eqn 1 For a deeper insight into the mechanism of microwave dielectric heating the review by Mingos et al is recommended tan 6 E Eqn 1 JPTIM ON However with any Biotage microwave synthesizer the problems of selecting the appropriate matrix is more or less invisible to the end user since these machines are capable of reliably heating a wide variety of substances and have both variable power output and temperature control Multimode vs Sin
81. aphic performance when preadsorbed samples are loaded The ZIF SIM unit is designed to eliminate dead space in the barrel by applying axial compression to the silica bed Axial compression eliminates any risk of sample dilution cracking of the bed and issues associated with sample remixing By utilizing zero insertion force technology insertion and removal of the head assembly is greatly simplified ZIF SIM10 ZIF SIM35 ZIF SIM60 120 FLASH 65i Ordering Information Item Flash 65i Compression Module Flash 65i Compression Module Upgrade 4 Liter Solvent Reservoir Injection Valve Air Regulator with Filter FLASH Start Up Kit Description Holds one 65 mm ID x 20 cm prepacked cartridge Flash 65i head assembly for use with older model FLASH barrel SS 4 Liter 9 23 cm ID x 14 36 cm high 3 way vertical injection valve with Luer Lok adapter for liquid injections Regulates air pressure for use with all air operated Biotage systems 0 60 psig contains particulate filter Frame and tubing Ordering Information Part Number 07868 08405 FN 004 04000 FIV VLV 1000 09350 07923 Biotage recommends annual replacement of Chemraz O rings to maintain FLASH 65i compression modules ZIF SIM Ordering Information ZIF SIM Zero Insertion Force Sample Injection Module Each ZIF SIM module comes with a ZIF head assembly 20 disposable sample loading ba
82. as you need Moving from left to right fill in the spaces below the table to create a custom part number example FS1 RD6 BF 117 118 FLASH Components and Accessories Item Injection Valve 1 Liter Solvent Reservoir 4 Liter Solvent Reservoir Air Regulator FLASH Start Up Kit FLASH 12i Compression Module FLASH 12 S Compression Module FLASH 12 M Compression Module FLASH 12 S Barrel FLASH 12 M Barrel FLASH 25 S Compression Module FLASH 25 M Compression Module FLASH 25 S Barrel FLASH 25 M Barrel FLASH 40i Compression Module FLASH 40 S Compression Module FLASH 40 M Compression Module FLASH 40 S Barrel FLASH 40 M Barrel FLASH Pac FLASH to ISCO Description 3 way vertical injection valve with Luer Lok adapter for liquid injections Stainless steel solvent reservoir ideal for FLASH 12 to 40 compression modules Larger stainless steel reservoir ideal when purifying multiple samples with the same eluent Also ideal for the FLASH 65i compression module Regulates air pressure for use with all air operated Biotage systems 0 60 psig contains particulate filter Frame and tubing For use with FLASH 12i and Samplet Cartridges For use with FLASH 12 S and Samplet Cartridges For use with FLASH 12 M and Samplet Cartridges For use with FLASH 12 S and Samplet Cartridges For use with FLASH 12 M and Samplet Cartridges For use with FLASH 25 S and Samplet Cartridges For use with FLASH 25 M a
83. bars included with vial order Accessories Microwave Vial Caps Qty 107 352298 Crimper 353671 Decapper 353913 Vial Adapter 0 2 0 5 mL Qty 10 355459 Vial Adapter 10 20 mL Qty 12 355367 O rings for Vial Adapters 10 20 mL Qty 10 354838 Initiator Cavity Air Guide 354974 Vial Rack Initiator 8 Vial 4 rack 0 2 5 mL 355391 Vial Rack Initiator 8 Vial 2 rack 10 20 mL 355390 Vial Rack Initiator 60 Vial 30 rack 0 2 5 mL 353478 Vial Rack Initiator 60 Vial 12 rack 10 20 mL 354836 Magnetic Stir Bars 10 20 mL Qty 5 353930 Biotage PathFinder Maximize the Benefits of Microwave Synthesis with Biotage PathFinder Web Biotage PathFinder Biotage PathFinder is the world s largest database of established methods for microwave synthesis The new Web based format offers chemists worldwide access to more than 3 800 diverse microwave methods More than 600 of the reactions are new Using simple keyword and or substructure search it is fast and easy to find microwave conditions for your reactions of interest along with experimental details and information needed to perform the reactions Additional Features Ask a Chemist In addition to the database Biotage Pathfinder includes an Ask a Chemist feature which allows chemists to have a dialog on microwave synthesis methods and get quick answers to their questions directly from a Biotage chemist experienced in microwave synthesis Vapor Pressure Calculator This software all
84. based fraction collection in an ergonomically designed format Figure 1 UV Detector e and Recorder TES Collector i Th Pm FLASH Compression Modules and Cartridges Pump M Figure 1 The Biotage Horizon HPFC Purification System includes a gradient pump UV detector and recorder Control is accomplished via single keyboard control Fast Solution Phase Synthesis of Fmoc Tyr t Bu Gly Gly Phe Leu NDMCP The desired polypeptide product was synthesized using the traditional solution phase synthesis technique that started with Fmoc Leu NDMCP The eight steps included four amino acid coupling reactions and four Fmoc deblocking reactions This work compares two sample workup methods 1 the traditional aqueous work up 2 and the in line HPFC technique for isolation of intermediates and final products Method 1 Aqueous Work Up Technique After reacting the Fmoc base peptide Fmoc Leu NDMCP with base and DBF dibenzofulvene the aqueous work up procedure requires two washes with NaCl three washes with buffer followed by drying with magnesium sulfate and then finally filtering the product and evaporating the excess solvent After the coupling reagent is added and the reaction occurs additional aqueous extractions are required again followed by magnesium sulfate drying steps Multiple aqueous washes with dilute acid or dilute base are utilized to remove excess reagents byproducts or excess reagents after each s
85. cartridge with the FLASH cartridge the highest purity 99 and the final product yield 93 was achieved Figure 4C This higher purity was achieved without sacrificing yield which is very important since the alkylation product is the starting material for additional reactions Figure 5 shows the purification chromatogram generated by purifying the reaction mixture with and without combining it with the Syntage KP PrSH Samplet cartridge Flash chromatography alone provided a higher purity product 8390 compared with the SPE cartridge The purity and yield comparison for the three different purification techniques are summarized in Table 1 Quad UV Pump and Cartridge Module Specifications Quadi Pump Quad1i2 25 Cartridge Module Quad4 40 Rack 136 Dimensions Flow Range Pressure Limit Power Requirements Delivery Volume Stroke Dimensions Flow Range Pressure Limit Power Requirements Number of Parallel Channels Cartridge Capacity Sample load Range Dimensions Number of Parallel Channels Cartridge Capacity 16 x 23 x 31 cm WxDxH 60 mL min 60 psig 30 psig compressed air or other inert gas 3 mL 18 x 23 x 48 cm W x D x H 60 mL min 60 psig 30 psig compressed air or other inert gas 4 Up to 4 FLASH 12 M or 25 M with or without Samplets 8 mg 1 5 g 36 x 16 x 28 cm W x D x H 4 Up to 4 FLASH 40 M or S Quad Specifications Quad3 Pu
86. ccur by solution phase synthesis the advantage of easily monitoring the reaction progress at any step by TLC or HPLC eliminates a major drawback compared to performing solid phase synthesis References 1 Morrissey M M Mohan R Xu W Tetahedron Lett 1997 38 7337 7340 2 a Lijima K Fukuda W Tomoi M Pure Appl Chem 1992 A29 3 249 261 b Organ M G Dixon C E Biotech Bioeng Comb Chem 2000 71 71 77 3 a Tomoi M Kato Y Kakiuchi H Makromol Chem 1964 185 2117 2124 b Tomoi M Watanabe T Suzuki T Kakiuchi H Makromol Chem 1985 186 2473 2481 4 Boisnard S Chastanet J Zhu J Tetahedron Lett 1999 40 7469 7472 74 Application Note 46 Rapid Synthesis and Clean up of an Array of Amines Using Pre packed Syntage KP PrSH silica Supported Thiol Cartridges By Shahnaz Ghassemi Ph D Product Syntage KP PrSH cartridges Abstract Fast efficient synthesis and purification of cyclic tertiary amines is important in drug discovery This work illustrates a new synthesis technique that produces a high purity array of cyclic tertiary amines using parallel solution phase synthesis by coupling Syntage KP PrSH Samplet cartridge with Flash chromatography KP PrSH is the silica bonded equivalent of 1 propanethiol Figure 1 designed to covalently scavenge alkyl benzyl and aryl halides as well as a variety of electrophiles including acid chlorides and isocyanates Addition
87. d 1 b OH re cy G 4 MA im gt Us Purification on Horizon System me Solvent A Hexane 20 80 Solvent B 0 5 TEA ethyl acetate Gradient Step 1 25 100 B for 10 CV Step 2 100 B for 2 CV HPLC analysis of purified fractions HPLC Purity 97 mv 170394908 800 00 2 FO K Calculated 308 22 Found M 1 309 24 200 00 0 00 4 L L 0 00 1 00 200 3 00 4 00 5 00 6 00 7 00 8 00 9 00 10 00 11 00 12 00 Minutes Figure 11 KMNO4 Representative Oxidation Procedure Flash purification of a synthetic aldehyde using Syntage 12 KMnO4 Samplet cartridge FLASH 12 Silica cartridge and a SP4 HPFC system Oxidized products were typically purified by elution starting from 1 4 to 1 1 dichloromethane hexane in 6 column volumes Syntage KP KMnO Samplet R oH Tag EP E gt 0 CH CL room temp 1 000 44 0 800 0 600 Starting Material 0 400 UV Abs AU Average 0 200 A typical purification profile of the oxidation of 4 tert butylphenethyl alcohol using the Biotage system 0 000 6 27 2003 6 27 2003 6 27 2003 6 27 2003 3 55 PM 4 00 PM 4 05 PM 4 10 PM Figure 12 KP NCO3 Representative Scavenging Example Scavenging Nitrophenol in a Mitsunobu Reaction Compound 5 was prepared via a Mitsunobu reaction DEAD was added to a solution of 9 fluorenemethylalcohol nitrophenol 1 8 eq and triphenylph
88. d a 4 liter solvent reservoir For low solubility samples mixtures can be dissolved in a polar solvent mixed with silica concentrated and then packed into the barrel of a ZIF SIM60 The ZIF SIM easily mounts between the solvent reservoir or HPFC pump and the FLASH 65i compression module to introduce the sample onto the cartridge Features and Benefits Easy insertion and removal of cartridges Low dead volume head assembly for improved sample injection efficiency All stainless steel flow path that simplifies cleaning between injections Less O ring wear A ZIF sealing mechanism that allows higher operating pressures and flow rates Even distribution of sample and solvent flow through each cartridge FLASH 65i compression module and cartridge FLASH 65i Specifications Cartridge Dimensions Weight Flow Rate Sample Size KP Sil 65 mm x 200 mm 350g 65 85 mL min 0 3 13 g KP C18 HS 65 mm x 200 mm 400g 65 85 mL min 0 03 1 3 g Please refer to page 121 for all FLASH 65i ordering information 119 ZIF SIM Zero Insertion Force Sample Injection Module Sample Injection Modules with Biotage s Patented Zero Insertion Force Design Improve Purification Performance The new ZIF SIM family of products is used with Biotage s FLASH 12 25 40 and 65i compression modules This new design utilizes our patented Zero Insertion Force ZIF technology This simplifies sample loading and improves chromatogr
89. d specified vial volumes If the volume is too low the temperature reading may be inaccurate and if the volume is exceeded insufficient space is left for pressure build up Biotage Microwave Vials Biotage Microwave Vials Biotage Microwave Vials Biotage Microwave Vials 0 2 0 5 mL 0 5 2 mL 2 5 mL 10 20 mL only with EXP systems only with EXP systems Features and Benefits Features and Benefits e All Biotage Microwave Vials withstand pressures of 20 bars ca 300 psi e With the Reseal design the septum can be resealed after it has been penetrated allowing repeated additions of reagents or insitu sampling e Manufactured from contaminant free microwave safe glass e Magnetic stirring promotes even temperature distribution throughout the entire reaction mixture including heterogeneous mixtures Available in four sizes 0 2 0 5 mL 0 5 2 0 mL 2 0 5 0 mL and 10 20 mL x Perform Microwave Synthesis in 4 simple steps E Step 1 Add stir bar Step 2 Add reagents and solvents 3 E 1y Step 3 Cap aN Step 4 Process Vial Ordering information Item Description Part Number Biotage Microwave Vials 0 2 0 5 mL Qty 100 355458 Biotage Microwave Vials 0 5 2 mL Qty 100 352016 Biotage Microwave Vials 0 5 2 mL Qty 3x100 354625 Biotage Microwave Vials 2 5 mL Qty 100 351521 Biotage Microwave Vials 2 5 mL Qty 3x100 354624 Biotage Microwave Vials 10 20 mL Qty 50 354833 Microwave vial caps and stir
90. dgements 2004 Biotage Syntage Sp4 FLASH HPFC Horizon Horizon Pioneer FLASH FLASH 12 FLASH 12i FLASH 25 FLASH 40 FLASH 40i FLASH 65i FLASH 75 FLASH 150 FLASH 400 KP Sil KP C18 HS KP C18 WP KP C4 WP KP Alumina SIM Production Scale FLASH Quad Quad3 Quad4 40 Samplet Quad3 CV Pump Quad UV Quad FLASH Collector Parallel FLASH Purification ZIF ZIF SIM Prepacked to Perfection EVALUPAK Proprep Parallex Parallex Flex Parallex Pilot Parallel HPLC Purification FLASH AC FlashElute MultElute 1 Point Support Initiator Emrys Creator Emrys Optimizer Emrys Advancer Emrys Process Vials Emrys Process Kits and Purification by Design are trademarks of Biotage AB Affinity by Design and Kiloprep is a registered trademarks of Biotage Diaion is a trademark of Mitsubishi Teflon and Chemraz are trademarks of E I Dupont FluoroFlash is a trademark of Fluorous Technologies Inc All other trademarks used within this publication are property of their respective companies www biotage com UNITED STATES and CANADA Biotage Tel 1 434 979 2319 Toll Free 1 800 446 4752 UNITED KINGDOM EIRE Biotage Tel 44 1992 501535 SWEDEN Biotage Tel 46 18 56 59 00 JAPAN Biotage Tel 81 422 281233 product_info biotage com
91. e 12 Port Manifold Installation and Training See page 6 in the Customer Support and Service Section for installation and training information Description Holds 4 FLASH 12 M cartridges in parallel Holds 4 FLASH 25 M cartridges Holds 4 FLASH 12 M or 25 M cartridges Holds 4 FLASH 12 M cartridges Holds 8 FLASH 12 M cartridges Holds 12 FLASH 12 M cartridges Adds 4 cartridge capacity to existing 4 or 8 pack module Pneumatic parallel 4 channel pump no upgrades Pneumatic parallel 4 channel Quad3 pump Pneumatic parallel 8 channel Quad3 pump Pneumatic parallel 12 channel Quad3 pump Adds 4 pump heads to existing 4 or 8 channel pump Fraction Collector Holds 2 test tube racks 4 trays pk 6 x 20 position rack for 13 x 100 mm test tubes 2 pk 4 x 20 position rack for 13 x 100 mm test tubes 2 pk 6 x 16 position rack for 16 x 150 mm test tubes 2 pk 4 x 16 position rack for 16 x 150 mm test tubes 2 pk 4 x 12 position rack for 18 x 150 mm test tubes 2 pk 4 x 12 position rack for 25 x 150 mm test tubes 2 pk 4 x 10 position rack for 20 mL scintillation vials 2 pk 6 x 5 position block for 13 mm test tubes 8 pk 4 channel UV monitor variable wavelength 210 320 nm Optional flow cell provides increased sensitivity 10 32 thread Holds 4 40 compression modules Includes a Quad 4 40 rack 4 40 M compression modules 4 40 S barrels and Quad tubing kit for channels A1 A2 A3 and A4 Includes a Quad 4 40 rac
92. e 316 SS air operated diaphragm pump e SS overbraided Teflon lined tubing e 316 SS valves with Teflon seats e Calibrated pressure gauges and pressure relief valves e Optional frame mounted hoist for loading and removing cartridge SS stainless steel pet M NETA DIAPHRAGM PUMP Hr yo 56 ON OFF Y v id l 12 OD 1 SS OB x INLET B 1 fi ty 1 2 OD MW SS OB E l VENTA G VACUUM BREAK aC RR SN FLOW PRODUCTION PG 2 V RATE FLASH e 100 PSI og ADJUSTMENT ENT AIR SUPPLY 406 MODULE CARTRIDGE Ps 100 PSI T MES TEC M 1 4 HDPE VENT s Y ey l n 9 DRAIN NITROGEN SUPPLY B T 100 PSI m 7 X L gt 1 VENT pk T VACUUM BREAK 1 4 HDPE 1 4 OD U ss uS 1 2 OD 5S OB MV MANUAL BALL VALVE TANK 12L PG PRESSURE GAUGE E PSV PRESSURE RELIEF VALVE prr P RESERVOIR OUTLET 1 OUTLET 2 Figure 1 FLASH 400 Process and Instrument Diagram 173 174 FLASH 400 Cartridges e FLASH 400 systems are designed for use with Biotage s family of 400 mm cartridges which are supplied with normal and reversed phase silica activated carbon polymeric adsorbents or custom packed media e Designed for both chromatography purification and adsorption p
93. e FLASH AC 12 cartridge 12 mm ID fits into a standard FLASH purification system See page 112 of this catalog for a complete description of FLASH 12 systems and options FLASH AC Convenience and Performance Batch vs Cartridge Convenience FLASH AC cartridges are easier to use than batch processes e Insert a prepacked FLASH AC cartridge into the compression module e Fill the pressure reservoir with solution e Apply gas pressure e Collect purified solution Clean up is even easier e Drain and blow down the system e Remove the top from the compression module e Remove the spent prepacked cartridge seal both ends send out for disposal e No reactors to clean e No exposure to potentially hazardous materials Batch vs Cartridge Performance Batch vs Cartridge The graph in Figure 2 clearly demonstrates that 120 FLASH AC cartridges are much more effective in removing impurities than a batch mode process 100 Even with a 24 hour contact time the batch process removed only 18 of the contaminant By E 80 contrast the FLASH AC cartridge removed 100 of o x the contaminant and did so in less than one tenth the time For complete details about this test O 60 A Pe 5 please see Biotage Application Note 12 on our Web E site www Biotage com E 40 2 o n 20 0 4 Batch Mode FLASH AC Cartridge Figure 2 Benefits of the FLASH AC cartridges Reduce process
94. e main benefit of microwave synthesis is shorter reaction times through an increase in reaction temperature Herein we describe how this affects chemistry development Chemistry like all sciences consists of never ending iterations of hypotheses and experiments with results 24 JPTIM ON guiding the progress and development of projects The shorter reaction times that microwave synthesis provides make it ideal for rapid reaction scouting and optimization allowing chemists to proceed very rapidly through the hypotheses experiment result iterations resulting in more decision points per time unit Figure 4 Result i gt ts K Result iD lt b Ht Fa LI i T gt Figure 4 Microwave assisted organic chemistry can be used g to speed up the chemistry development by allowing more Convention Chemistry Micrmwave Chemistry experiments to be conducted per time unit The only prerequisite for this approach to be productive is that reactions need to be successful enough times to make it worthwhile i e the molecules have to withstand the extreme thermal conditions for the short time that the reaction proceeds At a time when the trend in organic synthesis is moving toward using ambient conditions it may seem incredulous to heat reactions above 200 C nevertheless most reagents catalysts and substrates have been shown to survive extreme temperatures for short periods of time In order to fully benefit from this technique c
95. e same manner as the Quad UV system except that it is available with up to 12 independent flow streams for even higher productivity The Quad3 system is available in configurations of 4 8 or 12 channels It can be used with either a Quad3 cartridge module takes up to 12 FLASH 12 cartridges or one or two Quad4 40 modules which will meet growing productivity and throughput demands Other Options Continuous gradient module and an automatic step gradient module Features and Benefits e Pneumatic pump with parallel independent flow streams delivers consistent volume even if purified sample precipitates during purification e Patented ZIF Zero Insertion Force head assembly on the cartridge module pivots back for easy loading unloading of cartridges e Variable wavelength UV module tunes to optimal absorption e Multichannel UV monitoring of parallel flow streams facilitate compound location e Milligram to multigram purifications with patented FLASH cartridges and Samplets e Parallel independent no loss fraction collection accommodates up to 12 samples on one fraction collector e One button unattended operation simplifies use 135 2 bromo 4 methoxyacetophenone Product Starting material Impurities Figure 3 TLC and RP HPLC evaluation of the cyclic tertiary amine The yellow color is not visible in the HPLC trace but it is evident in the reaction mixture prior to puri
96. e solvent that you would normally use Polar solvents e g DMF NMP DMSO methanol ethanol and acetic acid couple well with microwaves due to their polarity i e the temperature will rise substantially with these solvents Non polar solvents e g toluene dioxane THF can be heated only if other components in the reaction mixture respond to microwave energy i e if the reaction mixture contains either polar reactants or ions When using less polar solvents more concentrated reaction mixtures are preferred Under such circumstances very high temperatures can be achieved Ionic liquids are reported as new environmentally friendly recyclable alternatives to dipolar aprotic solvents for organic synthesis The dielectric properties of ionic liquids make them highly suitable for use as solvents or additives in microwave assisted organic synthesis Ionic liquids absorb microwave irradiation extremely efficiently and they have a low vapor pressure enhancing the heating process Despite ionic liquids being salts they dissolve easily in a wide range of organic solvents and can be used to increase the microwave absorption of low absorbing reaction mixtures Solvents can behave differently at elevated temperatures and most solvents become less polar with increased temperature Water is the most interesting case At elevated temperatures the bond angle in water widens and its dielectric properties approach those of organic solvents Water a
97. eek from our field engineers who service the products and offer application support THE GOLD 1 Point Support Site Plan For customers with up to 75 FLASH and or Quad products and up to three Parallex Flex and or three Microwave Svnthesis svstems e Standard 1 Point Support e Spare parts e Visits three davs per week from our field engineers who service the products and offer application support THE PLATINUM 1 Point Support Site Plan For customers with up to 75 FLASH and or Quad products up to three Parallex Flex svstems and or five Microwave Svnthesis svstems e Standard l Point Support e Spare parts e Visits five davs per week from our field engineers who service the products and offer application support Repair and Return Policy Before Calling Us A 1 Point Support team member will be able to serve you more efficiently if you have the following information e Serial number and model number of each device involved e Concise list of symptoms e List of operating procedures and conditions you were using when the problem arose e List of other devices connected to the unit e List of other electrical connections in the room Warranty Repair Units covered under warranty will be repaired and returned to you at no charge If you have any questions about the applicability please contact your local 1 Point Support team Non Warranty Repair For out of warranty repairs contact your local 1 Point Support team A team member will d
98. elopment groups that plan to scale up further Kiloprep 100 System Configuration Component Solvent Reservoirs Inlet Header Sample Loading Instrumentation Flow Path Fraction Collection Peak detection Mode 182 Standard Configuration 2 37 L ASME A 4 B binary gradient and separate C inlet C inlet on main pump Dual channel variable wavelength UV detector Precolumn pressure transducer 3 head plunger pump Gradient mixer Waste or collect valve Time UV threshold UV slope Boolean combinations of parameters Options 3 60 L ASME Tertiarv gradient A B C Isolated sample pump Loop injector pH meter Conductivity meter Densitometer Thermometer Column delta pressure Column header with forward or reverse flow modes Recycle valve 12 port rotary valve Purge for high recovery Kiloprep 250 KILOPREP 250 The Kiloprep 250 is a compact skid mounted HPLC system designed for use in a pilot plant or full scale production environment It is available for use in areas that are nonhazardous or explosion proof as defined in the National Electrical Code Class I Division 1 Groups C amp D Kiloprep 250 systems are used to produce multigram to kilogram quantities of peptides oligonucleotides chiral enantiomers or natural products Kiloprep 250 System Configuration Component Standard Configuration Solvent Reservoirs 2 60 L ASME Inlet Header A B binary gradient and sepa
99. ent reservoir tubing grounding kit and manual FLASH 75L System 75L compression module air manifold SIM 500 SF 022 19071 12 L solvent reservoir tubing grounding kit and manual FLASH 75L Plus 75L Plus compression module interchangeable 75S SF 222 19071 System barrel air manifold SIM 500 including 25 frits 12 L solvent reservoir 10 FLASH 75L cartridges 800 g KP Sil 10 FLASH 75S cartridges 200 g KP Sil tubing grounding kit and manual UV Detector Recorder Free standing FLASH UV Detector Recorder Module 09043 with 0 1 mm path length flow cell and start up kit 144 Item FLASH 75 Compression Modules FLASH 75S Compression Module FLASH 75M Compression Module FLASH 75L Compression Module FLASH 75 Accessories FLASH 75S Barrel FLASH 75M Barrel FLASH 75L Barrel SIM 100 Sample Injection Module SIM 500 Sample Injection Module SIM Top Frits SIM Top Frits SIM Bottom Frit Holder SIM Bottom Frits Grounding Kit 3 way Injection Valve 4 L Solvent Reservoir 12 L Solvent Reservoir Air Manifold SIM 100 Start up Kit SIM 500 Start up Kit FLASH 75 Start up Kit Ordering Information Description Powder coated aluminum barrel with stainless steel endcaps V band clamps pressure indicator an ASME rated safety relief valve and mounting bars Powder coated aluminum barrel with stainless steel endcaps V band clamps pressure indicator an ASME rated safety relief valve and
100. fication Comparison of different purification techniques The alkylation reaction mixture is a yellow colored solution that included 43 of the desired product starting materials and some side products The reaction mixture was then purified utilizing three separate techniques 1 Syntage KP PrSH Samplet cartridge SPE alone Figure 4A 2 Flash chromatography alone Figure 4B 3 Syntage KP PrSH Samplet cartridge Flash chromatography Figure 4C Figure 4A indicates the SPE cartridge scavenged the excess alkyl halide starting material with an overall increase in purity to 62 but the remainder of the impurities including the yellow color eluted along with the product as verified by HPLC Flash chromatography alone was also used to purify the same reaction mixture but since the alkyl halide starting material eluted close to product the yield was sacrificed to achieve the desired purity level Figure 4b A KP PrSH SPE cartridge 62 pure MEN V ud J WAL a A AJ B Flash Cartridge 83 pure C Flash Cartridge plus Syntage KP PrSH Samplet cartridge 2 00 4 00 6 00 8 00 1000 1200 1400 Figure 4 Comparison of purified product by three different purification techniques Combination of Syntage KP PrSH cartridge and Flash cartridge provided the highest yield with the highest purity Finally by combining a Syntage KP PrSH Samplet
101. fies no void exist in the media bed Biotage does not warranty specific plate count peak symmetry or separation for any compounds For the Custom Packed Cartridges where the cartridge is packed with media provided by the Customer the Biotage warranty applies only to the plastic tube frits and labor required for packing of the cartridge Biotage will not be liable for any damage to media provided by the Customer that may be caused when Biotage packs such media in accordance with Biotage s standard operating procedures If the Custom Packed cartridge fails Biotage will make a reasonable effort to re pack the original media or pack a replacement cartridge with new media provided by the Purchaser Should any Special Request cartridges fail Biotage will make a reasonable effort to re pack the original media in a replacement cartridge Return of the Custom Packed or Special Request Cartridge is not permissible once the user applies a sample to the cartridge This is in compliance to government OSHA and TOSCA regulations KILOPREP CHROMATOGRAPHY SYSTEMS Biotage provides the limited warranty for each of its Kiloprep Chromatography Systems as is set forth in the applicable System quotation furnished to the Customer TERMS AND CONDITIONS 1 TERMS OF SALE Purchase of the products and services can only be made pursuant to the terms of the invoice that are set forth herein Receipt of the products or commencement of the services shall
102. formation Advancer 38 The batch microwave synthesizer for the scale up of organic synthesis The Advancer system is designed for reliable and reproducible batch scale up of microwave assisted synthesis handling reaction volumes from 50 to 300 mL at temperatures from 60 to 250 C and pressures of up to 20 bar Synthesis methods developed using other Biotage microwave synthesis systems can be directly transferred to the Advancer Purities and yields are comparable or even improved enabling chemists to safely and quickly translate optimized reactions to a larger scale With its 350 mL reaction vessel the Advancer serves the chemist s needs for scaffolds and intermediates or substance for early toxicological and formulation tests Features and Benefits Batch format microwave synthesizer Specifically designed so that methods developed in Biotage lab scale systems can be safely and predictably transferred to a larger scale without re optimization Powerful mechanical stirring up to 1000 rpm In addition to homogeneous reactions the system also supports heterogeneous reactions and the use of solid catalysts heavy slurries and crystallizing products Flash cooling The patent pending instant adiabatic cooling promotes high purity and yield For example 200 mL of EtOH can be cooled from 180 C to 65 C in just 30 seconds Efficient heating Powerful 0 1200 W focused magnetron rapidly heats large volumes Dyn
103. from the starting materials and side products Gradient flash conditions were used for this separation Product purity was confirmed by HPLC Conclusion Syntage KP TSOH Samplet cartridges easily and successfully integrate synthesis and purification Traditional work up steps are also eliminated when the Syntage Samplet cartridges are directly inserted into FLASH purification cartridges This integrated synthesis and purification procedure eliminates solution transfers and sample manipulations and it results in a higher yield of final product These on column reactions occur by solution phase synthesis allowing the reaction progress to be monitored at any step by TLC or HPLC eliminat ing a major drawback of solid phase synthesis References 1 Flynn D L Crish J Z Devraj R V Hockerman S L Parlow J J South M S Woodard S S J Am Chem Soc 1997 119 4874 2 Lawrence M R Biller S A Fryszman O M Poss M A Synthesis 1997 553 3 Liu Y S Zhao C Bergbreiter D E Romo d J Org Chem 1998 Vol 63 No 10 4 Boisnard S Chastanet J Zhu J Tetahedron Lett 1999 40 7469 7472 70 Application Note 45 On Column Clean Synthesis of an Array of a phenoxyacetophenones Using Syntage TBD Cartridges By Shahnaz Ghassemi Ph D Product Syntage TBD cartridges Introduction There is a need to develop a simple fast and flexible method to generate libraries of a phenoxyacetophenones
104. g a Kiloprep HPLC system compression module and prepacked cartridges Optimize your Optimizing FLASH Chromatography Method Development Tips for Better Purification Results This flash chromatography method development guide covers three specific types of flash purification methods Purification normal phase isocratic normal phase gradient and reversed phase These guidelines address important issues related to successful flash purification Normal phase e Converting TLC thin layer chromatography Rf retention factors to CV column volumes s Determining the best solvent selectivity using TLC e Determining the best solvent strength using TLC Determining the optimum cartridge size and sample load based on TLC data Reversed phase e Converting HPLC high performance liquid chromatography retention times and gradient methods to CV Determining the optimum cartridge size and sample load based on TLC data Normal phase Isocratic FLASH 1 Predicting compound retention and resolution using TLC For successful FLASH purification Biotage recommends method development using Biotage TLC plates Component retention on TLC plates is measured in terms of Rf retention factor In FLASH purification retention is usually measured in CV column volumes Methods developed using TLC are generally transferable to FLASH chromatography because the relationship between Rf and CV is reciprocal CV 1 Rf Figure 1
105. g compounds Development Scale Systems FLASH 75i and 150i Systems Save hours even days scaling up with the FLASH 75i Purify multigram quantities of synthetic compounds The FLASH 150i purifies kilogram quantities of promising drug candidates Provides capability for direct scale up from TLC FLASH AC Upgrade product quality by removing reaction byproducts color and other contaminants using activated carbon high performance adsorption Production Scale Systems FLASH 400 Production scale System Produce compounds with the highest purity yield and throughput at the kilogram scale with the FLASH 400 system and prepacked cartridges designed for purification according to the U S Code of Federal Regulations current Good Manufacturing Practices CGMP MP s000 Automate the FLASH 400 module with an MP 8000 pumping detection and fraction collection system SIM 10000 Increase cartridge lifetime when the crude product has limited solubility by pairing a SIM 10000 Sample Injection Module with the FLASH 400 system HPLC Systems for Scale up Parallex Flex The only HPLC preparative system available with one to four channels The V3 operating software is simple to use requiring only four clicks to load and run a sample Ideal for multiuser environments the Flex V3 offers reliable unattended parallel purification Kiloprep Purify grams to kilograms of peptides oligonucleotides enantiomers or natural products usin
106. ge extraction tool tubing user s manual and grounding kit 500 mL SIM a cartridge extraction tool tubing user s manual and grounding kit Cartridge extraction tool tubing user s manual and grounding kit Part Number FC 022 19161 FC 022 19041 FC 022 19071 FB 012 19160 FB 012 19040 FB 012 19070 SIM 0102 SIM 0502 FTF 0125 FTF 0225 FSS 0075 FBS 1025 FGD 15075 FIV 075 0000 FN 004 41201 FN 012 41201 AM 190 SU 275 0100 SU 275 0500 SU 075 2000 146 Syringe Nitrogen Cylinder UV MPLC MARS H AUTOFLASH 75 not included NI i T O lt J UL Fraction Collection UV Detector Figure 3 The Biotage AUTOFLASH 75 kit is designed for use with medium pressure liquid chromatography MPLC systems Medium Pressure Liquid Chromatography with AUTOFLASH 75 Compression Modules AUTOFLASH 75 compression modules are designed to provide FLASH 75 capacity and performance for your medium pressure liquid chromatography MPLC system AUTOFLASH 75 compression modules come with a pressure gauge one start up kit with all the tubing and fittings required to connect to an MPLC system and a four way valve with Luer Lok adapter for direct sample injection AUTOFLASH 75 compression modules use Biotage s standard FLASH 75 75 mm ID prepacked cartridges see page 141
107. ghput Sjaan Armentrout John Gu J Robert Bickler Abstract Normal phase flash purification is commonly used by organic chemists in pharmaceutical drug discovery and process development labs However for many synthesized products e g peptides nucleotides and basic drug 60 candidates purification on standard flash silica is not an option due to irreversible adsorption chemical interaction and or solubility issues Reversed phase flash purification is an excellent solution for these applications Yet this technique has been used sparingly because of perceived lower loading capacity higher operating pressures and a scarcity of publications addressing reversed phase flash chromatography Samples dissolved in organic solvents when injected into highly aqueous reversed phase solvent systems are typically not well retained by C 18 columns The injection solvent competes with the sample for access to the stationary phase causing early compound elution breakthrough and peak broadening The higher operating pressures required for reversed phase purification preclude the use of glass columns and cause leaks in many commercially available flash systems Research shows that system backpressure in psig when using reversed phased solvents is roughly four times the flow rate in mL min for 50 50 methanol water Figure 1 Pressure Increase with Flow Rate 50 50 methanol water 120 e eo e Pressure p
108. gle Mode Despite several subcategories there are two fundamentally different constructs of microwave heating devices namely multimode or single mode The main difference between the two is in the build up of the energy field within the systems In both cases microwaves are generated by a magnetron and led into the reaction chamber the cavity through a wave guide When the microwaves in a multimode apparatus enter the cavity they are reflected by the walls generating a three dimensional stationary pattern of standing waves within the cavity called modes Multimode microwaves are optimized to give high efficiency for 200 1000g loads and consequently they operate less reliably for smaller loads However for chemistries required on larger scales the multimode device is appropriate since single mode devices would be inefficient therefore this is what is used for our large scale microwave Advancer Ideally to obtain a well defined heating pattern for small loads a microwave apparatus utilizing a single mode cavity is preferred This type of cavity allows only one single mode to be present Much higher field strengths can be obtained giving rise to more rapid heating A properly designed cavity also prevents the formation of hot and cold spots within the sample resulting in a uniform heating pattern This is very important when microwave technology is used in organic chemistry since the heating pattern for small samples can be well controlled This
109. gure 3 After the reaction was complete the Syntage KP TsOH cartridge was directly inserted into FLASH cartridge eliminating the need for extra connections or sample handling The total sample is applied to the cartridge in a plug flow format with this sample introduction technique 4 The product was analyzed by HPLC and confirmed by FT IR H NMR and ES MS Figure 4 68 Figure 4 illustrates the results from the synthesis of one of the substituted benzofurans by starting with a phenoxyacetophonone that had been produced using the Syntage KP TBD Samplet cartridge The reaction yield was 89 and the purity of the final product was 99 HPLC Trace of Reaction Mixture and Final Product after Flash Purification Product Starting material E 2 00 N a 4 Crude AU 5 J reaction 1 00 4 J mixture 050 HPLC Conditions Wi ed er L Column C8 4 6 x 50 mm a 309r 7l 3E OU Mobile Phase A H20 ACN 95 5 2 00 Flash purified Minutes Purity 9996 0 1 TFA ag substituted Yield 86 B ACN H20 95 5 x l benzofuran 0 1 TFA 1 00 Gradient 5 100 B in 8 minutes hold 2 minutes s E Flow Rate 1 5 mL min 0 00 Detection 254nm 5 00 10 00 15 00 Minutes Figure 4 This is one example of the Flash purification and HPLC traces of the crude and isolated a phenoxvacetophenone product The Flash purification UV trace illustrates the a phenoxyacetophenone separation
110. han just a method for accelerating reactions Contemp Org Synth 1997 4 373 386 6 Gedye R N Wei J B Rate enhancement of organic reactions by microwaves at atmospheric pressure Can J Chem 1998 76 525 532 7 Pagnotta M Pooley C L F Gurland B Choi M J Phys Org Chem 1993 6 407 411 8 Perreux L Loupy A A tentative rationalization of microwave effects in organic synthesis according to the reaction medium and mechanistic considerations Tetraheron 2001 57 9199 9223 9 Hayes B L Microwave Synthesis Chemistry at the Speed of Light CEM Publishing Matthews NC 2002 10 Stuerga D Gonon K Lallemant M Tetrahedron 1993 49 6229 6234 11 Baghurst D R Mingos D M P Superheating effects associated with microwave dielectric heating J Chem Soc Chem Commun 1992 674 677 12a Dzierba C D Combs A P Microwave Assisted Chemistry as a Tool for Drug Discovery Annual Reports in Medicinal Chemistry 2002 37 247 256 b Santagada V Perissutti E Caliendo G The Application of Microwave Irradiation as New Convenient Synthetic Procedure in Drug Discovery Curr Med Chem 2002 9 1251 1284 C Wathey B Tierney J Lidstr m P Westman J The Impact of Microwave Assisted Organic Chemistry on Drug Discovery Drug Discovery Today 2002 7 373 380 d Wilson N S Roth G P Recent Trends in Microwave Assisted Synthesis Curr Opin Drug Disc amp Dev 2002 5 4 620 629 e Lar
111. hed M Hallberg A Microwave Assisted High Speed Chemistry A New Technique in Drug Disvovery Drug Discovery Today 2001 6 406 416 f Kappe C O High Speed Combinatorial Synthesis utilizing Microwave Irradiation Curr Opin Chem Bio 2002 6 3 314 320 g Larhed M Moberg C Hallberg A Microwave Accelerated Homogenous Catalysis in Organic Chemistry Acc Chem Res 2002 35 9 717 727 h Lidstr m P Tierney J Wathey B Westman J Microwave Assisted Organic Synthesis A Review Tetrahedron 2001 57 9225 9283 i Lidstr m P Westman J Lewis A Enhancement of Combinatorial Chemistry by Microwave Assisted Organic Synthesis Comb Chem High Throughput Screen 2002 5 441 458 13 Roth G Sarko C http www dddmag com feats 0109micro asp Discovery Developments Receive the Biotage Enewsletter Discovery Developments Discovery development Receive monthly updates on the latest technology and techniques in medicinal chemistry You will find articles by leaders in the fields of microwave synthesis and flash purification fast reactions and upcoming events To register go to www biotagedcg com and click on register to sign up for Discovery Developments and other E mail updates Initiator Microwave Synthesis Systems 30 The Initiator microwave synthesizer enables medicinal chemists to quickly synthesize compounds using microwave heating In addition to speed
112. hemists have to be ready to risk trying reactions at high temper atures and be prepared to fail or succeed While failure could cost a few minutes success would gain many hours or even days To gain the most from microwave synthesis it needs to be regularly used as the preferred technique for synthesis Simplicity Simplicity is one of the assets of modern microwave equipment Reactions are performed in glass vials sealed with crimp caps the vials are subsequently heated in the microwave cavity at a constant temperature for a set period of time The reaction can be analyzed by sampling through the septa With the availability of proper analytical facilities the time from the genesis of an idea to the result can be a matter of minutes enabling the chemist to rapidly test the feasibility of novel synthetic routes Optimize your Reaction be Perform Microwave Synthesis in 4 simple steps E Step 1 Add stir bar Step 2 Add reagents and solvents Step 3 Cap Step 4 Process Figure 5 The simple preparations needed to perform microwave syn thesis on a Biotage system Productivity Successful chemistry development relies on two basic foundations 1 the level of synthetic complexity and 2 the likelihood of a positive outcome Thus a synthetic procedure with a greater likelihood of success with the least possible effort usually becomes established and widely used As mentioned previously the synthetic
113. iator and Initiator EXP Y Y Y 2 355481 Platinum Initiator 60 and Initiator 60 EXP Y Y Y 2 P Y Y Y Support and 6 1 Point Support THE ANSWER TO ALL VOUR QUESTIONS Service Tiers Broad Support for Customers with Multiple Biotage Svstems In addition to the standard 1 Point Support plan automatically supplied with each Biotage system we offer you the option of further support designed to match the increased requirements at sites with numerous system and application demands We have created Bronze through Platinum 1 Point Support Site Plans to provide our customers with the exact level of service and support required As your business grows so too can the support you receive from Biotage Optimize the productivity of your group by allowing our dedicated team of more than 15 technical support engineers who have a combined total of over 100 years of support experience help you obtain the most from your equipment Learn more about our various extended 1 Point Support plans THE BRONZE 1 Point Support Site Plan For customers with up to 20 FLASH and or Quad products e Standard 1 Point Support e Spare parts A bimonthly visit from our field engineers who service the products and offer application support THE SILVER 1 Point Support Site Plan For customers with up to 50 FLASH and or Quad products and one Parallex Flex and or Microwave Synthesis svstem e Standard 1 Point Support e Spare parts e A Visit one day per w
114. imethylformamide DMF 153 250 4 o Dichlorobenzene 180 250 2 Tetrahydrofuran THF 65 110 3 Water 100 165 10 Xylenes 137 50 0 Some solvents can reach higher temperatures if they contain microwave absorbing material and are heated for a longer time For the very poorly microwave absorbing solvents much higher temperatures have been observed in various reactions for example xylene 150 C 1 4 dioxane 200 C water 220 C 1 2 dimethoxyethane 200 C and tetrahydrofuran 180 C Volume Do not exceed or fall below the vial s specified volumes Too low a volume will give an incorrect temperature measurement while too high a volume does not leave sufficient head space for pressure build up Since microwave heating is strongly dependent upon geometry and volume Biotage provides four different vial sizes to ensure similar performance and scalability throughout the entire volume range Figure 3 Figure 3 Biotage Microwave Vials For more information on Biotage s Microwave Vials go to page 44 21 22 JPTIM ON Concentration The concentration depends on the type of chemistry that is performed A unimolecular reaction is independent of concentration and can be performed in very diluted solutions Bi or tri molecular reactions on the other hand are highly dependent on the concentration a higher concentration gives a faster reaction The maximum obtainable concentration is dependent on the properties of the s
115. ing FLASH 12 Compression Head For use with direct dry loading into the 09547 Assembly FLASH 12 cartridges Dry Loading FLASH 25 Compression Head For use with direct dry loading into the 09543 Assembly FLASH 25 cartridges Dry Loading FLASH 40 Compression Head For use with direct dry loading into the 09535 Assembly FLASH 40 cartridges Dry Loading FLASH 12 S Compression Compression assembly with 09553 Module FLASH 12 S barrel Dry Loading FLASH 12 M Compression Compression assembly with 09554 Module FLASH 12 M barrel Dry Loading FLASH 25 S Compression Compression assembly with 09555 Module FLASH 25 S barrel Dry Loading FLASH 25 M Compression Compression assembly with 09556 Module FLASH 25 M barrel Dry Loading FLASH 40 S Compression Compression assembly with 09551 Module FLASH 40 S barrel Dry Loading FLASH 40 M Compression Compression assembly with 09552 Module FLASH 40 M barrel Dry Loading Frits 12 For use with dry loading in FLASH 12 09738 cartridges 20 package Dry Loading Frits 25 For use with dry loading in FLASH 25 09739 cartridges 20 package Dry Loading Frits 40 For use with dry loading in FLASH 40 09740 cartridges 20 package FLASH to ISCO Adapter Kit Adapter and tubing to allow 09335 use of FLASH or i style compression modules with ISCO systems ESS ax m d oe al ka gant es man mom es eee PUER ppan j iji SP Flash Purification System Fast Intuiti
116. ing purification and synthesis with Biotage systems and products are offered for new or advanced users and include hands on training with user supplied samples Instrument Training Plans Part Number Description Instrument SER FLEX TRAIN 3 day training course Parallex Flex systems SER FLEX INSTL Installation and Start up Parallex Flex systems SER QUAD3 INSTL Installation Training Quad 3 SER QUADUV INSTL Installation Training Quad UV INTR INT 000 Installation Training Initiator EXP INTR INT 0060 Installation Training Initiator 60 60 EXP INTR INT 1060 Installation Upgrade from Initiator 1 to Initiator 60 INTR SP1 0000 Installation Training SP1 353997 EWM Training Microwave Instrumentation G1 Point Support THE ANSWER TO ALL YOUR QUESTIONS Problem solving Techniques We can assist you in solving tough purification and synthesis problems to help you maximize your productivity Our customer support mission at Biotage is to offer superior service Our broad support superstructure is available through a single point of contact our 1 Point Support team Products and Plan Coverage Biotage 1 Point Support plans offer various levels of support for our purification and synthesis product lines In addition to the standard 1 Point Support provided to all Biotage customers as described in detail below you can purchase extended warranty plans for each product or broad support plans for your
117. iscuss service options with you and assist in arranging the return of the equipment for repair if necessary Biotage may utilize refurbished components when repairing units Return Procedure Contact your local Biotage 1 Point Support team see page 11 to obtain a Return Material Authorization RMA number before returning any Biotage system Carefully pack the system to prevent damage in transit Check with Biotage regarding proper method of shipment Indicate the RMA number on the carton and on the packing slip Biotage assumes no responsibility for damage caused by improperly packaged units Always insure for the replacement value of the system Include a description of symptoms your name address and telephone number and a purchase order to cover repair costs return and shipping charges if your company requires it In the U S and Canada In Europe ship to ship to Biotage AB Please contact your local 1 Point Support RMA Number representative for the correct return address 1725 Discovery Drive see contact information on page 11 Charlottesville VA 22911 In Japan ship to Biotage Japan Ltd RMA Number Medi Coop BLDG 8 5F 2 4 14 Kichijojihoncho Musashinoshi Tokyo Japan 180 0004 How to Place Your Order How to Use this Catalog How to Place Your Order Product information is displayed with easy to follow text and diagrams highlighting the system operation and application Sections are color coded
118. ity 90 particle size 35 70 um NH Cartridge Specifications surface area 230 m g porosity 100 particle size 40 75 um 8 17 01 4 05 PM Only 32 fractions UV Threshold 8 17 01 8 17 01 4 10 4 15 PM PM 8 17 01 4 20 PM Figure 6 Using a FLASH 65i a 3 component aniline mixture 10 g was successfully separated on a Horizon HPFC system with UV collection using a FLASH 65i compression module and 65i KP Sil cartridge KP Sil KP C18 KP NH Cartridge FLASH 12 S FLASH 12 M FLASH 25 S FLASH 25 M FLASH 40 S FLASH 40 M FLASH 65i FLASH 12 S FLASH 12 M FLASH 25 S FLASH 25 M FLASH 40 S FLASH 40 M FLASH 65i FLASH 12 S FLASH 12 M FLASH 25 S FLASH 25 M FLASH 40 S FLASH 40 M Dimensions 12x75 12 x 150 25x75 25 x 150 40 x 75 40 x 150 65 x 200 12x75 12 x 150 25x75 25 x 150 40 x 75 40 x 150 65 x 200 12 x 75 12 x 150 25 x 75 25 x 150 40 x 75 40 x 150 Load range mg 4 200 8 400 15 800 30 1600 40 2000 80 5000 300 20000 0 4 20 0 8 40 1 5 80 3 160 4 200 8 500 30 2000 2 100 4 200 8 400 15 800 20 1000 40 3000 FLASH Cartridge Ordering Information Additional Packing Media Specifications Packing Porosity Particle Size Application um Mitsubishi Diaion HP20 300 600 250 600 fermentation capture Mitsubishi Diaion HP20SS 300 600 75 150 fermentation capture KP C18 WPT 300 20 4
119. ivated carbon cartridges as indicated below FLASH AC Systems Ordering Information FLASH WAC Cartridge Model 12S 12M 75S 75M 75L 75S 75M 75L 150M 150L 400M 400L Compatible FLASH Purification System FLASH 121 FLASH 121 FLASH 75S FLASH 75M FLASH 75L AUTOFLASH 75S AUTOFLASH 75M AUTOFLASH 75L FLASH 150M FLASH 150L FLASH 400M FLASH 400L FLASH System Part Number SF 020 15024 SF 022 15024 SF 022 19161 SF 022 19041 SF 022 19071 AF 005 19160 AF 005 19040 AF 005 19070 SF 022 25071 SF 022 25151 SF 011 50070 SF 011 50150 Ordering Information FLASH 12i The Purification Setup FLASH WAC 12 Cartridges The FLASH 12i system includes one compression module FLASH 12Si 12 mm ID x 7 5 cm L one FLASH 12M barrel 12 mm x 15 cm interchangeable with the 12Si module one injection valve one solvent reservoir one ZIF SIM10 sample loading cartridge one stand a start up kit and a user s manual This system is also available without the ZIF SIM for a description of ZIF SIMs see page 118 of this catalog FLASH 12i Ordering information Item Description Part Number FLASH 12i with ZIF SIM Full system with ZIF SIM SF 022 15024 FLASH 12i Full system no ZIF SIM SF 020 15024 169 Production scale FLASH Systems 172 FLASH 200 FLASH 400 Systems The Biotage FLASH 400 is a complete skid mounted system designed for large scale flash chromatography
120. k fi H FLASH 12 S and 12 M columns head U assemblies FLASH 12 S and 12 M cartridges and Samplets Flash 12 Specifications Cartridge Dimensions Sample size Flow rate ZIF SIM Reservoir FLASH 12 S 12 mm ID x 75 mm 4 200 mg 10 15 mL min 10 1L FLASH 12 M 12 mm ID x 150 mm 8 400 mg 10 15 mL min 10 1L FLASH 124 Samplet 10 mm ID x 20 mm 1 mL SE Please refer to page 117 for all FLASH System ordering information Customize Your FLASH System The FLASH 25 Compression Module and 25 mm ID Prepacked Cartridges are Designed for Milligram to gram Scale FLASH Separations FLASH 25 M System with ZIF SIM10 ZIF SIM35 and ZIF SIM60 and test tube rack pt m P Li l r 1 FLASH 254S and 25 M columns Em mu head assemblies FLASH 25 S and 25 M cartridges and Samplets d Flash 25 Specifications Cartridge Dimensions Sample size Flow rate ZIF SIM Reservoir FLASH 25 S 25 mm ID x 75 mm 15 800 mg 20 50 mL min 10 or 35 1L FLASH 25 M 25 mm ID x 150 mm 30 1600 mg 20 50 mL min 35 1L FLASH 25 Samplets 23 mm ID x 20 mm 5mL sos aoe s353 Please refer to page 117 for all FLASH System ordering information 115 116 FLASH 40 M System with ZIF SIM10 ZIF SIM35 and ZIF SIM60 and test tube rack FLASH 40 S and 40 M columns head i assemblies FLASH 40 S and 40 M cartridges and Samplets Flash 40 Specifications Cartridge Dimensions Sample size Flow rate ZIF SIM Reservoir FLASH
121. k 4 40 M compression modules 4 40 S barrels and Quad tubing kit for channels B1 B2 B3 and B4 Single binary gradient module 6 step automatic gradient system requires manifold Required for use with Step Gradient Module Part Number QUD1 1200 0400 QUD1 0025 0004 QUD1 1225 0404 QUAD 0004 1000 QUAD 0008 1000 QUAD 0012 1000 QUAD 0004 0000 QUD1 0400 1000 07675 07677 07346 QUAD 0400 0000B QUD1 FRAC 2000 TRAY 04 QRAK 013 QRUV 013 QRAK 016 QRUV 016 QRAK 018 QRAK 025 QSNT 028 QRAK M13 09387 07529 08238 QPFP 440 2000A QPFP 440 2000B QCGM 1000 QSGM 1000 QMAN 1000 Development scale FLASH Chromatography Systems 142 Flash 75i for Reliable Scale up Purification Everything You Need to Begin FLASH 75i packages include all the components necessary to begin your purification a FLASH 751 radial compression module a fully integrated air manifold a solvent reservoir a sample injection module a start up kit with all necessary tubing a grounding kit and a user s manual FLASH 75i Uses Patented Radial Compression Biotage s patented radial compression technology reduces the chance of void and channel formation resulting in a higher bed density Compounds can then be collected in narrower bands for higher purity and yield High Flow Rate Operation withstand operating pressures up to 100 psig The FLASH 75 cartridges available in sizes containing 200 400 and 800 g
122. lections for e Forward or Reverse flow e Column 1 Column 2 e Bypass Recycle valve Heat exchanger 8 port rotary valve Purge for high recovery 185 186 Biotage Kiloprep cartridges are prepacked pretested and shipped with a certificate of compliance so they are ready to use in high resolution HPLC purifications We offer media from all the leading vendors covering the widest possible range of surface chemistries particle sizes and pore sizes Kiloprep cartridges are useful for all the following applications e Peptides e Oligonucleotides e Small synthetic molecules e Enantiomers e Natural products e Proteins Our cartridges offer the highest resolution and our dedicated packing facilitv ensures the highest possible reproducibilitv Biotage purchases media in large lot sizes We retain a sample of each lot to ensure consistencv and rapid deliverv and to offer complete analvtical HPLC laboratorv support to our preparative cartridge production team Kiloprep Cartridge Features e Prepacked pretested fully documented and ready to use e Maximum efficiency and peak symmetry Media fullv contained in cartridge so operators are never exposed to high activitv products or impurities e Radial compression technologv ensures a longer column life bv improving the column efficiencies e Cartridge changeover takes only minutes e Cartridges can be dedicated to an individual project then cleaned and reused Plates
123. lent of tosyl chloride media and are used for applications including scavenging alcohols amines si and other nucleophiles catch and release applications and synthesis tertiary amines esters and thioethers KP TBD 1 5 7 Triazabicyclo 4 4 0 Dec 5 ene silica Samplet cartridges are the silica bonded equivalent of 1 5 7 Triazabicylo ESS 4 4 0 Dec 5 ene media and are used for applications including scavenging of phenols and alkylation of phenols and amines LJ and esterification of carboxylic acids using alkyl halides KP NCO3 trimethylpropyl ammonium silica Samplet cartridges are silica bonded equivalent of trimethylpropyl ammonium N carbonate media and can be used for applications including s y 7 p neutralization of amine hydrochlorides and scavenging acids and acidic phenols KP KMNO4 potassium permanganate silica Samplet cartridges are silica absorbed with potassium permanganate T KMnO and can be used for applications including oxidation of alcohols FluoroFlash KP CF cartridges are for use in the separation and purification of perfluorinated compounds Fluorous compounds can be readily separated from nonfluorinated organic compounds or from each other based on fluorine content by chromatography using FluoroFlash fluorinated silica These separations capitalize on the unique feature of fluorous stationary phases to separate molecules primarily by fluorine content pe FLUOROUS Techn
124. let 5 Color Green Sp4B Sp1 Solvent Inlet 5 Color Gold Sp4B Sp1 Inlet Solvent for 3 16 In OD Tubing 20 um Porosity 316 SST 3 16 OD x 125 ID Teflon FEP Flangeless 3 16T x 5 16 24 Peek Natural Flangeless 3 16 Tubing Compression Module Outlet to Flow Path Module Inlet Sp4B Flow Path Module Outlet to Compression Module Inlet Sp4B 1 4 T X 1 8 T W Knurled Nut amp PFA Ferrules Flow Path Module UV Det Outlet to Flow Cell Inlet Sp4B 1 8 Flangeless 1 4 28 Peek Waste Outlet 6 Sp4B Set 1 8 316 SS 1 8 OD x 062 ID FEP 1 8inch Flangeless Fer 5 16 24 Peek Flangeless 1 8 Tefzel Collect Valve To Flow Cell Outlet amp Flow Path Module Waste Inlet Sp4B 1 8 T Short Flangeless 1 4 28 PEEK 1 8 Flangeless 1 4 28 Peek Blue 1 8 OD x 062 ID FEP 1 8 Flangeless 1 4 28 Peek 1 8inch Flangeless Fer 5 16 24 Peek Flangeless 1 8 Tefzel Part Number 08173 04794 04009 09321 09367 07482 07481 09653 09701 09472 05516 08192 06785 09470 09468 09664 01 09664 02 09664 03 09664 04 09376 09314 09311 09312 09665 09666 04144 09667 08399 09668 01037 00088 04795 04030 09669 08394 08372 00088 08399 04795 04030 SP4 Spare Parts continued Item Tube Assembly Ferrule Nut Tubing UV Detector Lamp Lamp 0 1 mm Flow Cell 0 3 mm Flow Cell Reducing Union Service Rates Item Hourly rate Off Site Service In House Service Daily rate Off Site Service Travel Charges Zone A Zone B Zone
125. ltiple samples and low solubility samples In a FLASH compression module Biotage FLASH cartridges tolerate liquid pressure up to 100 psig enabling higher flow rates the use of higher viscosity solvents such as those used in reversed phase chromatography and improved performance Biotage Packing Materials Biotage KP Sil Silica Features high surface area 500 m g moderate porosity 60 and a tight uniform distribution of particles sizes 40 63 um These three factors combined provide high loading capacity and efficiency KP Sil TLC Plates Biotage also offers KP Sil on TLC thin layer chromatography plates to assist in FLASH optimization and post chromatographic analysis Un LI di Mak a a a w E LE Pathe Doc gel Figure 1 Competitor s silica particle Figure 2 Biotage s KP Sil particle size size distribution distribution A more narrow particle distribution ensures consistent cartridge efficiency and performance FLASH Cartridge Packing Material Biotage KP C18 HS Features a high surface area 450 m g moderate porosity 90 A and a tight uniform distribution of particles sizes 35 70 um The media contains 18 carbon by weight and is end capped End capping is a silanization process designed to reduce the number of silanol sites that remain after bonding with C18 thus reducing irreversible binding interactions with polar compounds Biotage KP NH This is an a
126. lumes concentrations and phases and are characterized by reproducible results Because microwave synthesis occasionally uses uncommon methods the novel user might feel unsure of what conditions to use therefore Biotage has developed a database Biotage PathFinder which rapidly gives even novice users in the field a flying start toward the productivity increases inherent in microwave synthesis Biotage PathFinder is a Web based service featuring a unique microwave synthesis database including more than 3 800 carefully selected microwave reactions It gives chemists direct access to years of experience in microwave synthesis conducted on Biotage microwave systems delivered in a detailed easy to use and accessible format More information can be found in the Biotage PathFinder section on page 48 of this catalog or online at www biotagepathfinder com With more experience in microwave chemistry it is possible to translate conventional methods into microwave methods As previously mentioned reactions proceed faster using microwave synthesis simply because they are conducted at higher temperatures As a basic rule of thumb using the Arrhenius equation a 10 degree increase in reaction temperature doubles the reaction speed For example if your reaction took four hours at 60 C it will take approximately two hours at 70 C However keep in mind this new reaction temperature must be eval uated according to substrate and reagent stabilities as i
127. m KP PrNH 12 KP PrNH Samplet 25 KP PrNH Samplet 40 KP PrNH Samplet KP PrSH 12 KP PrSH Samplet 25 KP PrSH Samplet 40 KP PrSH Samplet KP TsOH 12 KP TSOH Samplet 25 KP TSOH Samplet 40 KP TSOH Samplet KP TsCl 12 KP TsCl Samplet 25 KP TsCI Samplet 40 KP TsCI Samplet KP KMn04 12 KP KMn04 Samplet 25 KP KMn04 Samplet 40 KP KMn04 Samplet KP TBD 12 KP TBD Samplet 25 KP TBD Samplet 40 KP TBD Samplet KP NCO3 12 KP NCO3 Samplet 25 KP NCO3 Samplet 40 KP NCO3 Samplet KP CF 12 KP CF Samplet 25 KP CF Samplet 40 KP CF Samplet Description 12 mm Syntage Samplet 24 case 25 mm Syntage Samplet 8 case 40 mm Syntage Samplet 4 case 12 mm Syntage Samplet 24 case 25 mm Syntage Samplet 8 case 40 mm Syntage Samplet 4 case 12 mm Syntage Samplet 24 case 25 mm Syntage Samplet 8 case 40 mm Syntage Samplet 4 case 12 mm Syntage Samplet 24 case 25 mm Syntage Samplet 8 case 40 mm Syntage Samplet 4 case 12 mm Syntage Samplet 24 case 25 mm Syntage Samplet 8 case 40 mm Syntage Samplet 4 case 12 mm Syntage Samplet 24 cse 25 mm Syntage Samplet 8 case 40 mm Syntage Samplet 4 case 12 mm Syntage Samplet 24 case 25 mm Syntage Samplet 8 case 40 mm Syntage Samplet 4 case 12 mm Syntage Samplet 24 case 25 mm Syntage Samplet 24 case 40 mm Syntage Samplet 24 case Part Number SAM PRNH12 SAM PRNH25 SAM PRNH40 SAM PRSH12 SAM PRSH25 SAM PRSH40 SAM TSOH12 SAM TSOH25 SAM TSOH40 SAM TSCL12
128. m x 30 cm KP Sil 150 mm x 60 cm KP C18 HS 75 mm x 9 cm KP C18 HS 75 mm x 15 cm KP C18 HS 75 mm x 30 cm KP C18 HS 150 mm x 30 cm 75 mm x 9 cm 75 mm x 15 cm 75 mm x 30 cm 75 mm x 9 cm 75 mm x 15 cm 75 mm x 30 cm 150 mm x 30 cm 150 mm x 60 cm 75 mm x 9 cm 75 mm x 15 cm 75 mm x 30 cm 150 mm x 30 cm 150 mm x 60 cm Qty Case 10 10 10 Ka a Ka a 10 10 10 PRP RP Ka Order FLASH Cartridges 150i Cartridge Ordering Information Part Number FKO 1107 19165 FKO 1107 19045 FKO 1107 19075 FKO 1107 19163 FKO 1107 19043 FKO 1107 19073 FKO 1107 25075 FKO 1107 25155 FLO 1118 19160 FLO 1118 19040 FLO 1118 19070 FLO 1118 25070 FT6 2030 19165 FT6 2030 19045 FT6 2030 19075 FT6 2030 19163 FT6 2030 19043 FT6 2030 19073 FT6 2030 25075 FT6 2030 25155 FT6 2530 19160 FT6 2530 19040 FT6 2530 19070 FT6 2530 25070 FT6 2530 25150 98 FLASH 75i and 150i Cartridges continued Cartridge C18 WP FLASH 75S FLASH 750M FLASH 75L FLASH 150M C4 WP FLASH 75S FLASH 75M FLASH 75L FLASH 150M Description 75 mm x 9 cm 75 mm x 15 cm 75 mm x 30 cm 150 mm x 30 cm 75 mm x 9 cm 75 mm x 15 cm 75 mm x 30 cm 150 mm x 30 cm Qty Case PPP Ka Ka Ka a Ka Part Number FV1 0906 19160 FV1 0906 19040 FV1 0906 19070 FV1 0906 25070 FV1 1104 19160 FV1 1104 19040 FV1 1104 19070 FV1 1104 25070 FLASH TLC Plates Ordering Information Cartridge KP Sil TLC Plates Glass
129. methods developed in other Biotage microwave synthesis systems can be directly transferred to the Advancer The product purities and yields are comparable or even improved This unique scalability enables the user to safely and rapidly translate optimized reactions to the larger scale With its 350 mL reaction vessel the Advancer serves all the chemist s needs for scaffolds and intermediates or substance for early toxicological and formulation tests Optimizing Microwave Synthesis Understanding Microwaves Microwave is a collective name for electro magnetic irradiation with frequencies in the range of 0 3 300 GHz To avoid interferences between the different applications it has been agreed that appliances for heating purposes will operate at 2 45 GHz corresponding to a wavelength of 12 2 cm Energy in the form of microwaves can be transferred to substances that are present in the beam line of the microwave radiation Absorption of the energy occurs when dipolar molecules rotate to align themselves with the fluctuating electric field component of the irradiation or when ions move back and forth by the same phenomenon Figure 1 VV ard Figure 1 Dipolar molecules and ions which try to move with an oscillating electric field It should be pointed out that the energy quantum of the microwave irradiation is totally inadequate for interacting directly with atom atom bonds or for exciting specific molecules When molecules rotate or move back
130. microwave synthesis offers other advantages such as simplicity and broader exploration Through superior heating features the Initiator is able to quickly achieve temperatures and pressures beyond the traditional reflux heating This allows chemists to perform complex reactions formerly not possible and is therefore the fastest growing technology in the pharmaceutical research laboratory Features and Benefits Compact footprint The Initiator is 45 smaller than its predecessor fitting easily into any standard fume hood space Touch Logic control Load and run your samples using a touch screen monitor for simple and intuitive navigation without the need for an external computer keyboard or mouse 0 2 to 20 mL without system modifications With the EXP function use four different vial sizes in any order or combination at any time without system modifications for greater flexibility and direct scale up of milligrams to grams Modular design Easily upgrade from a single sample manual format to an automated 8 or 60 position system Enhanced heating performance The new single mode applicator with the proven Dynamic Field Tuning feature offers faster heating of a broader range of solvents Best in class safety All Biotage microwave synthesizers are designed for safe operation at elevated temperatures and pressures The Initiator triple tier safety lock feature ensures maximum operator safety at all times 1 Point Support
131. mino functionalized silica with moderate surface area 230 m g moderate porosity 100 and uniform distribution of particles sizes 40 75 um This unique media accepts high sample loads of polar amine containing samples and minimizes amine silanol interaction FLASH Cartridge Features and Benefits e High surface area silica ensures high capacity and efficiency e Uniform particle size distribution generates narrow elution bands e High pressure 100 psig capability allows faster flow rates and provides increased throughput e Recessed inlet provides room for Biotage Samplet or Syntage cartridge and axial compression for higher throughput and resolution e Three diameters accommodate purification of milligram to multigram sample loads e Two cartridge lengths provide options for separating simple or complex mixtures e Three standard medias provide selectivity choices for optimal purification e FLASH silica available in TLC format improves optimization and analysis e Polyethylene cartridge barrels reduce product cost and are disposable e Cartridges meet 21 CFR 177 1520 regulations for extractables e Prepacked cartridges eliminate hazards of loose silica and are safer than glass columns 91 FLASH 40 Standard FLASH 40 Applications e Synthetic organic mixtures e Natural product isolation Minutes 4 e Purification of highly basic compounds e Protected peptides
132. mited instrumentation warranty is subject to the conditions set forth below 1 Biotage s obligation under this warranty is expressly limited at Biotage s sole discretion to the repair or replacement of the defective instrument part or item all other damage and claims statutory or otherwise being hereby expressly waived by the Customer Biotage may discharge this obligation in full either by repairing the instrument or by shipping F O B to the Customer the original site of delivery a similar instrument part or item provided however that as a condition precedent to this obligation the Customer must have notified Biotage during the warranty period of any alleged defect and provided further that Biotage must after examination of the part or item reasonably agree that the part or item is defective 2 This limited warranty is for the benefit of and applies only to the Customer 3 This warranty does not apply to any consumable items included in the System such as tubing fittings o rings and gaskets or any other part that comes in contact with the sample path 4 This warranty does not apply to any computer hardware delivered for use with the System as any such computer hardware will be subject to applicable manufacturers warranties 5 This warranty does not apply to failure or damage to the System incurred through neglect lack of maintenance misuse or accident or to improper installation runaway microwave reactions induced
133. mited to the implied warranties of merchantability fitness for a particular purpose and non infringement 5 LIMITATION OF LIABILITY Unless otherwise prohibited by applicable law Biotage s total liability whether for breach of warranty or contract tort including negligence or otherwise shall in no event exceed the amount paid by Customer to Biotage for the product or service alleged to have caused the damage to Customer In no event shall Biotage be liable to the Customer for any consequential or incidental damages including but not limited to damages due to loss of data or information or loss of use loss of profits or loss of revenue even if Biotage has been advised of the possibility of such damages nor shall Biotage have any liabil ity for any claim of any third party 206 6 SOFTWARE To the extent there is any software included with the Products the Software is being licensed to Customer not sold and all rights title and interest therein shall remain in Biotage or its licensors Use of the Software shall be in accordance with the software license delivered with the applicable Products 7 AGENTS STATEMENTS NOT BINDING No agent employee or other representative of Biotage has the right to modify or expand any of Biotage s standard warranties or to make any representations as to the Products or Services other than as may be set forth in any user documentation delivered with the Products and any such warranty or
134. mp and Cartridge Module Specifications Quad3 Pump Flow Range Pressure Limit Power Requirements Delivery Volume Stroke Quad3 Cartridge Module Dimensions Flow Range Pressure Limit Power Requirements Parallel Channels Cartridge Capacity Quad4 40 Rack Dimensions Parallel Channels Cartridge Capacity 60 mL min 60 psig 30 psig compressed air or other inert gas 3 mL 18 x 23 x 48 cm W x D x H 60 mL min 60 psig 30 psig compressed air or other inert gas 12 Up to 12 FLASH 12 M with or without Samplets 36 x 16 x 28 cm W x D x H 4 Up to 4 FLASH 40 M or S Quad UV Module and Quad Flash Collector Components Specifications Quad UV Dimensions Module UV Flow Cell Path Length Maximum Cell Pressure Power Requirements UV Source UV Wavelength Range UV Output Voltage Quad FLASH Dimensions Collector Power Requirements Quad4 40 Dimensions Rack Number of Parallel Channels Cartridge Capacity 32 x 54 x 33 cm W x D x H 0 1 mm 0 3 mm optional 150 psig 115 V AC 60 Hz or 220 V AC 50 Hz Deuterium Lamp 210 320 mm 1 AU full scale 48 x 46 x 33 cm W x Dx H 115 V AC 60 Hz or 220 V AC 50 Hz 36 x 16 x 28 cm WxDxH 4 Up to 4 FLASH 40 M or S Component Component Component Component Component Options l 2 3 4 5 For the Quad3 System A 13 mm Test A 13 mm Test Tube R
135. n 25 30 min 45 min 40 45 min 60 min Solvent A Solvent B VP gt 100mm Hg VP lt 10mm Hg Rules of thumb 1 Make sure samples will not degrade on silica or at selected temperature 2 Gradually increase vacuum without heat 1 No to low vacuum 760 650 mm Hg for VP gt 300 mmHg 2 Low to moderate vacuum 700 600 mm Hg for VP gt 100 mm Hg 3 Moderate to high vacuum 650 550 mm Hg with heat for VP lt 100 mm Hg 3 If heating set temp at 15 20 C below sol vent boiling point 4 Use Biotage heating blocks Syntage Cartridge Capacity Syntage Cartridge Capacity Samplet Cartridge Cartridge Vol mL Cartridge Flow Rate Loading mL min mmol cartridge KP PrNH 12 1 0 26 0 40 10 15 KP PrNH 25 5 1 3 2 1 20 50 KP PrNH 40 10 3 1 4 8 30 100 KP TsOH 12 1 0 26 0 40 5 10 KP TsOH 25 5 1 3 2 1 15 25 KP TSOH 40 10 3 1 4 8 30 50 KP PrSH 12 1 0 21 0 31 5 10 KP PrSH 25 5 1 1 1 6 15 25 KP PrSH 40 10 2 5 3 7 30 50 KP TsCL 12 1 0 26 0 40 5 10 KP TsCL 25 5 1 3 2 1 15 25 KP TsCL 40 10 3 1 4 8 30 50 KP TBD 12 1 0 63 0 77 5 10 KP TBD 25 5 3 51 4 29 15 25 KP NCO3 12 1 0 51 0 62 5 10 KP NCO3 25 5 2 81 3 63 15 25 KP NCO3 40 10 4 05 4 95 30 50 KP KMNO4 12 1 0 82 1 0 5 10 KP KMNO4 25 5 4 56 5 58 15 25 KP KMNO4 40 10 10 53 12 87 30 50 At 40 C 107 Syntage Chemistry Cartridges Ordering Information Ite
136. n all chemistry The reaction temperature and time can be easily converted using the prediction chart provided below in Table 1 Table 1 Time Prediction chart Change in field color represents change in unit hour minutes seconds relevant to your starting unit For example if your reaction took 6 hours at 100 C in this instance white hours it will take Prediction Chart approximately 5 minutes at 160 C blue minutes see red numbers in the table above 30 15 30 8 15 30 45 4 8 15 23 30 45 2 4 8 m 15 23 45 2 EE RES T1 23 45 2 3 a 6 11 23 45 6 11 23 40 1 3 6 11 20 1 3 6 10 1 3 5 it 53 4 1 3 0 26 53 1 7 13 26 40 53 3 7 13 20 26 40 2 3 7 10 13 20 40 1 2 3 5 7 10 20 40 1 2 2 3 5 10 20 40 1 1 2 2 5 10 20 35 1 1 2 5 10 18 1 1 2 5 9 1 2 4 With courtesy of David Rudge AstraZeneca Macclesfield UK 20 C PTIM ON Practical Tips and Tricks When Performing Microwave Synthesis Solvent Different solvents interact very differently with microwaves because of their diverse polar and ionic properties Acetonitrile DMF and alcohols are often used for microwave assisted organic synthesis You might not need to change from the solvent that is specified for the reaction under traditional chemistry conditions First try using th
137. nal Electrical Code requirements Class I Division 1or 2 Groups C amp D for explosion proof service e 316 SS ASME rated compression module and radial compression tank s 304 SS frame with industrial style casters e SS overbraided Teflon lined tubing e 316 SS valves with Teflon seats Calibrated pressure gauges and pressure relief valves SS stainless steel SIM 10000 Specifications 100 psig Flow 1 7 L min Temperature 5 80 C Weight 440 Ib 200 kg Dimensions 34 x 26 x 54 Lx W x H SIM 10000 Cartridges and Frits SIM 10000 Ordering Information Item Description Part number SIM 10000 Sample Injection Module SIM 10000 SIM 10000 Cartridge Includes two tubes with bottom frit FTF 10000 assemblies and four frits for packing 177 Kiloprep Preparative HPLC Systems Kiloprep Preparative HPLC Systems Kiloprep is a preparative HPLC system combined with prepacked cartridge technology to offer the highest resolution of any available preparative purification technique Biotage offers systems and cartridges that in a single run will purify quantities of material ranging from milligrams to kilograms Customers using solid phase synthesis to produce peptides or oligonucleotides will need this high resolution to reach their purity goals Customers who need chiral separations or have closely related small molecules from synthesis fermentation or natural products also can benefit from
138. nd Samplet Cartridges For use with FLASH 25 S and Samplet Cartridges For use with FLASH 25 M and Samplet Cartridges For use with FLASH 40i and Samplet Cartridges For use with FLASH 40 S and Samplet Cartridges For use with FLASH 40 M and Samplet Cartridges For use with FLASH 40 S and Samplet Cartridges For use with FLASH 40 M and Samplet Cartridges Set of 12 M 25 M 40 M Compression Modules and 12 S 25 S 40 S Barrels Adapters and tubing to allow use of FLASH or i style compression modules with ISCO systems Part Number FIV VLV 1000 FN 001 41201 FN 004 04000 09350 07923 FC022 15024 07857 07858 07395 07383 FC 022 16024 FC 022 16044 FB 022 16024 FB 022 16044 FC022 17024 07975 07976 07387 07388 08615 09335 FLASH 65i FLASH 65i Compression Module The FLASH 65i compression module for 65 mm cartridges now incorporates the Zero Insertion Force ZIF sealing mechanism which enables effortless insertion and removal of cartridges and reduces O ring wear The new ZIF head assembly improves bed stability efficiency and sample loading by applying axial compression to the cartridge ZIF technology also allows increased sealing pressures up to 100 psig providing faster flow rates and eliminating the leaks due to variations in bed heights The FLASH 65i compression module can be used with the Horizon HPFC system for automated purification or with our new FLASH stand an
139. ng a single paraben methyl ethyl propyl or butyl were prepared and loaded into alternate wells in two 96 well plates methyl and ethyl in plate one propyl and butyl in plate two Each plate was loaded onto a two channel Parallex Flex and the 1 mL samples from each well of the entire plate were injected Collected fractions were analyzed using an analytical HPLC system to determine the amount of cross contamination The results in Table 1 show cross contamination levels of less than 1 Compound Average Deviation Number of Samples Methyl Paraben 0 34 0 15 0 68 47 Ethyl Paraben 0 70 0 05 1 49 47 Propyl Paraben 0 49 0 35 0 76 47 Butyl Paraben 0 12 0 02 0 29 47 Table 1 Cross contamination levels for Parallex Flex HPLC Sample recovery was determined using a mix of ethyl and butyl parabens with threshold and slope settings of 0 01 The samples were separated using gradient elution on a 20 x 50 mm C18 column at 20 mL min Collected fractions were analyzed by analytical HPLC with the results listed in Table 2 Recovery levels exceeding 96 were achieved using the Parallex Flex Compound Average Standard Deviation Number of Samples Ethyl Paraben 96 4 0 07 24 Butyl Paraben 97 6 0 05 24 Table 2 Fraction recovery for the Parallex Flex HPLC Application Note 38 Continuous Gradient Purification of Closely Related Drug Intermediates Using FLASH Chromatography By Jack Liu
140. ng requirements of the National Electrical Code MP 8000 Features e Diaphragm pump 0 8 8 0 L min at 150 psig e 316 SS static mixer s 304 SS frame and NEMA X control cabinet explosion proof e Dual wavelength UV detector e 8 port fraction collector e 4 color touch panel and programmable logic controller e Optional Coriolis flow meter SS stainless steel MP 8000 Ordering Information Item Description Part Number MP 8000 Gradient MPLC System Custom Automation system for FLASH 400 Optional configurations for the MP 8000 XP include in line monitors for pH conductivity and or gradient density To discuss your specific automation needs for FLASH or High Pressure Liquid Chromatography call Biotage 1 Point Support US 1 800 446 4752 EU 46 18 56 59 11 JP 81 422 28 1233 SIM 10000 SIM 10000 Production scale Sample Injection Module The Biotage SIM 10000 is a skid mounted sample injection module designed for use with a FLASH 400 system The SIM 10000 module holds disposable 10 L cartridges to be filled with your media Sample injection modules are essential for the successful purification of low purity and or low solubility materials SIM 100005 are designed to ensure maximum recovery of your valuable compounds The SIM 10000 is built with industrial grade components that are appropriate for operations under U S Food and Drug Administration regulations for current Good Manufacturing Practices cGMP and meet Natio
141. nitrile to the cartridge Dry the Syntage Samplet using a Syntage heating block in a vacuum oven for five hours at 80 C Remove the cartridge from the oven and allow it to cool to room temperature before placing it into a FLASH 25 S KP Sil purification cartridge Elute the alkylated phenol using an appropriate elution profile o Kes C POE g TBD CY DN TA 80 C Vo JU 1000 ho FA Cartridge FLASH 25 S KP Sil 900 X3 Solvent A DCM 800 o Ne B DCM MeOH 9 1 700 I l J ma TS Gradient Program Step 1 0 50 B in 10CV 400 Step 2 50 100 B in 10CV 300 Flow Rate 25 mL min 200 100 m E T Collected Fraction 15 mL 0 0 il 2 Minutes 3 4 5 36i Conditions M Column YMC C8 S 3 120 4 6 x 50 mm 2 00 4 ME Injection 3 mL he j Mobile Phase M aet A H20 ACN 95 5 0 1 TFA E Purified B ACN H20 95 5 0 1 TFA Gradient B 5 100 in eight minutes hold two minutes Flow Rate 1 5 mL min Detection 254nm 6 00 7 00 8 00 9 00 10 00 Minutes Samplet chemistry cartridges liquid handler Features and Benefits Syntage heating blocks are designed to enhance solvent evaporation and cartridge based reactions with Syntage Syntage heating blocks improve heat transfer to the cartridge when used in a vacuum oven thereby decreasing solvent evaporation time These modular blocks are constructed of solid anodized aluminum for maximum heat retention and to pr
142. nsertion tool Chemraz 111 O ring Chemraz 109 O ring ZIF SIM 35 Spare Parts Item Frit Insertion Tool Upper O ring Lower O ring Description Polypropylene frit insertion tool Chemraz 208 O ring Chemraz 206 O ring ZIF SIM 60 Spare Parts Item Frit Insertion Tool Upper O ring Lower O ring Quad Spare Parts Item Filter Kit Reducing Union Inlet Filter Packs Quad Check Valve 1 8 Fitting Kit 1 8 PEEK Nut 1 8 TEFZEL Ferrule Description Polypropylene frit insertion tool Chemraz 211 O ring Chemraz 209 O ring Description Replacement filters 1 4 x 1 8 reducing union 4 required per set solvent inlet filters 12 1 4 x 1 8 reducing union 4 required per set Solvent inlet filters 12 Check disc 1 8 PEEK flangeless fittings 1 8 PEEK nut 1 8 TEFZEL ferrule 1 8 PEEK nut 1 8 TEFZEL ferrule Part Number 08771 08856 08857 04144 Part Number 03024 01903 Part Number 0087 00656 06176 Part Number 0088 06178 06177 Part Number 0184 08555 06221 Part Number QFILT 0000 04144 01801 05516 QFTT 4795 04795 04030 Quad Spare Parts continued Item 1 16 Fitting Kit 1 16 PEEK Nut 1 16 TEFZEL Ferrule Upper O Ring Kit Chemraz 108 O ring Chemraz 107 O ring Retaining Clip Lower O Ring Kit Description 1 16 PEEK flangeless fittings 1 16 PEEK nut 1 16 TEFZEL ferrule 1 16 PEEK nut 1 16 TEFZEL fer
143. nt method Software Features 129 Specifications Solvent delivery Flow rate Pressure limit Liquid sample loading Solid sample loading UV detection Flow cell path length Fractionation modes Collection vessels Power System control amp data management Dimensions Certifications Constant volume 3 mL electric HPFC pump 1 100 mL min 100 psi 7 bar Optional 3 way liquid injection valve Samplet sample loading cartridge ZIF SIM Choice of variable dual wavelength 200 320 nm or fixed 254 nm detector Choice of 0 1 mm or 0 3 mm Collect all UV threshold UV slope Test tubes mm 13 16 17 18 and 25 Bottles mL 120 and 240 110 120 VAC 60 Hz US Japan 220 240 VAC 50 Hz On board Pentium computer with 10 4 diagonal touch screen interface and Windows XP based software 19 x 23 x 24 WxDxH 48 cm x 58 cm x 61 cm CE CSA certified Ordering Information SP System and Product Components Ordering Information Systems SP1 Part Number Base System Country Certification SP1 X0A No detector US CSA SP1 XOB No detector UK CE SP1 X0C No detector EU CE SP1 A1A Fixed Wavelength 0 1 mm cell US CSA SP1 A1B Fixed Wavelength 0 1 mm cell UK CE SP1 A1C Fixed Wavelength 0 1 mm cell EU CE SP1 A2A Fixed Wavelength 0 3 mm cell US CSA SP1 A2B Fixed Wavelength 0 3 mm cell UK CE SP1 A2C Fixed Wavelength 0 3 mm cell EU CE SP1
144. oad 300 samples at 5 00 p m and return at 8 00 a m with assurance of completion The Flex V3 system can process from 1 mg to 400 mg per injection The automated loader can handle plates vials and tubes without reconfiguration Proven Reliability The Flex V3 system constantly monitors solvent levels and fraction collection tubes and is capable of full recovery after a power loss without losing any samples A robust intelligent UV guided fraction collection system can be tailored to minimize the number of fractions or to maximize yield Intelligent algorithms patent pending ensure fractionation of unresolved components thereby yielding higher purity fractions The Flex V3 system fully interfaces with the Water s ZQ for on line mass characterization Flex V3 Operating Software Simple Software The Flex V3 system is designed for walk up walk away operation The simple user interface enables a chemist to load and run a sample in four easy steps from a single software screen Figure 1 Simple yet robust this system software is suited for a multiuser environment where chemistry is the focus Figure 1 Load a sample in four easy steps Step 1 Click on the well Step 2 Choose your method Step 3 Name your sample Step 4 Click on the run button Figure 2 Single screen user interface representing the sample injector top left fraction collectors top right and selectable sample status table bottom
145. ologies Incorporated Fast Pure and Simple Under license from Fluorous Technologies Inc FluoroFlash is a trademark of Fluorous Technologies Inc Application Specifications Figure 7 KP PrNH Representative Scavenging Procedure Flash purification of a synthetic sulfonamide using Syntage PrNH25 Samplet cartridge FLASH 25 M silica cartridge and the following method on a Horizon HPFC system The PrNH cartridge removes excess Tosyl chloride simplifing the purification s SO CI LT 1 5 eq CI TEA DCM 074 is 7 NH N S o Impurities Product f J i Solvent A EtOAc Hexane 20 80 l Solvent B EtOAc B L im 1 Gradient 0 100 B in 500 mL linear S m Flow Rate 35 mL min 2 0 UV Absorbency o S o 10 20 30 40 50 60 7 0 80 Minutes Figure 8 KP TsOH Representative Catch and Release Procedure A four component mixture of differing pKa values was dissolved in DMF and injected directly onto a TsOH12 Samplet and FLASH 12 M cartridge combination using the gradient method shown below The elution order of the nitrogen containing compounds is consistent with increasing pKa values rather than polarity phenol is not bound to TsOH Increasing pKa Sample 0 25M total concentration of phenol 4 nitroaniline 4 chloroaniline and caffeine dissolved in DMF Cartridge FLASH 12 M KP Sil Solvent A Hexane Solvent B 1 TEA
146. olume you can manually collect fractions using the displayed UV response as your signal to start collecting a new fraction The UV module comes standard with a 0 1 mm path length flow cell and monitors absorbance at 254 nm The absorbance is displayed recorded and saved electronically and can be downloaded to a PC for archival purposes This free standing detector recorder module can be used with any flash purification system including Biotage FLASH 751 systems using the provided 1 8 tubing kit Simply attach the cartridge outlet to the flow cell inlet and connect the other tube between the flow cell outlet and the collection vessel no tools are required for installation Specifications Utilities 110 120 VAC 60 Hz US CSA certified 220 240 VAC 50 Hz CE certified Fuses 0 5 A 250 V main 230V input power 0 1 A 250 V main 100 115 V input power Agency certification CE and CSA UV lamp Mercury 254 nm Path length 0 1 mm standard 0 3 mm optional Cell pressure rating 300 psig 20 bars Recorder RAM 1MB Biotage FLASH UV Size 10 25 cm W x 19 48 cm D x 7 18 cm H detector recorder module FLASH 75 Ordering Information Item Description Part Number FLASH 75 Systems FLASH 75S System 75S compression module air manifold SIM100 SF 022 19161 4 L solvent reservoir tubing grounding kit and manual FLASH 75M System 75M compression module air manifold SIM500 SF 022 19041 12 L solv
147. on 150M compression module air manifold SIM1000 37 L solvent reservoir tubing and manual 150L compression module air manifold SIM2000 60 L solvent reservoir tubing and manual 150M powder coated aluminum barrel with stainless steel endcaps fitted on a portable base V band clamps ASME rated relief valve 150L powder coated aluminum barrel with stainless steel endcaps fitted on a portable base V band clamps ASME rated relief valve Powder coated aluminum barrel with pressure relief valve uses FLASH 150M cartridges Powder coated aluminum barrel with pressure relief valve uses FLASH 150L cartridges Stainless steel sample injection module ideal for loading low solubility samples or liquid volumes up to 1000 mL Stainless steel sample injection module ideal for loading low solubility samples or liquid volumes up to 2000 mL 25 x 3 frits for either the SIM1000 or SIM2000 25 frits and sealing rings for SIM1000 and SIM2000 Grounding kit includes Teflon coated wires and clips to dissipate static charges 3 way 1 4 injection valve for direct liquid injections complete with Luer Lok injection port 37 L stainless steel solvent reservoir ASME rated with solvent shut off and relief valves 60 L stainless steel solvent reservoir ASME rated with solvent shut off and relief valves Air manifold safely regulates the flow of air pressure and solvent s to FLASH 150 systems 1000 mL SIM cartridge e
148. on work up is a major bottleneck in the synthetic process Generally the work up steps of an 64 organic reaction take considerably longer than the actual synthesis The major impediment with solution phase synthesis is the multiple aqueous work up steps often necessary to remove excess reagent and byproducts when isolating desired compounds It is often common for some reactions to require four or five aqueous extractions followed by drying and filtration Although these extractions are easy to perform they require additional chemist interaction and delay the goal of obtaining pure compound for further study In addition the formation of emulsions during these extraction wash steps causes lower yields and increases purification time However solution phase synthetic techniques offer many advantages over solid phase approaches including unlimited scale easy manipulation and reduction in validation time By planning the solution phase synthesis to include a Flash chromatography purification step the need for a post reaction work up is eliminated To illustrate the advantages of incorporating high performance flash chromatography HPFC into a solution phase synthesis reaction procedure a polypeptide Fmoc Tyr t Bu Gly Gly Phe Leu NDMCP was synthesized by an eight step solution phase technique using the Horizon HPFC system inline for fast product isolation Biotage s Horizon HPFC system combines gradient elution capability and UV
149. onents to suit your requirements Easy Setup Setting up a FLASH system is fast and easy To begin your separation simply insert a FLASH cartridge into the chosen compression module and attach the patented Zero Insertion Force ZIF head assembly For liquid samples inject directly onto the head of the cartridge using a suitable sized syringe For low solubility samples or difficult separations use our Samplet sample loading cartridges see page 88 for full description or a Zero Insertion Force Sample Injection Module ZIF SIM see page 118 for full description Turn the valve to let the solvent flow and collect your fractions by placing a tube rack under the outlet line Applications FLASH systems simplify and accelerate the isolation of organic compounds Typical applications involve purification throughout multistep solution phase synthesis in which there is high probability of complex mixtures The scale of purification ranges from milligrams to grams for simple and complex mixtures see page 50 for more detailed application information Available as a Complete System or as Individual Components FLASH systems include everything except the cartridges a solvent reservoir a stand a ZIF SIM and a compression module with short and or medium barrel s Other options include upgrades to larger compression modules for scale up to gram scale purification and a pressure regulator for controlling flow rates
150. or USB port 18 32 C 64 4 89 6 F 20 95 RH 220 240 VAC 50 Hz 16 EU 208 VAC 50 60 Hz 2 US JPN gt 60 lit min 5 10 bar 1 3 lit min lt 450 kg 985 Ibs 55 x 26 x 73 WxDxH 140 cm x 65 cm x 185 cm Two standard Upchurch Scientific HPLC connections and one Swagelok connection Ordering Information The Biotage Advancer features several vessel entry points to facilitate in situ sampling and the addition of reagents during a run The Biotage Advancer features a Teflon 350 mL reaction vessel insuring no cross contamination and easy clean up Ordering Information Item Description Part Number Advancer Instrument 2 Reaction Vessels 50 300 mL with Cap thick walled 354853 Teflon vessel for repetitive usage Infrared Temperature Probe Temperature Probe Pocket Pressure Sensor Reaction Collection Package includes an electrically driven Collection Valve connecting Collection Tubing and Collection Vessel for receiving cooled reaction mixture HPLC port consisting of fittings tubing and a manual valve for facilitating the connection of external hardware that can aid in sampling from or adding content to the reaction vessel Biotage Advancer Software 1 4 license Pd patent License Advancer O ring Kit Overhead Propeller Stirrer Biotage Advancer Tool Kit Installation and training See page 11 for more information on how to request a quote
151. or for ordering instructions Biotage Microwave Vials 44 4 exclusively designed microwave vials support Microwave Synthesis from mg to grams without the need for re optimization Simplicity Simplicity is one of the benefits of modern microwave equipment Reactions are performed in glass vials sealed with caps and heated in the microwave cavity Chemists can quickly analyze reactions by sampling through a septum Biotage Microwave Vials are designed to support microwave synthesis from mg to g without the need for reoptimization With the Initiator EXP system methods that are run at a lower volume are directly trans ferrable across the entire volume range of 0 2 20 mL Magnetic Stir Bars The reaction mixture is continuously stirred by magnetic stirring during the heating process promoting even heating throughout Optimum Vial Sizes Each Biotage Microwave Vial has been designed for safe and efficient heating within its specified volume range Together the four vial types provide full scalability within the volume range of 0 2 20 mL Multi Gram Scale Migrate directly to multi gram scale without reoptimization using the 10 20 mL vials These larger vials can also be used for preparation of scaffolds and intermediates or for generating larger quantities of active compounds for testing Choose the vial size that works best for your reaction Biotage recommends that reaction volumes do not fall below or excee
152. osphine Ph3P in THF The reaction mixture was stirred for three hours at room temperature and was transferred to a Syntage 12 KP NCO3 cartridge dried on a Syntage Samplet heating block inserted into a FLASH 12 M silica cartridge and purified using the following method on a Horizon HPFC system OH PhsP DEAD OLD Ph P 0 Excess THF 9 NO Compound 5 NO OH Excess Flash Chromatography purification of Compound 5 with an 89 yield and HPLC purity of 98 Compound 5 Product Solvent A EtOAc Hexane 20 80 Condition Isocratic Flow Rate 12 mL min HPLC of reaction FI A i a et mixture and isolated product Ph PO So 1 Reaction mixture N HPLC conditions Dj Column PVA 4 6 x 150 i Eluent A EtOAc Hexane 20 80 2 Scavenging B EtOAc Gradient 0 100 B in 10 min lu i Flow Rate 2 mL min WW Detection UV 254 nm excess nitrophenol by KP NCO3 cartridge Lae 3 Scavenging flash chromatography l Application Specifications Figure 13 KP TBD Representative Scavenging Procedure Typical procedure for phenol alkylation Williamson ether syn thesis involves choosing the appropriate Syntage cartridge size such that the phenol is 1 equivalent of the loading capacity of the cartridge Dissolve 1 mmole of phenol in 0 5 mL toluene and transfer the dissolved phenol to the Syntage KP TBD 25 cartridge Apply 1 mmole of alkyhalide dissolved in 0 5 mL aceto
153. otage s standard operating procedures Should any Biotage consumable fail to meet the limited warranty above after being tested in accordance with the applicable Biotage standard operating procedures Biotage will provide at its sole option either a replace ment cartridge or reaction vial If a cartridge it will be packed with the original media or packed with new media at no cost to the Purchaser If such failed cartridge is packed with media provided by the Purchaser Biotage will make a reasonable effort to re pack the original media or pack a replacement cartridge with new media provided by the Purchaser Returns of any Consumable must be authorized in advance Please contact Biotage for a Return Authorization RA number and shipping instructions All claims must be made within thirty 30 days of shipment from Biotage or after the initial test of the cartridge at the Customer s facility whichever is first Special Request and Custom Packed Media Cartridges Biotage provides a limited warranty for each of its Custom Packed Cartridges and Special Request Cartridges as is set forth in the applicable custom order quotation furnished to the Customer Biotage warrants that Biotage Custom Packed cartridges and Special Request cartridges will be of good quality and workmanship Each cartridge is packed to a density specification unique to each media and Biotage war rants the cartridge dimensions and weight of the packed media and speci
154. ovid ed a faster process for a multiplestep synthesis that was easily completed in a single day Application Note 44 On Column Clean Synthesis of a Substituted Benzofuran Using Syntage TsOH Cartridges By Shahnaz Ghassemi Ph D Product Syntage KP TsOH cartridges Abstract There is a need to develop a simple fast and flexible method to generate substituted benzofuran libraries that are utilized as potential antioxidants and drug candidates This work illustrates an on column synthesis of substituted benzofuran using the Syntage KP TSOH Samplet cartridge for on column synthesis of a heterocyclic benzofuran ring system 2 starting with phenoxyacetophenones 1 Figure 1 O Rs cor bl gt R R 1 Rs Flash 1 chromatography Figure 1 Synthesis of Substituted Benzofuran using the Syntage KP TsOH cartridge A Syntage KP TsOH Samplet cartridge is a silica bonded p toluenesulfonic acid with a high surface area media that exhibits excellent retention of basic compounds This reagent is useful for cyclodehydration reactions as well as scavenging amines The Syntage KP TsOH Samplet cartridge is sufficiently acidic to close the heterocyclic ring of organic halides and for the high throughput synthesis of aryl triflates and aryl nonaflates Silica bound TsOH is also routinely applied in a catch and release protocol whereby basic species are caught by the sulfonic acid and then subsequently released during the
155. ovide uniform cartridge heating Each anodized block is Teflon coated for added chemical resistance Syntage heating blocks are designed for use within a standard vacuum oven or on top of a thermocouple controlled hot plate Each heating block has one thermometer and one thermocouple well to ensure accurate temperature control and monitoring Syntage heating blocks are machined to standard microtiter plate dimensions for easy integration with an automated e Anodized and Teflon coated solid aluminum blocks ensure uniform cartridge heating and chemical resistance e Improved heat transfer to Svntage cartridges to decrease drying time e Protective sleeves available to minimize block contamination Available in 12 25 and 40 sample loading cartridges Microtiter plate dimensions for easy integration with an automated liquid handler Recommended Drying Times Solvent Boiling Point Vapor Recommended C Pressure Drying mm Hg Temperature C Acetone 56 181 35 40 Acetonitrile 82 72 4 60 65 T Butyl methyl ether 52 249 30 35 Chloroform 61 155 1 40 45 Cyclohexane 81 78 60 65 DCM 40 341 3 20 25 Diethyl ether 35 434 4 15 20 Ethanol 78 41 4 55 60 Ethyl acetate 77 72 4 55 60 Hexane 69 129 3 50 55 Methanol 65 93 1 45 50 Methyl acetate 56 173 35 40 THF 66 129 3 45 50 Size Solvent A Solvent B 12 20 min 30 mi
156. ows chemists to automatically calculate the Vapor pressure for some common solvents at various temperatures Biotage Pathfinder Cookbook A free feature that introduces all users to a sample of the valuable information available on PathFinder Chemists may quickly browse through a selection of popular reactions and obtain reaction details om om e oe ee do d de Se 8 Ad E hte www biotagepathtinder com Pt en ST Bee m a BEI DDR Lond ee em m L uA mum mm ac as ga cm 1mm mam WWW biotagepathfinder com Data Content The data content is continually updated with new chemistries and currently contains reactions from Biotage s highly skilled internal application chemists and select contributions from our renowned Scientific Partnership Program SPP To experience the benefits first hand visit WWW biotagepathfinder com Figure 1 Enter a simple substructure search I i J Figure 2 Search results are displayed in an easy to browse format Figure 3 Reaction details needed to perform reactions are listed in an easy to read format Ordering information Item Description Part Number Biotage Pathfinder Web Single Seat Annual Subscription 355239 Biotage PathFinder Web Free 1 Month Trial of PathFinder 355241 49 Published Microwave Reactions Solid Phase Microwave Reactions pg 52 Solid Supported Reagents pg 53 Polymer Supported Micro
157. poorly Heptan amp ez resolved from its major impurities A and C eptane In dichloromethane the retention of impurities Isooctane A and C has been dramatically altered providing a better purification of B Table 2 Solvent selectivity chart 3 Solvent strength optimization When the correct solvents have been determined the next step is to adjust the solvent composition solvent strength so the compound of interest elutes within the Rf range 0 15 0 35 6 7 2 8 CV By adjusting solvent strength to provide elution within this window the chances for optimal purification are greatly enhanced L R Snyder and J J Kirkland Introduction to Modern Liquid Chromatography Wiley 1979 83 FLASH PURIFICATION As with selectivity each solvent has its own polarity Table 3 Each solvent mixture or mobile phase then has its own unique solvent strength Calculation of a solvent mixture s strength is useful for comparison to other solvent mixtures Solvent mixtures with the same strength but different selectivity can then be evaluated To bring the Rf of the compound of interest into the optimal range reduce the amount of polar solvent in the mobile phase As an example Figure 4 the results of a solvent selectivity study show a mobile phase of 50 hexane and 50 ethyl acetate solvent strength 0 30 provides adequate selectivity for a crude sample Figure 4 top The Rf for the compound of interest B is 0
158. port Telephone 81 422 28 1233 E mail 1 pointsupport biotage co jp Synthesis Solutions and Optimization Microwave Synthesis Solutions Biotage Microwave Vials Biotage exclusively designed microwave vials support microwave synthesis from mg to grams without the need for re optimization Biotage microwave vials are uniquely designed to fit into all Biotage Initiator microwave synthesis systems These vials allow safe convenient and reproducible syntheses from 0 2 mL to 20 mL Methods that are synthesized at a lower volume are directly transferable across the entire volume range Initiator Microwave Synthesis System The Initiator microwave synthesizer enables medicinal chemists to quickly synthesize compounds using microwave heating Through superior heating features the Initiator is able to quickly achieve temperatures and pressures beyond the traditional reflux heating The base system is easily upgradable to an 8 or 60 position sample bed that supports the production of focused libraries multiuser environments and scale out The Initiator 8 and 60 provide flexible operation that enables the use of both large and small vials in combination at any time and in any order without manual intervention Advancer Advancer is designed for reliable and reproducible scale up of microwave assisted synthesis handling reaction volumes from 50 mL to 300 mL at temperatures from 60 250 C and pressures of up to 20 bar Synthesis
159. rams of media are built to Routinely operating at a flow rate of 250 mL min the FLASH 75i systems and FLASH 75 cartridges allow you to scale up and quickly complete runs saving hours even days of purification time pera a orn On Fi E ED IL om 0 AT ver C nnllam Zs n mE AM 190 Air Manifold Syringe Injection House Air Solvent FLASH 751 Reservoir Radial 4 L or 12 L Compression Module SIM Injection 100 mL or 500 mL Collect il if Fractions Figure 1 Purify up to 40 grams of synthetic compound on our FLASH 75i system FLASH 75 Specifications Cartridge Dimensions Sample Size Flow RatE mm ID x cm L g mL min FLASH 75 S 75x9 0 2 10 250 FLASH 75 M 75 x 15 0 4 20 250 FLASH 75 L 75 x 30 0 8 40 250 SIM sample injection module for low solubility samples or viscous oils SIM Volume mL 100 500 500 Reservoir Volume L 4 12 12 Flash 75 Media Variety Safer than Glass Columns FLASH 75 cartridges are constructed of rugged medium density polyethylene and resist cracking and splitting There is no breakable glass and all of the silica is completely self contained eliminating any exposure to silica dust or contaminants Cartridges are Available with a Variety of Media s KP Sil 40 63 um 60 silica e KP C18 HS 40 63 um 90 C18 bonded silica e Mitsubishi Diaion HP20 and H
160. rate C inlet Sample Loading C inlet on main pump Main Pump Three head double diaphragm Instrumentation Dual channel variable wavelength UV detector Precolumn pressure transducer Flow Path Pulse dampener Gradient mixer Fraction Collection Waste or collect valve Peak detection Mode Time UV threshold UV slope Options 3 120 L ASME Tertiary gradient A B C Isolated sample pump Flow meter with feedback control pH meter Conductivity meter Densitometer Thermometer Column delta pressure Filter header with selections for e Filter or Bypass e Filter 1 or Filter 2 Column header with selections for e Forward or Reverse flow e Column 1 Column 2 e Bypass Recycle valve Heat exchanger 12 port rotary valve Purge for high recovery Boolean combinations of parameters 184 KILOPREP Kiloprep 2000 The Kiloprep 2000 is a high throughput skid mounted HPLC system designed for use in a pilot plant or full scale production environment It is available for use in areas that are nonhazardous or explosion proof as defined in the National Electrical Code Class I Division 1 Groups C amp D These systems are used to produce multigram to kilogram quantities of peptides oligonucleotides chiral enantiomers or natural products Kiloprep 3000 4000 The Kiloprep 3000 and 4000 are high throughput HPLC systems with a control skid and a pump skid designed for a full scale production environment They
161. re that provides the largest ACV between the compound of interest and all the impurities All solvents fall into a selectivity group Each group has a different impact on a sample component s relative retention to another compound selectivity In Table 2 the most frequently used flash solvents and their selectivity groups are shown When possible selectivity optimization should include mixtures of hexane with ethyl acetate VIa methylene chloride V toluene VII tetrahydrofuran III and ether I For more polar compounds and amines mixtures of methylene chloride V with methanol II or acetonitrile VIb should be evaluated These solvent combinations provide a broad range of separation selectivity and will help define the correct solvents for a sample s purification Figure 3 For more discussion regarding solvent selectivity in chromatography see Introduction to Modern Chromatography by L R Snyder and J J Kirkland Solvent Selectivity Group Solvent Front Solvent Front Ether I Methanol II Ethanol II A e E B Isopropanol II ec S 2 Tetrahvdrofuran III Dichloromethane V 7 e A Acetone VIa Ethyl acetate VIa Origin Origin Acetonitrile VIb Hexane EtOA c Vla 2 1 Dichlorome thane V Toluene VII Chloroform VIII Figure 3 Impact of solvent selectivity on a chromatographic separation In hexane ethyl Hexane acetate the compound of interest B is
162. reaction performed on a dry solid support or in a continuous flow system When trying to understand these questions the introduction of the so called cooling while heating technique is very misleading This technology proposes the reaction mixture is heated as the outside of the vessel is simultaneously cooled hypothetically increasing the energy input However since the temperature is normally read on the surface of the vial the accuracy of the temperature measurement is lost along with reproducibility controllability and predictability The temperature difference of measured and actual temperature is strongly dependent on the microwave absorbing properties of the sample the temperature of the cooling gas the velocity of the cooling gas the thickness of the vial wall etc and can easily be as much as 50 C Figure 2 Cooling on i Internal and outer surface vial temperature vs time Inner temp Temperature mot difference gt 40 C m 5 mL EtOH Surface temp Temperaturei C Figure 2 Difference of measured and actual temperature fina 7 in 5 mL of ethanol The actual temperature is measured via oO FI r7 TY 300 EC mw d fiber optic probe inserted directly into the solvent Time s Under some circumstances the rapid rate of microwave heating can produce heat profiles that are not easily accessible using traditional heating techniques In such cases experiments performed using microwave syn
163. representation if made may not be relied upon by Customer nor shall it form part of the transaction between Biotage and Customer for the Products or Services 8 EXPORT Customer shall comply fully with all applicable provisions of the United States Export Control Laws as may be in effect for any of the Products and shall seek whenever required and at its own expense export licenses from the United States Department of Commerce prior to any export of the Products and shall further assure compliance with all re export restrictions of such United States Export Control Laws 9 GENERAL These terms and conditions can only be modified by a written agreement duly signed by per sons authorized to sign agreements on behalf of Biotage and Customer Any variance from the terms and con ditions listed in any order or other written notification from the Customer will be of no affect If any provisions of these terms and conditions shall be held to be invalid illegal or unenforceable the validity legality and enforceability of the remaining provisions shall not in any way be affected or impaired thereby These terms and conditions will be governed by the laws of the Commonwealth of Virginia Returns Policy Returns of any product must be authorized in advance Please contact Biotage for a Return Authorization RA number and shipping instructions All claims must be made within thirty 30 days of shipment from Biotage or after the initial test at the P
164. rocessing of a variety of organic synthetic and natural products e Minimum exposure to highly active or toxic compounds e Ensure highest purity yield and throughput Cartridge Specifications FLASH 400M FLASH 400L Diameter mm 400 400 Bed Length cm 30 60 Flow Rate L min 1 7 1 7 Column Void Volume L 28 56 KP Sil Silica Packing Weight kg 20 40 FLASH AC Activated Carbon Packing Weight kg 13 26 Mitsubishi HP20 Packing Vol L 38 76 Ordering Information FLASH 400 Ordering Information FLASH 400 Systems Item Part Number FLASH 400M and FLASH 400L packages include a compression module with ASME U stamp air operated diaphragm pump solvent select valve A or B injection valve collect valve 1 or 2 radial compression reservoir 304 SS frame with casters start up kit and engineering documentation package Can be purchased with or without hoist FLASH 400M no hoist SF 011 50070 FLASH 400L no hoist SF 011 50150 FLASH 400M with hoist SF 511 50070 FLASH 400L with hoist SF 511 50150 Requires daily rate service off site SER 001 LBR FLASH 400 Cartridges Item Pkg Wt kg Qty Case Part Number KP Sil Silica 40 63 um 60 A nominally 500 m2 g silica for high capacity normal phase purification FLASH 400M 20 2 FKO 1107 50075 FLASH 400L 40 2 FKO 1107 50155 KP C18 HS 32 63 um 90 18 carbon endcapped for high capacity reversed phase purification FLASH 400M 33 2 F
165. rrels each containing a bottom frit top frits a frit insertion tool tubing kit and user s manual Item ZIF SIM10 Sample loading Module ZIF SIM35 Sample loading Module ZIF SIM60 Sample loading Module ZIF SIM Consumables Item ZIF SIM10 Barrels ZIF SIM35 Barrels ZIF SIM60 Barrels ZIF SIM Tubing Kit These consumables may also be used on Sample Loading Modules SLM for FLASH Elute Systems Description Holds 2 5 g max of silica Holds 10 g max of silica Holds 20 g max of silica Description 20 barrels with top and bottom frits 20 barrels with top and bottom frits 20 barrels with top and bottom frits Luer fitting with 1 8 tubing and 1 8 knurled nut connects ZIF SIM outlet to compression module Part Number FZIM 0010 FZIM 0035 FZIM 0060 Part Number SBF 0010 SBF 0035 SBF 0060 01903 Dry Loading Compression Head Assembly The Dry Loading Compression Head Assembly allows chemists to load precoated loose silica directly into a Biotage FLASH cartridge The new dry loading head is compatible with all Biotage FLASH compression modules and FLASH purification cartridges Biotage offers adapter kits to accommodate any flash system A counter lock design provides a liquid tight seal with an easy reverse turn of the upper knob This motion creates a tight Zero Insertion Force ZIF seal with the cartridge to prevent sample loss from leakage due to any drop in the
166. rtridge and are designed to improve loading peak shape and resolution Teflon coated metal heating blocks of the corresponding sizes are also available for easy drying of solvents in the Syntage Samplet in a vacuum oven prior to FLASH chromatography e Samplet cartridge design improves sample loading peak shape and resolution e Integrated technology allows chemists to purify sample loads from 5 mg 5 g using FLASH 12 to 40 compression modules and cartridges e Compression modules have improved resolution Syntage Media SO H 3 medium pressure 100 psig capacity allowing faster flow rates and Description KP PrNH 3 propylamino silica Samplet cartridges are the silica bonded equivalent of n propylamine SAX media and are used for applications including scavenging organic acids acid chlorides isocyanates and other electrophiles KP TSOH phenyl sulfonic acid silica Samplet cartridges are the silica bonded equivalent of p toluenesulfonic acid SCX media and are used for applications including scavenging bases or catch and release amine purification SH KP PrSH 3 mercaptopropy silica Samplet cartridges are the silica bonded equivalent of 1 propanethiol media and are used for applications including scavenging of alkyl halides and removal of transition metals Syntage Media Description SO cl KP TsCI phenylsulfonyl chloride silica Samplet cartridges are the silica bonded equiva
167. rule Upper O ring Kit Chemraz 108 and 107 O rings Retaining Clip Chemraz 108 O ring Chemraz 107 O ring Retaining clip Chemraz 108 O ring Retaining clip Parallex Flex Spare Parts Item Needle Rotor for Inject Valve 2 mL Loading Loop 5 mL Syringe 5 mL Loading Loop 10 mL Syringe Labels Switching valve UV Detector UV Lamp Gasket Set Pump Check Valve Plunger Belt 3 way valve Seal Seal Fraction Collector Ferrule Nut Ferrule Nut Seal Kit Fuses Fuse Fuse Fuse Description Tapered tip needle 6 port rotor 0 040 ID 1 16 OD x 0 040 ID SS Gas tight syringe 1 16 OD x 0 040 ID SS Gas tight syringe Labels for barcode printer 5 port loop switching valve Deuterium Gaskets offset flow cell UV4 Check valve assembly Sapphire plunger w holder Motor belt Gradient 3 way proportioning valve Seal Seal 1 8 Tube Short Tefzel 1 8 Tube x 1 4 28 Tefzel 1 16 Tube Short Tefzel 1 16 Tube x 1 4 28 Tefzel Valve Check 25 psi Kalrez 1A 10A 2A Part Number QFTT 4794 04794 04009 QUOR 001 02948 04593 03024 QLOR 001 Part Number 02922 03947 02789 02935 02788 02751 06461 02748 03945 03944 06403 06404 06407 06408 06405 06406 02727 02728 02729 05694 03451 03120 03188 03191 198 Parallex Flex Miscellaneous Spare Parts Item Tubing Reservoir Pressure Tubing PEEK Tubing Teflon Tubing General Purpose Waste Detector
168. s purified using normal phase chromatography with both a Biotage FLASH 12 system and a self packed glass column containing the same silica mass for comparison The results in the table below clearly show higher yield and purity of the target peptide in 1 24 the time with the FLASH 12 system Column Dimensions Separation Solvent Crude Load Peptide Mass Peptide Peptide mm Time min Consumed mL mg Recovered mg Yield 9o Purity Open Glass 12 x 150 240 236 300 154 60 78 FLASH 12 12 x 150 10 50 300 192 95 99 54 Application Note 21 Effects of Solvent Type and Strength in FLASH Separations By J Robert Bickler M S Product Quad3 Abstract Isolation of desired compounds from reaction mixtures and natural products by flash chromatography is not always a straightforward process A product and its analogs may not be separable without performing some type of method development and optimization Application 21 discusses the role of TLC thin layer chromatography as a flash purification method development and optimization tool in relation to the impact of solvent choice on compound retention and selectivity In this application a mixture of four dyes three hydrazine dye analogs and an anthraquinone dye was used as an example TLC was used to evaluate five solvent mixtures for selectivity ethyl acetate heptane acetone heptane isopropanol heptane toluene and methylene chloride TLC was also used
169. sed flash purification we have set the industry standard for speed safety and ease of use The new Initiator microwave synthesizer and SP1 flash purification systems from Biotage each perform significant roles in the drug discovery process Small enough to fit side by side in a fume hood these instruments work together to reduce drug discovery cycle times and improve success rates The Biotage consumable line includes microwave vials and flash chromatography cartridges custom engineered to deliver maximum performance Four optimum microwave vial sizes allow chemists to migrate from milligrams to grams without re optimization Durable and safe these vials are manufactured from contaminant free glass capable of withstanding pressures of 20 bars ca 300 psi Biotage FLASH cartridges are available in a variety of sizes and medias to provide selectivity choices for optimal purifications Patented design accommodates five different loading techniques and withstands pressures up to 100 psi for faster flow rates and increased throughput Biotage products range from discovery through clinical trials and large scale production We offer application expertise and personal customer support customizing solutions to meet the needs of customers Biotage will continue providing new and innovative tools to meet today s research and development challenges Customer Support and Service 1 Point Support The Answer to All Yo
170. sig ba o o Figure 1 Pressure increase with flow rate N e o 5 10 15 20 25 Flow Rate mL min In recent years Biotage has developed High Performance FLASH Chromatography HPFC systems capable of operation at 100 mL min and an operating pressure of up to 100 psig HPFC systems utilize Biotage FLASH cartridges and Samplets for sample purification Samplets are premade sample preconcentration cartridges designed to adsorb samples dissolved in strong solvents Once the majority of strong solvent has been removed from the Samplet solvent effects are dramatically reduced improving sample load and purification Samplet based HPFC products simplify reversed phase FLASH purification and reduce user error A comparison of Samplet and direct liquid injection loading techniques is made in this application note Each of these techniques is used to separate and purify several basic pharmaceutical compounds by reversed phase HPFC Effects of injection mass and flow rate on reversed phase FLASH purification cartridges are shown Procedure The experiments were conducted using three samples Samples one and two consisted of a three component equal weight mixture of 1 methylbenzimidazole brucine and promethazine dissolved in methanol at 150 mg mL and 300 mg mL respectively Sample three contained 100 mg mL of promethazine in methanol which oxidized rapidly to produce a deep blue color Each solution was adsorbed onto
171. silica supported cartridges are as follows 1 Combining scavenging and flash purification in one step provides compounds sufficiently pure for NMR IR and MS confirmation 2 Eliminates aqueous work up 3 Enables real time monitoring 4 Increases the reaction rate 5 Eliminates the need to filter or remove the solid support reagents 6 Decreases solvent consumption 7 Facilitates easy automation for parallel solution phase synthesis Phenol pKa Adsorption Adsorption Adsorption Adsorption in Toluene in THF in DCM in ACN o Nitrophenol 7 00 98 9 86 7 89 3 90 1 m Nitrophenol 8 15 98 6 98 8 99 9 99 2 p Nitophenol 6 85 99 9 95 5 99 9 99 9 o Cresol 10 20 87 2 48 1 59 3 49 0 m Cresol 9 70 79 6 51 2 96 7 72 5 Table 1 Phenol loading on Syntage KP TBD as a function of solvent the maximum loading capacity was 0 3 mmole 42 mg of O nitrophenol per gram Solvent Effect on Capacity Loading Levels of Phenols of KP TBD As with all chemical reactions optimizing the solvent choice is very important The impact of the solvent type on this reaction was determined by evaluating the loading capacity of various disubstituted phenols in common reaction solvents The contact time between KP TBD and phenol was 15 minutes For the phenols tested the highest loading capacity was achieved in toluene Table 1 After 15 minutes of contact time between the Syntage KP TBD and the phenol the maximum loading capaci
172. t 250 C actually has similar dielectric properties as acetonitrile at room temperature Thus water can be used as a pseudo organic solvent at elevated temperatures where organic molecules will dissolve not only because of the temperature but also because of the change in dielectric properties This makes some reactions that normally would not run in water perfectly feasible Solvents with low boiling points e g methanol dichloromethane and acetone give lower achievable temperatures due to the pressure build up in the vessel If a higher absolute temperature is desirable to achieve a fast reaction it is advisable to change to a closely related solvent with a higher boiling point e g dichloroethane instead of dichloromethane Solvent Table Table 2 Solvent table To show the responses of various solvents to microwave irradiation we measured the temperature and pres sure of pure solvents after 100 seconds of microwave irradiation Fixed Hold Time was set to Off Absorption Level was set to Normal and the temperature was set to 250 C Solvent Boiling Point Attained Temp Attained Pressure Volume 2 5 mL 1 atm C C bar 1 Methyl 2 pyrrolidinone NMP 202 250 1 1 2 Dichloroethane 83 180 5 1 2 Dimethoxyethane DME 85 130 3 1 4 Dioxane 100 56 0 Acetone 56 150 7 Acetonitrile 81 180 13 Dichloromethane 40 110 5 Dimethylsulfoxide DMSO 189 250 1 Ethanol 78 155 13 Methanol 65 145 17 N N D
173. t Labor Travel Parts PMs 354292 Gold Liberator N N N 1 353075 Gold Synthesizer N N N 1 354266 Gold Optimizer N N N 1 SER 100 OEXP Gold Optimizer EXP N N N 1 353083 Gold Creator N N N 1 SER 100 CEXP Gold Creator EXP N N N 1 SER 100 QUAD Gold Quad 3 N N N 1 SER 100 QDUV Gold Quad UV N N N 1 SER 100 PIONR Gold Pioneer N N N 1 SER 100 HRZN Gold Horizon N N N 1 SER 100 SP4 Gold SP4 N N N 1 SER 102 FLEX Gold 2 Channel Flex N N N 1 SER 104 FLEX Gold 4 Channel Flex N N N 1 SER MUX FLEX Gold 4 Channel Flex MUX N N N 1 SER PXSC 0110 Gold Parallex N N N 1 SER 100 PLLXMUX Gold Parallex MUX N N N 1 355478 Gold Initiator and Initiator EXP N N N 1 355480 Gold Initiator 60 and Initiator 60 EXP N N N 1 SER 100 SP1 Gold New SP1 N N N 1 SER 100 SP4 Gold New SP4 N N N T 353074 Platinum Synthesizer Y Y Y 2 354264 Platinum Optimizer Y Y Y 2 SER 200 O EXP Platinum Optimizer EXP Y Y Y 2 353076 Platinum Creator Y Y Y 2 SER 200 CEXP Platinum Creator EXP Y Y Y 2 SER 101 QUAD Platinum Quad 3 Y Y Y 1 SER 101 QUADUV Platinum Quad UV Y Y Y 1 SER 200 PIONR Platinum Pioneer Y Y Y 1 SER 200 HRZN Platinum Horzon Y Y Y 1 SER 200 SP4 Platinum SP4 Y Y Y 1 SER FLEX SCO2 Platinum 2 Channel Flex Y Y Y 1 SER FLEX SC04 Platinum 4 Channel Flex Y Y Y 1 SER 300 FLEX4MUX Platinum 4 Channel Flex MUX Y Y Y 1 SER 400 PLLX Platinum Parallex Y Y Y 1 SER 400 PLLXMUX Platinum Parallex MUX Y Y Y 1 354921 Platinum Liberator Y Y Y 2 355479 Platinum Init
174. tamination Upgradable An SP1 system can be upgraded to an SP4 just by replacing the SP1 flow path module with an SP4 flow path module no software changes required R Hp m lea EE ess liii m rera AT mi 128 Ease of use With Touch Logic Control purifications are easily programmed and executed from one single window Self guided software At any time a step by step wizard is available to guide users through the run setup The Previous or Next buttons toggle between screens allowing users to confirm that complete information has been entered Solvent Monitoring All solvents are entered into the system with their eluting strengths and vapor pressures to optimize ease of use and performance The system continuously keeps track of your solvent and waste reservoirs warning when they are running low or having a risk of overflowing Find a fraction All fractions can be easily correlated to the corresponding test tube at any time in either the status screen or the results screen Using the Peak Mode all fractions correlating to the same peak can be identified at once Automated gradient creation from TLC data Create a sample specific gradient method by simply selecting the TLC solvents from a preprogrammed list and entering their ratio Then enter your compound s Rf value choose a cartridge collection rack UV wavelength and fractionation mode and the system creates a unique gradie
175. te shows the comparison of silica to KP NH for the purification of basic pharmaceuticals tolperisone verapamil and nifedipine Results KP Sil strongly binds most organic amines used in this study only nifedipine contains a nitroaromatic ring was recovered from silica at high yield In Figure 1 the selectivity difference between the KP Sil and KP NH for sample three is shown KP NH separates and elutes each individual component with tolperisone eluting first verapamil second and nifedipine third KP Sil binds tolperisone and does not separate verapamil from nifedipine Silica TEPCT TTT RL 51 mig oa nem 1 J TT Figure 1 Comparison of KP NH left and silica purification results using a hexane ethyl acetate 0 100 gradient KP NH is able to separate and elute all three compounds tolperisone 1 verapamil 2 and nifedipine 3 Using silica tolperisone elutes last with lower yield while verapamil and nifedipine co elute Sample load was 300 mg Cartridge size 12 x 150 mm Purification system Biotage Horizon Conclusion Biotage KP NH cartridges offer definite benefits as an alternative to plain silica in medicinal chemistry FLASH purification applications Specifically nitrogen heterocycles and tertiary organic amines are purified with better purity and yield on KP NH than on plain silica using simple hexane ethyl acetate gradients Application Note 41 Improving Reversed Phase FLASH Purification Throu
176. temperatures that can be reached sometimes a less reactive but more temperature stable reagent can be used If you see decomposition of reactants reagents products Lower the temperature e Shorten the reaction time It may be that the desired product is actually formed but then decomposes rapidly at elevated temperatures In some cases it is therefore possible to trap the product by using a shorter reaction time Decrease the concentration s of reagent s e Change to a more temperature stable reagent How Microwave Synthesis Impacts Chemistry Research and Development Microwave synthesis can have a significant impact on chemistry research and development if its strengths namely speed and simplicity are capitalized upon Speed The main benefit of microwave synthesis is shorter reaction times through an increase in reaction temperature Herein we describe how this affects chemistry development Chemistry like all sciences consists of never ending iterations of hypotheses and experiments with results Optimize your Reaction Optimize Your Reaction With the speed and simplicity offered by microwave synthesis in general and Biotage microwave synthesizers in particular optimization has never been easier Optimizing a microwave synthesis is very similar to optimizing a conventional synthesis If your first reaction was not a success changing the target temperature and reaction time can cause significant improvement
177. tention time of at least five minutes On the HPLC the optimal load can be determined by increasing the sample amount until resolution has been lost Normal Phase Gradient FLASH To transfer the HPLC method to flash convert compound retention time Tr to column volume using the following equation Compound CV compound Tr To where To the void time about 1 min at 3 mL min Use the same formula to convert the gradient program from time to CV Gradient segment length time To flash segment length CV By using these formulas and the same solvents reproducible reversed phase flash gradients can be developed Flash Scale up Flash scale up is based on equalizing solvent linear velocity and relative sample load for cartridges of different sizes or volumes Flash purification performed on a small scale is easily scaled to larger cartridges using the scale up factors in Table 4 which take into account the cartridge differences To successfully scale a purification find your current FLASH cartridge in the left column Then read across that row until you find the number closest to the scale up factor for your larger sample Read up to find the appropriate FLASH cartridge for that scale factor For example if a 1 gram purification on FLASH 25 S requires scale up to 30 grams the appropriate scale up cartridge according to the table is a FLASH 75L Scale to 12 S 1124M 25 S 1
178. tep Gradient Module UV Module f upgraded Cartridges Fractions Includes a 12 Port Manifold 2 pk G Quad4 40 Cartridge Module 4 Channel N 4 Port Fraction Collector Bracket for Scintillation Vial H 13 mm 40 mm Cartridges Microtiter Racks 4 ZIF SIM35s Block and 6x5 P 4 Port Fraction Collector 4 Injection Valves Fractions Bracket for Microtiter 8 pk Blocks Part Number Q3 P Instructions Simply choose one component from each of the component columns and then as many options as you need 138 Moving from left to right fill in the spaces below the table to create a custom part number example Q3P 3C1A A requires Option N Quad Accessories Add on Components and Accessories Item Quad 12 25 Cartridge Modules Quad 12M Pack Quad 25M Pack Quad 12M 25M Pack Quad3 Cartridge Modules Quad3 4 Pack Quad3 8 Pack Quad3 12 Pack Quad3 Upgrade Quad Pump Modules Quadi Quad3 4 Pack Quad3 8 Pack Quad3 12 Pack Quad3 Upgrade Quad FLASH Collector Quad FLASH Collector Fraction Trav 13 mm Test tube Rack 13 mm Test tube Rack 16 mm Test tube Rack 16 mm Test tube Rack 18 mm Test tube Rack 25 mm Test tube Rack Scintillation vial Rack 13 mm Microtiter Block Quad UV Monitor Quad UV Monitor 0 3 mm Path Flow Cell Quad4 404 Rack Quad4 404 Rack Quad4 40 Kit A Quad4 40 Kit B Quad Gradient Modules Continuous Gradient Module Step Gradient Modul
179. that are precursors of substituted benzofurans This work illustrates a new on column synthesis of phenoxy acetophenones 3 using Syntage KP TBD Samplet cartridges by reacting bromoacetophenones 1 and a range of commercially available phenols 2 using the Williamson ether synthesis technique Figure 1 Syntage KP TBD is a silica supported strong hindered non nucleophilic tertiary base that will de protonate only moderately acidic hydrogen e g phenols and activated methylenes TBD has been used for the following 1 Alkylation of phenols and amines OH GX bai Esterification of carboxylic acids using alkyl halides R 1 D N Alkylation of activated methylene compounds C 5 2 3 4 De halogenation of organic halides 5 High throughput synthesis of aryl triflates and aryl nonaflates 6 Silica bound TBD may also be applied in a catch and OTO release protocol whereby the acidic species is caught by Ri R Rs the TBD in alkylation reactions and the addition of an electrophile results in nucleophilic substitution The product Figure 1 is subsequently released into the solution Figure 1 Williamson ether synthesis technique The use of solid supported reagents is a solution phase synthesis technique that allows easy monitoring of the reaction progress by LC MS TLC or NMR techniques and employs excess reagent without adding additional purification steps The advantages of using the Syntage
180. the Kiloprep technology Biotage Kiloprep HPLC systems are available with flow rate ranges from 25 mL min up to 15 L min and pressure as high a 4 000 psig Customers can begin with a bench top HPLC system the Kiloprep 100 and then move up to the pilot scale with a Kiloprep 250 and finally to production scale with the Kiloprep 2000 3000 or 4000 Features and Benefits e Fully integrated system with a wide range of options e Two system control options Desktop computer with Microsoft Windows NT compliant with U S FDA regulations for electronic records and signatures 21 CFR 11 PLC with touch screen interface offering full automation without a desktop computer e Complete cGMP documentation package with every system e Complete automation of loading elution and fraction collection e Patented gradient control algorithm for the greatest accuracy and reproducibility e Triple head pumps for minimum pulsation and maximum reliability e Explosion proof design with FM approved certification e Solvent reservoirs designed for safe operation at preparative flow rates 180 Kiloprep Specifications Kiloprep Systems Specifications Biotage Flow rate Range Maximum Pressure Suitable Column Environment Kiloprep System mL min psig bar Diameters mm ID KP 100 series KP124 25 250 4 000 270 25 75 Lab KP152 45 450 2 000 135 50 100 Lab KP192 90 900 2 000 135 75 150 Lab KP250 100 1 000 2 000 135 75 150 Pilot KP250EF 200
181. the compounds Rf Cartridge Dimensions Typical Column Silica Weight Difficult Typical Easy mm x cm Flow Rates Volume ACV 1 ACV 2 ACV 6 mL min FLASH 124S 12x 7 5 2 5 12 6mL 4 5g 4 20mg 20 100mg 100 200mg FLASH 12 M 12x15 2 5 12 12mL 9g 8 40mg 40 200mg 200 400mg FLASH 25 S 12 x 7 5 10 25 24mL 20g 15 80mg 80 400mg 400 800mg FLASH 25 M 12x15 10 25 48mL 40g 30 160mg 160 800mg 800 1600mg FLASH 40 S 40 x 7 5 25 50 66mL 50g 40 200mg 200 1000mg 1 2 FLASH 40 M 40 x 15 25 50 132mL 100g 80 400mg 400 2000mg 2 5g FLASH 65i 65 x 20 65 85 470mL 350g 300 1600mg 1 6 8g 8 20g FLASH 75S 75 x9 100 250 300mL 200g 160 800mg 0 8 4g 4 10g FLASH 75M 75x15 100 250 500mL 400g 400 2000mg 2 10g 10 20g FLASH 75L 75 x 30 100 250 1000mL 800g 800 4000mg 4 20g 20 40g FLASH 150M 150 x 30 500 1000 4 3L 2 5kg 3 16g 16 80g 80 160g FLASH 150L 150 x 60 500 1000 8 6L 5kg 6 32g 32 160g 160 320g FLASH 400M 400 x 30 7000 38L 20kg 0 03 0 25kg 0 25 1 0kg 1 2kg FLASH 400L 400 x 60 7000 76L 40kg 0 06 0 50kg 0 5 2 0kg 2 4kg Table 1 Biotage Cartridge Loading Table for Isocratic Purifications Optimize your 2 Optimizing selectivity Purification The first step in successful FLASH purification is maximizing ACV Accomplish this by evaluating various solvent mixtures by TLC Look for a binary mixtu
182. the high throughput process Data and material can be transferred easily between synthesis characterization and quantitation systems Its operating system is based on a Microsoft platform enabling data transfer in the form of Excel CSV and Access files Fraction information can be exported from the Flex to LC MS or MS systems and the data confirmation of molecular weight purity and quantity are then reimported to the Flex database The software s superior sample tracking features simplify data retrieval and management Upon completion of purification and fraction analysis Flex Pilot software can be used to select target fractions and if necessary combine and reformat the samples to the original plate format Fractions are selected using a simple wizard and transferred automatically using a Gilson 215 liquid handler Online Characterization Parallex Flex The high throughput purification process can be further refined by interfacing the Flex V3 system to a Waters ZQ mass spectrometer using MUX technology This interface combines robust UV triggered collection with essential mass spectral data to enable rapid identification of target compounds Figure 10 A single computer is used for loading samples selecting LC and MS methods and final data review ma ARMA RS LT ACH Bl x I RT E e a E T TI 1 Emi EN De zm br am ere Too SD E EYEM XI L eL LULa ia pes ee Nee A o
183. the products we build just as our customers do We have a group of chemists who study the science of flash purification and apply it to synthetic organic reaction mixtures natural product extracts and biochemicals such as peptides and nucleotides In this section we highlight the most popular of the application notes published on our Web site www biotage com Application Note 1 Purification of a Protected Peptide by FLASH Chromatography By Shahnaz Ghassemi Ph D Product FLASH Abstract Medical research involving peptides as pharmaceutical actives has increased over the last decade Peptide syn thesis requires the coupling of several different amino acids in sequential steps to obtain the desired product The efficacy and biological activity of peptides are dependent on their final purity Purity and yield are dramatically increased when purification is performed on smaller synthetic segments prior to segment coupling Historically peptide segments have been blocked from coupling using any number of bulky hydrophobic removable reagents such as Fmoc 9 fluorenylmethoxycarbonyl Boc tert butyloxy carbonyl trityl etc which bond to the peptide N terminus and C terminus The resulting protected peptides however are very hydrophobic and have been difficult to purify by reversed phase techniques In this application a solution containing 300 mg of a crude peptide protected with trityl and dimethylcyclopropyl amide groups wa
184. thesis may well result in a different outcome to conventionally heated reactions even if the final reaction temperature is the same Another phenomenon that might account for some of the claims of specific effects for reactions run under atmospheric pressure is the superheating effect Under microwave irradiation at atmospheric pressure the boiling point of solvents can be raised up to 26 C above their conventional values The enhanced boiling point can be maintained in pure solvents for as long as the microwave radiation is applied Substrates or ions JPTIM ON present in the solvent aid in the formation of a boiling nuclei and the rate at which the temperature of the mixture returns back to the normal boiling point is solvent dependent It is now accepted that the major part of rate enhancements observed with microwave synthesis is strictly due to thermal effects even though the unique temperature profiles accessible by microwave radiation may result in novel outcomes While the existence of a specific microwave effect cannot be completely ruled out the effect appears to be a rarity of marginal synthetic importance Where to Start with Microwave Synthesis Microwave synthesis is normally conducted under conditions that vary considerably from what is conventionally used in today s chemistry laboratories Biotage microwave systems support a wide variety of different reaction conditions accommodating different solvents vo
185. tion GO A CH N NO Tf 0 XIX Zz H CI a FT CNT F CHCl FT CNT F 80 C 94 Shackelford S A et al J Org Chem 2003 68 267 275 Amination conc HCI 1 05 equiv iPrOH or THF 125 C 15 min NH Br Br Organ M G et al J Combi Chem 2004 6 776 782 ween H 81 I Coupling 0 5 BE ST dimer R Bd ArBr a nnn ad K CO3 10 aq DMF o LiCl NaOAc Q 160 200 C 10 55 min 36 80 AT Larhed M et al J Org Chem 2004 69 3345 3349 Heterocycles RI el R CHO NH OAc AcOH N R2 ns SN 200 C 20 min N 17 63 NZ OH NZ Combs A P Sparks R B Org Lett 2004 6 14 2473 2475 AcO AcO CAN N N MeCN H 0 oO o 5 min 82 96 OMe OMe O N Bose A K et al Synthesis 2002 1578 1591 Br l RIK P PR 170 C 10 min X R Pd 10 ppm R H 0 TBAB Na CO 90 Leadbeater et al J Org Chem 2005 Article ASAP CN R HN Na NMS uw 2 CH CN HOAc SN RNH Ri R2 160 C 10 min R2 91 95 Han Y et al Org Lett 2004 6 25 4775 4778 TNL OH Polvmerization K N O S 0 TsO Qo f vos FX a TsO H H TNAO0 NO gt TA Initiati i O Te t i d nitiation Ly Propagation YAS ermination CoHio CoH C H 70 Schubert U S et al J Comb Chem 2005 7 10 13 CH Ca o Microwave Reactions Solid Supported Reagents 1 3 mol FibreCat RA OQ K CO EtOH 1
186. to fraction Collector Unions Y Union Tee Description 1 4 OD LDPE BLUE air line Sold by the foot 1 16 0 020 ID Sold by the foot 1 4 OD 0 040 wall 1 8 OD 1 16 OD connects to inlet valve Sold by the foot 1 4 Instan tite 1 4 316 SS union Part Number 01487 03263 00089 00088 02879 01478 01049 Parallex Flex Miscellaneous Spare Parts continued Item Ferrules and Nuts Ferrule Set Nut Ferrule Nut MVP Valve Connections Ferrule Nut VICI Valve Connections Nut Ferrule SS Stainless Steel SP1 Spare Parts Item Fraction Collector Guide UV collect valve Needle Nut Ferrule Valve Assembly Tube Assembly Ferrule Nut Tubing Fuse Adapter Adapter Description 1 4 316 SS 1 4 SS 1 16 SS 1 16 compression nut SS 1 16 w SS locking ring 1 16 Delrin 1 16 for SS tubing 1 16 SS Description Needle Horizon Flash Collector 1 6 inch OD x 030 Inch ID x 1 83L 1 6inch Flangeless Fer 1 4 28 Peek Flangeless 1 16 Tefzel UV Collect 3 Way 1 4 inch Trough Drain Tube Horizon Flash Collector Set 1 4 316 SS 1 4 Tube 316 SS 1 4 OD x 040 Wall FEP Teflon 5 X 20MM 2 5A 250V 40mm to 25mm Flash Module Rack 40mm to 12mm Flash Module Rack Part Number 01634 01635 02726 02725 02445 02446 03318 03319 Part Number 08072 08173 04794 04009 09341 08432 01634 01635 00089 09367 07482 07481 SP1i Spare Par
187. took 5 minutes at 160 C to provide the product at 80 yield Broader exploration using unconventional solvents O 0 0 0 S p Nah SO Na H O Temp Time Conventional reflux 16 hours Microwave 185 C 6 minutes Data courtesy of Michael H Howard DuPont Crop Protection Products Using microwave synthesis uncommon solvents such as water can be used When water is heated to a very high temperature the dielectric constant decreases as the temperature increases Water has a dielectric constant which decreases from 78 at 25 C to 20 at 300 C this latter value is comparable to that of solvents such as acetone at ambient temperature This molecular mutation promotes the solubility of organic substances in water not only because of the elevated temperatures but also due to the change in dielectric properties Therefore water acts as a pseudo organic solvent in the microwave synthesis process This example was performed at DuPont Crop Protection The product was used as a key intermediate in the syn thesis of potential metallo enzyme inhibitors While reflux in water gave lt 10 yield the microwave technique provided the desired product in 99 yield Specifications 34 Heating Process Temperature Temperature increase Pressure range Power range Reaction vials Reaction volumes Agitation System Requirements Temperature Humidity Electrical supply Maximum power consumed Cooling Weight
188. ts continued Item Operator s Manual Software Pump Valve Disk Lipseal Ring Tee Nut Ferrule Valve Nut Tube Assembly Tube Assembly Tube Assembly Tube Assembly Filter Reducing Union Tubing Nut Ferrule Tube Assembly Tube Assembly Union Tubing Nut Ferrule Tube Assembly Nut Nut Tubing Nut Ferrule UV Detector Lamp Lamp 0 1 mm Flow Cell 0 3 mm Flow Cell Description CD Spi Spi 10 psi Max NC Collar Mount Check 3 16 inch Balls Assembly Quad3 0 500 OD x 312 ID 302 SST Spring Wear 500 OD X 370 ID Quad1 Female Branch 1 8T x 1 8 FNPT x 1 8T 1 8inch Flangeless Fer 5 16 24 Peek Flangeless 1 8 Tefzel 150 psi Max NC Collar Mount 1 8 Flangeless 1 4 28 Peek Solvent Inlet 5 Color Red Sp4B Sp1 Solvent Inlet 5 Color Blue Sp4B Sp1 Solvent Inlet 5 Color Green Sp4B Sp1 Solvent Inlet 5 Color Gold Sp4B Spi Inlet Solvent for 3 16 In OD Tubing 20 um Porosity 316 SST 1 4 x 1 8 reducing union 4 required per set 3 16 OD x 125 ID Teflon FEP Flangeless 3 16T x 5 16 24 Peek Natural Flangeless 3 16 Tubing Compression Module Outlet to Flow Path Module Inlet Sp4B Flow Path Module Outlet to Compression Module Inlet Sp4B 1 4 T X 1 8 T W Knurled Nut amp PFA Ferrules 1 8 OD x 062 ID FEP 1 8inch Flangeless Fer 5 16 24 Peek Flangeless 1 8 Tefzel Collect Valve to Flow Cell Outlet amp Waste Sp1 1 8 T Short Flangeless 1 4 28 PEEK 1 8 Flangeless 1 4 28 Peek Blue 1 8 OD x 0
189. ty was 1 2 mmole of o nitrophenol per gram of Syntage KP TBD For the remainder of these experiments in this application note toluene was the solvent of choice for all of the reactions Synthesis procedure for x phenoxyacetophenones The following synthesis procedure was followed to produce the a phenoxyacetophenone array utilizing the Syntage KP TBD Samplet cartridges 1 1 2 mmole of each phenol was dissolved in 0 5 mL of toluene and transferred onto an individual FLASH 12 Syntage KP TBD Samplet cartridge Figure 2 2 These cartridges were placed in a vacuum oven for 30 minutes and then 1 2 mmole of alkyl bromide in 0 5 mL ACN was added to each cartridge and heated in a vacuum oven for five hours at 80 C Figure 2 Syntage KP TBD cartridge being loaded with phenol 3 After five hours the Syntage KP TBD Samplet cartridge was placed into the FLASH cartridge and purified using a Biotage automated FLASH system Figure 3 Figure 3 After the reaction was complete the Syntage KP TBD Samplet cartridge was directly inserted into FLASH cartridge eliminating the need for extra hardware connections or sample handling The total sample is applied to the cartridge in a plug flow format with this sample introduction technique Table 2 shows the a phenoxyacetophenones synthesized in this work illustrating the broad utility of this technique Each product was analyzed by HPLC and confirmed by FT IR H NMR and ES MS
190. ubstrates and reagents as well as the properties of the solvent s Phase In a Biotage microwave synthesizer all different phases can be used i e solution phase solid phase solid supported reagents solvent free and scavenger resins Please consider the difficulties in correctly measuring the temperature when solvent free techniques are used Stirring Always add a magnetic stirring bar to the process vial Stirring improves mass transport avoids tearing of solid materials and is beneficial for rapid heat distribution Inert atmosphere In general inert atmosphere is not initially employed in microwave chemistry and is often not needed even if the reaction is carried out in this way conventionally If needed flush the vial with an inert gas before capping Time As expected the reaction time will be a function of the reaction temperature and the thermal stability of substrates reagents and products A typical reaction normally requires two to 15 minutes of irradiation When trying out a new reaction for the first time three to five minutes of reaction time is normally used provided the required temperature is compatible with all of the ingredients Temperature All synthesis systems from Biotage work in a temperature range between 60 C and 250 C Optimally the reaction temperature should be as high as substrates and products allow before they start decomposing or as high as the reaction solvent allows whichever is lo
191. uirements The Quad UV and the Quad3 systems use the Quad FLASH Collector which provides no loss fractionation The Quad UV module has a true variable wavelength monochromator simultaneously monitoring and recording UV absorption of up to four samples A few simple keystrokes are all that is required to start a run Quad UV system with FLASH Collector Quad UV detector Quad 12 25 cartridge module and Quad1 pump The Quad UV 4 Channel Parallel Purification System This system simultaneously purifies up to four different reaction mixtures and collects each sample s fractions while recording the UV absorption of the compounds eluting from the FLASH cartridges Reaction mixtures from milligrams to grams can be purified by utilizing one of two cartridge modules the Quad12 25 or the Quad4 40 module The Quad 12 25 module holds up to four cartridges of either FLASH 12 M or FLASH 254 M The Quad4 40 rack holds four FLASH 40 compression modules A complete 4 channel Quad UV parallel purification system includes a Quadi pneumatic pump with four independent flow streams a multicartridge compression module a 4 channel variable wavelength UV monitor and a FLASH collector with parallel flow streams Quad3 Features The Quad3 includes R to L Quad34 Pump Quad3 Cartridge Module and Quad FLASH Collector The Quad3 Parallel Purification System with up to 12 Flow Streams The Quad3 system performs in th
192. ur Questions 6 1 Point Support THE ANSWER TO ALL VOUR QUESTIONS Biotage is readv to assist vou through specialists with vears of preparative chromatographv and svnthesis experience Our experts frequentiv visit customers and attend conferences to expand the Biotage knowledge base This gives us the abilitv to offer customized solutions in the following areas The Biotage 1 Point Support Philosophy Our service and support mission is to offer our customers a superior qualitv of service through our broad support superstructure which is reached through a single point of customer contact our 1 Point Support team This approach goes beyond simply providing product repair or replacement service It is a customer centered philosophv that provides the answer to all vour questions In fact each member of our support team is capable of supplving comprehensive solutions for vour questions concerning software hardware and applications issues when you contact our 1 Point Support Team your inquiries are resolved swiftly and efficiently saving you valuable time If we cannot answer your question immediately let us find solutions and quickly bring them to you Most importantly we are dedicated to building long term relationships with customers through quality service and timely customer support Scale up Our products and knowledge base support your progression from research to production scale Training and Workshops Courses on improv
193. urchaser s facility whichever is first Patent Indemnification Biotage warrants that the Goods as delivered to Buyer do not infringe any valid claim under any issued USA or EC patent Biotage will defend Buyer at Biotage s expense against any claim of infringement provided Buyer gives Biotage sole control in the defense and any related settlement negotiations and cooperates with Biotage in the defense of such claim In the event that the use of the Goods is enjoined Biotage at its sole option will either i procure a license to use ii or replace or modify the Goods with substantially equivalent non enjoined Goods iii or refund the full purchase price Biotage has no other liability in the event of a patent infringement claim BIOTAGE MAKES NO OTHER WARRANTY EXPRESSED OR IMPLIED INCLUDING MERCHANTABILITY OR SUITABILITY FOR ANY PURPOSE BIOTAGE WILL NOT BE LIABLE IN ANY EVENT FOR SPECIAL INDIRECT OR CONSEQUENTIAL DAMAGES WHETHER ARISING IN TORT UNDER ANY WARRANTY OR OTHERWISE Patent Acknowledgements U S Patents 4 900 537 5 089 124 5 112 492 5 139 661 5 167 822 5 186 838 5 200 069 5 223 142 5 242 586 5 601 708 6 019 897 6 068 766 6 090 278 6 074 556 6 117 329 6 132 605 6 136 157 6 139 733 6 149 403 6 168 400 6 200 113 6 221 252 6 294 087 6 403 939 6 398 953 6 436 284 6 551 467 6 596 130 6 614 010 6 629 626 6 630 654 6 649 051 6 652 746 6 720 540 and other patents pending Trademark Acknowle
194. ure pressure and power profile for the reaction being processed Edit the run on the fly ti Ez change parameters as needed bmi du ue 7509 eum 31 32 Advanced Edit Control pressure and power Set individual pressure and power thresholds for each reaction Build step or pulse sequences of up to 99 steps using all available control parameters time temperature pressure power fixed hold time and cooling Racks with Track Overlay Vial Type Check the specific parameters used and find the result for any given reaction on the tray Display the parameter of choice for all reactions and remove performed experiments Remote Viewer View the status of the instrument and the progress of your reaction from a remote location your office View save and print results R d eh 4 bep 77 P Representative Procedures Faster reactions and higher yields for Synthesis of 1 2 4 oxadiazoles l H N NaOMe EtOH 7 MZ NH T gt N 2 i O Temp Time Vield Conventional 78 9C 18 to 24 hours 7 to 63 Microwave 160 C 5 minutes 80 An early example from our in house work illustrates how reaction times can be shortened while improving yields Conventionally the condensation of aldoximes with esters required 18 to 24 hours at reflux to provide the 1 2 4 oxadiazoles in yields of 7 63 depending on what substrates were used Using microwaves it
195. ve Automated Flash Purification Reduces Bottlenecks and Increases Throughput Today s medicinal chemists are expected to create an increasing number of new chemical entities NCE each year Research shows purification to be the primary bottleneck in this drug discovery process Therefore chemists have turned to automated flash purification systems to purify crude samples faster using programmed elution gradients and UV fractionation However many of the interim steps needed to perform flash purification still remain manual and imprecise such as determining the elution gradient using thin layer chromatography TLC and choosing the right column As instrumentation and technology become more sophisticated they incorporate more and more of these manual steps further reducing the purification bottleneck Features and Benefits Touch Logic Control Load and run a sample in three simple steps using a 10 4 26cm touch screen a new graphical user interface and a built in Pentium computer TLC to gradient profile Automatically create a method and choose the optimal cartridge size based on the TLC data entered Change the gradient and fractionation parameters on the fly during operation Compact design The SP s compact footprint fits easily into a standard fume hood and accommodates the Biotage FLASH 12 40 and 65i cartridges Leak sensor optional Automatically detects fluid line leaks and pauses the system with
196. vers innovative solutions to streamLine your work process enabling to reach your purification goals faster and easier Cartridges and Samplets The Syntage Chemistry Samplet cartridges allow you to perform catalytic synthesis catch and release or scavenging of your reaction sample in the Samplet format Simply apply crude sample to the appropriate Syntage cartridge and insert it into the corresponding Biotage FLASH cartridges to integrate cleanup with downstream purification Discovery Scale Systems New SP1 automated FLASH purification system Our revolutionary touch screen operated single cartridge purification system automatically creates a gradient elution profile based on TLC data and will list cartridge sizes and recommended sample loads Upgradable to the SP4 for sequential purification of up to four samples Suitable for high throughput or multiuser needs FLASH 12 25 and 40 reservoir based FLASH purification systems Simplify and accelerate the isolation of your discovery scale compounds with our FLASH 12 25 and 40 reservoir based FLASH purification systems Quad Parallel Purification systems If you synthesize in parallel you can purify in parallel Available with four eight or 12 parallel flow paths the Quad system provides simple and efficient purification of multiple samples with the same or different solvent systems An optional four flow path variable wavelength UV monitor simplifies locatin
197. wave Reactions pg 53 Biotage Application Notes Application Note 1 Purification of a Protected Peptide by FLASH Chromatography pg 54 Application Note 21 Effects of Solvent Type and Strength in FLASH Separations pg 55 Application Note 28 Performance of Parallex Flex HPLC pg 56 Application Note 38 Continuous Gradient Purification of Closely Related Drug Intermediates Using FLASH Chromatography pg 57 Application Note 40 Purification of Heterocyclic Amines on a new FLASH Media KP NH Silica pg 58 Application Note 41 Improving Reversed Phase FLASH Purification Throughput pg 59 Application Note 42 Using TLC to Accurately Predict FLASH Purification Results pg 62 Application Note 43 Eliminating Aqueous Work Up in Multiple Step Solution Phase Synthesis using Flash Chromatography pg 63 Application Note 44 On Column Clean Synthesis of a Substituted Benzofuran Using TsOH Cartridges pg 67 Application Note 45 On Column Clean Synthesis of an Array of a phenoxyacetophenones Using Syntage TBD Cartridges pg 70 Application Note 46 Rapid Synthesis and Clean up of an Array of Amines Using Pre packed Syntage KP PrSH silica Supported Thiol Cartridges pg 74 Applications Corner In this section Biotage has hand selected published microwave reactions for your quick and easy reference All chemistries listed were created using Biotage systems Solution Phase Microwave Reactions Nitra
198. west Pressure The reactions can safely be performed at pressures of up to 20 bar If the pressure in a vial becomes higher the heating is automatically stopped and cooling begins For an indication of the expected pressure of a reaction please use the solvent table or the vapor pressure calculator at www biotagepathfinder com Optimize your Reaction Optimize Your Reaction With the speed and simplicity offered by microwave synthesis in general and Biotage microwave synthesizers in particular optimization has never been easier Optimizing a microwave synthesis is very similar to optimizing a conventional synthesis If your first reaction was not a success changing the target temperature and reaction time can cause significant improvement All remaining parameters e g concentration solvent reagent etc should be varied when applicable If the reaction is not proceeding at all or not going to completion e Increase the temperature As long as the reactants reagents can withstand the higher temperature the only limit is the pressure build up in the vial and the security limit of 250 C s Extend the reaction time e Increase the concentration s of reagent s e Change the solvent Some solvents e g water will behave differently at high temperatures as they become less polar This makes some reactions that normally would not work in polar solvents perfectly feasible Change the reagent s Due to the high
199. xtraction tool and all required tubing user s manual and grounding kit 2000 mL SIM cartridge extraction tool and all required tubing user s manual and grounding kit Cartridge extraction tool all required tubing user s manual and grounding kit Part Number SF 022 25071 SF 022 25151 FC 022 25071 FC 022 25151 FB 012 25070 FB 012 25150 SIM 1002 SIM 2002 FTF 0225 FBS 1025 FGD 15075 FIV 150 0000 FN 037 41200 FN 060 41200 AM 190 SU 150 1022 SU 150 2022 SU 150 0002 Parallex Flex HPLC Systems 154 The Flex V3 purification system is designed to meet the needs of discovery chemistry from lead discovery through lead optimization with simple software maximum versatility and proven reliability Simple Software The operating software for Flex V3 presents an intuitive graphical user interface This allows a chemist to load and run a sample with only four clicks of the mouse An email notification containing a full report with chro matogram and location of the target fractions is sent upon run completion Maximum Versatility A versatile purification tool the Flex V3 is the only preparative HPLC system available with one to four parallel flow streams It is also field upgradable as throughput needs increase A chemist can walk up with one or more vials and begin running the samples without disrupting another chemist s sequence In addition a core facility can l
200. ynthetic step multiple aqueous washes followed by drying filtration and evaporation are required Flow Chart 1 meets RSS ESS Repeat coupling and deblocking reactions Total of 6 times Fmoc Tyr t Bu Gly Gly phe leu NDMCP Flow Chart 1 Aqueous work up for removing excess reagents and byproduct Method 2 In Line HPFC Technique The new integrated technique incorporating Flash chromatography after each Fmoc deblocking step eliminates the aqueous wash protocol The crude reaction mixture is directly added to a Biotage FLASH 25 Samplet cartridge without any aqueous work up Samplet cartridges allow for quick direct loading of soluble and insoluble samples onto the FLASH cartridge The Samplet cartridge was inserted into the FLASH 25 cartridge and the compounds were purified using dichloromethanel methanol DCM MeOH gradients This integrated procedure reduces sample handling decreases the overall cycle time while providing higher yield with purer product Flow Chart 2 The collected fractions were tested by TLC and ninhydrin test Mass spectra running ESI mode were used to confirm identity of each isolated product after flash chromatography 65 66 Repeat coupling and deblocking reactions Total of 6 times Fmoc Tyr t Bu Gly Gly phe leu NDMCP Flow Chart 2 The new integrated technique incorporating Flash chromatography after each Fmoc deblocking step eliminates the aqueous wash protocol

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