BPG Columns - GE Healthcare Life Sciences
Contents
1. Index F ROW TOTE CUVE S oien ie Jted ow tat ntan ani n ans te ial ee N vt aches navn tetanus ie 12 G General grounding the column H HELP CGlCulationsiat Sacer 5 oe ced A Gre a Ree Otek oa E Se i ec 47 l installing the COLUMN weceeceeeessssssessessesecseeesesssssusutttuntusssssssessssessseseaseeceesesssissssuttuttunsssessssesseeeeeseesseeseee 17 L leckdge test sssrin A a EN nin Maan earned liane 32 M media QUantitieS oases caccee cscs ccncanc AE E A D AA AE EEEE EE 34 mounting an extension tube 100 140 and 200 COLUMNS ooeeeeeeeeesssssssssssssssssessessenstututtntuestnnssssssssssseeeeceeceeceeseeeessimutuussnsnnnessseeee 22 300 columns 450 columns N MOLES MOUCES zeae E AAEE E E E E E E E 4 P packing device location of components packing flow rate determination peak asymmetry calculations pressure LRA T E EE E E E E E E E E a annem caats 12 R recommended net sizes packing flow rates S safety EX marking 4 explosive atmospheres storage T test sample choice for COLUMNS ocecssssssesestesssssssssesessessssecseeceeseesessensunmutsstntssssussassessssessceseeeeseeseeseeniesatautee 46 testing for leaking COLUMN oc eescessssssssssssssesstesssssscssnsccscssceecneeceeceesessesustatmuttstnusnsssssassesasseeseeseeseseeseesuenisnatutee 17 torque wrench settings 100 140 and 200 columns al 300 columns 450 columns U UNPACKING thie COMMA aa a a A EAN 7 Ww warniho NOTES irern2. Installation and leakage test 30 Adaptor assembly 1 Place the new lubricated 20 ethanol O ring 32 between the inlet tube 31 and the adaptor 38 Tighten the Allen screws 33 2 Slide the spring 34 over the inlet tube 3 Carefully lower the adaptor screw 29 and lid 17 over the inlet tube 4 Screw the bolts 39 into the adaptor and tighten them The adaptor is now reassembled Mounting an extension tube packing device The packing device for BPG 450 is a stainless steel extension tube To mount the extension tube on a BPG 450 column proceed as follows 1 Remove the adaptor from the column and place the stainless steel extension tube onto the upper flange Make sure that the O ring is positioned properly in the flange Use the bolts delivered with the extension tube to secure the extension tube to the column The extended column is now ready for the first packing step in two step packing procedures Note Remove the extension tube after the initial sedimentation step Insert the adaptor and continue packing A tube length of 15 cm is required when installing the adaptor into the column tube if the packing extension is removed BPG Columns Instructions for use 18 1170 70 Edition AB Installation and leakage test 4 2 Connections Several different connectors are available to connect BPG columns to different types of tubing and systems see Chapter 3 in the BPG Columns Maintenance Guide The hygien
3. It may help to warm the filter net in water to facilitate mounting on the adaptor This completes the adaptor assembly procedure Mounting an extension tube packing device The extension tube is assembled when delivered Disassembly assembly of the extension tube is done the same way as for the column with the exception of the stand directives To mount an extension tube onto BPG 100 140 and 200 proceed as follows 1 Remove the adaptor from the column and place the extension tube onto the upper flange 10 so that the column rods fit into the open holes in the flange Fig 4 3 below on the extension tube Make sure that the thin O ring is positioned properly in the flange Fig 4 3 Packing device component location 22 BPG Columns Instructions for use 18 1170 70 Edition AB Installation and leakage test MMi 2 Use the washers and domed nuts from the column lid adaptor to secure the extension tube to the column The extended column is now ready for the first packing step in two step packing procedures The bolts and washers supplied with the extension tube are used to mount the adaptor onto the extension Note Do not pack the column to bed heights higher than the length of the column i e the adaptor should seal against the column wall NOT against the extension tube wall A tube length of 10 cm is required when installing the adaptor onto the column tube if the packing extension is removed
4. 4 1 2 BPG 300 Column disassembly To disassemble the BPG 300 column refer to Table A 2 and Figures A 3 and A 4 at the back of this manual and proceed as follows 1 Loosen one of the nuts 4 on the top plate clamp 1 that secures the adaptor assembly to the flange 5 and remove the clamp WARNING If the adaptor O ring seal is dry forced movement of the adaptor can damage it Wet the O ring with water or 20 ethanol solution DO NOT grease the O ring 2 Unscrew the seal adjuster 23 a few turns sufficient only to loosen the adaptor O ring seal 33 34 Remove the adaptor without letting the stainless steel touch the glass 3 Using a wrench and in the sequence shown in Fig 4 4 remove the nuts 2 and cup spring washers 3 that secure the support rods 8 to the flange 5 CAUTION Do not loosen each nut completely in one movement Loosen the nuts step by step in the sequence shown in Figure 4 4 Three to four turns per nut is enough to loosen and remove the nuts in the correct manner An adjustable spanner should be used on the flattened areas on the rods to prevent the rods from unscrewing BPG Columns Instructions for use 18 1170 70 Edition AB 23 Installation and leakage test 24 Fig 4 4 Nut removal and tightening sequence CAUTION When the flange is removed the column tube is unsupported Care should be taken to ensure that the column tube does not overbalance fall over Carefully rem
5. The figures in parentheses are achievable using a packing extension Maximum bed volume and bed height are based on a slurry concentration of 80 and a packing compression of 15 Maximum bed volume and bed height are based on a slurry concentration of 85 and a packing compression of 15 ao fF WN The operating temperature for BPG columns is 4 C to 40 C BPG Columns Instructions for use 18 1170 70 Edition AB 9 The BPG column 10 Table 3 2 Inlet and outlet dimensions on BPG columns Column size 100 140 200 1 0 dimensions mm 4 6 6 3 2 Column materials The materials used in the construction of BPG columns have been carefully selected for their compatibility with the solvents most commonly used in chromatography separations of biomolecules column maintenance and cleaning processes The polymeric materials have been tested for biological reactivity according to USP XXIII The materials have also been subjected to in vitro testing according to the Elution Test and in vivo testing according to the Systemic Injection Test and the Intracutaneous Test Extracts of 0 9 NaCl and sesame oil have been used as samples in the in vivo tests All materials meet the requirements as described in USP XXIII Class VI The components of the column may be considered to contain wet parts those coming into contact with process liquids and dry parts All materials of construction are made of high gra
6. e PTFE guide ring 3 e O ring 6 7 BPG Columns Instructions for use 18 1170 70 Edition AB 19 Installation and leakage test Fig 4 2 End piece component location on BPG 100 140 and 200 columns Secure the end piece 12 to the stand 15 using the bolts 13 and washers 14 Carefully place the glass tube 1 on the O ring 6 7 in the endpiece 12 Ensure that the glass tube does not touch any stainless steel Align the tube with the filter net 4 4 Screw the support rods 2 into the end piece 12 5 Position the flange 10 so that the support rods 2 fit into the appropriate holes Make sure that the O ring 6 7 on the flange is aligned with the glass tube The glass tube should not touch any stainless steel the glass tube may crack as a result Align the inner surface of the glass tube with the inner surface of the flange Mount the cup spring washers 9 and the nuts 8 on the support rods 2 and tighten them by hand 8 Seta torque wrench to the specifications set out in Table 4 1 below and tighten the nuts 8 and cup spring washers 9 to secure the support rods 2 to the flange 10 Follow the sequence shown in Figure 4 1 that is tighten the nuts in a diametrically opposite pattern WARNING The glass tube will crack if too much force is exerted when tightening the four nuts on the support rods the rods that secure the flange and the end piece to the glass column Always use the torque wrench
7. 2 8 8494 0 8 1113 06 8 8489 0 7 Flange O ring FEP X 1 8 0019 4 8 1113 07 8 0019 5 8 Nut M6 316 5 9 1582 0 Nut M8 316 5 7 9 0763 01 9 0763 0 9 Cup spring washer 316 4 8 8482 0 8 8482 01 8 8482 0 0 Flange 316 8 1134 2 8 1113 21 8 1134 22 1 Column cantering dowel PTFE 8 8 1134 2 8 1134 22 2 End piece 316L X 8 1144 18 8 1113 20 8 1112 62 3 Bolt M8 x 12 316 4 8 1113 28 8 1113 28 4 Washer 316 4 8 1143 26 8 1113 29 8 1113 29 5 Stand complete 316L 8 1031 10 8 1031 20 8 1031 20 6 Clamp 25 mm 304 8 1001 3 8 1001 31 8 1001 31 7 Adjusting knob PEEK 8 1109 27 8 1109 27 8 1109 27 8 Domed nut 316 4 8 1143 25 8 1113 27 8 1113 27 9 Adjuster nut body 316 1 0003 0 8 1113 26 8 1113 26 20 Bushing ring POM 2 8 0264 0 8 0267 0 8 0267 0 21 Top plate 316 8 1143 24 8 1113 24 22 Adjuster nut insert PEEK 8 0251 20 8 0252 20 8 0252 20 23 Allen screw M8 x 45 316 5 9 6375 0 9 6375 0 9 6375 0 24 Allen screw M4 x 14 316 5 9 6356 0 9 6356 0 9 6356 0 25 Stopper 316 8 1113 25 8 1113 25 26 Adaptor head screw 316 3 8 8457 0 8 8457 0 8 8457 0 27 Sealing unit 316L 8 1147 47 8 1113 3 8 1147 48 28 Adaptor O ring EPDM x 2 8 8475 0 8 1113 10 8 0275 0 29 Adaptor O ring FEP x 8 0019 40 8 1113 1 8 0019 50 30 Inner adaptor tube 316 8 1113 23 8 1113 23 8 1113 23 31 Outer adaptor tube 316 8 1134 83 8 1134 83 8 1134 83 32 Screw M6 x 12 316L PTFE 3 8 1145 08 8 1145 08 8 1145 08 33 Spring 316 PTFE 8 8491 01 8 8491 01 8 8491 01 34 Adaptor plate 31803 X
8. 42 by twisting a spatula or similar stainless blunt blade in the slot Remove the support net 40 Remove the stop screw 28 and locking nut 27 from the top plate 26 Release the sealing unit 30 and inner adaptor tube 35 by unscrewing the bolts 31 on the upper ring of the sealing unit Carefully remove the seal adjuster 23 releasing the tension on the spring 38 situated under the inner adaptor tube 35 Remove the top plate assembly 24 to 26 29 and 32 and separate it from the outer adaptor tube 36 by turning it in a clockwise direction Remove the sealing unit 30 and the adaptor O ring 33 34 Remove the outer adaptor tube 36 the inner adaptor tube 35 and spring 38 from the adaptor plate 39 by removing the screws 37 The top plate assembly 24 26 29 and 32 can be disassembled by removing the screws 32 This completes the adaptor disassembly procedure Adaptor assembly To assemble the adaptor on BPG 300 columns refer to Figure A 3 and proceed as follows Position the bushing rings 25 in the top plate 26 Using the screws 32 secure the adjuster nut insert 29 to the adaptor height adjuster body 24 through the top plate 26 so that the adaptor height adjuster body turns on the bushing rings 25 Slide the spring 38 inner adaptor tube 35 and outer adaptor tube 36 onto the adaptor plate 39 Fasten the outer adaptor tube 36 to the adaptor plate 39 using the scr
9. 8 1112 80 8 1113 30 8 1112 81 35 Support net adaptor PP X 2 8 1103 04 8 1112 99 8 0252 56 36 Net 54 um adaptor PP X 2 8 1126 96 8 1126 98 8 1127 00 Net 23 um adaptor PP X 2 8 1103 08 8 1113 01 8 9253 01 Net 10 um adaptor PA x 2 8 1103 05 8 1113 03 8 0252 76 Net 12 um adaptor PEEK X 2 8 1148 37 8 1148 39 8 1148 41 37 Gasket 25 mm EPDM X 2 8 0019 27 8 0019 27 8 0019 27 38 Feet stand 316 1 8 1126 93 8 1126 93 8 1126 93 39 Wheel 3 8 1001 09 8 1001 09 8 1001 09 40 4 port 2 way valve 316L PTFE X 1 8 5757 01 8 5757 01 8 5757 01 41 Tubing x 1 42 Washer PP 1 8 1142 79 NA NA 43 Washer PP 1 8 1142 80 8 1142 80 8 1142 80 See Table 3 3 in the BPG Maintenance Guide Tube i d 4 mm BPG 100 6 mm BPG 140 and 200 Comparable to 316L Table A 2 Components of BPG 300 columns Item numbers refer to Figures A 3 and A 4 Item Description Code number Material In contact with Qty no process liquid pk 1 Adaptor top plate 8 1012 48 316 1 2 Nut M12 8 1012 24 316 6 3 Cup spring washer 8 1012 25 316 6 4 Nut M10 8 1012 44 316 5 5 Flange 316 6 O ring 301 x 6 mm 8 1012 26 EPDM Xx 2 7 O ring 301 x 6mm 8 1012 27 FEP X 1 8 Rod for T300 500 8 1012 31 316 1 Rod for T300 750 8 1012 32 316 1 Rod for T300 9
10. assembled when delivered Disassembly assembly of the extension tube is done the same way as with the column with exception of the stand directives To mount an extension tube on the BPG 300 column refer to Figure A 4 and proceed as follows 1 Remove the adaptor from the column and place the extension tube onto the upper flange 5 Make sure that the thin O ring is properly positioned in the flange 2 Use the top plate clamp to tighten the extension tube to the column The extended column is now ready for the first packing step in two step packing procedures The extra clamp supplied with the extension tube is used to mount the adaptor on the extension Note Do not pack the column to bed heights higher than the length of the column i e the adaptor should seal against the column wall NOT against the extension tube wall A tube length of 10 cm is required when installing the adaptor into the column tube if the packing extension is removed BPG Columns Instructions for use 18 1170 70 Edition AB 27 Installation and leakage test 28 A 4 1 3 BPG 450 Column disassembly To disassemble the BPG 450 column refer to Table A 3 and Figure A 5 atthe back of this manual and proceed as follows 1 Loosen the adaptor sealing by turning the handle 24 WARNING If the adaptor O ring seal is dry forced movement of the adaptor can damage it Wet the O ring with water or 20 ethanol solution DO NOT grease the O ring 2 Move t
11. be dry and clean or filled with 20 ethanol in order to prevent bacterial growth Note Before using the column always check the performance of the packed column See instructions for this procedure in section 5 2 below 5 2 Column evaluation The efficiency of a column depends on how well it is packed A poorly packed column gives rise to uneven flow resulting in zone broadening and reduced resolution It is thus important to have a method by which the column can be tested before it is put into operation Such a method should be simple quantitative and should not introduce contaminating materials It is also an advantage if the same method can be used to monitor column performance over its working life so that it is easy to determine when the media should be re packed or replaced Methods that use colored compounds such as Blue Dextran should be avoided since they do not meet the above criteria and cannot be used with ion exchange and affinity chromatography media Experience has shown that the best method of expressing the efficiency of a column is in terms of the height equivalent to a theoretical plate HETP and the peak asymmetry factor As These values can be determined easily by applying a sample of a NaCl or acetone solution for example to the column It is important that the column is properly equilibrated before the packing is evaluated 2 column volumes It is preferable to run three test runs to see whether the test val
12. for the pressure to stabilize is dependent on the type of media it can take from a few minutes to a few hours f Continue to increase the pressure until a flow rate plateau is reached increased pressure does not give higher flow rates For some rigid gel media there will be no plateau In those cases use the flow rate at maximum allowable pressure for the column g When the maximum pressure has been reached the determination should be concluded BPG Columns Instructions for use 18 1170 70 Edition AB Operation STEP 8 Pressure flow rate curve gt Packing flow rate Flow rate g a oO o v 5 gt A a U x o O a 70 100 gt Pressure Fig 5 8 Optimum flow rate determination step 8 STEP 8 a Plot the pressure against the flow rate as indicated in the above example The optimal packing flow rate is 70 100 of the maximum flow rate For some very rigid media there will be no plateau in those cases use the flow rate at maximum allowable pressure for the column 5 1 4 Recommended packing flow rates for Amersham Biosciences media Recommended packing flow rates will be found in the instruction accompanying each delivery of media However for optimal packing we recommend a pressure flow curve determination be performed for each lot of media column or system configuration as described in section 5 1 3 5 1 5 Column packing method This packing method can be used with Bio
13. refer to Figure A 1 and Table 4 1 for torque wrench settings and proceed as follows 1 Position the bushing ring 20 in the recess in the top plate 21 2 Using the screws 23 secure the adjuster nut insert 22 to adjuster nut body 19 through the top plate 21 so that the adjuster nut body turns on the bushing ring 20 3 Slide the spring 33 inner adaptor tube 30 and outer adaptor tube 31 onto the adaptor plate 34 4 Fasten the outer adaptor tube 31 to the adaptor plate 34 using the screws 32 Slide the sealing unit 27 onto the adaptor tube 31 6 Ina counter clockwise direction screw down the top plate assembly 19 23 5 cm onto the outer adaptor tube 31 7 Turn the adjusting knob 17 clockwise onto the top of the adaptor until the holes in the inner adjusting tube 30 are level with the holes in the outer adaptor tube 31 3 BPG Columns Instructions for use 18 1170 70 Edition AB 21 Installation and leakage test 8 Finger tighten down the screws 26 into the upper half of the sealing unit 27 making sure they remain level and are aligned correctly with the holes until 1 2 mm the screw thread shows Finally tighten the screws with a ring spanner 9 Mount the stopper 25 onto the plate 21 with the screws 24 10 Mount the adaptor O ring 28 29 on the adaptor plate 34 11 Wet the filter net 36 with 20 ethanol Mount the support net 35 and the filter net 36 on the adaptor plate 34
14. should be as symmetrical as possible This is usually the case for gel filtration media but for certain ion exchange and affinity media it may be asymmetrical due to interaction with the media A change in the shape of the peak is usually the first indication of column deterioration The peak asymmetry factor should be as close as possible to 1 The peak asymmetry factor Ag is calculated from the graph in Fig 5 13 as follows A b a where a distance from peak apex to 10 of the peak height on the ascending side of the peak b distance from peak apex to 10 of the peak height on the descending part of the peak Note Measuring the HETP and A values is the best way to judge the condition of the packed column A packed column can look good but still need repacking for optimal performance Always check the column after packing and regularly between runs to ensure best column performance BPG Columns Instructions for use 18 1170 70 Edition AB 49 Operation 50 BPG Columns Instructions for use 18 1170 70 Edition AB 6 Abbreviations The abbreviations used in this manual are defined as follows As CIP EPDM FEP Abbreviations 6 Peak asymmetry factor Cleaning in place Ethylene propylene Fluroethenepropene Fast Flow Height equivalent to a theoretical plate High resolution Internal diameter lon exchange chromatography Bed height Number of theoretical plates Polyamide Polyetheretherketone Acetal plastic P
15. solution Fig 5 13 below shows a UV trace for acetone in a typical BPG column application and gives the calculated HETP and A values Column BPG 300 A Ve Media Sepharose 6 Fast Flow lt gt Bed height 57 5 cm a Bed volume 40 6 liters 2 Eluent Distilled water Sample 1 05 liters 2 1 acetone 1 NaCl Flow rate 19 cm h Vh Ve 18 8 Wh 0 9 50 HETP 0 024 cm a 0 90 a B b 0 85 10 b a 0 85 0 90 gt A 0 94 Volume Fig 5 13 A test chromatogram showing HETP and A calculations 5 2 2 HETP calculation The sample volume should be approximately 1 of the total bed volume and the concentration 1 0 v v NaCl or equivalent when using stronger buffer Alternatively 1 0 v v acetone can be used The linear flow rate should be 30 cm h for 34 um and 50 um media and 20 cm h for 90 um media To avoid dilution of the sample apply the sample as close to the column inlet as possible If an airtrap is included in the system it should be by passed during sample application to avoid back mixing Calculate the HETP value from the conductivity or UV curve as follows SS BPG Columns Instructions for use 18 1170 70 Edition AB 47 Operation SS HETP in its simplest terms is expressed as HETP L N where L Bed height cm N Number of theoretical plates N is defined by the equation N 5 54 Ve Wp where Ve Elution volume ml Wh Peak width at half height ml Ve is measur
16. very difficult to develop Most accurate pressure flow rate curves are obtained by experimental methods Figure 3 1 below shows the typical set up used in Amersham Biosciences laboratories to generate pressure flow rate curves To measurement Q z m gt cylinder or flow meter Manometer Safety valve 4 1 Fy oe Ms Pump Buffer Tank Notes 1 The maximum allowable length of the tube between the manometer and outlet is 1 5 m 2 The tube diameter is 10 mm Fig 3 1 Typical set up for determining pressure flow rate curves NOTE The column con tains only buffer The outflow must be at the same level as the manometer 12 BPG Columns Instructions for use 18 1170 70 Edition AB The BPG column eT 3 4 1 Pressure flow rate curves for empty columns Figures 3 2 and 3 3 show the pressure flow rate curves for empty BPG columns obtained using the method given in section 5 1 3 of this manual and the set up shown above ao BPG 100 3000 2500 BPG 140 2000 O K 5 1500 ee BPG 200 500 0 0 02 04 06 08 1 12 14 16 18 2 22 24 26 28 3 Pressure in bar Fig 3 2 Examples of pressure flow rate curves for empty BPG 100 140 and 200 columns Flow rate cm h 1000 BPG 300 800 BPG 450 600 400 200 0 0 0 05 01 015 02 025 03 O35 04 O45 05 Pressure in bar Fig 3 3 Pressure flow rate curves for empty BPG 300 and 450 columns Ga BPG Columns Instructions
17. 50 8 1012 33 316 1 9 Column tube T300 500 8 1012 28 Borosilicate glass x 1 Column tube T300 750 8 1012 29 Borosilicate glass x 1 Column tube T300 950 8 1012 30 Borosilicate glass x F 0 Net 23 um endpiece 8 1012 34 PP x 2 1 Net 10 um endpiece 8 1012 35 PA x 2 Net 12 um endpiece 8 1148 44 PEEK x 2 Net 54 um endpiece 8 1127 03 PP X 2 2 Support net endpiece 8 1012 36 PP x 2 3 Distribution plate 8 1012 37 PP x 2 4 Endpiece 8 1140 47 316L x 5 Gasket 25 mm i d 10 mm 8 0200 00 EPDM X 5 6 Gasket 25 mm i d 10 mm 8 1012 40 PTFE X 5 7 Connector 25 mm 10 mm PP X 2 8 4 port 2 way manual valve 8 1012 56 316L PTFE X 1 9 Stand complete 8 1012 38 316L 20 Washer 8 1012 61 316 6 21 Bolt M12 x 25 8 1012 65 316 6 22 Wheel 8 1001 09 7 1 23 Seal adjuster 8 1012 41 PEE 1 24 Adaptor height adjuster 8 1128 31 316 1 25 Bushing ring 8 1012 42 POM 2 26 Adaptor top plate 8 1129 06 316L 27 Nut M10 8 1012 44 316 5 28 Stop screw complete 8 1012 43 316 POM 1 29 Adjuster nut insert 8 1012 45 PEE 1 30 Sealing unit 8 1147 50 316L 31 Adaptor head screw M8 8 1012 46 316 3 32 Allen screw M8 x 60 8 1012 47 316 5 33 O ring 274 x 8mm 8 1012 51 EPDM X 2 34 O ring 274 x 8 mm 8 1012 52 FEP X 1 35 Inner adaptor tube 8 1147 83 316 36 Outer adaptor tube 8 1142 93 316L 1 37 Screw M8 x 16 8 1145 10 316L PTFE 5 38 Spring 8 1012 50 316 PTFE 1 39 Adaptor plate 8 1112 82 31803 X 1 40 Support net adaptor 8 1012 53 PP X 2 41 Net 23 um adaptor 8 1012 54
18. 6 316 52 Wheel support 8 1107 46 POM 316 53 Wheel 8 1177 75 Eye bolt 8 1109 64 54 Adaptor stopper 8 1115 18 316 Index A adaptor assembly 100 140 0nd 200 COUM NS ik 5 Ahk a Me ee Behe il saya als 21 300 columns 450 columns adaptor disassembly 100 140 and 200 columns 300 columns 450 columns approved operators Cc caution notices CE marking chemical resistance code numbers column tubes and standsS i2 fcsssdscassesdescocdessodszeecgsucctcsas assess ysedstssnsanb ossocsbecssascdncenjenivaieaiseectetssebsseesela 7 column assembly and disassembly An DOSIG COMPONENTS Aecicss sssscssssscacessocesceesecieccanic taste EAE E AE A 1 choice of test sample disposal evaluation grounding installation leakage materials packing principles of operation quantity of supplied components suggested packing method testing for leaks tube and stand code numbers unpacking column assembly 100 140 and 200 columns 300 columns 450 columns column disassembly 100 140 and 200 columns 300 columns 450 columns connectors D disposal of COI pan einen aa aapea i iae a io iaieiiea 5 E extension tube mounting on 100 140 and 200 columns mounting on 300 columns mounting on 450 columns BPG Columns Instructions for use 18 1170 70 Edition AB 1
19. 80 Fax 60 3 8024 2090 e Spain Tel 93 594 49 50 Fax 93 594 49 55 e Sweden Tel 018 612 1900 Fax 018 612 1910 e Switzerland Tel 0848 8028 12 Fax 0848 8028 13 e UK Tel 0800 616928 Fax 0800 616927 e USA Tel 800 526 3593 Fax 877 295 8102 imagination at work User Manual 18 1170 70 AB Elanders Tofters 2005
20. GE Healthcare BPG Columns Instructions for Use Important user information All users must read this entire manual along with the BPG Columns Maintenance Guide to fully understand the safe use of BPG columns Amersham Biosciences AB which was previously known as Amersham Pharmacia Biotech AB changed its name to the current name on 17th October 2001 CE Certifying This product meets all requirements of applicable CE directives The CE symbol and corresponding declaration of conformity is valid for the instrument when it is used as a stand alone unit or connected to other CE marked Amersham Biosciences instruments or connected to other products recommended or described in this manual and used inthe same state as it was delivered from Amersham Biosciences except for alterations described in this manual Declaration of conformity A Declaration of conformity stating European directives and harmonized standards valid for this type of equipment is included separately with each unit Safety Precautions eret Pia eS IMPORTANT Whenever the column is used always keep the Instructions for Use manual for the column on hand IMPORTANT The end user must ensure that all installation maintenance operation and inspection is carried out by qualified personnel who are adequately trained and understand the operating instructions WARNING Always use protective clothing appropriate with current
21. PP X 2 42 Net 10 um adaptor 8 1012 55 PA X 2 Net 12 um adaptor 8 1148 43 PEEK X 2 Net 54 um adaptor 8 1127 02 PP Xx 2 43 Tubing PVC X 44 Clamp 25 mm 8 1001 31 304 1 45 Wheel support 8 1033 33 POM 316 1 See Table 3 4 in the BPG Maintenace Guide Tube i d 10 mm Comparable to 316L Table A 3 Components of BPG 450 columns Item numbers refer to Figure A 5 Item Description Code number Material In contact with Qty no process liquid pk 1 Endpiece 8 1109 67 316L x 2 Support net endpiece 8 1104 35 PP x 3 Flange 8 1109 68 PP 316 X 4 U shaped seal 8 1104 40 EPDM x U shaped seal 8 1117 55 PFR x 5 Net 54 um endpiece 8 1127 05 PP x Net 23 um endpiece 8 1103 19 PP x Net 10 um endpiece 8 1103 18 PA x Net 12 um endpiece 8 1148 46 PEEK X 6 Flange O ring 446 x 6 8 1105 33 EPDM X 2 Flange O ring 446 x 6 8 1117 67 FEP X 7 Allen screw M8 x 50 8 1109 40 316 5 8 Rod BPG 450 500 8 1106 18 316 Rod BPG 450 750 8 1103 09 316 Rod BPG 450 1000 8 1103 10 316 9 Sealing unit 8 1109 69 316L 0 Pin bolt 8 3001 07 316 3 1 Stop screw 9 4363 01 316 2 2 Glass tube BPG 450 500 8 1103 14 Borosilicate glass x Glass tube 450 750 8 1103 15 Borosilicate glass x Glass tube 450 1000 8 1103 16 Borosilicate glass x 3 Revolving stop cap 8 1107 43 POM 4 Revolving stop 8 1109 41 316L 5 Screw M8 x 50 8 1109 40 316 5 6 Space
22. Process Media and is based ona constant flow rate being applied through the column To prepare for the column packing procedure proceed as follows A N WARNING Always use a safety valve connected directly after the pump BPG Columns Instructions for use 18 1170 70 Edition AB 41 Operation 42 STEP 1 Confirm that the optimum flow rate for the combination of equipment column type of media and quantity has been established refer to section 5 1 3 STEP 2 Check that there is sufficient packing buffer available A typical packing buffer is 0 1 M NaCl However always check the media instructions for packing buffers STEP 3 Wet the nets with 20 ethanol to eliminate trapped air in the nets STEP 4 Adjust the pump flow rate to the packing flow rate without connecting the column to the pump Record the pump speed in order to start the packing at the correct flow rate STEP 5 If the media is already in the column lift up the adaptor carefully stir the media and repeat steps 5 and 6 in section 5 1 3 If the media is notin the column repeat steps 1 6 of section 5 1 3 STEP 6 eee ST 7 Manometer Fig 5 9 Column packing method step 6 BPG Columns Instructions for use 18 1170 70 Edition AB Operation zy STEP 6 a Start the pump b Remove air bubbles in the tubing from the pump by pumping with a low fl
23. and soft thermo plastics e Composting material all materials matter that is converted to soil in the near future e Depositing material a small number of materials matter that are not suitable for recycling must be deposited Places for depositing can be located by contacting local authorities 1 6 2 BPG columns Before the column equipment is disposed of the following steps must be carried out e The column should be emptied cleaned and sterilized e For non flammable metals such as stainless steel contact a well recognized local scrapping company 1 7 Purpose of this manual This manualis designed as a general introduction and guide to column operation Specifically the manual will cover e Introduction Chapter 1 e Unpacking the column Chapter 2 e The BPG column Chapter 3 e Installation and leakage test Chapter 4 e Operation Chapter 5 Abbreviations Chapter 6 The accompanying manual BPG Columns Maintenance Guide covers the following topics Maintenance and service Chapter 1 Trouble shooting Chapter 2 e Ordering information Chapter 3 BPG Columns Instructions for use 18 1170 70 Edition AB 2 Unpacking the column Unpacking the column Unpack the equipment carefully and check it for signs of damage that may have occurred during transportation Table 2 1 below gives the code numbers for the column tubes and stands for the different column diameters check the code nu
24. application to ensure personal safety during operation WARNING Pay extra attention when working close to the column WARNING There should NEVER be air or gas under pressure in the column WARNING The working pressure of the column should never exceed the design pressure otherwise there is a risk of personal injury and damage to the column A suitable safety device must be fitted WARNING The working temperature range of the column should never fall outside the design temperature range WARNING Do not use other chemicals than those proven not harmful to the wetted parts of the column WARNING The wheel brakes must be activated at all times during operation or storage BPG Columns Instructions for use 18 1170 70 Edition AB WARNING Pay extra attention when moving the column to ensure that the column does not tip CAUTION Ensure that any water used with the column is particulate free since particles can block and damage the meshes BPG Columns Instructions for use 18 1170 70 Edition AB Contents 1 Introduction dlslbaa iJ 9 100 Reber ence tier OREN rene een NERY te ern Rea RE 1 1 2 Labelling of the COMIN cuencws cena uae eae 3 1 3 Safe tls aaa atcmcnieicnnenttansivie O E A mania 3 13 1 GENea a Rn ore SERENE een A 3 F32 WOlDINGS wrssscsscecsccticces eccrine E N AEA 4 L33 Caution DOUCES oser 4 DBA NOS nen A e EA 4 1 3 5 Chemical CSISEONICE ss cschccueiertuase c
25. atia cece adeabaneeel 4 1 3 6 Use in potentially explosive atmosphere waeeeccsssssssssssseeeee 4 1 4 Approved ODEFOLONS a2 ciccuntitiaidnsanuntatetaguamtanimanien 5 15 Store a A E A EAE RA AENOR 5 1 6 Disposal and recycling s ss sssssssssisssrsirsrissrrssrrssrrserssrrsserssrrssns 5 1 6 1 Materials and MAttEr ss 1 91191191101151101101010101s1strstrstrntrs0ts0est 5 162 BPOC UMN S a AR 6 1 7 Purpose of this Manual cesepoee dace el cheers eyes beovisteara sarin 6 2 Unpacking the column ccccssesesssesscscssscessesseseceeeseceeseseeeeees 7 3 The BPG column 3 1 Specifications and characteristics Se snnmorscucnecntneneerd 9 3 2 Column maternal Sunn ieee 10 3 3 Chemical resistance sc ucicccanainaandiaadadiadariadaclanerian 10 3 4 Pressure flow rate CURVES 2 8 sccacteiasadeccodicoraticairsecteniebisir 12 3 4 1 Pressure flow rate curves for empty Columns 13 3 4 2 Pressure flow rate curves for packed columnS 14 4 Installation and leakage test 4 1 Disassembly and assembly ssdansassvcsrnnssantaniconabanen 17 4 1 1 BPG 100 140 and 200i 17 42a BROGI niran n TER 23 gI ABROAS O e N E A N 28 4 2 GORNNECHONS mar esa AE Ta 31 4 3 Grounding the COIMMN az cosvecanistaerneunauaauneiuan 31 Ole kegkage testien en A ARES 32 Operation Bel SCIPIO OC KING sci sscrarlatons anomilat ncumenieeene 33 5 1 1 Media quantities required per cm bed height 34 5 1 2 Recommended net SIZE Soe ccswrsceainieist
26. ation and leakage test Fig 4 6 End piece component location for BPG 450 columns 2 Carefully place the column tube 12 on the U shaped seal 4 Ensure that the flange 3 is centered on the U shaped seal 3 Secure the tube to the end piece with the bolts 44 Tighten the bolts in cross wise sequence with the torque wrench set on 8 Nm Each bolt has to be tightened 3 times WARNING The column tube will crack if too much pressure is exerted when tightening the bolts which secure the flange 3 and the rods 8 to the column tube Always use the torque wrench set on 4 Nm when tightening these bolts See Table 4 3 below for torque wrench settings on the BPG 450 column Table 4 3 Torque wrench settings for BPG 450 columns Item Refer to Fig A 5 for location Torque wrench setting Lid and end piece bolts 26 amp 27 8 Nm Glass tube to flanges 3 and rods 8 4 Nm Adaptor disassembly To replace the O ring 32 To replace the O ring on BPG 450 columns refer to Table A 3 and Figure A 5 and proceed as follows also see torque wrench settings in Table 4 3 1 Remove the adaptor screw bolts 39 2 Lift off the lid 17 and adaptor screw 29 Take care not to bend the inlet tube 31 3 Remove the spring 34 4 Remove the screws 33 holding the inlet tube 5 Remove the old O ring 32 and discard it This completes the adaptor disassembly BPG Columns Instructions for use 18 1170 70 Edition AB 29
27. being unintentionally moved or run into Be careful not to step on or fall over hoses placed on the floor 4 1 Disassembly and assembly Disassemble and assemble the columns and adaptors in accordance with the instructions in the sections below Refer to Tables A 1 through A 3 and Figures A 1 through A 5 for item identification these tables and figures are located at the back of this manual Numbers in parentheses throughout the sections below refer to item numbers on the associated figures and tables at the back of this manual WARNING After reassembling the column ensure that the column is tight and not leaking 4 1 1 BPG 100 140 and 200 Column disassembly To disassemble the BPG 100 140 and 200 columns refer to Table A 1 and Figures A 1 and A 2 at the back of this manual and proceed as follows 1 Remove the domed nuts 18 and washers 14 that secure the adaptor to the flange 10 WARNING If the adaptor O ring seal is dry it may be damaged by forced movement of the adaptor Wet the O ring with water or 20 ethanol solution Do NOT grease the O ring BPG Columns Instructions for use 18 1170 70 Edition AB 17 Installation and leakage test 2 Unscrew the adjusting knob 17 several turns to loosen the adaptor O ring seal 28 29 and remove the adaptor 3 Using a socket wrench and in the sequence shown in Fig 4 1 remove the nuts 8 and cup spring washers 9 that secure the su
28. de materials which meet the requirements of the industrial production environment Tables A 1 A 2 and A 3 in the Appendix show the components of the columns and identify the materials from which the wet and dry parts of each component are manufactured 3 3 Chemical resistance Table 3 3 is intended as a guide to the solvent resistance of the materials used in BPG columns The information has been compiled from published material from several sources not from individual tests on the components of the column Consequently the operator should use this list only as a guide to the level of chemical resistance exhibited by each material It should be noted that the effects of a chemical will be more severe at higher temperatures and pressures and that combined effects have not been taken into consideration In general the use of the following chemicals in BPG columns should be avoided Extreme oxidizers such as peroxides e Halogenated compounds e Chlorinated solvents such as methylene chloride e Esters such as acetates e Aromatic hydrocarbons such as toluene BPG Columns Instructions for use 18 1170 70 Edition AB The BPG column SLU e High concentration of strong acids e Chloroform e THF e Toluene e Methylenedichloride Note Avoid salt in combination with pH below 4 Always wash the column with at least two column volumes ofpH neutral sol
29. den GE Healthcare Amersham Biosciences Europe GmbH Munzinger Strasse 9 D 79111 Freiburg Germany GE Healthcare Amersham Biosciences UK Ltd Amersham Place Little Chalfont Buckinghamshire HP7 9NA UK GE Healthcare Amersham Biosciences Corp 800 Centennial Avenue PO Box 1327 Piscataway NJ 08855 1327 USA GE Healthcare Amersham Biosciences KK Sanken Bldg 3 25 1 Hyakunincho Shinjuku ku Tokyo 169 0073 Japan Asia Paci c Tel 852 2811 8693 Fax 852 2811 5251 Australasia Tel 61 2 9899 0999 Fax 61 2 9899 7511 e Austria Tel 01 57606 1619 Fax 01 57606 1627 Belgium Tel 0800 73 888 Fax 03 272 1637 Canada Tel 800 463 5800 Fax 800 567 1008 e Central East amp South East Europe Tel 43 1 982 3826 Fax 43 1 985 8327 e Denmark Tel 45 16 2400 Fax 45 16 2424 e Finland amp Baltics Tel 358 0 9 512 39 40 Fax 358 0 9 512 39 439 France Tel 01 69 35 67 00 Fax 01 69 41 96 77 e Germany Tel 0761 4903 490 Fax 0761 4903 405 Italy Tel 02 27322 1 Fax 02 27302 212 e Japan Tel 81 3 5331 9336 Fax 81 3 5331 9370 e Latin America Tel 55 11 3933 7300 Fax 55 11 3933 7304 Middle East amp Africa Tel 30 210 9600 687 Fax 30 210 9600 693 e Netherlands Tel 0165 580 410 Fax 0165 580 401 e Norway Tel 815 65 555 Fax 815 65 666 e Portugal Tel 21 417 7035 Fax 21 417 3184 e Russia amp other C I S amp N I S Tel 7 095 23 2 0250 956 1137 Fax 7 095 230 6377 e South East Asia Tel 60 3 8024 20
30. e seal adjuster knob The stand is bolted to the column end piece and is fitted with wheels with foot operated breaks Note BPG 100 columns have adjustable feet Wheels can be ordered as an option BPG columns must not be used for any other purpose for example e Storage tank for chemicals etc e General pressure vessel e Fermentation vessel e Gas storage 2 BPG Columns Instructions for use 18 1170 70 Edition AB Introduction 1 2 Labelling of the column The CE mark on the identification label shows that the column fulfills the requirements of the European pressure equipment directive PED 97 23 EC and or the directive for equipment in potentially explosive atmospheres ATEX100 94 9 EC Columns where the product of volume and allowed pressure is too small must not be CE marked according to PED Equipment delivered outside the European Community might not be CE marked a N ANN Tm W BPG 100 500 Max column volume V 4 Code no 18 1103201 Design pressure PS be g Year of manufacturing 200 Test pressure PT 11 5 ber g date Tare mass 15 kg Maximum filing mass 6 w Operating tamp TS 44 c PED fluid group cat 1NA Amersham Biceciences AB T N Bitoean30 L Amersham C Ex II 3G S 751 84 Uppsala VW Biosciences amp Ye o Fig 1 1 The BPG column rating plate 1 3 Safety 1 3 1 General Information given in this Instructions for Use is suggested best working practice and shall
31. e sure that the O ring 6 7 on the flange is aligned with the glass tube The glass tube should not touch the stainless steel the glass tube may crack as a result 6 Align the inner surface of the glass tube with the inner surface of the flange WARNING The glass tube will crack if too much pressure is exerted when tightening the four nuts on the support rods the rods that secure the flange i and the end piece to the glass column Always use the torque wrench when tightening these nuts Refer to Table 4 2 for torque settings 7 Locate the cup spring washers 3 and the nuts 2 on the support rods 8 and finger tighten the nuts 8 Set the torque wrench in accordance with Table 4 2 below and tighten the nuts 2 in the sequence shown in Fig 4 4 Tighten the nuts stepwise to avoid uneven mechanical pressure on the glass tube Three to four turns per nut are recommended BPG Columns Instructions for use 18 1170 70 Edition AB 25 Installation and leakage test 26 Table 4 2 Torque wrench settings for BPG 300 columns Item Refer to Fig A 4 for location Torque wrench setting Rod nuts 2 Top plate clamp nuts 4 6Nm 6 5 Nm The column is now ready for use and the adaptor can be inserted Adaptor disassembly To disassemble the adaptor on BPG 300 columns refer to Figure A 3 and proceed as follows Locate the small slot at the base of the filter net ring Remove the filter net 41
32. eancensrnvereatareis nt 34 5 1 3 Optimum packing flow rate determination 35 5 1 4 Recommended packing flow rates for Amersham Biosciences media oaesssssssssssssssssssssessesssesseessnnnee 41 5 1 5 Column packing INOTNOG scss i cieressvisandisdeiindeersearssnceanchivle 41 5 2 Column evaluat oNaserseninn 46 5 2 1 Choice of test sample for COlUMNS ssssasssescesccstecsossesssoues 46 5 22 ETP calculation assener 47 5 2 3 Peak asymmetry factor calculation eccesesssssssssssesseseseeeee 49 6 ABBDEVIGUONS vesisicccicsisssecssvecessicsedastsnmrasteotsaiatsnduarsavonseeneaseantene 51 Introduction 1 Introduction 1 1 General BPG columns are intended for industrial applications that demand high standards of hygiene The columns are available for chromatographic separation media volumes up to 131 liters and permit the use of high flow rates When BPG columns are used in conjunction with BioProcess Media such as Sepharose Fast Flow and Sephacryl High Resolution the most stringent regulatory demands can be satisfied The materials from which BPG columns are manufactured have been carefully selected to resist sanitizing and cleaning agents such as sodium hydroxide and ethanol The pressure specifications and low flow resistance make these columns suitable for use with BioProcess Media in ion exchange hydrophobic interaction affinity and gel filtration chromatography Desalting with Sephadex G 25 Coarse Medium Fine and SuperFine grad
33. ed as the volume passed through the column from the middle of the sample application to the peak maximum Wpis measured as the width at the half height of the peak from the base to the top From the example in Fig 5 13 the HETP value can be calculated from the chromatogram as follows Table 5 3 HETP calculations Ve ml Wh ml N N m HETP cm Acetone 18800 900 2417 4203 0 024 Well packed columns will have low HETP values It is however only possible to compare columns that have been packed with the same type of media and that have been tested under identical conditions As a general rule of thumb a good HETP value is approximately two to four times the mean bead diameter of the media in question provided that the sample is not interacting with the media In practice the correlation between the HETP value and the column performance can only be assessed by the column operator Once this has been established a standard can be set to judge the acceptability of a column packing 48 BPG Columns Instructions for use 18 1170 70 Edition AB Operation For example the column operator may know from experience that a column packed with Sephadex G 25 Media with HETP values above 0 05 cm does not give the required separation Consequently the column operator will set this value to the maximum permissible i e the minimum acceptable quality 5 2 3 Peak asymmetry factor calculation The shape of the peak
34. ee ea a e kaa a e a E fae 4 BPG Columns Instructions for use 18 1170 70 Edition AB Figure A 1 BPG 100 140 and 200 column parts identification Figure A 2 BPG 100 140 and 200 column parts identification continued Figure A 3 BPG 300 column parts identification Figure A 4 BPG 300 column parts identification continued Fig A 5 BPG 450 column parts identification BPG Sepharose Sephacryl Sephadex Superdex and Drop Design are www ch romatog ra ph y rademarks of Amersham Biosciences Limited Amersham and Amersham amershambiosciences com Biosciences are trademarks of Amersham plc Teflon is a trademark owned by E l du Pont de Nemours and Co Triton is a trademark owned by Rohm amp Haas ne GE Hea Ithcare All goods and services are sold subject to the terms and conditions of sale of i he company unless otherwise agreed to in writing within the Amersham Amersha m Biosciences AB Biosciences group that supplies them A copy of these terms and conditions is 10 available on request Bj rkgatan 30 Should you have any comments on this product we will be pleased to receive 751 84 Uppsala hem at Sweden 2005 General Electric Company All rights reserved Amersham Biosciences AB a General Electric company going to market as GE Healthcare GE Healthcare Amersham Biosciences AB Bjorkgatan 30 751 84 Uppsala Swe
35. ent on temperature type of media batch and quantity Consequently for each individual system the optimum flow rate must be determined empirically by producing a pressure flow rate curve The recommended methods are described comprehensively in sections 5 1 3 and 5 1 5 BPG Columns Instructions for use 18 1170 70 Edition AB 33 Operation 5 1 1 Media quantities required per cm bed height The media quantities required per cm bed height for BPG columns are given below Table 5 1 Media quantity requirements for BPG columns Amount of media per Amount of media per centimeter of packed bed Media liter of packed column r BPG 100 BPG140 BPG 200 BPG300 BPG 450 DEAE Sepharose Fast Flow 25 98 m 92m 393 m 882 m 97 Sepharose Fast Flow 25 98 m 92 m 393 m 882 m 97 CM Sepharose Fast Flow 25 98 m 92 m 393 m 882 m 97 S Sepharose Fast Flow 25 98 m 92 m 393 m 882 m 97 Phenyl Sepharose Fast Flow 25 98 m 92 m 393 m 882 m 97 Sephadex G 25 C and M grades 250g 19 6g 38 2 g 78 5 g 176g 394g Sephadex G 25 SF 280g 22 09 42 99 88 0g 198g 443 g Sepharose 4 and 6 Fast Flow 20 94m 83m 377m 847m 89 Sephacryl High Resolution 25 98 m 92 m 393 m 882 m 97 Superdex 75 and 200 prep grade 20 94m 83m 377m 847m 89 Q and SP Sepharose High Performance 20 94m 83m 377m 847m 89 Phenyl Sepharose High Performance 20 94m 83m 377m 847m 89 5 1 2 Rec
36. es can also be used effectively The flow distribution system has been developed to match the high performance of Amersham Biosciences media All BPG columns are supplied complete with an adaptor Packing devices are available for applications requiring extended bed heights Column assembly and column packing protocols for BioProcess Media are also described in this manual A special section in the BPG Columns Maintenance Guide Chapter 1 is dedicated to column maintenance protocols applicable to individual process requirements The three major assemblies that comprise the column adaptor column components and stand are shown in the fold out exploded diagrams atthe back of this manual The basic components of the column are e Glass tube e Flange e O rings e Filter nets e Support nets e End piece e Rods BPG Columns Instructions for use 18 1170 70 Edition AB 1 Introduction The rods hold the flange and end piece to the glass tube which is sealed by O rings The O rings effectively isolate the glass tube from direct contact with the stainless steel components The adaptor top plate assembly and adjuster nut are clamped or bolted to the column flange The adjuster nut is allowed to rotate on a bushing ring in the top plate altering the height of the adaptor plate in the column tube The adaptor O ring forms a seal between the adaptor and the glass column when it is compressed by the sealing unit This is controlled by th
37. ews 37 Slide the sealing unit 30 onto the outer adaptor tube 36 In a counter clockwise direction screw down the top plate assembly 24 to 26 29 and 32 approximately 5 cm onto the outer adaptor tube 36 BPG Columns Instructions for use 18 1170 70 Edition AB Installation and leakage test MM 7 Turn the seal adjuster 23 clockwise onto the top of the adaptor until the holes in the inner adaptor tube 35 are in line with the holes in the outer adaptor tube 36 8 Finger tighten the screws 31 into the upper half of the sealing unit 30 making sure they remain level and aligned correctly with the holes until 1 2 mm of the screw thread shows Finally tighten the screws with a ring spanner 9 Locate the stop screw 28 and locking nut 27 in the top plate 26 Turn the stop screw until it engages the groove in the outer adaptor tube 36 preventing sideways movement of the tube without preventing vertical movement Lock the stop screw 28 in position with the locking nut 27 10 Mount the adaptor O ring 33 34 on the adaptor plate 39 11 Wet the filter net 41 42 with 20 ethanol to remove all air bubbles trapped in the filter net If the filter net is tight warm it in 40 C water 12 Mount the distribution plate 13 support net 40 and the filter net 41 42 on the adaptor plate 39 This completes the BPG 300 adaptor assembly procedure Mounting an extension tube packing device The extension tube is
38. for use 18 1170 70 Edition AB 13 The BPG column 3 4 2 Pressure flow rate curves for packed columns Figures 3 4 to 3 6 show the pressure flow rate curves for packed BPG columns obtained using the method given in section 5 1 5 and the set up shown in Fig 3 1 Flow rate in cm h BPG 100 70 60 5 BPG 200 BPG 300 40 30 20 10 0 0 0 1 0 2 0 3 0 4 0 50 6 0 Pressure in bar Fig 3 4 Pressure flow rate curve for Superdex 200 prep grade in BPG columns Bed height 60 cm Flow rate in cm h 300 250 200 150 100 50 0 0 0 0 5 1 0 1 5 2 0 2 5 3 0 3 5 4 0 Pressure in bar Fig 3 5 Pressure flow rate curve for Q Sepharose High Performance in BPG 300 500 Bed height 10 cm 14 BPG Columns Instructions for use 18 1170 70 Edition AB The BPG column Flow cm t 400 300 200 100 0 1 0 2 0 3 0 4 0 5 0 6 0 7 Pressure in i bar Fig 3 6 Pressure flow rate curve for a BPG 450 column packed with Q Sepharose Fast Flow to a bed height of 15 cm BPG Columns Instructions for use 18 1170 70 Edition AB 15 The BPG column 16 BPG Columns Instructions for use 18 1170 70 Edition AB Installation and leakage test 4 Installation and leakage test All BPG columns have sanitary clamp fittings as connectors Caution Only use hoses and clamps approved for the maximum pressure of the column and resistant to the chemicals intended to be used Caution Place the column in an out of the way area to protect it from
39. he adaptor down a few millimeters to loosen the O ring from the glass wall Then move it up as far as it will go 3 Remove the bolts 26 and washers 27 Loosen the lid 17 and lift out the adaptor Note The adaptor on BPG 450 columns is heavy For safety reasons it is recommended that the adaptor is lifted with a ceiling hoist connected to the column lid eye bolts 4 Loosen the bolts 44 to separate the column tube 12 including flanges 3 from the end piece 1 Carefully lift off the column tube If the O ring adheres to the glass tube or the end piece wet the O ring with 20 ethanol and then remove it 5 To change the top and bottom flange O rings loosen the Allen screws 7 and carefully lift off the top flange Remove the glass tube and replace the bottom O ring Put back the glass tube and fit the new top O ring Finally fit the top flange Be careful to align the glass tube with the flanges This completes the adaptor disassembly procedure Column assembly To assemble the BPG 450 column refer to Figure A 5 and Table A 3 and proceed as follows 1 Referring to Figure 4 6 below ensure that the following items are positioned correctly in the end piece e Distribution plate 41 e Snap plug 36 e Support net 2 e Filter net 5 e U shaped seal 4 mounted on the filter net Note Wet the filter net with 20 ethanol to eliminate trapped air BPG Columns Instructions for use 18 1170 70 Edition AB Install
40. ic sanitary clamp fittings on BPG columns permit cleaning in place Other connectors are made of polypropylene with good chemical resistance The outer diameter on all BPG clamp fittings is 25 mm A 51 mm clamp is used with the manometer kits and the safety valve 4 3 Grounding the column To avoid discharge from static electricity the column can be properly grounded earthed Note The equipment must be grounded when it is to be used in a potentially explosive atmosphere For this purpose a suitable grounding kit is available as an accessory and a grounding connection terminal is provided on the column stand as shown below Grounding terminal cable on BPG 100 140 200 and 450 columns Grounding terminal cable on BPG 300 columns Fig 4 7 Grounding connection terminal on the column stand To ground the column connect one end of the grounding cable to an earth terminal in the plant grounding network and the other end to the ground terminal located on the column stand The column ground terminal has an M6 thread BPG Columns Instructions for use 18 1170 70 Edition AB 31 Installation and leakage test 4 4 Leakage test To test for leakage across the adaptor 1 Fill the column with water or sanitizing solution and then position the adaptor in the column to the same height as you intend to run the column Seal the adaptor with the seal adjuster Remove the air in the column 4 Close the column outlet valve lt Colu
41. in no way take precedence over individual responsibilities or local regulations Great effort has been placed on the design and manufacture of the various parts of the equipment so thot it will comply with all applicable safety aspects for this type of equipment During the operation and during other work with a pressurized vessel it is always each individual s responsibility to consider e Their own and others personal safety e The safety of the equipment through correct use in accordance with the descriptions and instructions given in this Instructions for Use BPG Columns Instructions for use 18 1170 70 Edition AB 3 Introduction 1 3 2 Warnings WARNING The Warning symbol highlights instructions that must be strictly followed in order to avoid personal injury Do not proceed until the instructions are clearly understood and all stated conditions are met 1 3 3 Caution notices CAUTION The Caution sign highlights instructions or conditions that must be followed to avoid damage to the product or to other equipment Do not proceed until the instructions are clearly understood and all stated conditions are met 1 3 4 Notes Note The Note signis used to indicate information important for trouble free and optimal use of the product 1 3 5 Chemical resistance Specifications including chemical resistance are included in Chapter 3 of this Instructions for Use 1 3 6 Use in potentially explosive atmosphere BPG co
42. lumns do not contain any electrical circuits However the following precautions should always be observed e Avoid discharge from static electricity by grounding the column Use grounding earthing wire connected to the plant grounding network A grounding kit is available as an accessory e Only tools for use in potentially explosive atmospheres should be used for operation and maintenance e Always use suitable clothing when working in potentially explosive environments EX marking The EX code on the identification label defines the level of explosion protection of the equipment The Ex category of the equipment must be in conformity with the protection level of the room where it is to be used Ex Il3G Equipment in this category is designed to be capable of functioning in conformity with the operating parameters established by the manufacturer and ensuring a normal level of protection Equipment in this category is intended for use in areas in which explosive atmospheres caused by gases vapors or mists are unlikely to occur or if they do occur are likely to do so only infrequently and for a short period only BPG Columns Instructions for use 18 1170 70 Edition AB Introduction 1 4 Approved operators A BPG column is a piece of equipment that can be compared with a pressure vessel with a variable volume Operators should have experience of working with pressure vessels as well as with equipment that provides pressurized media fo
43. m of buffer remaining Valve Closed vy Fig 5 2 Optimum flow rate determination step 2 STEP 2 a Continue to pump buffer and air until there is only 2 3 cm of buffer remaining in the column b Use aspirit level to check that the column is standing horizontally Correct with the adjustable feet wheels as necessary ee 36 BPG Columns Instructions for use 18 1170 70 Edition AB Operation es STEP 3 Fig 5 3 Optimum flow rate determination step 3 STEP 3 a Mix the packing buffer with the media to form a 75 slurry i e packed volume slurry volume 0 75 Note This slurry volume is appropriate for BioProcess Media For other chromatography media check recommended slurry volume with your vendor STEP 4 Valve Closed vy Fig 5 4 Optimum flow rate determination step 4 STEP 4 a Carefully pour the homogenous slurry into the column Make sure that no air bubbles are trapped within the slurry PCs BPG Columns Instructions for use 18 1170 70 Edition AB 37 Operation ee STEP 5 Valve Closed Fig 5 5 Optimum flow rate determination step 5 STEP 5 Note This step should be completed quickly so that the media does not sediment completely a Insert the adaptor b Use the four bolts BPG 100 140 and 200 or top plate adaptor clamp BPG 300 and 450 to secure the column adaptor in place c Use the height adjuster handle to position the adap
44. mbers of the new equipment to assure that it matches Table 2 1 Code numbers for column tubes and stands Column Column tube length mm diameter Stand mm 500 750 950 100 18 1103 01 18 1103 02 18 1103 03 18 1031 10 140 18 1113 08 44 7320 47 18 1113 09 18 1031 20 200 18 1103 11 18 1103 12 18 1103 13 18 1031 20 300 18 1103 21 18 1103 22 18 1103 23 450 18 1103 71 18 1103 72 18 1103 73 The tube height of the BPG 450 column is 1000 mm Note Both the stand and the column must be ordered to obtain a complete column set up for BPG 100 140 and 200 Check that all of the components listed in Table 2 2 below are present Table 2 2 Quantity of components supplied with BPG columns Quantity Designation BPG BPG BPG BPG BPG 100 140 200 300 450 Blind flange with gasket EPDM 2 2 2 2 2 Clamp 23 mm 51 mm 2 2 2 2 2 Stand 1 1 23 um nets 2 2 2 2 2 Bolts for attaching the 6 4 stand to the column Instructions for use Maintenance Manuals each 1 1 1 1 1 eC BPG Columns Instructions for use 18 1170 70 Edition AB 7 Unpacking the column 8 BPG Columns Instructions for use 18 1170 70 Edition AB The BPG column Ea 3 The BPG column This chapter provides a general background on the specifications and characteristics of BPG columns 3 1 Specifications and characteristics Table 3 1 Primary specifications and characteri
45. mn outlet valve Sy Pressure gauge Adaptor BPG Column Safety valve Column inlet valve Fig 4 8 Schematic representation of the column valves and pump for leakage testing Raise the pressure to maximum working pressure Wait about 15 minutes until the pressure has stabilized Check the pressure drop on the pressure gauge The pressure drop should not exceed 0 1 bar during 5 minutes A recorder linked to a pressure controller gives more precise figures This completes the leakage test 32 BPG Columns Instructions for use 18 1170 70 Edition AB Operation 5 Operation WARNING Please handle the column with care since the pressure specifications are only valid if the glass tube is undamaged and free from scratches Never blow air or gas into the column Note Before packing the column for the first time we recommend that the column and tubing are flushed with CIP solution e g NaOH to ensure that all surfaces are clean Refer to section 1 3 in the BPG Column Maintenance Guide For optimum performance from BPG columns we recommend the column packing protocols outlined in the paragraphs below Before using the columns it is recommended that you read the complete chapter 5 1 Column packing There are many different packing methods each dependent on the column and type of media used For BPG columns we recommend the packing method described in sub section 5 1 5 The optimum packing flow rate is depend
46. olypropylene Polytetrafluoroethane Polyvinyl chloride Tetrahydrofurane Ultra violet Elution volume Peak width at half peak height Per fluor rubber Fluor carbon rubber BPG Columns Instructions for use 18 1170 70 Edition AB 51 6 Abbreviations 52 BPG Columns Instructions for use 18 1170 70 Edition AB Table A 1 Components of BPG 100 140 and 200 columns Item numbers refer to Figures A 1 and A 2 Item Description Material In contact with Qty BPG 100 BPG 140 BPG 200 no process liquid pk Code no Code no Code no 1 Column tube 500 Borosilicate glass x 8 0251 0 8 1112 95 8 1152 01 Column tube 750 Borosilicate glass x 8 0251 02 8 1152 02 Column tube 950 Borosilicate glass x 8 0251 03 8 1112 96 8 1152 03 2 Rod for tube 500 316 8 1105 80 8 1105 83 8 1105 83 Rod for tube 750 316 8 1105 81 8 1105 84 Rod for tube 950 316 8 1105 82 8 1105 85 8 1105 85 3 Guide ring PTFE 8 0251 22 8 1112 97 8 0252 22 4 Net 54 pm end piece PP X 2 8 1126 97 8 1126 99 8 1127 0 Net 23 um end piece PP X 2 8 9252 0 8 1113 00 8 9254 0 Net 10 um end piece PA x 2 8 0251 77 8 1113 02 8 0252 77 Net 12 um end piece PEEK X 2 8 1148 38 8 1148 40 8 1148 42 5 Support net end piece PP X 2 8 0251 55 8 1112 98 8 0252 55 6 Flange O ring EPDM x
47. ommended net sizes It is important to select the appropriate adaptor and end piece nets Nets are available in 10 12 23 and 54 um porosities Improperly sized nets can lead to back pressure problems or media leakage Recommended net sizes are as follows Table 5 2 Recommended net sizes for BPG column Net size um Material Bead size um 10 PA lt 70 12 PEEK2 lt 70 23 PP3 FF media 54 PP3 BB media t Polyamide 2 Polyetherether ketone 3 Polypropylene 34 BPG Columns Instructions for use 18 1170 70 Edition AB Operation a 5 1 3 Optimum packing flow rate determination To establish the optimum packing flow rate for BPG columns proceed as follows STEP 1 gt CN gt DA Valve Closed Y Fig 5 1 Optimum flow rate determination step 1 WARNING Always try to keep hazardous fluids in closed containers to avoid gas or spillage in the working area STEP 1 a Wet the column nets with 20 ethanol to eliminate trapped air b Fill the column with 10 15 cm of packing buffer c Drain the tubes to remove all air bubbles d Connect a tube to the suction side of a pump e Start the pump place the tube onto the bottom net and extract any remaining air bubbles trapped under the net BPG Columns Instructions for use 18 1170 70 Edition AB 35 Operation ee STEP 2 2 to 3 c
48. ove the flange 5 Using an adjustable spanner unscrew the support rods 8 from the end piece 14 Lift off the glass tube 9 Carefully place the glass tube on a secure surface If the O ring adheres to the glass tube or the end piece wet the O ring with 20 ethanol solution and then remove it Using a ring spanner remove the bolts 21 and washers 20 securing the end piece 14 to the stand 19 Separate the end piece and stand Column assembly To assemble the BPG 300 column refer to Figure A 4 and proceed as follows Note Wet the net with 20 ethanol to eliminate trapped air 1 Referring to Figure 4 5 ensure that the following items are positioned correctly in the end piece 14 Distribution plate 13 The profiled side should face the end piece and the flat side should face the support net Support net 12 Net 10 11 O ring 6 7 BPG Columns Instructions for use 18 1170 70 Edition AB Installation and leakage test Fig 4 5 End piece component location for BPG 300 columns 2 Secure the end piece 14 to the stand 19 using the bolts 21 and washers 20 3 Carefully place the glass tube 9 on the O ring 6 7 in the endpiece 14 Ensure that the glass tube does not touch the stainless steel Align the glass tube with the bottom filter net 10 11 4 Screw the support rods 8 into the end piece 14 Position the flange 5 so that the support rods 8 fit into the appropriate holes Mak
49. ow rate c While pumping at the low flow rate connect the manometer between the pump and the column then connect the tubing to the top of the column d Open the bottom valve e Adjust the flow rate to the predefined rate f Measure the flow rate at regular intervals and adjust if necessary The back pressure of the bed will increase and reduce the flow rate as the media packs g Ensure that the pressure displayed on the manometer does not exceed the maximum pressure of the media refer to user manual for each media or the column STEP7 ofety valve SQ E Pump ie Y Manometer En Mark top of bed gt cS Valve By Fig 5 10 Column packing method step 7 STEP 7 a When the bed has settled mark the top of the bed b Stop the pump c Close the bottom valve d Loosen the adaptor O ring seal by turning the adjuster knob BPG 100 140 and 200 or the seal adjuster handle BPG 300 and 450 on top of the column counter clockwise PCs BPG Columns Instructions for use 18 1170 70 Edition AB 43 Operation ee STEP 8 Safety valve Pump Ha Manometer a A Valve o Closed o Fig 5 11 Column packing method step 8 STEP 8 a Lower the adaptor quickly by turning the height adjuster handle clockwise The buffer between the sedimented bed and the adaptor passes between the adaptor O ring and the column wall During this operation the bed will start to ri
50. pport rods 2 to the flange 10 WARNING Do not loosen each nut completely in one movement Loosen the nuts step by step in the sequence shown in Figure 4 1 Three to four turns per nut is enough to loosen and remove the nuts in the correct manner A crescent wrench should be used on the flattened areas on the rods to prevent the rods from unscrewing 18 BPG Columns Instructions for use 18 1170 70 Edition AB Installation and leakage test Fig 4 1 Nut removal and tightening sequence WARNING When the flange is removed the column tube is unsupported A N Care should be taken to ensure that the column tube does not tip over 4 Carefully remove the flange 10 Using an adjustable wrench unscrew the support rods 2 from the end piece 12 6 Liftoff the glass tube 1 Carefully place the glass tube on a secure surface If the O ring adheres to the glass tube or the end piece wet the O ring with 20 ethanol and remove it 7 Using a wrench remove the bolts 13 and washers 14 securing the end piece 12 to the stand 15 Separate the end piece and stand Column assembly To assemble BPG 100 140 and 200 columns refer to Table A 1 and Figure A 2 at the back of this manual Proceed as follows Note Wet the net with 20 ethanol to eliminate trapped air 1 Referring to Figure 4 2 below ensure that the following items are positioned correctly in the end piece 12 e Support net 5 e Filter net 4
51. r axial compression packing control To use the column safely and correctly the operator must have read understood and be fully acquainted with the Instructions for Use in general and the safety instructions in particular Improper operation of the column can result in property damage as well as personal injury 15 Storage The column must be stored at design specified temperature and must not be exposed to extreme temperature fluctuations 20 ethanol is a recommended storage buffer 1 6 Disposal and recycling When the column has reached the end of it s life cycle it should be handled according to current laws and regulations regarding environmental aspects of recycling and disposal of industrial equipment This is the responsibility of the end user or equivalent Contact your local authorities if you have any questions regarding the environmental aspects or the laws in force for recycling or disposing of industrial equipment of this type 1 6 1 Materials and matter For disposal purposes materials are generally divided into the following categories e Flammable e Wood e Compound e g coated with plastic or metal foil e Paper e Non flammable e Metal Note Metals that are hazardous or contain hazardous matter e g lead cadmium mercury etc must be separated and disposed of in a controlled way BPG Columns Instructions for use 18 1170 70 Edition AB 5 Introduction e Plastics separated into hard hardened
52. r 8 1109 42 316L vj Lid 8 1109 70 316 8 Bushing ring 8 1109 43 PP 2 9 Adaptor height adjuster 8 1109 44 POM 20 Handle 8 1109 45 316 21 Nut flange 8 1109 46 PEEK 23 Seal adjuster 8 1109 47 PEEK 24 Handle 8 1109 48 316 25 nob 8 1109 49 PP 26 Screw M10 x 65 8 1109 50 316 2 27 Washer 10 5 x 20 8 1109 51 316 0 28 Domed nut M10 and washer 8 1103 20 316 29 Adaptor screw 8 1109 54 316L 30 Adjustment tube 8 1109 55 316 31 Inlet tube i d 12 mm 8 1109 53 316L X 32 O ring 12 3 x 2 4 8 1103 70 EPDM X 2 O ring 12 3 x 2 4 8 1117 65 FEP x 33 Screw M4 x 16 8 1109 56 316 2 34 Spring 8 1107 44 316 PTFE 35 Screw M5 x 10 8 1109 57 316 2 36 Snap plug 8 1107 45 PP x 2 38 Adaptor plate 8 1109 71 31803 x 39 Screw M8 x 25 8 1145 12 316L PTFE 3 40 O ring 419 x 10 8 1017 47 EPDM x O ring 419 x 10 8 1117 66 FEP X 41 Distributor plate 8 1106 04 PP x 2 Comparable to 316L continued on next page Components of BPG 450 columns continued Item Description Code number Material In contact with Qty no process liquid pk 42 Adaptor support net 8 1104 34 PP x 43 Net 54 um adaptor 8 1127 04 PP x Net 23 um adaptor 8 1001 62 PP x Net 10 um adaptor 8 1017 46 PA x Net 12 um adaptor 8 1148 45 PEEK X 44 Screw M10 x 80 8 1109 59 316 2 45 Screw M10 x 16 8 1109 60 316 2 46 Bracket 8 1109 61 316 47 Screw M5 x 16 8 1109 62 316 2 48 Bottom inlet tube 8 1109 63 316L X 50 Allen screw M10 x 40 8 1109 65 316 2 51 Leg 8 1109 6
53. se Stop lowering the adaptor when it is 0 5 to 1 cm above the bed E 44 BPG Columns Instructions for use 18 1170 70 Edition AB Operation STEP 9 ea ome aa Manometer Safety valve V o O Fig 5 12 Column packing method step 9 STEP 9 a Seal the adaptor O ring by turning the knob BPG 100 140 and 200 or the seal adjuster handle BPG 300 and 450 clockwise b Open the bottom valve c Start the pump Adjust the flow rate to the packing flow rate Check the manometer to ensure that the pressure does not exceed the maximum pressure for the media refer to user manual for each media The bed will be compressed again to the level of the mark on the column made in step 7 e Repeat steps 7 and 8 until the adaptor reaches the mark on the column Note When there is a maximum of 1 cm of liquid remaining do not open the adaptor O ring Instead use the height adjuster handle to screw the adaptor down the last remaining centimeter Never lower the adaptor more than a maximum of 5 mm into the bed Disconnect the tubing on the top of the column so that excess buffer can be removed via the top of the adaptor BPG Columns Instructions for use 18 1170 70 Edition AB 45 Operation 46 This concludes the column packing procedure Wash out any remaining buffer or slurry from the top of the adaptor The space between the adaptor head and the top of the column should
54. stics of BPG columns Column Cross Tube Bed Bed Max Total Adaptor Overall diameter sectional height height 4 volume 14 pressure weight weight dimensions mm area cm cm liters bar kg kg cm cm min max min max DxWxH 00 79 50 O 26 48 0 0 2 0 3 8 8 15 7 48x48x127 00 79 75 25 41 66 2 0 3 2 5 2 8 16 7 48x48x152 00 79 95 45 54 83 4 3 5 42 6 6 8 17 7A 48x48x172 40 154 50 O 26 48 0 0 40 7 4 6 25 1 59x59x127 40 154 75 25 41 66 3 9 63 10 2 6 26 a 59x59x152 40 154 95 45 54 83 6 9 83 12 8 6 27 1 59x59x172 200 314 50 O 26 48 0 0 82 15 1 6 34 3 59x59x127 200 314 75 25 41 66 7 3 12 9 20 7 6 36 3 59x59x152 200 314 95 45 54 83 4 14 1 16 9 26 1 6 39 3 59x59x172 296 688 50 O 26 48 0 0 17 9 33 0 4 68 29 69x69x133 296 688 75 25 41 66 17 2 28 7 45 4 4 73 29 69x69x158 296 688 95 45 54 83 4 31 0 37 2 57 1 4 78 29 69x69x178 446 1562 50 3 23 42 4 7 35 9 65 6 2 5 200 100 80x80x140 446 1562 75 22 39 58 34 4 60 9 90 6 25 215 100 80x80x165 446 1562 100 47 58 83 4 73 4 90 6 131 25 2 5 230 100 80x80x190 1 Maximum bed volumes and bed heights are based on a slurry concentration of 75 and a packing compression of 15 The compression is the difference in volume between a sedimented bed and a bed under pressure Use a manometer to monitor the pressure to order see Chap 3 in the BPG Maintenance Guide
55. tor 1 5 mm below the top surface of the liquid ee 38 BPG Columns Instructions for use 18 1170 70 Edition AB Operation le STEP 6 Valve Closed Fig 5 6 Optimum flow rate determination step 6 a Sealthe adaptor O ring by turning the seal adjuster knob BPG 100 140 and 200 or handle BPG 300 and 450 on the top of the adaptor clockwise b With the O ring sealed lower the adaptor filling it with buffer to expel any air in the adaptor tube The adaptor tube is full when the buffer creates a meniscus and no further air bubbles appear E BPG Columns Instructions for use 18 1170 70 Edition AB 39 Operation 40 STEP 7 ee 2 EN Manometer Fig 5 7 Optimum flow rate determination step 7 STEP 7 WARNING Always use a safety valve connected directly after the pump a Start the pump and remove air in the tubing from the pump by pumping with a low flow rate b While pumping at the low flow rate connect a manometer between the pump and the column and then connect the tubing to the top of the column Open the bottom valve Establish a pressure flow rate curve by beginning with a low flow rate The media will begin to settle leaving a clear space between the bed and the adaptor Do NOT adjust the adaptor e Slowly increase the pressure in increments and record the flow rate when the pressure has stabilized The time taken
56. ues are stable If a poor result improves during this test the reason can be that the column was not properly equilibrated To check that the bed is stable run the column at 70 packing pressure for 20 hours and test it again preferably three test runs Note that pressure spikes may cause poor packing cracking If this happens an air trap and a pressure relief valve are needed between the pump and the column The pressure relief valve should be placed between the air trap and column 5 2 1 Choice of test sample for columns The most appropriate material for column testing is of course the sample that is to be run in the application but this is not always practical or economical As an alternative a solution of either NaCl or acetone can be used to give a good indication of the column packing quality The eluate is monitored by measuring BPG Columns Instructions for use 18 1170 70 Edition AB Operation the conductivity or UV absorption the resulting elution profile is used to calculate the HETP value The advantages of using a NaCl solution are that it is readily available and can be used safely to test all columns One disadvantage is that NaCl may interact with the media matrix especially ion exchanger matrices and thus give erroneous results Acetone however does not interact with the matrix and is detected by UV absorption at 280 nm Alternatively the running buffer concentration can be increased 10 fold and used as a test
57. ution between use of salt and low pH buffers Table 3 3 Chemical resistance table of materials in BPG columns Substance Concentration by 60 90 days volume Acetic acid 10 see note 7 Acetic acid 25 see note 7 Acetonitrile 5 see note 2 Acetonitrile 50 see notes 3 4 Acetone 10 OK Cyclohexane 100 see notes 3 4 Ethanol 100 see note 2 Ethyl acetate 100 see note 4 Ethylene glycol 50 OK Glycerol 100 OK exane 100 see notes 2 7 ydrochloric acid 0 1M see notes 6 7 sopropul alcohol 100 see note 2 ethanol 100 OK itric acid 0 1M see note 7 n Propanol 100 OK Sodium chloride 2M see note 5 Sodium hydroxide 2M OK Trifluoroacetic acid 0 1 see notes 7 8 Triton X 100 100 OK Tween Tri n butyl phosphate 1 0 3 OK UREA 8M OK 1 The test does not include PVC tubing 2 EPDM rubber changes characteristics For repetitive and long term use use a FEP PFR PTFE O rings and gaskets BPG Columns Instructions for use 18 1170 70 Edition AB 11 The BPG column 3 Change to FEP PFR PTFE O rings and gaskets Polypropylene plastic resistance is fair 5 Can be used under normal running conditions Do not use NaCl in storage solutions Please note that NaCl can cause corrosion on stainless steel in acid solutions pH below 4 0 6 Not longer than 4 hours 7 Not recommended to be used with PA nets 8 FEP changes characteristics Thus use EPDM rubber 3 4 Pressure flow rate curves In general theoretical pressure flow rate curves are
58. when A N tightening these nuts 20 BPG Columns Instructions for use 18 1170 70 Edition AB Installation and leakage test E Table 4 1 Torque wrench settings for BPG 100 140 and 200 columns BPG Column 100 140 200 Moment Nm 4 5 5 9 The column is now ready for use and the adaptor can be inserted Adaptor disassembly To disassemble the adaptor on BPG 100 140 and 200 columns refer to Table A 1 and Figure A 1 and proceed as follows 1 Remove the filter net 36 by bending it carefully Remove the support net 35 Be careful not to scratch the adaptor as this may cause leakage 2 Remove the stopper 25 from the top plate 21 by removing the bolts 24 3 Release the sealing unit 27 and inner adaptor tube 30 by unscrewing the bolts 26 on the upper ring of the sealing unit 4 Carefully remove the adjusting knob 17 This releases the tension on the spring 33 situated under the inner adaptor tube 30 5 Remove the top plate assembly 19 23 and separate it from the outer adaptor 31 by turning it in a clockwise direction Remove the sealing unit 27 and the adaptor O ring 28 29 Remove the outer adaptor tube 31 the inner adaptor tube 30 and spring 33 by removing the screws 32 8 The top plate assembly 19 23 can be disassembled by removing the screws 23 This completes the adaptor disassembly procedure Adaptor assembly To assemble the adaptor on BPG 100 140 and 200 columns
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