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OQ and PQ Operating Instructions

1. Settings specific for the Dionex RF2000 Emission Sensitivity Med 10 Emission Autozero 0 5 Emission EmWavelength 380 0 000 Inject Emission AcgOn 0 250 Emission EmWavelength 381 0 500 Emission EmWavelength 382 0 750 Emission EmWavelength 383 1 000 Emission EmWavelength 384 1 250 Emission EmWavelength 385 1 500 Emission EmWavelength 386 1 750 Emission EmWavelength 387 2 000 Emission EmWavelength 388 2 250 Emission EmWavelength 389 2 500 Emission EmWavelength 390 2 750 Emission EmWavelength S91 3 000 Emission EmWavelength 392 3 250 Emission EmWavelength 393 3 500 Emission EmWavelength 394 3 750 Emission EmWavelength 395 4 000 Emission EmWavelength 396 4 250 Emission EmWavelength 397 4 500 Emission EmWavelength 398 4 750 Emission EmWavelength 399 5 000 Emission EmWavelength 400 5 250 Emission EmWavelength 401 5 500 Emission EmWavelength 402 5 750 Emission EmWavelength 403 6 000 Emission EmWavelength 404 6 250 Emission EmWavelength 405 6 500 Emission EmWavelength 406 6 750 Emission EmWavelength 407 7 000 Emission EmWavelength 408 7 250 Emission EmWavelength 409 7 500 Emission EmWavelength 410 8 000 Emission AcqOff End HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 Page 87 of 95 OQ and PQ Operating Instructions 7 11 Baseline Noise and Drift of the RI Detector Noise and Drift
2. cccccccececeeeeeeeceeeceeeeeeeeeceeeaaeaeeeeeeesenecnueaeeeeeeeteeenaea 39 3 5 4 Device Names E E E ei ae hacen end rae cede eae erat leliaat aaa rats 40 3 6 Performing the CHECKS zeiciccsccecsseccesscectcedeesecensecsvedcesseestaednetecenssvevecesevusacesseesteecpsverecesesesacesteeee 43 Sif CHECK Oa e ra ct cee E aa a Aa a aTa ect ecpevsesueu tine Shing Ea aa Kr oap 44 3 8 Evaluating the SeqQuences seesccsseeneseseeeseeeseeeeseeeeneesesesneesesesneeseseseeesesesneesesesseenseseseenseneenen 44 3 9 Repeating Checks cciciciseccesestcedevecdensdesmectdeeeBecegas a E decenss evening E E 44 4 Special Test Procedures for Individual MOdUIeS sceeeeeeeseeeeeeeeeeees 45 ALA anoet o dko n A ice ereenceist A a Wht need devees sau s diate A AE 45 4 2 Dionex VWD 3x00 Detectors Noise and Drift with Dummy Flow Cells seseeseeeeee 45 HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 Contents of IV OQ and PQ Operating Instructions 4 3 Dionex Autosamplers Sample Temperature ACCUIaCy cc seeceeeeeeeeeeeeseneeeeeeeeeeeeseeenes 45 4 3 1 Test Procedure vite tii tiie Hats Gi E Hate ee a tet tee eee 45 4 3 2 Connecting and Configuring the System cece eeeeeeeeeeeneeeeeeaeeeeeeaeeeeeeaaeeeeseneeeeneaas 46 4 3 3 Performing the Check sciatica conve trinii einari te edn ia riia dies 47 4 3 4 DUratlON eese a eei a Taa ne ee es EA 47 5 Proc dures sstecctietienieats eee ee
3. Hj HHHH 4 i en tA jatit Hjt Fide Ha 4 j tf ftn AEEA EA E TA j ii I AL TTE ee Tia 10 0 i tit Talat erin TT I T 1 y1 l l l l l l SEINI pawa oia P es a EEL ual Xai J boii tt ui JJ ULL bi 2k Lit C 5 0 Ee ees ee ERNER ee ee Hot fe Hid HI i4 J44H Hj HHHH Hi 4 Ne i HHHH H Hj HHHH t H tF FT 7 i t Hl AI i4 tit Fist Fr Fie t I Ha 4 7 j EPEAT ENS ATERA ERAT sy roa j 0 04 Uae TAUT ENIR ARLEN TTY Emi f toc EEEE AT Tt ik Poot ron FAT Cat ea E uit pawai owa P eepo e L ME 5 eae wa min 0 00 0 50 1 00 1 50 2 00 2 50 3 00 3 50 4 00 4 50 5 00 5 50 6 00 6 50 7 00 7 50 8 00 Figure 20 Chromatogram for defining the relative maximum of the emission spectra between 380 nm and 410 nm 5 12 2 Remarks on the Manufacturer Specification It is only possible to check the manufacturer specification of 2 nm for the excitation and the emission wavelengths by using a special flow cell and a mercury lamp For OQ and PQ the instrument should preferably be checked with the components used for the measurements Page 60 of 95 HPLC_OQ _PQ_E Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions 5 13 Baseline Noise and Drift of the RI Detector 5 13 1 Theory Drift and baseline noise are important specifications for a detector Increased baseline noise considerably reduces the dete
4. Pump specific settings Settings specific for the Dionex P580 SA Type Automatic Flow 0 300 UV_VIS_1 Wavelength 272 UV VIS _1 Step 0 20 Page 74 of 95 HPLC _OQ_PQ_E_Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions UV_VIS_1 Average On Detector specific settings Settings specific for the Dionex UVD 340S 3DFIELD Step 0 2 UV_VIS_1 Bandwidth 1 Uv_VIS_1 RefWavelength 600 UV_VIS_1 RefBandwidth Sampler specific settings Settings specific for the Dionex ASI 100 Sampler InjectMode Normal Sampler UpSpeed 10 0 Sampler DownSpeed 10 0 Sampler SampleHeight 0 50 Sampler WashSpeed 50 00 Sampler DispSpeed 50 00 Sampler DrawSpeed 25 00 Sampler SyringeDelay 5 0 000 UV Autozero Detector specific settings Inject UV_VIS_1 AcqOn 3 000 UV VIS_1 AcqOff HPLC_OQ_PQ_E_Manual doc Version 6 8 dated Oktober 2008 Page 75 of 95 OQ and PQ Operating Instructions 7 5 Carry over by the Autosampler and Linearity of the Injection r r r r r r Volume Injector Linearity and Carry over PGM Version 22 05 2001 Restriction capillary PEEK ID 0 13 mm Length 13 m Back Pressure 80 bars Solvent A Water HPLC quality Solvent degassed via online degasser HPLC System Pump specific
5. EY DIONEX Operational Quallification Performance Qualification for HPLC Instruments Operating Instructions Version 6 8 Date Oktober 2008 2008 DIONEX Doc HPLC_OQ_PQ_E_Manual doc OQ and PQ Operating Instructions OQ and PQ Operating Instructions Table of Contents 1 How to Use this Manual cccccceeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeees 1 2 TPVER OCC THON ia aa aaa aeaa aaa opa Aaaa eaaa apaia OEA aapa Eaa aE Aa 3 26 DevtirninG the Limits riae AA A 3 2 1 1 Operational Qualification OQ usiana E EEE AE 3 2 1 2 Performance Qualification PQ eorr a ar a E A E EE NAE 3 2 1 3 System Suitability Check SSC also System Suitability Test SST eeeeeeeeeeeeeeeees 3 2 2 Basic Requirements for Successful OQ and PQ ecceeeceeeeeeeeeeeeeeeeeesenseeaeeeceeeeeseeseaneeeeee 4 23 Overview of the CHECKS 26 s22 ccccecasscasnaccctscceasncoasaccestinnsendeansctenecarauaccapeneecetenseznccanndescetanenssanteuests 6 2 3 1 UV Detectors Using Analytical Flow CellS 0 cceccceeeee tenes erties ee eiieeeeetieeeeeeteeeestneeeeee 6 2 3 2 UV Detectors Using Non Analytical Flow CellS 0 eccccceeeeeeeeeeenneeeeeenaeeeeeeenaeeeseenaeeeenenas 11 2 3 3 Autosamplersiisiccistchiciand a a a aa a des a Gl Be eee 13 2 3 4 PUMPS 23 se ecient etait ceed eed a a tele ee ete 18 2 3 5 Thermostatted Column Compartments and Column OVENS ccceceeeeeeceeeeeeeeeeteees 22 2
6. Exchange the lamp If necessary close the windows and shield the instrument from the air conditioning system Lamp intensity The lamp is too old The flow cell is incorrectly installed Exchange the lamp Install the flow cell correctly Detector linearity The lamp is too old The concentration of standards is not correct The peak height of the sample with the highest concentration is not in the linearity range specified for the detector i e usually gt 1500 mAU for Shimadzu detectors and Dionex VWD 3100 and VWD 3400RS detectors gt 2500 mAU Exchange the lamp Use new standards Reduce the injection volume for all samples used for the detector linearity check so that the peak height of the sample with the highest concentration is in the linearity range of the detector i e usually lt 1500 mAU HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 Page 63 of 95 OQ and PQ Operating Instructions 6 2 2 Autosampler Check Precision of injection volume Reason The autosampler draws air from the vial There are air bubbles in the syringe The autosampler is leaking The injection valve is leaking Remedial Action Either there is too little sample volume in vial or the value set for the Needle Depth parameter is too low Prime the syringe Autosampler Manual Autosampler Manual Linearity of Injection Volume The dete
7. HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 Page 9 of 95 OQ and PQ Operating Instructions Instrument Parameter Description Limits OQ TSP UV2000 Baseline Noise Pure water is pumped through the 0 05 mAU 0 10 mAU flow cell The flow rate is 1 ml min 0 5 mAU h 1 0 mAU h Wavelength 254 Lamp Intensity ere ee Not Not checked checked Wavelength Pyrene is injected using methanol 1 0nm 1 0nm Accuracy as solvent The flow rate is 1 ml min The characteristic maximum of pyrene is determined at 239 nm and compared to its theoretical value Linearity Five caffeine solutions are injected r2 99 90 r2 99 90 in different concentrations RSD lt 5 IRSD lt 5 Concentration and peak area are represented in a graph The a an y A regression coefficient of the resulting line and the deviations from it indicate the linearity TSP UV3000 Baseline Noise Pure water is pumped through the 0 08 mAU 0 15 mAU flow cell The flow rate is 1 ml min Drift 0 5 mAU h 1 0 mAU h Wong 4 an 0 5 mAU h 1 0 mAU h Lamp Intensity Not Not checked checked Wavelength Pyrene is injected using methanol 1 0nm 1 0nm Accuracy as solvent The flow rate is 1 ml min The characteristic maximum of pyrene is determined at 333 nm and compared to its theoretical value Linearity Five caffeine solutions are injected r2 99 90 r2 99 90 in different concentrations RSD lt 5 IRSD lt 5 Concentration
8. UltiMate 3000 Dionex VWD 3100 UltiMate 3000 Dionex VWD 3400RS UltiMate 3000 Dionex PDA 3000 UltiMate 3000 Dionex PDA 100 Dionex PDA 100U Dionex AD25 Dionex UVD 340U Dionex UVD 170U Dionex UVD 340S Dionex UVD 170S Dionex UVD 160S Dionex UVD 320S Agilent 1100 1200 series G1315A Agilent 1100 1200 series G1315B Agilent 1100 1200 series G1315C Agilent 1100 1200 series G1315D Agilent 1100 1200 series G1314A Agilent 1100 1200 series G1314B Agilent 1100 1200 series G1314C Agilent 1100 1200 series G1365A Agilent 1100 1200 series G1365B Agilent 1100 1200 series G1365C Agilent 1100 1200 series G1365D Waters PDA996 Diode Array Detector Waters PDA2996 Diode Array Detector Waters 2487 Dual Lambda Absorbance Detector Page 4 of 95 HPLC_OQ_PQ_E_ Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions Instrument Supported Model UV Detectors TSP UV1000 Single Lambda Detector Cont d TSP UV2000 Dual Lambda Detector TSP UV3000 analog and digital data acquisition TSP UV6000 PDA Shimadzu LC 2010 SPD Shimadzu SPD 10Avp Shimadzu SPD 10AVvp Samplers Dionex ACC 3000 T UltiMate 3000 Dionex WPS 3000 T RS UltiMate 3000 Dionex WPS 3000 T SL UltiMate 3000 Dionex WPS 3000 T PL UltiMate 3000 Dionex WPS 3000TBPL Analytical UltiMate 3000 Dionex ASI 100 Dionex GINA 50 Dionex GINA 160 Agilent 1100 1200 series G1313A Agilent 1100 1200 series G1329A Agilent 1100 12
9. 20 and 25 ul Dionex WPS 3000 T SL Micro Standard 3 1 5 10 15 and 20 ul with 250 ul injection volume kit Standard 2 10 20 40 80 and 160 ul Dionex WPS 3000 T PL sample loop volume 20 ul Standard 3 1 3 6 9 and 12 ul sample loop volume 21 99 ul Standard 3 1 5 10 15 and 20 ul Dionex WPS 3000TBPL Analytical Standard configuration Standard 3 5 10 15 20 und 25 ul Large Volume configuration Standard 2 20 50 80 110 und 140 u Dionex WPS 3000 T RS Micro Option Standard 3 1 5 10 15 und 20 ul Also see section 3 1 Area mAU min gt N N WwW fF on ono GOGO Hn Oo i 1 1 1 fi fi 1 oO oO N oO 30 40 50 60 70 80 90 Injection Volume pl Figure 18 Linearity of the injection volume HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 Page 55 of 95 OQ and PQ Operating Instructions 5 7 Sample Temperature Accuracy of Autosamplers 5 7 1 Theory The sample temperature accuracy mainly depends on the cooling and heating accuracy of the autosampler the insulation of the sample compartment and the thermal transfer from the carousel to the vial 5 7 2 Performing the Check The sample temperature accuracy is determined with the help of an external thermometer The temperature sensor is placed in a standard vial 1 8 ml filled with water The carousel cover must be closed during the test The autosampler temperature is set to a nominal temperatur
10. 99 900 Result ok Reviewer s signature Date Operator s signature Date Chromeleon c DIONEX 2008 PQ_OQ _Report_6_8 RI_LLINEARITY Version 6 80 SR5 Build 2413 137116 Printed 27 10 2008 2 54 PM Seq DQ_PQ_OQ_FOQ PQ_OQ HPLC DEMO_for_Manual HPLC_OQ_DEMO_RUNS OQ_SAMPLER_LIN_CO Smp Injector linearity_5 Page 1 of 2 Runtime 14 Apr 2004 00 43 39 EE DIONEX Operational Qualification e Injector Linearity Instruments and Fluidics Instrument Name Pump Autosampler Column Oven UV Detector Chromeleon Datasystem Accessories Back Pressure Device Sample 2 Solvent A Supplier s Nam Serial Number P680 LPG DIONEX 1920401 ASI 100 DIONEX 1860410 TCC 100 DIONEX 1850409 UVD 340U DIONEX 1830402 V 6 60 Build 1428 DIONEX 11 Capillary L 15 m ID 0 18 mm Caffeine in Water 10 pg ml Water HPLC Grade e Additional Information Customer Operator Execution Date Next Qualification Customer s Name Operator s Name Operator s Jobtitle Apr 14 04 Oct 04 e Limits Values and Test Results Injector Linearity Corr Injector Linearity Observed Value Result 99 99000 99 99979 Test passed 0 500 RSD 0 238 RSD Test passed Reviewer s signature Date Chromeleon c DIONEX 2008 Version 6 80 SR5 Build 2413 137116 Operator s signature Date PQ_OQ_Report_6_8 INJ_LINEARITY Printed 27 10 2008 3 11 PM Seq DQ_PQ_OQ_FOQ PQ_OQ HPLC DEMO_for_Manual HPLC_OQ_DEMO_
11. D lt 0 07 or SD lt 0 02 min RSD lt 0 07 or SD lt 0 02 min Ten injections are analyzed When the Dionex GINA 50 or GINA 160 autosampler is used six injections are analyzed All of them contain the same standard sample The relative standard deviation and the standard deviation of the retention times indicate the flow precision The greater value is the valid limit A step gradient of two channels is programmed and measured three times The deviation between the measured and theoretical signal heights indicates the accuracy with which the pump forms the gradient The deviations between the three gradient runs indicate the gradient precision For channel A water is used as solvent for channel B water with 0 1 Vol acetone is used For each step of the above gradient runs the ripple is determined relative to the absorption of solvent B G1311A lt 1 5 G1312A B lt 0 7 G1311A lt 1 5 G1312A B lt 0 7 Pump module of Flow Precision the Waters Alliance 2690 Separation Module Gradient Accuracy Gradient Precision Ten injections are analyzed When the Dionex GINA 50 or GINA 160 autosampler is used six injections are analyzed All of them contain the same standard sample The relative standard deviation and the standard deviation of the retention times indicate the flow precision The greater value is the valid limit A step gradient of two channels is programmed and
12. DIONEX 2008 PQ_OQ_Report_6_8 INJ_REPRO_AND_RET_REPRO Version 6 80 SR5 Build 2413 137116 Printed 27 10 2008 2 54 PM Seq DQ_PQ_OQ_FOQ PQ_OQ HPLC DEMO_for_Manual HPLC_OQ_DEMO_RUNS OQ_INJECTOR_FLOW_REPRO Page 3 of 3 Smp Injector and flow reproducibility_ 10 Runtime 14 Apr 2004 00 20 19 e Charts for Injector and Flow Reproducibility Test Area Deviation from Mean Value Sample No Retention Time Deviation from Mean Value Sample No Reviewer s signature Date Operator s signature Date Chromeleon c DIONEX 2008 PQ_OQ_Report_6_8 INJ_ REPRO_AND_RET_REPRO Version 6 80 SR5 Build 2413 137116 Printed 27 10 2008 2 54 PM Seq DQ_PQ_OQ_FOQ PQ_OQ HPLC DEMO_for_Manual HPLC_OQ_DEMO_RUNS OQ_UV_LINEARITY Page 1 of 2 Smp Detector linearity_5 Runtime 14 Apr 2004 00 33 46 EE DIONEX Operational Qualification e UV Detector Linearity Instruments and Fluidics Instrument Name Supplier s Name Serial Number Pump P680 LPG DIONEX 1920401 Autosampler ASI 100 DIONEX 1860410 Column Oven TCC 100 DIONEX 1850409 UV Detector UVD 340U DIONEX 1830402 Chromeleon Datasystem V 6 60 Build 1428 DIONEX 11 Accessories Name Back Pressure Device Capillary L 15 m ID 0 18 mm Sample 2 Caffeine in Water 10 pg ml Sample 3 Caffeine in Water 60 pg ml Sample 4 Caffeine in Water 140 ug ml Sample 5 Caffeine in Water 220 ug ml Sample 6 Caffeine in Water 300 ug ml Solvent A Water HPLC Grade Additional Inform
13. P580 LPG ASI 100 UVD 340S STH 585 RI 101 RF 2000 HPLC_OQ_PQ_E_Manual doc Version 6 8 dated Oktober 2008 Page 95 of 95 Seq DQ_PQ_OQ_FOQ PQ_OQ HPLC DEMO_for_Manual HPLC_OQ_DEMO_RUNS OQ_WARM_UP Page 1 of 3 Smp Warm up Runtime 14 Apr 2004 14 36 35 2 DIONEX Operational Qualification Instruments Instrument Name Supplier s Name Serial Number Pump P680 LPG DIONEX 1920401 Autosampler ASI 100 DIONEX 1860410 Column Oven TCC 100 DIONEX 1850409 UV Detector UVD 340U DIONEX 1830402 Fluorescence Detector Other DIONEX not available RI Detector Other Shodex not available ELS Detector Other DIONEX not available Chromeleon Datasystem V 6 60 Build 1428 DIONEX 11 Additional Information Customer Customer s Name Operator Operator s Name Operator s Jobtitle Execution Date Apr 14 04 Period between two Qualifications 6 months Next Qualification Oct 04 Reviewer s signature Date Operator s signature Date Chromeleon c DIONEX 2008 PQ_OQ_Report_6_8 SPECIFICATION Version 6 80 SR5 Build 2413 137116 Printed 27 10 2008 3 24 PM Seq DQ_PQ_OQ_FOQ PQ_OQ HPLC DEMO_for_Manual HPLC_OQ_DEMO_RUNS OQ_WARM_UP Page 2of3 Smp Warm up Runtime 14 Apr 2004 14 36 35 e Accessories Accessories Exp Date Back Pressure Device Capillary L 15 m ID 0 18 mm Sample 1 Pyrene in Methanol 3 ug ml LA 91960 Sample 2 Caffeine in Water 10 ug ml LA 91955 Sample 3 Caffeine in Water 60 ug ml LA 91956 Sample 4 Caff
14. Precision of Injection Ten injections are analyzed Each of them contains 5 ul of the same Kit for 250 ul syringe Volume Linearity of Injection Volume standard The relative standard deviation of the peak areas indicates the precision of the injection volume r2 99 99 RSD lt 0 5 r2 99 90 RSD lt 1 0 Five injections of the same standard sample are analyzed The injection volumes are different The injection volume and the peak area are represented in a graph The regression coefficient of the resulting line and the deviation from it indicate the linearity Carry Over lt 0 05 After a highly concentrated test sample has been injected a blind sample is injected The blind sample contains only solvent The peak area measured in this chromato gram indicates the carry over Page 14 of 95 HPLC_OQ_PQ_E_Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions sl Gell acca tlt lt WPS 3000 T Temperature only with er Accuracy Kit for 250 ul syringe Cont d WPS 3000TBPL Analytical Standard and Large Volume configuration ASI 100 250 ul syringe Precision of Injection Volume Linearity of Injection Volume Carry Over Temperature Accuracy Precision of Injection Volume Linearity of Injection Volume Carry Over The sample temperature is measured using an external thermometer in a standard vial The st
15. RI Noise ELS Adjusted Limits 0 030 mAU 0 80 mAU h 500000 counis s 0 75 nm 99 980 5 000 RSD 0 300 RSD 0 0100 min SD 0 050 RSD 0 100 99 99000 0 500 RSD n a 1 000 0 500 STD 0 500 1 0 C 0 30 mV 40 00 mV 80 00 mV 10 nm 50 0 nRIU 500 0 nRIU h 99 900 0 3 mV Reviewer s signature Date Chromeleon c DIONEX 2008 Version 6 80 SR5 Build 2413 137116 Page 3 of 3 Runtime 14 Apr 2004 14 36 35 Limits with Optimized Conditions 0 030 mAU 0 80 mAU h 500000 counts s 0 75 nm 99 980 5 000 RSD 0 300 RSD 0 0100 min SD 0 050 RSD 0 100 99 99000 0 500 RSD n a 1 000 0 500 STD 0 500 1 0 C 0 30 mV 40 00 mV 80 00 mV 10 nm 50 0 nRIU 500 0 nRIU h 99 900 0 3 mV Operator s signature Date PQ_OQ_Report_6_8 SPECIFICATION Printed 27 10 2008 3 24 PM Seq DQ_PQ_OQ_FOQ PQ_OQ HPLC DEMO_for_Manual HPLC_OQ_DEMO_RUNS OQ_DAD_ WAVELENGTH Page 1 of 2 Smp Detector wavelength accuracy_DAD Runtime 13 Apr 2004 19 32 27 EE DIONEX Operational Qualification e Wavelength Accuracy of the DAD Instruments and Fluidics Instrument Name Supplier s Name Serial Number Pump P680 LPG DIONEX 1920401 Autosampler ASI 100 DIONEX 1860410 Column Oven TCC 100 DIONEX 1850409 UV Detector UVD 340U DIONEX 1830402 Chromeleon Datasystem V 6 60 Build 1428 DIONEX 11 Accessories Name Back Pressure Device Capillary L 15 m ID 0 18
16. 2003 16 10 23 EE DIONEX Operational Qualification RI Detector Noise and Drift Instruments and Fluidics Instrument Name Supplier s Name Serial Number Pump P680 HPG DIONEX 1920105 Autosampler ASI 100 DIONEX 9932012 Column Oven TCC 100 DIONEX Demo RI Detector RI 101 Shodex not available Chromeleon Datasystem Version 6 80 SR5 Build DIONEX 11 Accessories Back Pressure Device Capillary L 15 m ID 0 18 mm Solvent A Water HPLC Grade Additional Information Customer Customer s Name Operator Operator s Name Operator s Jobtitle Execution Date Jan 31 03 Next Qualification Jul 03 Limits Values and Test Results Observed Value Result Noise RI 50 0 nRIU 26 6 nRIU Test passed Drift RI 500 0 nRIU h 321 8 nRIU h Test passed Remark Noise and drift are measured dynamically with a floated cell The limits are different from published specifications because they are valid for static conditions only empty cell R er Reviewer s signature Date Operator s signature Date Chromeleon c DIONEX 2008 PQ_OQ_Report_6_8 RI_NOISE_AND_DRIFT Version 6 80 SR5 Build 2413 137116 Printed 27 10 2008 2 55 PM Seq DQ_PQ_OQ_FOQ PQ_OQ HPLC DEMO_for_Manual HPLC_OQ_DEMO_RUNS OQ_RI_NOISE_DRIFT Page 2 of 3 Smp Rl Detector noise drift Runtime 31 Jan 2003 16 10 23 e Data for Detector Noise and Drift Test Segment No Noise nRIU Amount of Drift nRIU h 173 299 231 224 282 414 839 815 4
17. 3 6 Fluorescence Detectors ccccccceseececceceeeeeeeeeenaececeeeeeesaceaeaneeceeeseseeaeaeaecesesesesseneeeeeeeeess 24 2 3 7 Refractive Index Detectors ccccccccceceeeeeeeeecceeeceeeee cece caaeeeceeeeeceseaeaeeeseeeeeeegeensieeeeeeeeteees 25 2 3 8 Evaporative Light Scattering Detectors 0 ee ceeeeeeeeneeeeeeeeneeeeeeaeeeeeeaeeeeeenaeeeeeenaeeeeneaas 25 3 PROCES S A 27 3 1 General Test Procedure ea rr czas Aaa a aa aa eaaa a a adaa p Aaaa ahaa AEEA sA apra aae haaa Daai aade 27 3 2 Test Procedure for Single Wavelength and VWD 3400RS Detectors ccssseseseereeeeees 29 3 3 Connecting and Configuring the System s sssssssseensnnnunnnnnnnnnnnnunnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnn 29 3 3 1 SYSTEM COMMECUONS oie aS indies daceaddincd gnaia ces indy de A A TA 29 S29 2 COMMOUTAUON PAPETA PE P E N E E E P T E E PE E 31 m R ME PRE oF TK Lae I AO EET trenchassaunceenstaaectsanactt sees 34 3 4 1 Preparing the HPLC System oeenn eid ea ee i ea 34 3 4 2 Checking the FIUIGICS eopardi i aane een eee deed dees 34 3 5 Preparing Chromeleom 1cc cc c cc ccccscis ctevscceceeastac ceteedee eit saceeceessavegtesdeeesvestaeeetesteeeressavesterenarentesdee 35 3 5 1 Template Str ct r ssir eea eel aval vat ETa AE AE AEE Aa bees cep a aaa a RAE aa aeaaaee 35 3 5 2 Creating the Sequence Templates ccccccceesececceceeeeeseeeneaeceeeeeeesenecneaeeeeeeeeeseesnaeeeees 36 3 5 3 Adapting the Report and Method
18. 35 mg ml r r Pump specific settings Log Pump ModelNo Log Pump ModelVariant Log Pump SerialNo Sampler specific settings Log Sampler ModelNo Log Sampler SerialNo Column Oven specific settings UV Detector specific settings Log UV ModelNo Fluorescence detector specific settings RI Detector specific settings Log RI ModelNo Page 90 of 95 HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions Pressure LowerLimit 10 bar Pressure UpperLimit 300 bar sA Equate Water Pump specific settings Settings specific for the Dionex P580 r r A Type Automatic Flow 1 000 RI_1 Step 0 2 RI_1 Average On Detector specific settings Settings specific for the Shodex RI 101 RI Temperature Nominal 35 Purge off Recorder Range 512 00 Integrator Range 500 Rise Time 0 50 Polarity plus Baseline Shift 0 Sampler specific settings r Settings specific for the Dionex AS Sampler InjectMode Normal Sampler UpSpeed 10 0 Sampler DownSpeed 10 0 Sampler SampleHeight 0 50 Sampler WashSpeed 50 00 Sampler DispSpeed 50 00 Sampler DrawSpeed 25 00 Sampler SyringeDelay 5 0 000 Detector specific settings RI Autozero Settings specific for complete systems Pump specific settings Vait Sampler Ready Oven specific
19. 386 214 676 204 259 953 907 494 897 344 469 47 102 923 453 4 237 221 655 181 431 3 639 185 468 545 040 163 138 417 141 Average 26 6 nRIU 321 8 nRIU h Limit 50 0 nRIU 500 0 nRIU h Result ok ok o oo N DOD oa fF ON Oo 3a amp amp s amp s om O Oo ON O o A wo N O Reviewer s signature Date Operator s signature Date Chromeleon c DIONEX 2008 PQ_OQ_Report_6_8 RI_NOISE_AND_DRIFT Version 6 80 SR5 Build 2413 137116 Printed 27 10 2008 2 55 PM Seq DQ_PQ_OQ_FOQ PQ_OQ HPLC DEMO_for_Manual HPLC_OQ_DEMO_RUNS OQ_RI_NOISE_DRIFT Page 3 of 3 Smp Rl Detector noise drift Runtime 31 Jan 2003 16 10 23 Charts for Noise and Drift Test Detector Noise I 10 Segment No Amount of Drift 1000 00 800 00 600 00 400 00 200 00 0 00 10 Segment No Reviewer s signature Date Operator s signature Date Chromeleon c DIONEX 2008 PQ_OQ_Report_6_8 RI_NOISE_AND_DRIFT Version 6 80 SR5 Build 2413 137116 Printed 27 10 2008 2 55 PM Seq DQ_PQ_OQ_FOQ PQ_OQ HPLC DEMO_for_Manual HPLC_OQ_DEMO_RUNS OQ_RI_LINEARITY Page 1 of 2 Smp RI_Detector linearity_5 Runtime 24 Feb 2003 09 45 59 EZ DIONEX Operational Qualification RI Detector Linearity e Instruments and Fluidics Instrument Name Supplier s Name Serial Number Pump P680 HPG DIONEX 1920105 Autosampler ASI 100 DIONEX 9932012 Column Oven TCC 100 DIONEX Demo RI Detector RI 101 Shodex not
20. 49 5 1 Baseline Noise Drift and Lamp Intensity of the UV Detectot cceeeseseesseeeeeeeeees 49 5 1 1 TG ORY 2 26 E citer sre mete de ten E E EEE EN TETA EAA EE T E E 49 5 1 2 Performing and Evaluating the CheckS sssseessrresssrresserreerrnnsstennnarrennasttnnaatennsetennnaninaeanna 49 5 2 Wavelength Accuracy of the UV Detector 0 ccccccesseneseseeeeeeesneeeeeesnenseeeesaeeeeeseenseeeseeeseeeeees 50 5 2 1 FPerronmong the CHECK efra a a S 50 5 2 2 Evaluating the Check for the UV Detector eee cceeeneeeeenneeeeeeeeeeeeeaeeeeeteaeeeeeenaeeeeneaas 50 5 2 3 Evaluating the Check for the Photodiode Array Detector eeeeeeeeeeeeeeeeeeeeenneeeeeneaas 50 5 2 4 Evaluating the Check for Two Channel Detectors and the UVD 3208 eee 51 5 2 5 Evaluating the Check for Single Wavelength and VWD 3400RS Detectors ee 51 5 3 Linearity of the UV Detector ccccsssesceseseeeceeesneneeeeseenseeesenseeeeseaeseseseeeseseseanseseseaeseeseseaneeeeeaes 51 5 3 1 THEORY kee anette ee ees ari ete te eds ee eee 51 5 3 2 Performing and Evaluating the Check recesie eaaa EEE EA EAS 52 5 4 Precision of the Injection Volume sssssssesnunsnennnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnn nnn 52 5 4 1 THEO ireen T E A A A T ted Seo noosa 52 5 4 2 Performing Me GHEREA aA AEE ARAE TE SAARA A E EE ARERR 52 5 5 Carry over by the Autosampler sssssusnsennnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnn
21. Apr 14 04 Next Qualification Oct 04 e Limits and Test Results Observed max Deviation Result of all Steps Step Accuracy Test passed Step Ripple Test passed Reviewer s signature Date Operator s signature Date Chromeleon c DIONEX 2008 PQ_OQ_Report_6_8 PUMP_GRADIENT Version 6 80 SR5 Build 2413 137116 Printed 27 10 2008 3 15 PM Seq DQ_PQ_OQ_FOQ PQ_OQ HPLC DEMO_for_Manual HPLC_OQ_DEMO_RUNS OQ_STD_GRAD Page 2 of 3 Smp STD Gradient_1 Runtime 14 Apr 2004 10 10 40 e Chromatogram of STD Gradient_1 OQ_STD_GRAD 2 STD Gradient_1 uv_VIS_1 mAU WVL 265 nm Flow ml min Reviewer s signature Date Operator s signature Date Chromeleon c DIONEX 2008 PQ_OQ_Report_6_8 PUMP_GRADIENT Version 6 80 SR5 Build 2413 137116 Printed 27 10 2008 3 15 PM Seq DQ_PQ_OQ_FOQ PQ_OQ HPLC DEMO_for_Manual HPLC_OQ_DEMO_RUNS OQ_STD_GRAD Page 3 of 3 Smp STD Gradient_1 Runtime 14 Apr 2004 10 10 40 Data of STD Gradient_1 Expected Calculated Abs Critical Calculated Value Value Deviation Deviation Result Observed Value mAU 0 000 1 118 49 725 99 017 100 000 Ripple of STD Gradient_1 Z Ripple Calculated Critical Step mAU Ripple Ripple 1 00 0 007 0 003 0 500 50 00 0 018 0 008 0 500 99 00 0 053 0 025 0 500 Reviewer s signature Date Operator s signature Date Chromeleon c DIONEX 2008 PQ_OQ_Report_6_8 PUMP
22. PQ Operating Instructions 4 3 2 Connecting and Configuring the System e System connections e Connect the type K temperature sensor to the thermometer and make the settings that is adapt the sensor type and calibration values as described in the instructions for the thermometer e Fill an open standard glass vial 1 8 ml with water and place it at sample position RC8 e Insert the temperature sensor into the vial at a right angle until the tip touches the vial bottom Figure 13 Temperature sensor inserted into the vial e Rotate the carousel until the carousel cover closes completely Page 46 of 95 HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions Configuration Connect the thermometer to a free COM port on the Chromeleon Server PC Install the Dostmann Thermometer P500 P600 in the Chromeleon Server Configuration program On the General page select the COM port to which the thermometer is connected In addition install a virtual channel Device Name VirtualChannels_01 Signal Name TemperatureOVEN Preparing Chromeleon To qualify the sample temperature accuracy select the following sequence SAMPLER_TEMP_ACC 4 3 3 Performing the Check Set the sample temperature to 10 C 15 C for the ACC 3000T When the nominal temperature is reached the external thermometer is used to record the sample temperature over a period of 30 minutes i Tip Do not perform any a
23. Performance Qualification kit part no 4832 5000A Article No Description Quantity All kits 709 7021 10 ul sample loop 1 16 1 2261 0102 VS 2 VA 1 16 connecting unit 1 2200 5502 Single part hand tight fitting 2 3323 0010 Standards kit caffeine and Pyrene 1 709 6000 066 Finger tight 33 mm fitting set 5 sets ferrule and fitting screws 0 2 Kits with pressure regulator available until spring 2007 also include 2251 6010A Pressure regulator complete 1 Kits with restriction tubing available since spring 2007 also include 5035 3000 Restriction tubing ID 0 18 mm length 15 m 1 2261 0102 connecting unit VS 2 VA 1 16 1 2251 6001 PEEK tubing ID 0 25 mm length 0 10 m 1 For a kit without sample loop order part no 4832 5010A The Standards kit part no 3323 0010 contains the seven required caffeine pyrene standards The standard at position RA8 or 8 contains water as solvent Due to legal shipping restrictions the pyrene standard is shipped in solid form Before you can use the standard dissolve the solid pyrene in ml of methanol HPLC grade Unscrew the cap from the 2 ml vial labeled 3 ug Pyrene Add about 1 ml of methanol HPLC grade which is about half the vial volume Screw the cap onto the vial Make sure that the cap seals tight Shake the vial for about 10 seconds to dissolve the solid pyrene Place the vial at the appropriate positi
24. Pump P580 Fluorescence detector specific settings Settings specific for the Dionex RF2000 Detector Fluorescence Detector RF2000 Pump specific settings Log Pump ModelNo Log Pump ModelVariant Log Pump SerialNo Sampler specific settings Log Sampler ModelNo Log Sampler SerialNo Column Oven specific settings Log ColumnOven ModelNo UV Detector specific settings Log UV ModelNo Fluorescence detector specific settings Log Emission ModelNo Pressure LowerLimit 10 Pressure UpperLimit 300 SA Equate Water Pump specific settings Settings specific for the Dionex P580 SA Type Automatic Flow 1 000 1 00 Emission ExWavelength 350 Emission EmWavelength 450 Emission Gain 4 0 Emission Response HARNS Emission Step 1 Emission Average Off Detector specific settings Page 84 of 95 HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions Settings specific for the Dionex RF2000 mission Sensitivity Med 0 50 Emission Autozero 0 000 Emission AcqOn Inject 1 000 Emission EmWavelength 394 17 000 Emission AcqOff End HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 Page 85 of 95 OQ and PQ Operating Instructions 7 10 Wavelength Accuracy of the Fluorescence Detector Wavelength accuracy for fluor
25. Runtime 13 Apr 2004 23 12 48 EE DIONEX Operational Qualification e UV Detector Noise and Drift Instruments and Fluidics Instrument Name Supplier s Name Serial Number Pump P680 LPG DIONEX 1920401 Autosampler ASI 100 DIONEX 1860410 Column Oven TCC 100 DIONEX 1850409 UV Detector UVD 340U DIONEX 1830402 Chromeleon Datasystem V 6 60 Build 1428 DIONEX 11 Accessories Name Back Pressure Device Capillary L 15 m ID 0 18 mm Solvent A Water HPLC Grade Additional Information Customer Customer s Name Operator Operator s Name Operator s Jobtitle Execution Date Apr 13 04 Next Qualification Oct 04 e Limits Values and Test Results Observed Value Result Noise UV 0 030 mAU 0 021 mAU Test passed Drift UV 0 80 mAU h 0 44 mAU h Test passed Lamp Intensity UV 500000 counts s 1026240 counts s Test passed Remark Noise and drift are measured dynamically with a floated cell The limits are different from published specifications because they are valid for static conditions only empty cell E a Reviewer s signature Date Operator s signature Date Chromeleon c DIONEX 2008 PQ_OQ_Report_6_8 DET_NOISE_AND_DRIFT Version 6 80 SR5 Build 2413 137116 Printed 27 10 2008 3 16 PM Seq DQ_PQ_OQ_FOQ PQ_OQ HPLC DEMO_for_Manual HPLC_OQ_DEMO_RUNS OQ_UV_NOISE_DRIFT Page 2 of 3 Smp Detector noise drift and lamp intensity Runtime 13 Apr 2004 23 12 48 Data for Detector Noise Test Segment No No
26. T SL PL WPS 3000TBPL Analytical WPS 3000 T RS and ACC 3000 T For the five standards required for qualifying the RI detector order part no 3325 0010 Sample Position Substance Concentration Checks ASI 100 WPS 3000xx Any Sampler RAY RB1 9 Glycerin in water 5 mg ml RI detector linearity RA10 RI detector linearity RA11 RB3 RI detector linearity RA12 Glycerin in water 25 mg ml RI detector linearity 13 RA13 RBS Glycerin in water 35 mg ml RI detector linearity xx WPS 3000 T SL PL WPS 3000TBPL Analytical WPS 3000 T RS und ACC 3000 T In addition the following solvents are required Solvent Quantity Checks Methanol HPLC grade Approx 100 ml Wavelength accuracy of UV Detector exceptions Channel A see section 3 2 Water HPLC grade Approx 600 All tests except wavelength accuracy of the UV Channel A 1200 ml detector exceptions see section 3 2 Water HPLC grade with Approx 300 ml Gradient accuracy gradient precision and ripple 0 1 Vol acetone Channel B For qualifying the column compartment a calibrated thermometer is required The thermometer is provided in the Column Thermostat PQ kit part no 5705 0050A For qualifying the column compartment of the ACC 3000 T a flexible temperature sensor Tempera ture Sensor Type K for Thermometer P600 part no 6820 0010 is required in addition to the Column Thermostat PQ kit Page 28 of 95 HPL
27. compartment install the temperature sensor as shown in Figure 4 Be sure to use the type K temperature sensor and not the sensor from the Column Thermostat PQ Kit Install the temperature sensor behind the left capillary clip from a vertical point of view in the center of the oven and 2 cm away from the right edge of the heat conductive pad Page 30 of 95 HPLC_OQ_PQ_E_ Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions Figure 4 ACC 3000 T Position of the temperature sensor 3 3 2 Configuration RF2000 fluorescence detector Before you connect the RF2000 fluorescence detector with Chromeleon set the ZWAVE parameter to 1 You can set this value only on the instrument e Disable the keyboard interlock by simultaneously pressing lt Shift gt and lt CE gt e Press lt func gt until the ZWAVE command appears on the display e Press lt 1 gt on the number keypad and confirm with lt Enter gt e Enable remote operation To do so press lt func gt until the RS232 command appears on the display Confirm with lt Enter gt and then press lt func gt On the display the reading is CONNECT Confirm with lt Enter gt You can then connect the instrument with Chromeleon AD25 UV detector On the Signals page for the AD25 in the Server Configuration program change the unit to mAU instead of AU and the factor to 1000 instead of 1 000 WPS 3000 T PL autosampler For successful quali
28. for RI Detectors PGM Version December 20 2002 Restriction capillary PEEK 5 ID 0 13 mm Length 13 m A Back Pressure 90 bar Eluent A Water HPLC quality Solvent degassed via online degasser HPLC System Pump specific settings Pump Dionex P580 Pump Sampler specific settings Sampler Dionex ASI 100 T Autosampler Detector specific settings Detector Dionex UVD 340S PDA Detector specific settings Detector Shodex RI 101 Pump specific settings Log Pump ModelNo Log Pump ModelVariant Log Pump SerialNo Sampler specific settings Log Sampler ModelNo Log Sampler SerialNo Column Oven specific settings RI Detector specific settings Log RI ModelNo Pump specific settings Pressure LowerLimit 10 bar Pressure UpperLimit 300 bar sA Equate Water Flow 1 000 Pump specific settings Settings specific for the Dionex P580 SA Type Automatic Page 88 of 95 HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions RI 1 Step 1 50 RI_1 Average On Detector specific settings Settings specific for the Shodex RI 101 RI Temperature Nominal 35 Purge off Rise Time 0 50 Polarity Minus Baseline Shift 0 Sampler specific settings Settings specif
29. in the Dionex TCC 100 TCC 3100 TCC 3200 B or TCC 3000RS SD For a list of the supported valves see section 3 5 For information about the fluid connections see the images below e Case a 6 port 2 position valve Position A or 1 Position B or 2 r sunny Waste Waste Left A EFump unit Pump Unit Perry 7 o lgs essure pace Regulater Detector gt Waste Figure 2 Fluid connection for testing the Dionex P680 DGP and DGP 600A using a 6 port 2 position valve Left The valve is in position A or 1 depending on the valve type Right The valve is in position B or 2 depending on the valve type e Case b 10 port 2 position valve Position A Position B Waste Waste Left Pump Unit Waste Waste Figure 3 Fluid connection for testing the Dionex P680 DGP and DGP 600A using a 10 port 2 position valve Left The valve is in position A Right The valve is in position B e Manual injection valve Verify that the injection valve is fitted with a 10 ul sample loop e Autosampler Position the standards as shown in the tables on page 27 and the following pages e Column compartment When qualifying the column compartments the temperature sensor of the thermometer must be securely attached to the heating block e On the supported Shimadzu column compartments loosen a fastening screw insert the sensor between the screw and the metal block and carefully retighten the screw e When qualifying the Dionex ACC 3000 T column
30. measured minimum and maximum value The noise is the distance between these two lines The calculated values are averaged for all 20 intervals to establish the final value Page 62 of 95 HPLC_OQ_PQ_E_Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions 6 Troubleshooting 6 1 General Notes e A system pressure that is well above 130 bar for a flow rate of 1 ml min solvent water after the pressure regulator has been connected indicates that a capillary is contaminated Inspect and exchange the capillaries and the pressure regulator to ensure that OQ and PQ are correctly performed e f problems occur during the checks that cannot be solved observing the notes below also refer to the respective sections in the instruments manuals 6 2 Failure of Individual Checks 6 2 1 UV Detector Check Wavelength accuracy Reason Spectrum calibration was not successful during connect in Chromeleon Increased drift Remedial Action First disconnect and then reconnect the detector See below Baseline noise Drift The solvent is contaminated The lamp is too old There are air bubbles in the flow cell The detector is not yet warmed The system is not equilibrated The lamp is defective There are fluctuations in the ambient temperature Exchange the solvent Exchange the lamp Prime the flow cell Allow the detector sufficient time to warm up Rinse the system
31. measured three times The deviation between the measured and theoretical signal heights indicates the accuracy with which the pump forms the gradient The deviations between the three gradient runs indicate the gradient precision For channel A water is used as solvent for channel B water with 0 1 Vol acetone is used STD lt STD lt 0 5 0 5 The ripple is determined relative to lt 0 5 0 5 the absorption of solvent B for each step of the above gradient runs Page 20 of 95 HPLC_OQ_PQ_E_Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions Instrument Parameter Description Limits OQ PQ P2000 Flow Precision P4000 Gradient Accuracy Gradient Precision Shimadzu Flow Precision LC 10ADvp Gradient Accuracy Gradient Precision Ten injections are analyzed When the Dionex GINA 50 or GINA 160 autosampler is used six injections are analyzed All of them contain the same standard sample The relative standard deviation and the standard deviation of the retention times indicate the flow precision The greater value is the valid limit A step gradient of two channels is programmed and measured three times The deviation between the measured and theoretical signal heights indicates the accuracy with which the pump forms the gradient The deviations between the three gradient runs indicate the gradient precision For channel A water is used a
32. mm Sample 1 Pyrene in Methanol 3 ug ml Solvent A for Wavelength Accuracy Methanol HPLC Grade Additional Information Customer Customer s Name Operator Operator s Name Operator s Jobtitle Execution Date Apr 13 04 Next Qualification Oct 04 Limits Values and Test Results Obs Value Result Wavelength Accuracy at 272 1 nm 0 75 nm Test passed Wavelength Accuracy at 333 3 nm 0 75 nm Test passed Reviewer s signature Date Operator s signature Date Chromeleon c DIONEX 2008 PQ_OQ_Report_6_8 DET_WAVELENGTH_DAD Version 6 80 SR5 Build 2413 137116 Printed 27 10 2008 2 53 PM Seq DQ_PQ_OQ_FOQ PQ_OQ HPLC DEMO_for_Manual HPLC_OQ_DEMO_RUNS OQ_DAD_WAVELENGTH Page 2 of 2 Smp Detector wavelength accuracy_DAD Runtime 13 Apr 2004 19 32 27 e Data for the Wavelength Accuracy Test DAD Observed Wavelength Expected Abs Critical Calculated Pyrene Maximum 1 271 88 272 10 0 75 0 22 ok Pyrene Maximum 2 333 28 333 30 0 75 0 02 ok e Pyrene Spectrum for the Wavelength Accuracy Test DAD 60 0 PE 100 at 0 31 min 50 05 40 05 30 05 20 05 10 04 Reviewer s signature Date Operator s signature Date Chromeleon c DIONEX 2008 PQ_OQ_Report_6_8 DET_WAVELENGTH_DAD Version 6 80 SR5 Build 2413 137116 Printed 27 10 2008 2 53 PM Seq DQ_PQ_OQ_FOQ PQ_OQ HPLC DEMO_for_Manual HPLC_OQ_DEMO_RUNS OQ_UV_NOISE_DRIFT Page 1 of 3 Smp Detector noise drift and lamp intensity
33. one minute interval is defined for each step For each interval Chromeleon uses the data to calculate a regression line based on the method of least squares Parallel to the regression line two lines are drawn through the measured minimum and maximum value The noise is the distance between these two lines This is an indication for the ripple Page 58 of 95 HPLC_OQ_PQ_E_Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions 5 10 Temperature Accuracy of the Column Compartment 5 10 1 Theory Depending on the type of application temperature fluctuations of the solvent and especially of the column can result in considerable retention time fluctuations In addition to the precision of the temperature achieved with the column compartment the accuracy is important as well Only high accuracy allows transferring applications to different systems 5 10 2 Performing the Check Four measuring points are used to check the temperature accuracy of the column compartment The check is performed with the Column_Oven sequence The achieved temperature is measured with an external calibrated thermometer The achieved temperatures are compared to the set values The difference indicates the temperature accuracy Exception e Itis not possible to set the temperature on the column compartment module of the Waters Alliance 2690 Separation Module or on the Shimadzu column compartments LC 2010 CTO CTO 10Avp CTO 10ACvp u
34. peak area and concentration are represented in a graph and the regression line is determined The regression coefficient of this line indicates the linearity HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 Page 61 of 95 OQ and PQ Operating Instructions 5 15 Baseline Noise of the ELS Detector 5 15 1 Theory Baseline noise is an important specification for a detector Increased baseline noise considerably reduces the detection sensitivity as it is not possible to distinguish between small signals and noise The baseline noise of the evaporative light scattering detector mainly depends on the lamp There is a considerable increase in noise if an old lamp with poor light intensity is used The evaporator temperature and carrier gas flow also affect the noise Therefore make sure that the measuring and ambient conditions are kept constant 5 15 2 Performing and Evaluating the Check The ELS NOISE sequence is used for the noise test Water is pumped through the detector at a flow rate of 1 ml min The conditions for recording the ELS signal are as follows e Nebuliser temperature 50 C e Evaporator temperature 90 C e Carrier gas flow 1 6 SLM 4 1 bar To calculate drift and noise the measuring signal is split into 20 intervals of 1 minute each For each interval Chromeleon calculates a regression based on measured values using the method of least squares Parallel to the regression line two lines are drawn through the
35. settings Settings specific for the Dionex P580 Pump P580 Sampler specific settings Settings specific for the Dionex ASI 100 Sampler ASI 100 Detector specific settings T Settings specific for the Dionex UVD 340S Detector Diodearray UVD 340S Samples Caffeine 10 ug ml Linearity Caffeine 300 ug ml Carry over Water Carry over Pump specific settings Log Log Log Sampler specific settings Log Log Column Oven specific settings Log UV Detector specific settings Log Pump ModelNo Pump ModelVariant Pump SerialNo Sampler ModelNo Sampler SerialNo ColumnOven ModelNo UV ModelNo Fluorescence detector specific settings Log Pressure LowerLimit Pressure UpperLimit SA Equate Pump specific settings CT Ct Emission ModelNo 10 300 Water Page 76 of 95 HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions r r r Settings specific for the SA Type Flow UV_VIS_1 Wavelength UV_VIS_1 Step UV_VIS_1 Average Detector specific settings fos Settings specific for the 3DFIELD Step UV_VIS_1 Bandwidth UV_VIS_1 Ref UV_VIS_1 Vavelength RefBandwidth Pa Sampler specific settings 0 000 1 250 Settings specific for the Sampler InjectMode Sampler UpSpeed Sam
36. settings Jait RI Ready Inject RI_1 AcqOn Column 1 Acgoff 100 HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 Page 91 of 95 OQ and PQ Operating Instructions 7 13 Baseline Noise of the ELS Detector Noise for ELS Detectors PGM Version January 08 2007 Pressure regulator Pressure 103 25 bar Flow 0 3 2 ml min Eluent A Water HPLC quality Solvent degassed via online degasser HPLC System Pump specific settings Pump Dionex P680 Pump Sampler specific settings Sampler Dionex ASI 100 T Autosampler Detector specific settings Detector not specified ELS Detector specific settings Detector Polymer Laboratories ELS 2100 Detector Pump specific settings Log Pump ModelNo Log Pump ModelVariant Log Pump SerialNo Sampler specific settings Log Sampler ModelNo Log Sampler SerialNo Column Oven specific settings UV Detector specific settings Fluorescence detector specific settings RI Detector specific settings ELS Detector specific settings Log ELSD ModelNo Pump Pressure LowerLimit 10 bar Pump Pressure UpperLimit 300 bar Pump A Equate Water Pump B 0 0 Pump C 0 0 Pump D 0 0 Pump specific settings Pump Flow 1 000 Page 92 of 95 HPLC_OQ_PQ_E Manual d
37. that the peak height of the sample with the highest concentration is in the linearity range of the tested detector i e usually below 1500 mAU As an exception a linearity range of up to 2500 mAU is specified for the supported Shimadzu detectors and the Dionex VWD 3100 and VWD 3400RS detectors with an analytical flow cell installed The Dionex VWD 3100 and VWD 3400RS detectors with a semimicro or micro analytical flow cell installed have a specified linearity range of up to 1700 mAU If an autosampler is used when the Dionex PDA 100 or PDA 3000 detectors are tested only 8 ul of sample will be injected by default However when qualifying the Dionex VWD 3100 and VWD 3400RS detectors using an autosampler 13 ul of sample will be injected 5 4 Precision of the Injection Volume 5 4 1 Theory The precision of the injection volume is mainly influenced by the quality of the autosampler syringe and the syringe volume that has been adjusted to the injection volume In addition the mechanics for the syringe movement is a decisive factor for the accuracy and precision of the injection volume Especially when you use a manual injection valve verify that there are no air bubbles in the sample If a manual injection valve is used inject at least five times the sample loop volume i e inject at least 50 ul Varying injection volumes affect the peak areas even if the same standard is injected 5 4 2 Performing the Check With the Injector_Flow_Repro s
38. ug ml Solvent A Water HPLC Grade e Additional Information Customer Customer s Name Operator Operator s Name Operator s Jobtitle Execution Date Apr 14 04 Next Qualification Oct 04 e Limits Values and Test Results Observed Value Result Injector Presicision Area 0 300 RSD 0 188 RSD Test passed 0 050 RSD 0 104 RSD Flow Presicision Ret Time Test passed 0 0100 min SD 0 0011 min SD Reviewer s signature Date Operator s signature Date Chromeleon c DIONEX 2008 PQ_OQ_Report_6_8 INJ_ REPRO_AND_RET_REPRO Version 6 80 SR5 Build 2413 137116 Printed 27 10 2008 2 54 PM Seq DQ_PQ_OQ_FOQ PQ_OQ HPLC DEMO_for_Manual HPLC_OQ_DEMO_RUNS OQ_INJECTOR_FLOW_REPRO Page 2 of 3 Smp Injector and flow reproducibility __10 Runtime 14 Apr 2004 00 20 19 e Data for Injector and Flow Reproducibility Test Volume 5 0 ul Sample Name Ret Time Area min mAU min Caffeine Caffeine UV_VIS_1 UV_VIS_1 Injector and flow reproducibility __1 Injector and flow reproducibility _2 Injector and flow reproducibility _3 Injector and flow reproducibility _4 Injector and flow reproducibility _5 Injector and flow reproducibility _6 Injector and flow reproducibility _7 Injector and flow reproducibility _8 Injector and flow reproducibility _9 Injector and flow reproducibility __10 Average RSD RSD Limit SD SD Limit Result Reviewer s signature Date Operator s signature Date Chromeleon c
39. up to up to 1 7 AU 1 7 AU Linearity Five caffeine solutions are injected in different concentrations Concentration and peak area are represented in a graph The regression coefficient of the resulting line and the deviations from it indicate the linearity Pure water is pumped through the flow cell The flow rate is 1 ml min Wavelength 254 nm Lamp Intensity gt 125000 gt 100000 counts s counts s Wavelength Accuracy Pyrene is injected using methanol as solvent The flow rate is 1 ml min The characteristic maximum of pyrene is determined at 333 nm and compared to its theoretical value Five caffeine solutions are injected in different concentrations Concentration and peak area are represented in a graph The regression coefficient of the resulting line and the deviations from it indicate the linearity Pure water is pumped through the flow cell The flow rate is 1 ml min Wavelength 254 nm Lamp Intensity r2 99 98 r2 99 90 RSD lt 5 RSD lt 5 up to up to 1 5 AU 1 5 AU Linearity gt 125000 counts s gt 100000 counts s Wavelength t 0 75 nm Accuracy Pyrene is injected using methanol as solvent The flow rate is 1 ml min The characteristic maximum of pyrene is determined at 333 nm and compared to its theoretical value r2 99 90 r2 99 90 RSD lt 5 RSD lt 5 Linearity Five caffeine solutions are injected in different concentrations Concentrat
40. 00 series G1329B Agilent 1100 1200 series G1367A Agilent 1100 1200 series G1367B Agilent 1100 1200 series G1367C Sampler module of the Waters Alliance 2690 Separation Module Waters WISP 717plus TSP AS3000 AS3500 Shimadzu LC 2010 autosampler Shimadzu SIL HTA Shimadzu SIL HTC Shimadzu SIL 10ADvp Column Compartments Dionex ACC 3000 T UltiMate 3000 Dionex TCC 3000RS UltiMate 3000 Dionex TCC 3000SD UltiMate 3000 Dionex TCC 3000 UltiMate 3000 Dionex TCC 3100 UltiMate 3000 Dionex TCC 3200 B UltiMate 3000 Dionex STH 585 Dionex TCC 100 Agilent 1100 1200 series G1316A Agilent 1100 1200 series G1316B Column compartment module of the Waters Alliance 2690 Sep Module TSP AS3000 AS3500 optional Shimadzu LC 2010 column compartment Shimadzu CTO 10Avp Shimadzu CTO 10ACvp Shimadzu CTO 10ASvp Fluorescence Detectors Dionex RF2000 Dionex RF1002 Refractive Index Shodex RI 101 Detectors Agilent 1100 1200 series G1362A Evaporative Light Polymer Laboratories ELS2100 Scattering Detector If you use other instruments or a different system configuration adapt the procedures correspondingly gt section 3 5 When qualifying systems that include a Dionex FLM 3x00 Flow Manager use the NANO_CAP_LC_Templates not the HPLC_TEMPLATES In addition Chromeleon gt 6 50 SP10 6 60 SP6 or 6 70 SP3 is required HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 Page 5 of 95 OQ and PQ Operati
41. 001 The program start time is undefined SOLVENT_CHANGE 91 SOLVENT_CHANGE Sampler Missing inject command Pump Eluent A changed from methanol to water Is this correct OQ_COLUMN_OVEN 64 TemperatureOven Setting of property Average overrides channel type default Page 42 of 95 HPLC_OQ_PQ_E_Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions When qualifying a column compartment with manual data acquisition with Chromeleon 6 50 or later you can use sequence templates that were created with Chromeleon lt 6 50 only after you have deleted the STH_manual connect line from the COLUMN_OVEN program file Use the above names in the Chromeleon Server Configuration program also when you use instruments other than those listed If you do not use these names the system cannot automatically select the checks to be performed and the appropriate sequences It may then be necessary to adapt the program files manually 3 6 Performing the Checks To copy the template section 3 5 2 follow the steps below e Inthe Browser open the Qualification menu e Select Instruments OQ or Instruments PQ A Wizard guides you through copying the sequences Clicking Next gt takes you to the next Wizard steps e Select the timebase for which you want to perform OQ or PQ Enter the name of the computer on which the timebase is installed e Select the source directory of the template to b
42. 008 OQ and PQ Operating Instructions Sampler module of the Waters Alliance 2690 Separation Module Cont d Waters WISP 717plus TSP AS3000 3500 Shimadzu SIL 10Advp Precison of Injection Volume Linearity of Injection Volume Precision of Injection Volume Linearity of Injection Volume Precision of Injection Volume After a highly concentrated test lt Sa jea 1 lt 75 1 sample has been injected a blind sample is injected The blind sample contains only solvent The peak area measured in this chromatogram indicates the carry over Ten injections are analyzed Each of them contains 5 ul of the same standard The relative standard deviation of the peak areas indicates the precision of the injection volume Five injections of the same standard r2 99 90 r2 99 90 sample are analyzed The injection Rsp lt RSD lt volumes are different 5 ul to 80 ul 4 9 1 0 The injection volume and the peak area are represented in a graph The regression coefficient of the resulting line and the deviation from it indicate the linearity After a highly concentrated test lt 0 1 sample has been injected a blind sample is injected The blind sample contains only solvent The peak area measured in this chromatogram indicates the carry over Ten injections are analyzed Each of them contains 5 ul of the same standard The relative standard deviation of the peak areas indicate
43. 53 PM Seq DQ_PQ_OQ_FOQ PQ_OQ HPLC DEMO_for_Manual HPLC_OQ_DEMO_RUNS OQ_FLUORESCENCE Page 2 of 2 Smp Fluorescence_Detector_Wavelength Runtime 30 May 2001 00 54 06 e Data for the Wavelength Accuracy Test Fluorescence Detector Obs Wavelength Exp Wavelength e Chromatogram of Wavelength Accuracy Test Fluorescence Detector The emission wavelength is changed from 380 nm to 410 nm in steps of 1 nm per 15 sec The maximum of the emission spectrum is determinated as maximum of the signal of this chromatogram OQ FLUORESCENCE 2 Emission mV EM 380 nm Reviewer s signature Date Operator s signature Date Chromeleon c DIONEX 2008 PQ_OQ_Report_6_8 RF_DET_WAVE Version 6 80 SR5 Build 2413 137116 Printed 27 10 2008 2 53 PM Seq DQ_PQ_OQ_FOQ PQ_OQ HPLC DEMO_for_Manual HPLC_OQ_DEMO_RUNS OQ_STD_GRAD Page 1 of 3 Smp STD Gradient_1 Runtime 14 Apr 2004 10 10 40 EE DIONEX Operational Qualification e Step Accuracy of the STD Gradient_1 e Instruments and Fluidics Instrument Name Supplier s Name Serial Number Pump P680 LPG DIONEX 1920401 UV Detector UVD 340U DIONEX 1830402 Chromeleon Datasystem V 6 60 Build 1428 DIONEX 11 Accessories Name Back Pressure Device Capillary L 15 m ID 0 18 mm Solvent A Water HPLC Grade Solvent B for Gradient Water 0 1 Acetone e Additional Information Customer Customer s Name Operator Operator s Name Operator s Jobtitle Execution Date
44. 8 COLUMN OVEN Version 6 80 SR5 Build 2413 137116 Printed 27 10 2008 2 51 PM Seq DQ_PQ_OQ_FOQ PQ_OQ HPLC DEMO_for_Manual HPLC_OQ_DEMO_RUNS OQ_COLUMN_OVEN Page 2 of 2 Smp Oven Temperature Runtime 14 Apr 2004 11 29 06 e Data for Temperature Accuracy Setpoint Measured Deviation Result Temperature Temperature C C Obs max Deviation Limit If the setpoint temperature of 10 C cannot be reached the reason can be that the ambient temperature is too high Nevertheless the test is passed if the measured temperature is at least 15 C below ambient Reviewer s signature Date Operator s signature Date Chromeleon c DIONEX 2008 PQ_OQ_Report_6_8 COLUMN OVEN Version 6 80 SR5 Build 2413 137116 Printed 27 10 2008 2 51 PM
45. Accuracy for detectors UVD 340S UVD 170S PDA 100 PGM Version 09 02 2000 Restriction capillary 1 13 m ID 0 13 mm PEEK Solvent Methanol degassed via online degasser 7 HPLC System Pump specific settings Settings specific for the Dionex P580 Pump P580 Sampler specific settings Settings specific for the Dionex ASI 100 Sampler ASI 100 T Detector specific settings Settings specific for the Dionex UVD 340S Detector Diodearray UVD 340S Sample Pyrene 3yg ml Pump specific settings Log Pump ModelNo Log Pump ModelVariant Log Pump SerialNo Sampler specific settings Log Sampler ModelNo Log Sampler SerialNo Column Oven specific settings Log ColumnOven ModelNo UV Detector specific settings Log UV ModelNo Fluorescence detector specific settings Log Emission ModelNo Pressure LowerLimit 10 Pressure UpperLimit 300 SA Equate Methanol Pump specific settings Settings specific for the Dionex P580 SA Type Automatic UvV_VIS_1 Wavelength 331 HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 Page 67 of 95 OQ and PQ Operating Instructions UV_VIS_1 Bandwidth 1 UV VIS 1 Step 0 20 UV VIS 1 Average On UV VIS 2 Wavelength 333 UV_VIS_ 2 Bandwidth iL UV VIS 2 Step 0 20 UV VIS 2 Average On UV_VIS_ 3 Wavelen
46. C_OQ_PQ_E_Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions 3 2 Test Procedure for Single Wavelength and VWD 3400RS Detectors Some important steps in the test procedure for single wavelength detectors and Dionex VWD 3400RS detectors differ from the steps described in section 3 1 The Standards kit part no 3323 0010 includes seven caffeine and pyrene standards However as caffeine is used for the wavelength accuracy check you do not have to prepare that is dissolve the pyrene standard Sample position RA1 or 1 is not used In addition the solvent used for the wavelength accuracy check is water not methanol Therefore only the following solvents are required Solvent Quantity Checks Water HPLC grade Approx 700 1300 ml All Channel A Water HPLC grade with 0 1 Approx 300 ml Gradient accuracy gradient Vol acetone Channel B precision and ripple 3 3 Connecting and Configuring the System The steps below describe the fluid connections of the HPLC system and all configuration settings required for OQ and PQ in Chromeleon Server Configuration Program or on the instrument Perform all steps for each module in the system 3 3 1 System Connections e System Remove the column from the system In both positions of the motorized switching valve of the autosampler thoroughly rinse all fluid components of the autosampler and injection valve with water R
47. Detector Noise Instruments and Fluidics Instrument Name Supplier s Name Serial Number Pump P580 LPG DIONEX 9730 028 Autosampler ASI 100 DIONEX 9932012 Column Oven STH 585 DIONEX not available Fluorescence Detector RF2000 DIONEX not available Chromeleon Datasystem Version 6 80 SR5 Build DIONEX 11 Accessories Back Pressure Device Capillary L 15 m ID 0 18 mm Solvent A Water HPLC Grade Additional Information Customer Customer s Name Operator Operator s Name Operator s Jobtitle Execution Date May 30 01 Next Qualification Nov 01 Limits Values and Test Results Observed Value Result Noise RF 0 30 mV 0 10 mV Test passed Signal RF min 40 00 mV 42 97 mV Test passed Signal RF max 80 00 mV 43 10 mV Test passed Reviewer s signature Date Operator s signature Date Chromeleon c DIONEX 2008 PQ_OQ_Report_6_8 RF_DET_NOISE Version 6 80 SR5 Build 2413 137116 Printed 27 10 2008 2 53 PM Seq DQ_PQ_OQ_FOQ PQ_OQ HPLC DEMO_for_Manual HPLC_OQ_DEMO_RUNS OQ_FLUORESCENCE Page 2of3 Smp Fluorescence_Detector_Noise Runtime 30 May 2001 00 36 00 e Data for RF Detector Noise Segment No Noise mV 1 2 3 4 5 6 7 8 Average Limit Result Reviewer s signature Date Operator s signature Date Chromeleon c DIONEX 2008 PQ_OQ_Report_6_8 RF_DET_NOISE Version 6 80 SR5 Build 2413 137116 Printed 27 10 2008 2 53 PM Seq DQ_PQ_OQ_FOQ PQ_OQ HPLC DEMO_for_Manual HPLC
48. Detectors Noise and Drift with Dummy Flow Cells For qualifying the Dionex VWD 3100 and VWD 3400 detectors with dummy flow cell you have to add the UV_NOISE_DRIFT_VWD3x00 sequence manually to the batch list This sequence can be used only for the above detectors and the test procedure requires that the flow cell be changed twice The following table shows the drift and noise limits for dummy flow cells The specifications have to be entered into the report manually oer ea OQ VWD 3100 Baseline Noise Measured with the dummy flow 0 010 mAU 7 T mAU VWD 3400RS cell included in the shipment 1 OQ limits with optimum measuring conditions recommended PQ limits 4 3 Dionex Autosamplers Sample Temperature Accuracy This section describes how the sample temperature accuracy is determined for the following Dionex autosamplers WPS 3000TRS WPS 3000TPL WPS 3000TBPL Analytical WPS 3000TSL and ACC 3000T For this test only the autosampler is required The other modules of the HPLC system are not required 4 3 1 Test Procedure The following table lists the materials required for performing the test Part No Description Quantity 6820 0010 Type K temperature sensor for P600 thermometers 1 5705 0050A Column Thermostat PQ Kit 1 In addition a standard glass vial 1 8 ml is required Fill the vial with water do not seal HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 Page 45 of 95 OQ and
49. ERMHENE_localAuditsiC UltiMate3000 Figure 11 Entering the location for saving the template Page 38 of 95 HPLC_OQ_PQ_E_ Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions e A list of sequences is displayed based on the timebase configuration in the Chromeleon Server Configuration program Instruments Operational Qualification Setup Ea Checks Select the checks to be installed H Wavelength O0_UVY_Noise_Dnift VJOG_Injector_Flow_FRepro VjOQ_UV_Linearity iv Y 00_SAMPLER_LIN_CO 00_STD_GRAD 00_LONG_GRAD 00_MICRO_GRAD OG_sToP Figure 12 List of sequences for the timebase When Chromeleon cannot automatically determine the pump type or the pump variant you can adapt the sequences as required This applies to the Dionex M480 pump that is the HPG_M480 sequence for the M480 HPG and LPG_M480 sequence for the M480 LPG It also applies to the mixing chamber size of the Dionex P680 and UltiMate 3000 pumps except for the low pressure micro pumps volume of the installed mixing chamber STD_GRAD sequence for the standard mixing chamber Micro_Grad sequence when the MicroFlow Kit is installed and LONG_GRAD sequence when a mixing chamber extension is installed In all other cases the selection is read only Select the sequences required for the checks that you want to perform section 4 The selected sequences are automatically copied to the corresp
50. G1315A flow cell The flow rate is 1 ml min G1315B Wavelength 254 nm G1315C Wavelength Pyrene is injected using methanol 2 nm 2nm G1315D Accuracy as solvent The flow rate is 1 ml min G1365A The characteristic maximum of G1365B pyrene is determined at 333 nm and G1365C compared to its theoretical value G1365D G1314A Baseline Noise Pure water is pumped through the 0 04 mAU 0 04 mAU G1314B flow cell at a flow rate of 1 0 ml min G1314C Wavelength 254 nm Wavelength Caffeine is injected using water as 2 0nm 2 0nm Accuracy solvent The flow rate is 1 ml min The characteristic maximum of caffeine is determined at 272 5 nm and compared to its theoretical value Page 8 of 95 HPLC_OQ_PQ_E_Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions Instrument Parameter Description Limits OQ Waters PDA996 Baseline Noise Pure water is pumped through the 0 10 mAU 0 10 mAU Waters PDA2996 flow cell The flow rate is 1 ml min 1 0 mAU h 1 0 mAU h Wavelength 254 Lamp Intensity ere ee Not Not checked checked Wavelength Pyrene is injected using methanol 1 0 nm 1 0nm Accuracy as solvent The flow rate is 1 ml min The characteristic maximum of pyrene is determined at 333 nm and compared to its theoretical value Linearity Five caffeine solutions are injected r2 99 90 r2 99 90 in different concentrations RSD lt 5 IRSD lt 5 Concentration and peak area are repres
51. HPLC Grade e Additional Information Customer Customer s Name Operator Operator s Name Operator s Jobtitle Execution Date Jan 26 07 Next Qualification Jul 07 Limits Values and Test Results Limit Observed Value Result Carry Over Area 0 050 no data Test passed Reviewer s signature Date Operator s signature Date Chromeleon c DIONEX 2008 PQ_OQ_Report_6_8 INJ_CARRY_OVER Version 6 80 SR5 Build 2413 137116 Printed 27 10 2008 3 11 PM Seq DQ_PQ_OQ_FOQ PQ_OQ HPLC DEMO_for_Manual HPLC_OQ_DEMO_RUNS OQ_SAMPLER_LIN_CO Smp Inject solvent e Chromatogram for Carry Over Test Page 2 of 2 Runtime 26 Jan 2007 15 30 06 UV_VIS_1 OQ_SAMPLER_LIN_CO 9 Inject solvent MAU 2 Caffeine 0 567 WVL 272 nm e Data for Carry Over Test Sample Name Ret Time min Caffeine UV_VIS_1 Inject solvent_Reference Carry over_Reference Carry over Inject solvent Carry over Limit Result Area mAU min Caffeine UV_VIS_1 0 132 4 842 447 221 0 093 no data 0 050 ok Reviewer s signature Date Chromeleon c DIONEX 2008 Version 6 80 SR5 Build 2413 137116 Operator s signature Date PQ_OQ_Report_6_8 INJ_CARRY_OVER Printed 27 10 2008 3 11 PM Seq DQ_PQ_OQ_FOQ PQ_OQ HPLC DEMO_for_Manual HPLC_OQ_DEMO_RUNS OQ_FLUORESCENCE Page 1 of 3 Smp Fluorescence_Detector_Noise Runtime 30 May 2001 00 36 00 ZI DIONEX Operational Qualification e Fluorescence
52. I detector e 0 5 more hours when checking the ELS detector e 2 more hours when checking a ternary high pressure gradient system channels C and B After the wavelength accuracy of the UV detector has been checked that is after approximately 15 min or 3 h 15 min you are prompted to change the solvent for channel A from methanol to water If necessary connect the fluid components of the RI detector to the system If an autosampler is installed OQ PQ will then run automatically Exemption It is not necessary to change the solvent manually when qualifying systems in which the UV detector is a Dionex VWD 3400RS or a single wavelength detector 3 8 Evaluating the Sequences To evaluate the detector linearity enter the actual concentrations for the used standards into the amount columns of the QNT file The master sequence on the Chromeleon CD and thus all copies made from it for OQ and PQ are linked to the corresponding report Do not change this report for exceptions gt section 3 5 3 In the report many references link the separate data sheets When lines or columns are inserted or deleted the references are lost and thus the calculations will be wrong To ensure that the data are correctly read and processed in the report print the report as Batch Report from the Browser Select the sequence for which you want to print the report Verify that no sample is selected On the File menu select Batch Report and then click OK to start
53. IS_1 UV_VIS_1 UV_VIS_1 da 250 MAU 200 1504 1004 WVL 265 nm Data of three Gradients Calculated Calculated Calculated Expected Value Gradient_1 Gradient_2 Gradient_3 STD Value Value Value Critical Calculated STD Result Reviewer s signature Date Operator s signature Date Chromeleon c DIONEX 2008 PQ_OQ_Report_6_8 PUMP_GRADIENT_REPRO Version 6 80 SR5 Build 2413 137116 Printed 27 10 2008 3 15 PM Seq DQ_PQ_OQ_FOQ PQ_OQ HPLC DEMO_for_Manual HPLC_OQ_DEMO_RUNS OQ_COLUMN_OVEN Page 1 of 2 Smp Oven Temperature Runtime 14 Apr 2004 11 29 06 EE DIONEX Operational Qualification e Temperature Accuracy of the Column Oven Instruments and Fluidics Instrument Name Supplier s Name Serial Number Pump P680 LPG DIONEX 1920401 Autosampler ASI 100 DIONEX 1860410 Column Oven TCC 100 DIONEX 1850409 UV Detector UVD 340U DIONEX 1830402 Chromeleon Datasystem V 6 60 Build 1428 DIONEX 11 Accessories Thermometer SN 43077 Temperature Sensor SN 111988 Additional Information Customer Customer s Name Operator Operator s Name Operator s Jobtitle Execution Date Apr 14 04 Next Qualification Oct 04 e Limits Values and Test Results Obs max Deviation Result Temperature of Column Oven Test passed Reviewer s signature Date Operator s signature Date Chromeleon c DIONEX 2008 PQ_OQ_Report_6_
54. O_CAP_LC_TEMPLATES Demo and Reports The HPLC_TEMPLATES directory contains all master sequences required for OQ or PQ of a common HPLC configuration This directory has a SPECIAL HPLC TEMPLATES subdirectory for special tests gt section 4 When creating the sequence templates the Wizard provides only those sequences that match the timebase For IC and BioLC systems the Wizard provides the sequences from the IC_TEMPLATES directory For systems with electrochemical detector the sequences from the ED_TEMPLATES directory are provided For nano cap and micro systems the Wizard provides the sequences from the NANO_CAP_LC_TEMPLATES directory and for HPLC systems the Wizard provides the sequences from the HPLC_TEMPLATES directory The Dionex PDA 100 and PDA 3000 detectors are included in the master sequences of the IC_TEMPLATES and HPLC_TEMPLATES directories These OQ PQ operating instructions refer only to the sequences of the HPLC_TEMPLATES directory HPLC_OQ_PQ_E_Manual doc Version 6 8 dated Oktober 2008 Page 35 of 95 OQ and PQ Operating Instructions i Tip z PQ_OO DEMO E ED_Templates HPLC_Templates SPECIAL_HPLC_TEMPLATES E IC_Templates E NANO_CAP_LC_Templates H E Reports Figure 6 PQ_OQ directory structure on the CM_CD The sequences of the HPLC_TEMPLATES directory do not support the qualification of systems that include a Dionex FLM 3x00 Flow Manager Sequences for qualifying these systems are availab
55. PQ setup wizard welcome page e Select the timebase for which you want to perform OQ or PQ Enter the name of the computer on which the timebase is installed Instruments Operational Qualification Setup My Computer C_UltiMate3000 _ C_UltiMates000 Favorites C GERMHENE My Computer Network Neighborhood Figure 9 Selecting a timebase HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 Page 37 of 95 OQ and PQ Operating Instructions e Select the source directory of the master sequences Select PQ_OQ Instruments Operational Qualification Setup xi Insert the Chromeleon CD Insert the Chromeleon CD in the drive indicated below or select another drive C Get the files from CD Drive D m Get the files from another location Source CM_CD PO_og Use this option if you have copied the PG 0G master template from the Chromeleon CD to a network datasource to specify the folder which contains the PQ 0G master template Figure 10 Selecting the source directory e Enter a unique name for the timebase The directory that contains all sequence templates is then saved under this name Instruments Operational Qualification Setup xi Store Template Please specify the desired location of your customized template Name Directory LENU3000500_Templates hd Browse Note The recommended location for the templates is the subdirectory 0Q_Templates inside the timebase s directory C G
56. RUNS OQ_SAMPLER_LIN_CO Page 2 of 2 Smp Injector linearity_5 Runtime 14 Apr 2004 00 43 39 e Calibration Curve Caffeine External UV_VIS_1 Area mAU min e Data for Injector Linearity Test Sample Name Ret Time i Vol Area min mAU min Caffeine Caffeine UV_VIS_1 UV_VIS_1 Injector linearity_1 Injector linearity_2 Injector linearity_3 Injector linearity_4 Injector linearity_5 Cal Type Number of Points Offset Slope a VIS_1 a VIS_1 ca VIS_1 aa VIS_1 a ee cores 184 os 447 99 99979 99979 0238 238 as ee 99000 ae 500 Result Reviewer s signature Date Operator s signature Date Chromeleon c DIONEX 2008 PQ_OQ_Report_6_8 INJ LINEARITY Version 6 80 SR5 Build 2413 137116 Printed 27 10 2008 3 11 PM Seq DQ_PQ_OQ_FOQ PQ_OQ HPLC DEMO_for_Manual HPLC_OQ_DEMO_RUNS OQ_SAMPLER_LIN_CO Page 1 of 2 Smp Inject solvent Runtime 26 Jan 2007 15 30 06 EBV DIONEX Operational Qualification Injector Carry Over Instruments and Fluidics Instrument Name Supplier s Name Serial Number Pump LPG 3400M DIONEX 1340410 Autosampler WPS 3000PL DIONEX 7 Column Oven TCC 3200 DIONEX Demo UV Detector UVD 340U DIONEX 4310409 Chromeleon Datasystem V 6 80 SP2 Build 2258 Be DIONEX 11 Accessories Name Back Pressure Device Capillary L 15 m ID 0 18 mm Sample 2 Caffeine in Water 10 ug ml Sample 7 Caffeine in Water 2000 ug ml Sample 8 Water HPLC Grade Solvent A Water
57. TNGOPY A E E eves A A E eee E ned dines 59 5 102 Performing the Check vise ereader rereinen asain andrei idi oei ienei aiii eei aerea 59 5 11 Baseline Noise and Signal Height of the Fluorescence Detector usssssusseunnnrnnnnnnnnnnnnnnn 59 OSTA gt TMC a E P EA ANS E P E E P A PE E E E PE P E E E 59 5ST Performing the Che k iummieniias niiae aa e ein a ee ei 60 Contents II of IV HPLC_OQ_PQ_E_Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions 5 12 Wavelength Accuracy of the Fluorescence Detector cccccesseccceseereeeesneeeesesneeeseeeseenseeeses 60 SIAT Performing the Checka aeir aa haan eect Sar ie a ae en 60 5 12 2 Remarks on the Manufacturer Specification ccccccccccceeeeeeeeeeceeeeeeeeeesecnaeeeeeseeeeeeeennaees 60 5 13 Baseline Noise and Drift of the RI Detector c c eccceeceeeeeeeeeeeeeeeeeeeeeeeaneeeeeeeeeeeseeesneeeeeees 61 Sae INEI aea ees sae duetasde dtc as shediaadeudntess saaodelasBect fetusuahte es danedelastanteavasanivassaabtessiatanteadaoteens 61 5 13 2 Performing and Evaluating the Check ccceccceeeeeeee ee eeeeeeeeeeeeeeeeeseeeaeeeseeeaeeeseeaaeeeeenaaees 61 5 14 Linearity of the RI Detectors cccc tccccceseecivcitescienciesccecetsscceceessttiecctasccecntsstenesessitertissecedearsstcncee eere 61 SIAT TROON shiccvcsssecvdlabs pevedencdticns E recie abhccesdtagieesddah ccedian AE 61 5 14 2 Performing and Evaluating the Check c ccceee
58. There are air bubbles in the flow cell Remedial Action Rinse the sample and reference cells for up to one hour using degassed water flow rate 1 ml min Repeatedly press the Purge key If necessary repeat the procedure with methanol Drift The solvent is contaminated There are fluctuations in the ambient temperature There are air bubbles in the flow cell Use new solvent Position the detector at a location with few temperature fluctuations Rinse the sample and reference cells with degassed water see above Detector linearity The concentration of the standard is not correct Use fresh standards 6 2 6 ELS Detector Check Reason Remedial Action Baseline noise The pump pulsation is too high Purge the pump and all channels if necessary Baseline spikes The gas supply is contaminated Replace the gas supply HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 Page 65 of 95 OQ and PQ Operating Instructions Page 66 of 95 HPLC_OQ_PQ_E_Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions 7 PGM Files As described in section 3 5 the OQ and PQ Setup Wizards adapt the program files to the instruments installed in the related timebase The section lists the PGM files for the instruments of a Dionex HPLC system 7 1 Wavelength Accuracy of the Photodiode Detector Wavelength
59. UV Detector ecceeeceeeeseeeeereeeeees 70 7 3 Linearity of the UV Detector ccccsssesecesseeeeeeeeeneeeeseeseseseeeseeeesneeseseseaeseseseanseseseeeseeeesenseeneaes 72 7 4 Precision of the Injection Volume sssssussesnunsnennnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnn nnn 74 7 5 Carry over by the Autosampler and Linearity of the Injection Volume cseseeereeeeees 76 7 6 Sample Temperature ACCUIaCy r aaa re aae aa E ee aaa a a aaa E aem taa EEK ataa aaa aE aan 78 7 7 Solvent Composition of a Gradient Pump Accuracy Precision and Ripple 80 7 8 Temperature Accuracy of a Column Compartment c eceeceeeeeeeeeeeeeneeeeeeeeseeseeneeeeeeees 82 7 9 Baseline Noise and Signal Height of the Fluorescence Detector usssssussennnnrnnnnnrnnnnnnnnn 84 7 10 Wavelength Accuracy of the Fluorescence Detector ccccesseccceseeneeeeseenseesseeeneeeseeeneeeeses 86 7 11 Baseline Noise and Drift of the RI Detector cc eccecececeeeeeeeeeeeeeeeeeeseeaneeseeeeeeseneeesneeeeeees 88 7 12 Linearity of the RI Detector 0 ccccecseeeeeeseeeeeeeeeeeeeeeeeeeseeeseeeeseeeseeeseseseaeeeseseaeseseseaeeeseneeneeeeanes 90 7 13 Baseline Noise of the ELS Detector ccccccceceeeeeeeee cece eee eeeeeeeeeeeeeee see snaeeeseseeeeseeeeesneeeeeees 92 8 Example Report sc ccevss cor este ew tether oe co eee eat ee we tae 95 HPLC_OQ_PQ_E Manual doc
60. Version 6 8 dated Oktober 2008 Contents III of IV OQ and PQ Operating Instructions Contents IV of IV HPLC_OQ_PQ_E_ Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions 1 How to Use this Manual The layout of this manual is designed to provide quick reference to the sections of interest to the user However we recommend that you review the manual thoroughly before starting Operational or Performance Qualification in order to obtain full understanding of the procedure This manual is provided as is Every effort has been made to supply complete and accurate information and all technical specifications and programs have been developed with the utmost care However Dionex assumes no responsibility and cannot be held liable for any errors omissions damage or loss that might result from any use of this manual or the information contained therein We appreciate your help in eliminating any errors that may appear in this document At various points throughout the manual message of particular importance are indicated by the following symbols whose relevance is as follows a Tip Indicates general information to help obtain optimum performance of the instrument The information contained in this document is subject to change without notice CHROMELEON Summit HPLC and UltiMate 3000 are registered trademarks of Dionex Corp Any other mentioned trade or company names are subject to the copyright an
61. _GRADIENT Version 6 80 SR5 Build 2413 137116 Printed 27 10 2008 3 15 PM Seq DQ_PQ_OQ_FOQ PQ_OQ HPLC DEMO_for_Manual HPLC_OQ_DEMO_RUNS OQ_STD_GRAD Page 1 of 3 Smp STD Gradient_2 Runtime 14 Apr 2004 10 36 48 EE DIONEX Operational Qualification e Step Accuracy of the STD Gradient_2 e Instruments and Fluidics Instrument Name Supplier s Name Serial Number Pump P680 LPG DIONEX 1920401 UV Detector UVD 340U DIONEX 1830402 Chromeleon Datasystem V 6 60 Build 1428 DIONEX 11 Accessories Name Back Pressure Device Capillary L 15 m ID 0 18 mm Solvent A Water HPLC Grade Solvent B for Gradient Water 0 1 Acetone e Additional Information Customer Customer s Name Operator Operator s Name Operator s Jobtitle Execution Date Apr 14 04 Next Qualification Oct 04 e Limits and Test Results Observed max Deviation Result of all Steps Step Accuracy Test passed Step Ripple Test passed Reviewer s signature Date Operator s signature Date Chromeleon c DIONEX 2008 PQ_OQ_Report_6_8 PUMP_GRADIENT Version 6 80 SR5 Build 2413 137116 Printed 27 10 2008 3 15 PM Seq DQ_PQ_OQ_FOQ PQ_OQ HPLC DEMO_for_Manual HPLC_OQ_DEMO_RUNS OQ_STD_GRAD Page 2 of 3 Smp STD Gradient_2 Runtime 14 Apr 2004 10 36 48 e Chromatogram of STD Gradient_2 OQ_STD_GRAD 3 STD Gradient_2 UV_VIS_1 JmAU WVL 265 nm Flow ml min Reviewer s signature Date Operator s signature Date Chro
62. _OQ_DEMO_RUNS OQ_FLUORESCENCE Page 30f3 Smp Fluorescence_Detector_Noise Runtime 30 May 2001 00 36 00 Charts for RF Detector Noise Test RF Detector Noise 15 Segment No Reviewer s signature Date Operator s signature Date Chromeleon c DIONEX 2008 PQ_OQ_Report_6_8 RF_DET_NOISE Version 6 80 SR5 Build 2413 137116 Printed 27 10 2008 2 53 PM Seq DQ_PQ_OQ_FOQ PQ_OQ HPLC DEMO_for_Manual HPLC_OQ_DEMO_RUNS OQ_FLUORESCENCE Page 1 of 2 Smp Fluorescence_Detector_Wavelength Runtime 30 May 2001 00 54 06 FE DIONEX Operational Qualification e Fluorescence Detector Wavelength Accuracy Instruments and Fluidics Instrument Name Supplier s Name Serial Number Pump P580 LPG DIONEX 9730 028 Autosampler ASI 100 DIONEX 9932012 Column Oven STH 585 DIONEX not available Fluorescence Detector RF2000 DIONEX not available Chromeleon Datasystem Version 6 80 SR5 Build DIONEX 11 Accessories Back Pressure Device Capillary L 15 m ID 0 18 mm Solvent A Water HPLC Grade e Additional Information Customer Customer s Name Operator Operator s Name Operator s Jobtitle Execution Date May 30 01 Next Qualification Nov 01 Limits Values and Test Results Obs Deviation Result Wavelength Accuracy RF Test passed Reviewer s signature Date Operator s signature Date Chromeleon c DIONEX 2008 PQ_OQ_Report_6_8 RF_DET_WAVE Version 6 80 SR5 Build 2413 137116 Printed 27 10 2008 2
63. adient Pump PGM Version 22 05 2001 Restriction capillary PEEK ID 0 13 mm Length 13 m Back Pressure 80 bar Solvent A Water HPLC quali ct ct Solvent B Water 0 1 SO HPLC System Pump specific settings fo 2 ty Acetone both HPLC quality lvents degassed via online degasser Settings specific for the Dionex P580 Pump P58 0 Sampler specific settings Settings specific for the Dionex ASI 100 Sampler ASI 100 Detector specific settings Settings specific for the Dionex UVD 340S Diodearray UVD 340S Detector T Pump specific settings Log Log Log Sampler specific settings Log Log Pump ModelNo Pump ModelVariant Pump SerialNo Sampler ModelNo Sampler SerialNo ColumnOven ModelNo UV ModelNo Emission ModelNo Column Oven specific settings Log UV Detector specific settings Log Fluorescence detector specific settings Log 2 000 Pressure LowerLimit GUE Pressure UpperLimit SA Equate SB Equate Pump specific settings Settings specific SA Type B Type oO 10 300 Water Water with 0 1 Acetone for the Dionex P580 Automatic Automatic Page 80 of 95 HPLC_OQ_PQ_E_Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Ope
64. alibrated thermometer is used to measure the temperature that is reached If technically possible the data are automatically read by Chromeleon Else enter the values manually during the measurement A 4 step temperature gradient is programmed An externally calibrated thermometer is used to measure the temperature that is reached If technically possible the data are automatically read by Chromeleon Else enter the values manually during the measurement A 4 step temperature gradient is programmed An externally calibrated thermometer is used to measure the temperature that is reached If technically possible the data are automatically read by Chromeleon Else enter the values manually during the measurement 2 C 3 C Measured Measured at at 35 C 35 C 40 C 40 C 1 C Measured 2 C Measured Measured at 5 C 20 C 60 C 85 C 2 C Measured at 5 C 20 C 60 C 80 C Page 22 of 95 HPLC_OQ_PQ_E_Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions Instrument Parameter Description Limits OQ PQ Column compartment module of the Waters Alliance 2690 Separation Module Column oven of the TSP AS3000 AS3500 Autosamplers Shimadzu Column Compartments LC 2010 CTO 10ASvp Shimadzu Column Compartment CTO 10Avp Shimadzu Column Compartment CTO 10ACvp Temperature Accu
65. alify a gradient pump except the Dionex UltiMate 3000 pumps the STD_GRAD sequence is required For the Dionex M480 an M300 pumps separate sequences are available which are automatically selected by the Wizard For the ternary high pressure gradient systems from Shimadzu the Wizard automatically selects TERN_GRAD_C_B sequence This sequence is used to determine the accuracy and the precision of the gradient and the ripple between the solvent channels C and B HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 Page 57 of 95 OQ and PQ Operating Instructions 5 9 3 Performing the Checks for the Dionex P680 and UltiMate 3000 Pumps Five sequences are provided for qualifying the Dionex LPG 3400A B DPG 3600A B HPG 3200A HPG 3200M HPG 3200RS HPG 3400A HPG 3400M HPG 3400RS and P680 pumps STD_GRAD MICRO_GRAD LONG_GRAD STD_GRAD_P680DGP_Left and LONG_GRAD_P680DGP_Left The MICRO_GRAD sequence is used to qualify a pump with MicroFlow kit e g an HPG 3200M or HPG 3400M The gradient program corresponds to the program of the STD_GRAD sequence However the mixing volume of a pump with MicroFlow kit is lower than the volume of a pump with standard mixing chamber That is why the detector signal is detected earlier This fact is considered for the evaluation of the check The LONG_GRAD sequence is used if a mixing chamber extension 600 ul or 1200 ul for UltiMate 3000 pumps or 1250 ul for the P680 pump is installed A h
66. alue of the drift Parallel to the regression line two lines are drawn through the measured minimum and maximum value The noise is the distance between these two lines The calculated values are averaged for all 20 intervals to establish the final value 5 14 Linearity of the RI Detector 5 14 1 Theory The detector linearity mainly depends on the optical and electronic systems With electronic systems non linearity is caused by dark current and dark current drift Dark measurements can be used to compensate the influence of these factors However with decreasing light intensity caused by lamp ageing or absorption of the solvent or sample the influence of the dark current on the linearity increases As water is the solvent in both the sample and reference cells the influence of the solvent is negligible The influence of the sample is fully used to determine the detector linearity Consider that the resulting deviations of the linear behavior are only important with high sample concentrations and high absorption rates gt 600 RIU 5 14 2 Performing and Evaluating the Check To establish the detector linearity the RI_LINEARITY sequence is required The detector linearity is determined using five glycerin standards set concentrations 5 mg ml 10 mg ml 15 mg ml 25 mg ml and 35 mg ml dissolved in water the actual concentrations are entered into the QNT file and taken into account Water is used as solvent The flow rate is 1 ml min The
67. and peak area are up to up to represented in a graph The 1 5 AU 1 5 AU regression coefficient of the resulting line and the deviations from it indicate the linearity TSP UV6000 Baseline Noise Pure water is pumped through the 0 06 mAU 0 10 mAU flow cell The flow rate is 1 ml min Drift 2 0 mAU h 4 0 mAU h Veena 2s an 2 0 mAU h 4 0 mAU h Lamp Intensity Not Not checked checked TSP UV6000 Wavelength Pyrene is injected using methanol 1 0nm 1 0nm Cont d Accuracy as solvent The flow rate is 1 ml min The characteristic maximum of pyrene is determined at 333 nm and compared to its theoretical value Linearity Five caffeine solutions are injected r2 99 90 r2 99 90 in different concentrations RSD lt 5 IRSD lt 5 Concentration and peak area are represented in a graph The ve AG ve AG regression coefficient of the resulting line and the deviations from it indicate the linearity Page 10 of 95 HPLC_OQ_PQ_E_Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions Parameter Description Limits OQ Shimadzu Baseline Noise Pure water is pumped through the 0 05 mAU 0 10 mAU LC 2010 SPD flow cell The flow rate is 1 ml min SPD 10Avp Drift Wavelength 254 nm 0 8 mAU h 2 0 mAU h A SPD 10AVvp Lamp Intensity l Not Not checked checked Wavelength Pyrene is injected using methanol 1 0 nm t 1 0 nm Accuracy as solvent The flow rate is 1 ml min The ch
68. and the deviations from it indicate the linearity Pure water is pumped through the 0 03 mAU 0 10 mAU flow cell The flow rate is 1 ml min Wavelength 254 nm gt 1300000 gt 1000000 0 counts s 0 counts s Pyrene is injected using methanol 1 0nm 1 0nm as solvent The flow rate is 1 ml min The characteristic maximum of pyrene is determined at 333 nm and compared to its theoretical value Five caffeine solutions are injected r2 99 90 r2 99 90 in different concentrations RSD lt 5 IRSD lt 5 Concentration and peak area are up to up to represented in a graph The 1 5 AU 1 5 AU regression coefficient of the resulting line and the deviations from it indicate the linearity Pure water is pumped through the 0 03 mAU 0 04 mAU flow cell The flow rate is 1 ml min 0 2 mAU 0 2 mAU A a ae 02mAU 02mAU Not Not checked checked Caffeine is injected using water as t 2 0 nm t 2 0 nm solvent The flow rate is 1 ml min The characteristic maximum of caffeine is determined at 272 5 nm and compared to its theoretical value Five caffeine solutions are injected r2 99 90 r2 99 90 in different concentrations RSD lt 5 RSD lt 5 Concentration and peak area are represented in a graph The A AU ve ve regression coefficient of the resulting line and the deviations from it indicate the linearity HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 Page 7 of 95 OQ and PQ Operating Instr
69. andard vial is filled with water Measured at 10 C Ten injections are analyzed Each of them contains 5 ul Standard Con figuration or 20 ul 250 ul injection volume kit of the same standard The relative standard deviation of the peak areas indicates the precision of the injection volume Five injections of the same standard sample are analyzed The injection volumes are different The injection volume and the peak area are represented in a graph The regression coefficient of the resulting line and the deviation from it indicate the linearity After a highly concentrated test sample has been injected a blind sample is injected The blind sample contains only solvent The peak area measured in this chromato gram indicates the carry over The sample temperature is measured using an external thermometer in a standard vial The standard vial is filled with water Measured at 10 C Ten injections are analyzed Each of them contains 5 ul of the same standard The relative standard deviation of the peak areas indicates the precision of the injection volume Five injections of the same standard sample are analyzed The injection volumes are different 5 ul to 80 ul The injection volume and the peak area are represented in a graph The regression coefficient of the resulting line and the deviation from it indicate the linearity After a highly concentrated test sample has been injected a blind sample is injecte
70. aracteristic maximum of pyrene is determined at 333 nm and compared to its theoretical value Linearity Five caffeine solutions are injected r2 99 90 r2 99 90 in different concentrations RSD lt 5 RSD lt 5 Concentration and peak area are represented in a graph The e i e i regression coefficient of the resulting line and the deviations from it indicate the linearity 1 2 OQ limits with optimum measuring conditions recommended PQ limits When qualifying a detector with a non analytical flow cell such as a micro nano or dummy flow cell you have to enter the corresponding specifications manually into the report The reason is that automatic recognition of micro and nano flow cells is not supported For information about the limits for a non analytical flow cell refer to the table in section 2 3 2 3 The lamp intensity is measured only for controlled detectors 2 3 2 UV Detectors Using Non Analytical Flow Cells When qualifying a detector with a non analytical flow cell such as a micro or nano flow cell you have to enter the corresponding specifications manually into the report The reason is that automatic recognition of micro and nano flow cells is not supported or not implemented The table lists the limits that apply for non analytical flow cells Instrument Parameter Description VWD 3100 Baseline Noise Pure water is pumped through the 0 035 mAU 0 070 mAU VWD 3400RS far ila cell The flow rate is 1 m
71. ate Chromeleon c DIONEX 2008 PQ_OQ_Report_6_8 PUMP_GRADIENT Version 6 80 SR5 Build 2413 137116 Printed 27 10 2008 3 15 PM Seq DQ_PQ_OQ_FOQ PQ_OQ HPLC DEMO_for_Manual HPLC_OQ_DEMO_RUNS OQ_STD_GRAD Page 1 of 2 Smp STD Gradient_3 Runtime 14 Apr 2004 11 02 57 EE DIONEX Operational Qualification e Reproducibility of the Stand Gradient Instruments and Fluidics Instrument Name Supplier s Name Serial Number Pump P680 LPG DIONEX 1920401 UV Detector UVD 340U DIONEX 1830402 Chromeleon Datasystem V 6 60 Build 1428 DIONEX 11 Accessories Name Back Pressure Device Capillary L 15 m ID 0 18 mm Solvent A Water HPLC Grade Solvent B for Gradient Water 0 1 Acetone Additional Information Customer Customer s Name Operator Operator s Name Operator s Jobtitle Execution Date Apr 14 04 Next Qualification Oct 04 e Limits and Test Results Observed max STD Result of all Steps Gradient Presicision Test passed Reviewer s signature Date Operator s signature Date Chromeleon c DIONEX 2008 PQ_OQ_Report_6_8 PUMP_GRADIENT_REPRO Version 6 80 SR5 Build 2413 137116 Printed 27 10 2008 3 15 PM Seq DQ_PQ_OQ_FOQ PQ_OQ HPLC DEMO_for_Manual HPLC_OQ_DEMO_RUNS OQ_STD_GRAD Smp STD Gradient_3 e Overlay of three Gradients lt d 0Q_STD_GRAD 4 STD Gradient_3 lt 2 OQ_STD_GRAD 2 STD Gradient_1 4 3 OQ_STD_GRAD 3 STD Gradient_2 Page 2 of 2 Runtime 14 Apr 2004 11 02 57 UV_V
72. ation Customer Customer s Name Operator Operator s Name Operator s Jobtitle Execution Date Apr 14 04 Next Qualification Oct 04 Limits Values and Test Results Observed Value Result Detector Linearity Corr 99 980 99 999 Test passed Detector Linearity 5 000 RSD 0 422 RSD Test passed Reviewer s signature Date Operator s signature Date Chromeleon c DIONEX 2008 PQ_OQ_Report_6_8 DET_LINEARITY Version 6 80 SR5 Build 2413 137116 Printed 27 10 2008 3 16 PM Seq DQ_PQ_OQ_FOQ PQ_OQ HPLC DEMO_for_Manual HPLC_OQ_DEMO_RUNS OQ_UV_LINEARITY Page 2 of 2 Smp Detector linearity_5 Runtime 14 Apr 2004 00 33 46 e Data for Detector Linearity Sample Name Amount Area ppm mAU min Caffeine Caffeine UV_VIS_1 UV_VIS_1 Detector linearity_1 4 258 Detector linearity_2 25 036 Detector linearity_3 i 58 740 Detector linearity _4 92 094 Detector linearity_5 l 124 911 e Calibration Curve Caffeine External UV_VIS 1 Area mAU min Cal Type Number of Points Offset Slope UV_VIS_1 UV_VIS_1 UV_VIS_1 UV_VIS_1 99 999 0 422 a 99 980 5 000 Result Reviewer s signature Date Operator s signature Date Chromeleon c DIONEX 2008 PQ_OQ_Report_6_8 DET_LINEARITY Version 6 80 SR5 Build 2413 137116 Printed 27 10 2008 3 16 PM Seq DQ_PQ_OQ_FOQ PQ_OQ HPLC DEMO_for_Manual HPLC_OQ_DEMO_RUNS OQ_RI_NOISE_DRIFT Page 1 of 3 Smp Rl Detector noise drift Runtime 31 Jan
73. available Chromeleon Datasystem Version 6 80 SR5 Build 24 DIONEX 11 Accessories Back Pressure Device Capillary L 15 m ID 0 18 mm Sample 9 Glycerine in Water 5 mg ml Sample 10 Glycerine in Water 10 mg ml Sample 11 Glycerine in Water 15 mg ml Sample 12 Glycerine in Water 25 mg ml Sample 13 Glycerine in Water 35 mg ml Solvent A Water HPLC Grade e Additional Information Customer Customer s Name Operator Operator s Name Operator s Jobtitle Execution Date Feb 24 03 Next Qualification Aug 03 Limits Values and Test Results Observed Value Result Detector Linearity Corr Rl 99 900 99 999 Test passed Reviewer s signature Date Operator s signature Date Chromeleon c DIONEX 2008 PQ_OQ _Report_6_8 RI_LINEARITY Version 6 80 SR5 Build 2413 137116 Printed 27 10 2008 2 54 PM Seq DQ_PQ_OQ_FOQ PQ_OQ HPLC DEMO_for_Manual HPLC_OQ_DEMO_RUNS OQ_RI_LINEARITY Page 2 of 2 Smp RI_Detector linearity_5 Runtime 24 Feb 2003 09 45 59 e Data for Detector Linearity Sample Name Amount Area Height HRIU min RIU Glycerine Glycerine Glycerine RI_1 RL1 RL1 RI_Detector linearity_1 6 536 Rl_ Detector linearity_2 10 00 12 223 144 5 Rl_Detector linearity_3 15 00 17 967 210 4 Rl_Detector linearity_4 25 00 29 214 343 2 Rl_ Detector linearity _5 35 00 40 476 473 2 Glycerine External JArea LRIU min Cal Type Number of Points Offset Slope RI RI RI RI Oe Correlation Coefficient Limit
74. c Flow 1 000 UV_VIS_1 Wavelength 254 UV VIS _1 Step 1 0 UV_VIS_1 Average On Po Detector specific settings Settings specific for the Dionex UVD 340S 3DFIELD Step I UV VIS 1 Bandwidth 1 UV VIS 1 RefWavelength 600 UV_VIS_1 RefBandwidth Sampler specific settings Settings specific for the Dionex ASI 100 Sampler InjectMode Normal Sampler UpSpeed 10 0 Sampler DownSpeed 10 0 Sampler SampleHeight 0 50 Sampler WashSpeed 50 00 Sampler DispSpeed 50 00 Sampler DrawSpeed 25 00 Sampler SyringeDelay 5 0 000 UV Autozero Detector specific settings Inject UV_VIS_1 AcqOn 1 000 Log LampiIntensity Protocol The lamp intensity is given at 254 nm in counts s 20 000 UV VIS 1 AcqOff HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 Page 71 of 95 OQ and PQ Operating Instructions 7 3 Linearity of the UV Detector UV Detector Linearity PGM Version 22 05 2001 Restriction capillary PEEK 5 ID 0 13 mm Length 13 m A Back Pressure 80 bar Solvent A Water HPLC quality Solvent degassed via online degasser HPLC System Pump specific settings Settings specific for the Dionex P580 Pump P580 Sampler specific settings Settings specific for the Dionex ASI 100 Sampler ASI 100 T Detecto
75. ction sensitivity as it is not possible to distinguish between small signals and noise With increased drift it is more difficult to integrate the signals correctly because the less stable the baseline is the more inaccurate is integration The baseline noise of the detector mainly depends on the lamp There is a considerable increase in noise if an old lamp with poor light intensity is used This is also true when the reference and or sample cell is dirty In addition make sure that the measuring and ambient conditions are constant and that the flow cell is free from gas bubbles To measure the drift of a RI detector make sure that the measuring and ambient conditions are constant In addition it is very important that the lamp has been burning for several hours and that the flow cell has been rinsed sufficiently 5 13 2 Performing and Evaluating the Check The RIi_NOISE_DRIFT sequence includes both the checks of the noise and the drift Water is pumped through the sample cell at a flow rate of 1 ml min the reference cell too has been rinsed with water before The RI signal is recorded at a temperature of 35 C To calculate drift and noise the measuring signal is split into 20 intervals of 1 minute each For each interval Chromeleon calculates a regression based on measured values using the method of least squares The slope of the curve indicates the drift of the measured signal the absolute value of the slope indicates the absolute v
76. ctor linearity check failed The syringe is old See above Exchange the syringe 6 2 3 Pump Check Flow precision Reason The autosampler draws air from the vial There are air bubbles in the syringe The autosampler is leaking The injection valve is leaking Remedial Action Either there is too little sample volume in vial or the value set for the Needle Depth parameter is too low Prime the syringe Autosampler Manual Autosampler Manual Gradient accuracy There is air in the system The system is not equilibrated The composition of solvent B is not correct Prime the system Rinse the system Make sure that the solvent composition is correct Gradient precision There is air in the system The system is not equilibrated Prime the system Rinse the system Ripple There is air in the system The system is not equilibrated Prime the system Rinse the system 6 2 4 RF2000 Fluorescence Detector Check Wavelength accuracy Reason The Raman peak of water is not visible because the instrument performs an Autozero whenever the wavelength is changed Remedial Action On the instrument set the ZWAVE parameter to 1 gt section 3 4 1 Page 64 of 95 HPLC_OQ_PQ_E_Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions 6 2 5 RI Detector Check Baseline noise Reason
77. ctor specific settings Inject UV_VIS 1 AcqOn 1 250 1 AcqOff m c Dp al 272 0 20 On Dionex UVD 3408S 0 2 600 Dionex ASI 100 Normal 10 0 10 0 0i 50 50 00 50 00 25 00 5 HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 Page 73 of 95 OQ and PQ Operating Instructions 7 4 Precision of the Injection Volume Injector and Flow Reproducibility PGM Version 22 05 2001 Restriction capillary PEEK 5 ID 0 13 mm Length 13 m A Back Pressure 80 bar Solvent A Water HPLC quality Solvent degassed via online degasser HPLC System Pump specific settings Settings specific for the Dionex P580 Pump P580 Sampler specific settings Settings specific for the Dionex ASI 100 Sampler ASI 100 T Detector specific settings Settings specific for the Dionex UVD 340S Detector Diodearray UVD 340S Sample Caffeine 140 pg ml Pump specific settings Log Pump ModelNo Log Pump ModelVariant Log Pump SerialNo Sampler specific settings Log Sampler ModelNo Log Sampler SerialNo Column Oven specific settings Log ColumnOven ModelNo UV Detector specific settings Log UV ModelNo Fluorescence detector specific settings Log Emission ModelNo Pressure LowerLimit 2 Pressure UpperLimit 300 SA Equate Water
78. d The blind sample contains only solvent The peak area measured in this chromatogram indicates the carry over r2 99 99 RSD lt 0 5 r2 99 90 RSD lt 1 0 lt 0 03 lt 0 05 RSD lt 0 3 r2 99 99 RSD lt 0 5 RSD lt 0 5 r2 99 90 RSD lt 1 0 lt 0 1 HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 Page 15 of 95 OQ and PQ Operating Instructions conic Gel a ol OQ PQ GINA 50 GINA 160 250 ul syringe Precision of Injection Volume Linearity of Injection Volume Carry Over Six injections are analyzed Each of RSD lt RSD s them contains 10 ul of the same 0 4 0 5 standard sample The relative standard deviation of the peak areas indicates the precison of the injection volume Five injections of the same standard r2 99 99 r2 99 90 sample are analyzed The injection volumes are different 10 ul to 80 ul Injection volume and peak area are represented in a graph The regression coefficient of the resulting line and the deviations from it indicate the linearity After a highly concentrated test sample has been injected a blind sample is injected The blind sample contains only solvent The peak area measured in this chromatogram indicates the carry over Agilent 1100 1200 G1313A G1329A G1329B G1367A G1367B G1367C Precision of Injection Volume Linearity of Injection Volume Carry Ove
79. d at 20 C 40 C 60 C 80 C 3 C Measured at 20 C 40 C 60 C 3 C Measured at 35 C 60 C 80 C 3 C Measured at 20 C 60 C 80 C 1 OQ limits with optimum measuring conditions recommended PQ limits 2 C Measured Measured at 25 C 35 C 45 C 60 C 3 C Measured at 35 C 45 C 60 C 3 C Measured at 25 C 45 C 60 C 2 It is not possible to set the temperature on the column compartment module when the retention time is negative The first measurement reading is 10 minutes after the sample has been started At this time equilibration of the column compartment may not be complete Therefore the same temperature is set also for the second measuring point The column compartment module has passed the check even if the target temperature is reached only for the second measuring point 3 According to the specification of the column compartment module only target temperatures above ambient are permitted That is why measuring points below 35 C are not evaluated HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 Page 23 of 95 OQ and PQ Operating Instructions 2 3 6 Fluorescence Detectors RF 2000 Baseline Noise Pure water is pumped through the lt 7 30 mV lt z 30 mV flow cell The flow rate is 1 ml min Excitation wavelength 350 nm emission wavelength 394 nm Signal Pure water
80. d at 333 nm and compared to its theoretical value Five caffeine solutions are injected in different concentrations Concentration and peak area are represented in a graph The regression coefficient of the resulting line and the deviations from it indicate the linearity Pure water is pumped through the flow cell The flow rate is 1 ml min Wavelength 254 nm Pyrene is injected using methanol as solvent The flow rate is 1 ml min The characteristic maximum of pyrene is determined at and 333 nm and compared to its theoretical value r2 99 97 r2 99 90 RSD lt 3 RSDS3 up to up to 2 5 AU 2 5 AU 0 03 mAU 0 05 mAU 0 8 mAU h 2 0 mAU h gt 500000 gt 400000 counts s counts s 0 75nm 0 75nm r2 99 98 RSDsS5 up to 1 5 AU r2 99 90 RSD lt 5 up to 1 5 AU 1 0 mAU h 1 0 mAU h Page 6 of 95 HPLC_OQ_PQ_E_ Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions OQ PQ DAD 3000 RS MWD 3000 RS analytical flow cell Cont d PDA 3000 PDA 100 PDA 100U PDA 3000 PDA 100 PDA 100U Cont d Baseline Noise Lamp Intensity Wavelength Accuracy Lamp Intensity Wavelength Accuracy Linearity Five caffeine solutions are injected r2 99 95 r2 99 90 in different concentrations RSD lt 5 IRSD lt 5 Concentration and peak area are represented in a graph The ve AD ve a regression coefficient of the resulting line
81. d the property and trademark rights of the respective companies All rights reserved including those for photomechanical reproduction and storage on electronic media Without the written permission of Dionex no part of this publication may be reproduced in any form by means of photocopy microfilm or any other process for any purpose or processed copied transmitted or distributed in any other form independent from the means electronic or mechanical that is used HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 Page 1 of 95 OQ and PQ Operating Instructions Page 2 of 95 HPLC_OQ_PQ_E_Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions 2 Introduction The increasing number of standards and official regulations provide evidence that it is extremely important to monitor the used instruments and to make sure that they work as intended if you want to achieve reliable analytical results To make the results transparent quality management according to ISO 9000 and following monitors and documents the quality of the equipment at different times For a description of the Operational Qualification OQ and Performance Qualification PQ procedures refer to the sections below 2 1 Defining the Limits According to The development and application of guidance on equipment qualification of analytical instruments of P Bedson and M Sargent Accred Qual Assur 1996 1 265 274 the follo
82. detector linearity Consider that the resulting deviations of the linear behavior are only important with high absorption rates gt 1 5 AU 300 Peak area mAU min 0 50 100 150 200 250 300 350 Concentration ppm Figure 17 Linearity of the detector signal depending on the peak area HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 Page 51 of 95 OQ and PQ Operating Instructions 5 3 2 Performing and Evaluating the Check The detector linearity is established with the UV_Linearity sequence The detector linearity is determined at 272 nm using five different caffeine standards set concentrations 10 ug ml 60 ug ml 140 ug ml 220 ug ml and 300 ug ml dissolved in water the actual concentrations are entered into the QNT file and taken into account Water is used as solvent The flow rate is 1 ml min The peak area and concentration are represented in a graph and the regression line is determined The regression coefficient and the relative standard deviation of this line indicate the linearity i Tip Depending on which injection module you use it may happen that the peak height of the sample with the highest concentration increases 1500 mAU This is usually not within the linearity range of UV detectors Thus the limits for the regression coefficient and the relative standard deviation may not be met In this case reduce the injection volume for all samples used for the linearity check so
83. e Autosampler User defined sample Sample loop volume loop volume setting WPS 3000 T PL 20 39 ul 20 ul 40 124 ul 50 ul gt 125 ul 125 ul WPS 3000 T SL 20 39 ul 20 ul WPS 3000 T RS 40 129 pl Micro meaa a gees pete hee gt 130 ul Analytical With 250 ul injection volume kit 344 ul 344 ul ACC 3000 T 21 49 ul 20 ul 51 199 ul 50 ul gt 200 ul 200 ul Page 32 of 95 HPLC_OQ_PQ_E_Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions e Qualifying the column compartment Option A Using the Column Thermostat PQ kit Connect the thermometer to a free COM port on the Chromeleon server PC Install the Dostmann Thermometer P500 P600 in the Chromeleon Server Configuration program On the General tab page select the COM port to which the thermometer is connected In addition install a virtual channel Device name VirtualChannels_017 signal Name TemperatureOVEN i Tip When changing the temperature sensor you may have to adapt the calibration values and sensor type setting of the thermometer Otherwise the thermometer may show the wrong temperature This is especially important when qualifying the column compartment of the ACC 3000 T which is qualified using a type K temperature sensor Option B Automatic data acquisition as analog signal In the Chromeleon Server Configuration program install the analog output of the exte
84. e e g 10 C depending on the autosampler type When the nominal temperature is reached the sample water temperature is recorded over a period of 30 minutes Within the 30 minutes the sample temperature reaches a stable value The temperature accuracy is the temperature difference between the sample temperature and the nominal autosampler temperature 5 8 Flow Precision 5 8 1 Theory The flow precision can be determined very exactly by weighing out which quantity of solvent is delivered over a specific period For statistic evaluation of the results repeat this measurement several times However this requires a lot of work The measuring time must be at least five minutes if it is not electronically linked to the weighing process Otherwise inaccuracies in the timing affect the results An additional disadvantage is that the procedure cannot be automated and that the used scales must be very exact As an alternative the flow precision can be determined by injecting the same sample standard multiple times The flow precision primarily affects the precision of the retention time This method is used during automated OQ and PQ 5 8 2 Performing the Check The precision of the flow and the precision of the injection volume are established with the Injector_Flow_Repro sequence Standard 4 is injected ten times using an injection volume of 5 ul for each injection deviations see the table in section 5 4 2 The relative standard deviation
85. e pump forms the gradient The deviations between the three gradient runs indicate the gradient precision For channels A and C water is used as solvent for channel B water with 0 1 Vol acetone is used For each step of the above gradient runs the ripple is determined relative to the absorption of solvent B A step gradient of two channels is programmed and measured three times The deviation between the measured and theoretical signal heights indicates the accuracy with which the pump forms the gradient The deviations between the three gradient runs indicate the gradient precsion For channels A and C water is used as solvent for channel B water with 0 1 Vol acetone is used O lt 1 5 lt 2 0 Gradient ot checked Not checked Precision Ripple Flow Precision For each step of the above gradient lt 0 5 runs the ripple is determined relative to the absorption of solvent B Six injections are analyzed All of them contain the same standard sample The relative standard deviation and the standard deviation 0 02 min of the retention times indicate the flow precision The greater value is the valid limit HPLC_OQ_PQ_E_Manual doc Version 6 8 dated Oktober 2008 Page 19 of 95 OQ and PQ Operating Instructions PQ Agilent 1100 1200 Flow Precison G1310A G1311A G1312A G1312B Gradient Accuracy Agilent 1100 1200 G1311A G1312A G1312B Gradient Precision
86. e used e Enter a unique name for saving the copy default template name date e A list is displayed containing all sequences of the corresponding template Click to select the sequences required for the checks gt section 4 After the sequences have been copied the batch list of the corresponding timebase is automatically opened Start the batch to process the sequences The checks are processed in the following order 1 Fluid preparation of the system Warm up sequence 2 Temperature accuracy of the column compartment for manual data acquisition Column Oven sequence 3 Wavelength accuracy of the UV detector Wavelength sequence Baseline noise drift and lamp intensity of the UV detector UV Noise Drift sequence Precision of injection volume and flow njector Flow Repro sequence and Injector Flow Repro_P680DGP_Left sequence if necessary Linearity of the UV detector UV Linearity sequence Linearity of the injection volume Sampler Lin CO sequence Carry over by the autosampler Sampler Lin CO sequence Baseline noise signal height and wavelength accuracy of the fluorescence detector Fluorescence sequence 10 Baseline noise and drift of the RI detector RI_Noise_Drift sequence 11 Linearity of the RI detector R _Linearity sequence 12 Baseline noise of the evaporative light scattering detector ELS Noise sequence 13 Solvent composition of gradient pumps accuracy precision and ripple LPG_M480 or HPG_M480 se
87. ecause the pump does not support a gradient program with more than 9 steps HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 Page 21 of 95 OQ and PQ Operating Instructions 2 3 5 Thermostatted Column Compartments and Column Ovens Instrument Parameter Description Limits OQ PQ Column compartment module of the ACC 3000 T autosampler TCC 3000RS TCC 3000SD STH 585 Agilent 1100 1200 G1316A G1316B Temperature Accuracy Temperature Accuracy Temperature Accuracy Temperature Accuracy Temperature Accuracy Temperature Accuracy A 3 step temperature gradient is programmed An externally calibrated thermometer is used to measure the temperature that is reached If technically possible the data are automatically read by Chromeleon Else enter the values manually during the measurement A 4 step temperature gradient is programmed An externally calibrated thermometer is used to measure the temperature that is reached If technically possible the data are automatically read by Chromeleon Else enter the values manually during the measurement A 4 step temperature gradient is programmed An externally calibrated thermometer is used to measure the temperature that is reached If technically possible the data are automatically read by Chromeleon Else enter the values manually during the measurement A 4 step temperature gradient is programmed An externally c
88. ected The blind sample contains only solvent The peak area measured in this chromatogram indicates the carry over 0 02 lt 0 02 1 OQ limits with optimum measuring conditions recommended PQ limits 2 3 4 Pumps Instrument Parameter Description Limits OQ PQ ISO 3100A LPG 3400A B LPG 3400M B DGP 3600A B DGP 3600M B HPG 3200RS HPG 3200A HPG 3200M HPG3400RS HPG 3400A HPG 3400M P680 and P580 with analytical pump heads LPG 3400A B LPG 3400M B DGP 3600A B DGP 3600M B HPG 3200A HPG 3200M HPG 3200RS HPG 3400A HPG 3400M HPG 3400RS P580 HPG and LPG and P680 HPG LPG and DGP all mixing chamber types all pumps with analytical pump heads Flow Precision Gradient Accuracy Gradient Precision Ripple Ten injections are analyzed When the Dionex GINA 50 or GINA 160 autosampler is used six injections are analyzed All of them contain the same standard sample The relative standard deviation and the standard deviation of the retention times indicate the flow precision The greater value is the valid limit A step gradient of two channels is programmed and measured three times The deviation between the measured and theoretical signal heights indicates the accuracy with which the pump forms the gradient The deviations between the three gradient runs indicate the gradient precision For channel A water is used as solvent for channel B water
89. ector Turn on the detector lamp Allow the lamp to be on for at least six hours before you start the check When you use one of the following detectors just turn on the UV lamp DAD 3000 RS MWD 3000 RS PDA 100 PDA 3000 VWD 3100 VWD 3400RS AD25 TSP UV1000 TSP UV2000 TSP UV3000 TSP UV6000 or Shimadzu SPD 10AVvp Refractive Index Detector When you use an RI detector turn on the instrument at least one hour before you start the check Rinse the reference cell and the sample cell at a flow rate of 10 0 ml min mobile phase water If you check the wavelength accuracy of the UV detector using methanol all detectors except the Dionex VWD 3x00 detectors and all single wavelength detectors disconnect the fluid components of the RI detector from the HPLC system after you have rinsed the cells with water Fluorescence Detector Turn on the detector lamp Allow the lamp to be on for approximately 30 minutes before you start the check Evaporative Light Scattering Detector Turn on the detector lamp Allow the lamp to be on for approximately 30 minutes before you start the check Autosampler Before you start the check rinse the autosampler thoroughly with water To do so inject 250 ul of water at least five times If the allowed maximum injection volume of the autosampler is smaller inject five times the largest possible volume Make sure that the fluid components and the syringe are free of air bubbles Note Although methanol is us
90. ed as solvent for the first OQ and PQ check rinse the autosampler with water as water is the solvent for all successive checks Automatically rinsing the system after the wavelength accuracy check ensures that the fluid system is sufficiently prepared Pump When qualifying an RI detector rinse the entire HPLC system with water If you want to check the wavelength accuracy for the UV detector disconnect the fluid components of the RI detector from the HPLC system before you rinse channel A with methanol When qualifying Dionex VWD 3400RS detectors and all single wavelength detectors or if the wavelength accuracy is not checked use water to rinse channel A In this special case you need not disconnect the fluid components of the RI detector from the HPLC system For gradient pumps use water 0 1 Vol acetone to rinse channel B For a ternary high pressure gradient system or an M480 low pressure gradient pump use water to rinse channel C 3 4 2 Checking the Fluidics Injection Valve and Autosampler Verify that there are no pressure fluctuations when the valve switches from Load to Inject and vice versa Pressure fluctuations indicate system leakage or contamination Eliminate any leaks and contamination before you start the check Page 34 of 95 HPLC_OQ_PQ_E_Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions 3 5 Preparing Chromeleon 3 5 1 Template Structure To prepare Chromeleon for Operationa
91. eine in Water 140 pg ml LA 91957 Sample 5 Caffeine in Water 220 ug ml LA 91958 Sample 6 Caffeine in Water 300 pg ml LA 91959 Sample 7 Caffeine in Water 2000 ug ml LA 91959 Sample 8 Water HPLC Grade Sample 9 Glycerine in Water 5 mg ml Sample 10 Glycerine in Water 10 mg ml Sample 11 Glycerine in Water 15 mg ml Sample 12 Glycerine in Water 25 mg ml Sample 13 Glycerine in Water 35 mg ml Sample 14 Water Uncapped Vial Solvent A Water HPLC Grade Solvent A for Wavelength Accuracy Methanol HPLC Grade Solvent B for Gradient Water 0 1 Acetone Solvent C for Gradient Water HPLC Grade Thermometer SN 43077 Temperature Sensor SN 111988 Reviewer s signature Date Operator s signature Date Chromeleon c DIONEX 2008 PQ_OQ_Report_6_8 SPECIFICATION Version 6 80 SR5 Build 2413 137116 Printed 27 10 2008 3 24 PM Seq DQ_PQ_OQ_FOQ PQ_OQ HPLC DEMO_for_Manual HPLC_OQ_DEMO_RUNS OQ_WARM_UP Smp Warm up e Limits Criterion Noise UV Drift UV Lamp Intensity UV Detector Wavelength Accuracy Detector Linearity Corr UV Detector Linearity Injector Presicision Area Flow Presicision Ret Time Carry Over Area Injector Linearity Corr Injector Linearity Temperature of Injector Gradient Accuracy Gradient Presicision Pump Ripple Temperature of Column Oven Noise RF Signal RF min Signal RF max Wavelength Accuracy RF Noise RI Drift RI Detector Linearity Corr
92. emperature is 2 C 4 C measured using an external thermometer in a standard vial The standard vial is filled with water measured at 15 C Ten injections of the same standard RSD lt RSD s are analyzed The injection volume 0 3 0 5 is as follows 5 ul analytical autosampler 2 ul micro autosampler and 10 ul autosampler with 250 ul injection volume kit The relative standard deviation of the peak areas indicates the precision of the injection volume Five injections of the same standard r2 99 99 r2 99 90 sample are analyzed The injection Rsp lt RSD lt volumes are different Analytical 0 5 1 0 autosampler 5 ul to 80 ul Micro option 1 ul to 20 ul The injection volume and the peak area are represented in a graph The regression coefficient of the resulting line and the deviation from it indicate the linearity After a highly concentrated test 0 01 lt 0 01 sample has been injected a blind sample is injected The blind sample contains only solvent The peak area measured in this chromato gram indicates the carry over The sample temperature is 2 C 4 C measured using an external thermometer in a standard vial The standard vial is filled with water measured at 10 C HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 Page 13 of 95 OQ and PQ Operating Instructions sel dela all OQ WPS 3000 T Micro and analytical autosampler versio
93. ented in a graph The a an y A regression coefficient of the resulting line and the deviations from it indicate the linearity Waters 2487 Dual Baseline Noise Pure water is pumped through the 0 05 mAU 0 05 mAU Lambda flow cell The flow rate is 1 ml min 0 5 mAU h 10 5 mAU h Wavelenath 254 bri Absorbance Detector Lamp Intensity Not Not checked checked Wavelength Pyrene is injected using methanol 1 0 nm 1 0nm Accuracy as solvent The flow rate is 1 ml min The characteristic maximum of pyrene is determined at 239 nm and compared to its theoretical value Linearity Five caffeine solutions are injected r2 99 90 r2 99 90 in different concentrations RSD lt 5 IRSD lt 5 Concentration and peak area are up to up to represented in a graph The 1 5 AU 1 5 AU regression coefficient of the resulting line and the deviations from it indicate the linearity TSP UV1000 Baseline Noise Pure water is pumped through the 0 50 mAU 0 10 mAU flow cell The flow rate is 1 ml min 0 5 mAU h 1 0 mAU h Dee an 0 5 mAU h 1 0 mAU h Lamp Intensity Not Not checked checked Wavelength Not Not Accuracy checked checked Linearity Five caffeine solutions are injected r2 99 90 r2 99 90 in different concentrations RSD lt 5 IRSD lt 5 Concentration and peak area are up to up to represented in a graph The 1 5 AU 1 5 AU regression coefficient of the resulting line and the deviations from it indicate the linearity
94. equence a caffeine standard solvent water at a flow rate of 0 3 ml min wavelength 272 nm is injected six or ten times The autosampler type determines the injection volume standard and the number of injection see the table The relative standard deviation of the peak areas of the six or ten injections indicates the precision of the injection volume Page 52 of 95 HPLC_OQ_PQ_E_Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions Autosampler Standard used Injection volume Number of injections Other Standard 4 5 ul 10 Dionex Gina 50 Standard 3 10 ul 6 Dionex Gina 160 Dionex ACC 3000 T 10 Sample loop volume 200 ul Standard 3 20 ul Dionex WPS 3000 T SL Micro Standard 5 2 ul 10 with 250 ul injection volume kit Standard 3 10 ul 10 Dionex WPS 3000TBPL Analytical Large Volume configuration Standard 3 20 ul 10 Dionex WPS 3000 T RS Micro option Standard 5 2 ul 10 Also see section 3 1 HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 Page 53 of 95 OQ and PQ Operating Instructions 5 5 Carry over by the Autosampler 5 5 1 Theory After a highly concentrated sample a sample containing only solvent is injected Ideally only the signal for the solvent is displayed in the chromatogram However if a signal for the sample is displayed this indicates the carry over by the autosampler As the highly concentrated sample e
95. escence detector 7 PGM Version 09 02 2000 Restriction capillary PEEK 5 ID 0 13 mm Length 13 m A Back Pressure 90 bar Solvent A Water HPLC quality Solvent degassed via online degasser 7 HPLC System Pump specific settings Settings specific for the Dionex P580 Pump P580 Fluorescence detector specific settings Settings specific for the Dionex RF2000 Detector Fluorescence Detector RF2000 Pump specific settings Log Pump ModelNo Log Pump ModelVariant Log Pump SerialNo Sampler specific settings Log Sampler ModelNo Log Sampler SerialNo Column Oven specific settings Log ColumnOven ModelNo UV Detector specific settings Log UV ModelNo Fluorescence detector specific settings Log Emission ModelNo Pressure LowerLimit 10 Pressure UpperLimit 300 SA Equate Water Pump specific settings Settings specific for the Dionex P580 SA Type Automatic Flow 1 000 2 00 Emission ExWavelength 350 Emission EmWavelength 450 Emission Gain 4 0 Emission Response HARNS Emission Step 1 Emission Average Off Page 86 of 95 HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions Detector specific settings
96. fication of the Dionex WPS 3000 T PL autosampler make sure that the Upgrade Kit for a 250 ul syringe is installed WPS 3000TBPL Analytical autosampler In order to ensure a successful qualifcation the Dionex WPS 3000TBPL Analytical autosampler must be equipped with the standard or large volume configuration In addition it must be activated in the Server Configuration on the Options page of the WPS 3000TBPL Analytical driver HPLC_OQ_PQ_E_Manual doc Version 6 8 dated Oktober 2008 Page 31 of 95 OQ and PQ Operating Instructions WPS 3000 Autosampler General Sharing Segments Pump Link Options Relays Inputs Error Levels Needle Size C 24 10 3 pl 154 C 30y C Other size pl Syringe Size C 25ul f 100p 250 pl C 500p C 1000p v Thermostatting option installed Micro Fraction Collection option installed Jv WPS 3000TBPL Analytical v Automatically Wash after Abort errors Loop Size C et C 20 fe 50 pl C Other size pl Buffer Tubing Size C 50 pl fe 500 pl Warning 100 pl 125 ul C 250p C 500p m C 10004 C 2000p Ensure thatthe settings on this page match the installed items Beeman Hilfe e Dionex UltiMate 3000 autosamplers with user defined sample loop volume The sample loop volume must be at least 20 ul In addition you must set the sample loop volume in the Server Configuration program to a value predefined by Chromeleon see tabl
97. gOft AcgOft AcqOff HPLC_OQ_PQ_E_Manual doc Version 6 8 dated Oktober 2008 Page 69 of 95 OQ and PQ Operating Instructions 7 2 Baseline Noise Drift and Lamp Intensity of the UV Detector Noise and Drift PGM Version 09 02 2000 Restriction capillary PEEK 5 ID 0 13 mm Length 13 m A Back Pressure 90 bar Solvent A Water HPLC quality Solvent degassed via online degasser HPLC System Pump specific settings Settings specific for the Dionex P580 Pump P580 Sampler specific settings Settings specific for the Dionex ASI 100 Sampler ASI 100 T Detector specific settings Settings specific for the Dionex UVD 340S Detector Diodearray UVD 3408S Pump specific settings Log Pump ModelNo Log Pump ModelVariant Log Pump SerialNo Sampler specific settings Log Sampler ModelNo Log Sampler SerialNo Column Oven specific settings Log ColumnOven ModelNo UV Detector specific settings Log UV ModelNo Fluorescence detector specific settings Log Emission ModelNo Pressure LowerLimit 10 Pressure UpperLimit 300 SA Equate Water Pump specific settings Settings specific for the Dionex P580 Page 70 of 95 HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions SA Type Automati
98. gth 335 UV_ VIS 3 Bandwidth 1 UV VIS 3 Step 0 20 UV VIS 3 Average On UV_VIS_ 4 Wavelength 250 UV_ VIS 4 Bandwidth L UV_VIS_ 4 Step 0 20 UV VIS 4 Average On Detector specific settings Settings specific for the Dionex UVD 3408S 3DFIELD MaxWavelength 350 0 3DFIELD MinWavelength 250 0 3DFIELD BunchWidth 1 9 3DFIELD Step 0x2 3DFIELD RefWavelength 600 3DFIELD RefBandwidth 19 UV VIS_1 RefWavelength 600 UV VIS_1 RefBandwidth 1 UV VIS 2 RefWavelength 600 UV_VIS_2 RefBandwidth 1 UV VIS_3 RefWavelength 600 UV VIS_3 RefBandwidth J UV VIS 4 RefWavelength 600 UV VIS 4 RefBandwidth 1 tosampler specific settings ttings specific for the Dionex ASI 100 Sampler InjectMode Normal Sampler UpSpeed 10 0 Sampler DownSpeed 10 0 Sampler SampleHeight 0 50 Sampler WashSpeed 50 0 Sampler DispSpeed 50 00 Sampler FillSpeed 25 0 SyringeDelay 5 Flow 1 000 0 000 UV Autozero Detector specific settings r Inject 3DFIELD AcqOn UV_VIS_1 AcqOn UV_VIS_2 AcqOn Uv_VIS_3 AcqOn UV_VIS_4 AcqOn 1 250 3DFIELD AcqOff Page 68 of 95 HPLC_OQ_PQ_E_Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions End UV VI UV VI UV VI UV VIS Yr U U V BwWNE AcgOft Ac
99. ic for the Dionex ASI 100 Sampler InjectMode Normal Sampler UpSpeed 10 0 Sampler DownSpeed 10 0 Sampler SampleHeight 0 50 Sampler WashSpeed 50 00 Sampler DispSpeed 50 00 Sampler DrawSpeed 25 00 Sampler SyringeDelay 5 40 000 Detector specific settings RI Autozero Settings specific for complete systems Wait Sampler Ready Column Oven specific settings Pump specific settings Wait RI Ready Purge on 39 500 Purge off 39 000 Purge On 38 500 Purge Off 38 000 Purge On 20 000 Purge Off 3 000 RI Autozero 0 000 Pump specific settings Inject RI_1 AcqOn 20 000 RI _1 AcqOff End HPLC_OQ_PQ_E_Manual doc Version 6 8 dated Oktober 2008 Page 89 of 95 OQ and PQ Operating Instructions 7 12 Linearity of the RI Detector RI Detector Linearity PGM Version December 20 2002 Restriction capillary PEEK A ID 0 13 mm Length 13 m Back Pressure 80 bar Eluent A Water HPLC quality Solvent degassed via online degasser 7 HPLC System Pump specific settings Pump Dionex P580 Pump Sampler specific settings Sampler Dionex ASI 100 T Autosampler Detector specific settings Detector Dionex UVD 340S PDA Detector specific settings Detector Shodex RI 101 r Samples Glycerine 5 mg ml 10 mg ml 15 mg ml 25 mg ml
100. igher mixing chamber volume increases the equilibration time of the gradient The gradient program has been adapted accordingly and evaluation of the check considers this For DGP pumps P680 and UltiMate 3000 DGP 3600A B qualification is performed for both pump units The sequences STD_GRAD and LONG_GRAD are used to check the right pump unit The sequences STD_GRAD_P680DGP_Left and LONG_GRAD_P680DGP_Left are used to check the left pump unit For the UltiMate 3000 LPG 3400M B and DGP 3600M B micro pumps only the LONG_GRAD and LONG_GRAD_P680DGP_Left sequences are available 5 9 4 Evaluating the Check To facilitate the comparison the absorption values are converted and expressed as B To compensate the detector drift the absorption of the pure solvent A is measured at the beginning and at the end of the gradient These values are the basis for the regression line that is used to correct the baseline of the entire chromatogram To define the gradient accuracy the measured step height is compared to the height that must theoretically result from the solvent composition To define the precision three gradients are recorded The standard deviations of the step heights indicate the precision The gradient precision is not determined for M480 pumps For M480 pumps a second gradient is measured in addition to the gradient described above For the second gradient channel C delivers water as solvent The ripple is determined for all steps A
101. ine in water c 60 ug ml at a flow rate of 1 ml min As water is used as solvent it is not necessary to change the solvent manually 5 2 2 Evaluating the Check for the UV Detector The signals are recorded at 331 nm 333 nm and 335 nm A parabola is calculated from the signal heights of the pyrene signal and the wavelengths The parabola maximum is determined and compared to the theoretical value of the spectral maximum for pyrene 333 3 nm 5 2 3 Evaluating the Check for the Photodiode Array Detector The UV spectrum for pyrene is recorded between 250 nm and 350 nm The spectral maxima between 250 nm and 290 nm and between 330 nm and 350 nm are determined by Chromeleon and compared to their theoretical values 272 1 nm and 333 3 nm FER Pise 100 50 0 a ia 317 8 X OTT T T T T T T T T T 2494 260 0 270 0 280 0 290 0 300 0 310 0 320 0 330 0 340 0 348 2 Figure 16 UV Spectrum of pyrene in methanol Page 50 of 95 HPLC_OQ_PQ_E_Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions 5 2 4 Evaluating the Check for Two Channel Detectors and the UVD 320S e For the following detectors the signals are recorded at 235 nm 240 nm and 245 nm Dionex UVD 160 UVD 160S UVD 320 UVD 320S detector the TSP UV2000 detector and the Waters 2487 detector A parabola is calculated from the signal heights of the pyrene signal and the wavelengths The maximum of the parabola is determined and compared t
102. inse also the pump thoroughly with water Only then connect the pressure regulator or restriction tubing from the Performance Qualification kit to the injection valve and the UV detector using the fitting screws shipped with the kit Use the 33 mm finger tight fitting for the injection valve and the single part hand tight fitting for the UV detector With the restriction tubing install the PEEK tubing from the Performance Qualification kit between the injection valve and the restriction tubing using the connecting units from the kit if necessary Figure 1 Single part hand tight fitting 2200 5502 Union 2261 0102 Injection Valve Finger tight fitting set 33 mm 709 6000 066 Column Compartment optional Restriction tubing 5035 3000 Detector E Figure 1 Restriction tubing installed between injection valve and detector If the system includes several detectors that are connected in series connect the pressure regulator or the restriction tubing to the detector that was connected to the column HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 Page 29 of 95 OQ and PQ Operating Instructions e Qualifying a Dionex P680 DGP DGP 3600A B or DGP 3600M B pump To qualify both pumps of a Dionex dual gradient pump with the same autosampler pressure regulator or restriction tubing and detector you have to use an external motorized switching valve such as a valve
103. ion and peak area are up to up to represented in a graph The 1 0 AU at 1 0 AU at regression coefficient of the 8 ul 8 ul resulting line and the deviations from it indicate the linearity 1 OQ limits with optimum measuring conditions recommended PQ limits Page 12 of 95 HPLC_OQ_PQ_E_ Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions 2 3 3 Autosamplers OQ PQ ACC 3000 T WPS 3000 T RS Analytical version and micro option Precision of Injection Volume Linearity of Injection Volume Carry Over Temperature Accuracy Precision of Injection Volume Linearity of Injection Volume Carry Over Temperature Accuracy Ten injections are analyzed Each of RSD lt RSD s them contains 5 ul of the same 0 5 1 0 standard The relative standard deviation of the peak areas indicates the precision of the injection volume Five injections of the same standard r2 99 95 r2 99 90 sample are analyzed The injection volumes are different see section 5 6 2 The injection volume and the peak area are represented ina graph The regression coefficient of the resulting line and the deviation from it indicate the linearity After a highly concentrated test 20 02 lt 0 02 sample has been injected a blind sample is injected The blind sample contains only solvent The peak area measured in this chromato gram indicates the carry over The sample t
104. ions required for that application e g solvents column material and temperature must be taken into account The check can be developed by the supplier on request However it is not part of the test procedures below Do not use limits that are more restrictive than those used for Performance Qualification are HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 Page 3 of 95 OQ and PQ Operating Instructions 2 2 Basic Requirements for Successful OQ and PQ As described in section 2 1 OQ and PQ are system specific procedures The procedures described below apply to the following instruments Instrument Supported Model Pumps Dionex ISO 3100A UltiMate 3000 Dionex LPG 3400A B UltiMate 3000 Dionex LPG 3400M B UltiMate 3000 Dionex DGP 3600A B UltiMate 3000 Dionex DGP 3600M B UltiMate 3000 Dionex HPG 3200A UltiMate 3000 Dionex HPG 3200M UltiMate 3000 Dionex HPG 3200RS UltiMate 3000 Dionex HPG 3400A UltiMate 3000 Dionex HPG 3400M UltiMate 3000 Dionex HPG 3400RS UltiMate 3000 Dionex P680 Dionex P580 Dionex M480 Dionex M300 Agilent 1100 1200 series G1310A Agilent 1100 1200 series G1311B Agilent 1100 1200 series G1311A Agilent 1100 1200 series G1312A Agilent 1100 1200 series G1312B Pump module of the Waters Alliance 2690 Separation Module TSP P2000 TSP P4000 Shimadzu LC 2010 pump Shimadzu LC 10ATvp Shimadzu LC 10ADvp UV Detectors Dionex DAD 3000 RS UltiMate 3000 Dionex MWD 3000 RS
105. is pumped through the gt 40 mV gt 40 mV Minimum flow cell The flow rate is 1 ml min Signal Excitation wavelength 350 nm Itis lt 80 mV lt 80 mV Maximum observed how the signal changes when the emission wavelength changes from 450 nm to 394 nm Wavelength Pure water is pumped through the 10 nm 10 nm Accuracy flow cell The flow rate is 1 ml min At the excitation wavelength of 350 nm the emission wavelength is varied from 380 nm to 410 nm in 1 nm increments The relative signal maximum is compared to the theoretical maximum RF 1002 Baseline Noise Pure water is pumped through the lt 0 60 mV lt 0 60 mV flow cell The flow rate is 1 ml min Excitation wavelength 350 nm emission wavelength 394 nm Signal Pure water is pumped through the gt 40 mV gt 40 mV Minimum flow cell The flow rate is 1 ml min Signal Excitation wavelength 350 nm Itis lt 80 mV lt 80 mV Maximum observed how the signal changes when the emission wavelength changes from 450 nm to 394 nm Wavelength Pure water is pumped through the 10 nm 10 nm Accuracy flow cell at 1 0 ml min At an excitation wavelength of 350 nm the emission wavelength is varied from 380 nm to 410 nm in 1 nm increments The relative signal maximum is compared to the theoretical maximum 1 OQ limits with optimum measuring conditions recommended PQ limits 2 The manufacturer specification of 2 nm for the excitation and emission wave
106. ise mAU o ON DOD oa A WO DP Oo 3a s amp s s amp s s amp s wo O Oo ON Oo A O N O Average 0 021 mAU Limit 0 030 mAU Result ok Reviewer s signature Date Operator s signature Date Chromeleon c DIONEX 2008 PQ_OQ_Report_6_8 DET_NOISE_AND_DRIFT Version 6 80 SR5 Build 2413 137116 Printed 27 10 2008 3 16 PM Seq DQ_PQ_OQ_FOQ PQ_OQ HPLC DEMO_for_Manual HPLC_OQ_DEMO_RUNS OQ_UV_NOISE_DRIFT Page 3 0f3 Smp Detector noise drift and lamp intensity Runtime 13 Apr 2004 23 12 48 Chart for Noise Test Detector Noise eoogoceseoso S0000000002 SANBDRADINBWGSO I 10 Segment No Reviewer s signature Date Operator s signature Date Chromeleon c DIONEX 2008 PQ_OQ_Report_6_8 DET_NOISE_AND_ DRIFT Version 6 80 SR5 Build 2413 137116 Printed 27 10 2008 3 16 PM Seq DQ_PQ_OQ_FOQ PQ_OQ HPLC DEMO_for_Manual HPLC_OQ_DEMO_RUNS OQ_INJECTOR_FLOW_REPRO Page 1 of 3 Smp Injector and flow reproducibility_10 Runtime 14 Apr 2004 00 20 19 FZ DIONEX Operational Qualification e Injector and Flow Reproducibility Instruments and Fluidics Instrument Name Supplier s Name Serial Number Pump P680 LPG DIONEX 1920401 Autosampler ASI 100 DIONEX 1860410 Column Oven TCC 100 DIONEX 1850409 UV Detector UVD 340U DIONEX 1830402 Chromeleon Datasystem V 6 60 Build 1428 DIONEX 11 Accessories Name Back Pressure Device Capillary L 15 m ID 0 18 mm Sample 4 Caffeine in Water 140
107. l Qualification or Performance Qualification follow the steps below The Wizard generates sequence templates from the master sequence of the Chromeleon CD providing only sequences that match the timebase In addition the Wizard adapts the programs automatically to the devices installed in the timebase Figure 5 step 1 For each check that is performed on the same system a separate copy of the sequence template is made Figure 5 step 2 OQ PQ is then performed with the sequences of the copied template section 3 6 for OQ PQ In this way you may need to adapt the sequence templates only once to the device configuration to be checked Chromeleon CH_CD PQ_OQ Browser File Edit View Workspace Qualification Batch Tools Window Help D i l E EB BEE C 7 E N i Instruments PO ENa e CM_CD Chromeleon IQ Chromeleon OG Instruments IQ Instruments OO Name J cM_og DEMO H E DEMO ED_Templates ey IQ HPLC_Templates LIB IC_Templates Nano LC Appii NANO _CAP_MIC_LC_ Templates mPOA OQ Reports f master pgm HPLC_Templates IC_Templates NANO_CAP_MIC_LC_Templates Reports QUERY Figure 5 Performing OQ PQ The PQ_OQ directory on the Chromeleon CD has the following subdirectories gt Figure 5 ED_TEMPLATES HPLC_TEMPLATES IC_TEMPLATES and NAN
108. l min micro flow cell Ort Wavetenat 254 nm a measuring wavelength of 230 nm Wavelength Caffeine is injected using water as 2 0nm 2 0nm Accuracy solvent The flow rate is 1 ml min The characteristic maximum of caffeine is determined at 272 5 nm and compared to its theoretical value Linearity Five caffeine solutions are injected r2 99 95 r2 99 90 in different concentrations RSD lt 3 IRSD lt 3 Concentration and peak area are up to up to represented in a graph The 1 7 AU 1 7 AU regression coefficient of the resulting line and the deviations from it indicate the linearity HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 Page 11 of 95 OQ and PQ Operating Instructions VWD 3100 VWD 3400RS semi micro flow cell UVD 340S UVD 170S UVD 340U UVD 170U micro flow cell UVD 340S UVD 170S UVD 340U UVD 170U nano flow cell 0 035 mAU 0 070 mAU Baseline Noise Pure water is pumped through the Drift flow cell The flow rate is 1 ml min 0 3 mAU h 0 3 mAU h Wavelength 254 nm Lamp Intensity The lamp intensity is determined at gt 50 gt 40 a measuring wavelength of 230 nm Wavelength Caffeine is injected using water as 2 0 nm 2 0nm Accuracy solvent The flow rate is 1 ml min The characteristic maximum of caffeine is determined at 272 5 nm and compared to its theoretical value r2 99 95 r2 99 90 RSD lt 3 RSD lt S3
109. le in the NANO_CAP_LC_TEMPLATES directory 3 5 2 Creating the Sequence Templates To install the sequences required for your system follow the steps below e Insert the Chromeleon CD or verify that you can access the PQ_OQ directory e From the Browser open the Qualification menu Chromeleon CH_CD PQ_0 Browser File Edit View Workspace Qualification Batch Tools Window Help D we bed E EY Chromeleon IG BEBE Sic dbp eju mp JE CM_CD PQ_OQ Brows Dee t A a G Nano LC Application Templates 1D eee i Chromeleon OG Instruments IQ 3 Instruments PQ PO Setup Instrument OG C GERKLENZEN_local Name y W CM Seminar OQ Setup Installs Edits Operational Qualification Templates CM_CD DatasoureoRG ED_Templates CM_0Q Fecal D HPLC_Templates DEMO IC_Templates ar NY NANO_CAP_MIC_LC_Templates ESG E Reports OQ or PQ Setup f master pgm DEMO ED_Templates HPLC_Templates IC_Templates NANO_CAP_MIC_LC_Templates E Reports Figure 7 Selecting OQ or PQ setup Page 36 of 95 HPLC_OQ_PQ_E_Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions e Select OQ Setup or PQ Setup A Wizard assists you in copying the sequences Clicking Next gt takes you to the next Wizard step Instruments Operational Qualification Setup Figure 8 OQ
110. lengths can be checked only using a special flow cell and a mercury lamp For OQ and PQ the instrument should preferably be checked with the components used for the measurements Page 24 of 95 HPLC_OQ_PQ_E_Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions 2 3 7 Refractive Index Detectors PQ Shodex RI 101 Baseline Noise Pure water is pumped through the zy nRIU 50 nRIU Agilent 1100 1200 i flow cell The flow rate is 1 ml min 500 nRIU 2500 nRIU h G1362A Temperature 35 C h Linearity Five glycerin solutions are injected r gt 99 9 r gt 99 9 in different concentrations Concentration and peak area are represented in a graph The regression coefficient of the resulting line and the deviations from it indicate the linearity 1 OQ limits with optimum measuring conditions recommended PQ limits 2 3 8 Evaporative Light sisal Detectors Gaia ial acc OE OQ Polymer Baseline Noise Pure water is pumped through the 0 3 mV 7 mV Laboratories flow cell The flow rate is 1 ml min ELS 2100 1 OQ limits with optimum measuring conditions recommended PQ limits HPLC_OQ_PQ_E_Manual doc Version 6 8 dated Oktober 2008 Page 25 of 95 OQ and PQ Operating Instructions Page 26 of 95 HPLC_OQ_PQ_E_Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions 3 Process 3 1 General Test Procedure All required materials are provided in the
111. meleon c DIONEX 2008 PQ_OQ_Report_6_8 PUMP_GRADIENT Version 6 80 SR5 Build 2413 137116 Printed 27 10 2008 3 15 PM Seq DQ_PQ_OQ_FOQ PQ_OQ HPLC DEMO_for_Manual HPLC_OQ_DEMO_RUNS OQ_STD_GRAD Page 3 of 3 Smp STD Gradient_2 Runtime 14 Apr 2004 10 36 48 Data of STD Gradient_2 Expected Calculated Abs Critical Calculated Value Value Deviation Deviation Result Observed Value mAU 0 000 1 150 49 788 98 991 100 000 e Ripple of STD Gradient_2 Z Ripple Calculated Critical Step mAU Ripple Ripple 1 00 0 033 0 015 0 500 50 00 0 036 0 017 0 500 99 00 0 012 0 006 0 500 Reviewer s signature Date Operator s signature Date Chromeleon c DIONEX 2008 PQ_OQ_Report_6_8 PUMP_GRADIENT Version 6 80 SR5 Build 2413 137116 Printed 27 10 2008 3 15 PM Seq DQ_PQ_OQ_FOQ PQ_OQ HPLC DEMO_for_Manual HPLC_OQ_DEMO_RUNS OQ_STD_GRAD Page 1 of 3 Smp STD Gradient_3 Runtime 14 Apr 2004 11 02 57 EE DIONEX Operational Qualification e Step Accuracy of the STD Gradient_3 e Instruments and Fluidics Instrument Name Supplier s Name Serial Number Pump P680 LPG DIONEX 1920401 UV Detector UVD 340U DIONEX 1830402 Chromeleon Datasystem V 6 60 Build 1428 DIONEX 11 Accessories Name Back Pressure Device Capillary L 15 m ID 0 18 mm Solvent A Water HPLC Grade Solvent B for Gradient Water 0 1 Acetone e Additional Information Customer Customer s Name Opera
112. nalytical UltiMate 3000 Dionex ASI 100 Dionex GINA 50 Dionex GINA 160 HP1100 G1313A Agilent 1100 1200 series G1329A Agilent 1100 1200 series G1329B Agilent 1100 1200 series G1367A Agilent 1100 1200 series G1367B Agilent 1100 1200 series G1367C Sampler Samplers Cont d Sampler module of the Waters Alliance 2690 Separation Module TSP AS3000 AS3500 Shimadzu LC 2010 sampler Shimadzu SIL HTA Shimadzu SIL HTC Shimadzu SIL 10ADvp Sampler Column thermostats Dionex ACC 3000 T UltiMate 3000 Dionex TCC 3000RS UltiMate 3000 Dionex TCC 3000SD UltiMate 3000 Dionex TCC 3000 UltiMate 3000 Dionex TCC 3100 UltiMate 3000 Dionex TCC 3200 B UltiMate 3000 Dionex TCC 100 Dionex STH 585 Agilent 1100 1200 series G1316A Agilent 1100 1200 series G1316B Column compartment module of the Waters Alliance 2690 Separation Module Shimadzu LC 2010 column compartment Shimadzu CTO 10Avp Shimadzu CTO 10ACvp Shimadzu CTO 10Asvp ColumnOven thermometer Column oven of the TSP AS3000 AS3500 Autosampler Sampler Signal name of TemperatureOV the external EN Device name of the virtual channel VirtualChannels 01 Fluorescence Dionex RF2000 Emission detectors Dionex RF 1002 RI detector Shodex RI 101 RI Agilent 1100 1200 series G1362A ELS detector Polymer Laboratories ELS2100 ELSD x Tip When you start the batch the following warnings may appear SOLVENT_CHANGE 91 Warning P0
113. nd CTO 10Asvp when the retention time is negative The first measurement reading is 10 minutes after the sample has been started At this time equilibration of the column compartment may not be complete Therefore the same temperature is set for the second measuring point 50 C too The column compartment module has passed the check even if the target temperature is reached only for the second measuring point This means that evaluation is performed for three measuring points only e Due to its small temperature range evaluation for the Dionex ACC 3000 T is performed for three measuring points only i Tip With the column compartment of the Waters Alliance 2690 Separation Module the set temperature can be changed during a sample only when the autosampler is injecting That is why 1ul of water is injected for qualifying the column compartment of the Waters Alliance 2690 Separation Module 5 11 Baseline Noise and Signal Height of the Fluorescence Detector 5 11 1 Theory Drift and baseline noise are important specifications for the detector Increased baseline noise considerably reduces the detection sensitivity as it is not possible to distinguish between small signals and noise The baseline noise of the detector mainly depends on the lamp There is a considerable increase in noise if an old lamp with poor light intensity is used In addition contamination in the flow cell leads to an increase noise level Also make sure that the mea
114. ng Instructions 2 3 Overview of the Checks The following tables provide an overview of the parameters to be checked and list the recommended PQ limits for each HPLC module 2 3 1 UV Detectors Using Analytical Flow Cells Instrument Parameter Description Limits OQ VWD 3100 VWD 3400RS analytical flow cell UVD 340S UVD 170S UVD 340U UVD 170U analytical flow cell DAD 3000 RS MWD 3000 RS analytical flow cell Baseline Noise Pure water is pumped through the flow cell The flow rate is 1 ml min Wavelength 254 nm 0 025 mAU 0 050 mAU 0 3 mAU h 0 3 mAU h Lamp Intensity The lamp intensity is determined at gt 50 gt 40 a measuring wavelength of 230 nm Wavelength Accuracy Lamp Intensity Wavelength Accuracy Baseline Noise Wavelength Accuracy Caffeine is injected using water as solvent The flow rate is 1 ml min The characteristic maximum of caffeine is determined at 272 5 nm and compared to its theoretical value Five caffeine solutions are injected in different concentrations Concentration and peak area are represented in a graph The regression coefficient of the resulting line and the deviations from it indicate the linearity Pure water is pumped through the flow cell The flow rate is 1 ml min Wavelength 254 nm Pyrene is injected using methanol as solvent The flow rate is 1 ml min The characteristic maximum of pyrene is determine
115. nnnnnnnnnnnnnnnnnnnn nnmnnn nnn 54 5 5 1 TROO ori raae EE EE E E ORA 54 5 5 2 Performing the Chetki arsinenieedeitiein opier eei areir hidey ine eased 54 5 6 Linearity of the Injection VOIUMEC ccceceseccceeeeeeeeeeeeneeeeseenseeeeeeeeseeesenseeeseaeseeeseanseeeseaeseeesees 54 5 6 1 Hii Ee a EE IE A E E E E S T A AE S O A A A pele etait 54 5 6 2 Performing the Cheikerrasneennnaeera e peed adi n 55 5 7 Sample Temperature Accuracy of AutosamplerS sssssssssessunnnnunnnnnnnnnnnnnnnnnnnnnnnnnnnnnnn nnmnnn 56 5 7 1 TRBOY 22th tantie cont haan nadie eed art eae eee aaa eld a a 56 5 7 2 Performing the Chetk oies teinar aeisi nin eevee EAEE NEETA ae tueels 56 5 8 Flow Precision cneA a cevescucedeu suds dveste eteududes dvs sudeedvesteredeuctesdeeduars 56 5 8 1 TREO ienaa er renee epeesrcerrececer cree ere rer etrecr rrceree cece error reer rpeesrcercrt tee a a a a 56 5 8 2 Performing the Gneckiesiztccucieticteesdacccit aadcceubsasictenact tet R EAE AS 56 5 9 Solvent Composition of the Gradient Pump Accuracy Precision and Ripple 57 5 9 1 TING ORY winter nite i A Cn ata 57 5 9 2 Performing the Checks oaie ices cess ek TEE ET dha has dena T ee 57 5 9 3 Performing the Checks for the Dionex P680 and UltiMate 3000 PUMPS 58 5 9 4 Evaluating the Gneck orrainn AE ARAE ANEA RER N AA ETNAS EE tua en ade te 58 5 10 Temperature Accuracy of the Column Compartment cccceeeeeseeeeeeeeeeeeeeeseeeneeeeenees 59 9210 1
116. ns and 250 ul injection volume kit WPS 3000 T SL Micro and analytical autosampler versions and Precision of Injection Volume Linearity of Injection Volume Carry Over Temperature Accuracy Ten injections of the same standard are analyzed The injection volume is as follows 5 ul analytical autosampler 2 ul micro autosampler and 10 ul autosampler with 250 ul injection volume kit The relative standard deviation of the peak areas indicates the precision of the injection volume Five injections of the same standard sample are analyzed The injection volumes are different Analytical autosampler 5 ul to 80 ul Micro autosampler 1 ul to 20 ul autosampler with 250 ul injection volume kit 10 ul to 160 pl The injection volume and the peak area are represented in a graph The regression coefficient of the resulting line and the deviation from it indicate the linearity After a highly concentrated test sample has been injected a blind sample is injected The blind sample contains only solvent The peak area measured in this chromato gram indicates the carry over The sample temperature is measured using an external thermometer in a standard vial The standard vial is filled with water measured at 10 C RSD s 0 3 r2 99 99 lt 0 01 reo 0 5 r2 99 90 lt 0 01 250 ul injection volume kit WPS 3000 T PL only with Upgrade aa RSD lt RSD lt 0 3 0 5
117. nto 20 intervals of 1 minute each For each interval Chromeleon calculates a regression based on measured values using the method of least squares Parallel to the regression line two lines are drawn through the values with maximum distance from this regression line The calculated values are averaged for all 20 intervals to establish the final value To calculate the drift Chromeleon calculates a regression line from all data points within a range of 1 to 21 minutes based on the method of least squares The slope of the regression line is the calculated drift 5 2 Wavelength Accuracy of the UV Detector 5 2 1 Performing the Check For UV detectors without PDA option the wavelength accuracy is established with the UV_Wavelength sequence For photodiode array detectors use the DAD_Wavelength sequence For single wavelength detectors and the Dionex VWD 3400RS detector use the Wavelength_Single sequence Separate sequences are available for the following detectors Dionex UVD 160 UVD 160S UVD 320 UVD 320S detetors the TSP UV 2000 detector and the Shimadzu LC 2010 SPD SPD 10Avp and SPD 10AVvp detectors The Wizard uses the appropriate sequence automatically when the corresponding detector is installed In all cases wavelength accuracy is determined using pyrene in methanol c 3 ug ml at a flow rate of 1 ml min However for single wavelength detectors and the Dionex VWD 3400RS detector the wavelength accuracy is determined using caffe
118. o 1 on the instrument see section 3 4 1 Water is pumped through the flow cell at a flow rate of 1ml min For an excitation wavelength of 350 nm the emission wavelength changes in 1nm increments from 380 nm to 410 nm The relative signal maximum is compared to the theoretical maximum 40 g_ OQ_FLUORESCENCE 2 Fluorescence_Detector_Wavelength Emission mV ie ao LE ON de ua jeusi Aye B OR Oe Qype Ai e Om e a E380 nm D R A OO eR Red fH 8 O H HO iS we Se Se Sys ys Li Li 1i ai 1i i 1i i ii i ii i Ti Ti Li ii Li Li W Li Li Li Li i 1i i i i 1i Li E pel He pelhe f e Hel e Helg Hiel e Hele e ete eee ee pg Het relHet pe ltet y e Hel e wW j wW W uH JU UH U H H g WW ug wW wW W A M A HM Hd H wW Ww wW U UH f U uH ug 35 0 t t t j t t t t t t t 7 f Pritt tmm CASAR ridin t rit I Primos tari a r TT ATN mT I I TATT yi a TN MII nT 30 0 ji A ars mf A N uij Loins youd fate Lote eee ut Liu tt unis f AL LAAU Lit N Lit LLa p Ee j fa eo AE O E Lit 4 b Hit HI i4 f J44IH LII4HE IGH TER 25 0 HeH f HEHH H 4 i tmd Hed eee H R Fit er FI t kE Ho j FEAT EAS AEEA EA ty rq j PEMAI TWEET TT mT 20 0 rmi mT HEMD eta eel avit e Th Tht bLiuit 4d J DLL L ut iui 4 1 sO eiL ui Lou Hf JJ ULL EAA L i MER dl Loi tf ui ee ge eg L iui 4 15 0 H H PHHH HHHH IH i HHH H
119. o the theoretical value of the spectral maximum for pyrene 239 4 nm e For the Shimadzu detectors the signals are recorded at 333 nm and 335 nm A parabola is calculated from the signal heights of the pyrene signal and the wavelengths The maximum of the parabola is determined and compared to the theoretical value of the spectral maximum for pyrene 333 3 nm 5 2 5 Evaluating the Check for Single Wavelength and VWD 3400RS Detectors For the following detectors the signals are recorded at 270 nm 272 nm and 274 nm Dionex VWD 3100 VWD 3400 and AD25 detectors and the G1314A G1314B and G1314C detectors of the Agilent 1100 200 series A parabola is calculated from the signal heights of the caffeine signal and the wavelengths The maximum of the parabola is determined and compared to the theoretical value of the spectral maximum for caffeine 272 5 nm 5 3 Linearity of the UV Detector 5 3 1 Theory The detector linearity mainly depends on the optical and electronic systems With electronic systems non linearity is caused by dark current and dark current drift Dark measurements can be used to compensate the influence of these factors However with decreasing light intensity caused by lamp ageing or absorption of the solvent or sample the influence of the dark current on the linearity increases As water is the solvent for this check the influence of the solvent is negligible The influence of the sample is fully used to determine the
120. oc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions iS_1 Average On LS_1 Step 0 10 s Ei m Detector specific settings Settings specific for the Polymer Laboratories ELS 2100 Detector ELSD Standby NoStandby ELSD LightSourcelIntensity 90 ELSD PMTGain 1 0 ELSD SmoothWidth 1 ELSD EvaporatorTemperature 90 C ELSD NebuliserTemperature 50 LG ELSD CarrierFlow 1 60 silm Sampler specific settings Settings specific for the Dionex ASI 100 Sampler InjectMode Normal Sampler UpSpeed 10 0 Sampler DownSpeed 10 0 Sampler SampleHeight 02 50 Sampler WashSpeed 50 00 Sampler DispSpeed 50 00 Sampler DrawSpeed 25 00 Sampler SyringeDelay 5 0 000 Detector specific settings ELSD Autozero Settings specific for complete systems Pump specific settings Jait Sampler Ready Column Oven specific settings Vait ELSD Ready Sampler Inject ELS _1 AcqOn 21 000 ELS 1 AcgOff End HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 Page 93 of 95 OQ and PQ Operating Instructions Page 94 of 95 HPLC_OQ_PQ_E_Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions 8 Example Report For an OQ example report refer to the following pages The report was generated for the following system configuration
121. og Sampler ModelNo Log Sampler SerialNo Column Oven specific settings Log ColumnOven ModelNo UV Detector specific settings Log UV ModelNo Fluorescence detector specific settings Log Emission ModelNo 35 000ColumnOven Temperature 10 00 0 000 Inject ColumnOven Temperature 10 00 TemperatureOVEN Step 1 00 TemperatureOVEN Average On TemperatureOVEN AcqgOn 9 900 ColumnOven Temperature 10 00 10 000 ColumnOven Temperature 20 00 49 900 ColumnOven Temperature 20 00 50 000 ColumnOven Temperature 60 00 89 900 ColumnOven Temperature 60 00 90 000 ColumnOven Temperature 80 00 130 000 ColumnOven Temperature 80 00 Page 82 of 95 HPLC_OQ PQ _E Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions 130 100 TemperatureOVEN AcgqOff ColumnOven Temperature 25 00 End HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 Page 83 of 95 OQ and PQ Operating Instructions 7 9 Baseline Noise and Signal Height of the Fluorescence Detector Noisetest for fluorescence detector PGM Version 09 02 2000 Restriction capillary PEEK 5 ID 0 13 mm Length 13 m A Back Pressure 90 bar Solvent A Water HPLC quality Solvent degassed via online degasser 7 HPLC System Pump specific settings Settings specific for the Dionex P580
122. on in the autosampler i Tip The pyrene standard is used for checking the wavelength accuracy of all UV detectors except the Dionex VWD 3100 and VWD 3400 detectors the AD25 detector and any other supported single wavelength detectors see Section 3 2 Concentrations deviating by 30 do not affect the test results Sample Position Concentration Checks 3 ug ml Wavelength accuracy of UV methanol detector see section 3 2 for a list of excluded detectors Caffeine in water 10 pg ml Linearity of injection volume for sample loop volumes gt 50 ul Detector linearity Carry over by the autosampler HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 Page 27 of 95 OQ and PQ Operating Instructions Sample Position Concentration Checks Dionex Sampler Caffeine in water 60 ug ml Wavelength accuracy VWD 3400RS single wavelength detectors Precision of injection volume and flow Gina 50 ACC 3000 and WPS 3000 with sample loop volumes gt 200 ul Linearity of injection volume for sample loop volumes lt 50 ul Detector linearity Caffeine in water Precision of injection volume and flow not for Gina 50 Detector linearity Caffeine in water Precision of injection volume and flow WPS 3000 T SL Micro e Detector linearity 300 ug ml e Detector linearity 2000 ug ml e _Carry over by the autosampler Water solvent gt e __Carry over by the autosampler Dionex autosamplers ASI 100 T WPS 3000
123. onding datasource When installation is complete the report opens on the Specification page i Tip If you use the TSP UV1000 UV detector you can select the sequences only if the UV lamp is turned on in the detector 3 5 3 Adapting the Report and Method e Disable writing protection on the Edit menu click Layout Mode and then enter the e charge number expiry date and actual concentration of the standard e customer name and tester name e component that generates the backpressure default capillary L 15 m ID 0 18 mm For all devices listed in the table below the instrument names and limits recommended by Dionex are automatically entered into the report only when you open the report after the Warm up sample The information is not yet entered when the sequence is copied Do not fill in the report for the supported devices see below The limits are listed from line 154 on Change the limits only if you do not want to use the limits recommended by Dionex HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 Page 39 of 95 OQ and PQ Operating Instructions i Tip When you use a Dionex UVD 170S UVD 340S UVD 170U UVD 340U VWD 3100 or VWD 3400RS detector with a non analytical flow cell you have to enter the specifications listed in section 2 3 2 or 4 2 manually into the report The reason is that automatic recognition of flow cell is not supported or not implemented The serial number is entered automatically for
124. onex HPG 3400RS UltiMate 3000 Device Name Pump Module SETETE EEEE Pump Device Name Pump Dionex ISO 3100A UltiMate 3000 Pump Dionex LPG 3400A B UltiMate 3000 Dionex HPG 3200A UltiMate 3000 Dionex HPG 3200M UltiMate 3000 Dionex HPG 3400A UltiMate 3000 Dionex HPG 3400M UltiMate 3000 Dionex P680 all models except P680A DGP 6 Dionex P580 Dionex M480 Dionex M300 Agilent 1100 1200 series G1310A Agilent 1100 1200 series G1311A Agilent 1100 1200 series G1312A Agilent 1100 1200 series G1312B Pump module of the Waters Alliance 2690 Separation Module TSP P2000 TSP P4000 Shimadzu LC 2010 pump Shimadzu LC 10ATvp Shimadzu LC 10ADvp Dionex LPG 3400M B UltiMate 3000 MicroPump Page 40 of 95 HPLC_OQ_PQ_E_ Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions Device Supported Model Name Pumps Dionex DGP 3600A B UltiMate 3000 Cont d Dionex P680 DGP 6 Left pump unit PumpLeft Right pump unit PumpRight Dionex DGP 3600M B UltiMate 3000 Left pump unit Right pump unit LoadingPump MicroPump Column switching valve for qualifying Dionex DGP pumps MSV of the Dionex TCC 3000RS UltiMate 3000 MSV of the Dionex TCC 3000SD UltiMate 3000 MSV of the Dionex TCC 3100 UltiMate 3000 MSV of the Dionex TCC 3200 B UltiMate 3000 MSV of the TCC 100 as device name ColumnOven Reodyne RV EV Valve Reodyne LabPro Valve Valco Multi Posi
125. or the standard deviation of the retention times of the six or ten injections indicates the flow precision The larger of the values is the valid limit Page 56 of 95 HPLC_OQ_PQ_E_Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions 5 9 Solvent Composition of the Gradient Pump Accuracy Precision and Ripple 5 9 1 Theory If the gradient pump composes the solvent inaccurately this will mainly effect the retention times To keep the measuring effort low different compositions are checked based on the ASTM instructions Use 100 water for solvent A Solvent B is a mixture of water and acetone 0 1 Vol Acetone is highly absorbing in the range of 265 nm The gradient can be observed in a chromatogram There are no injections required 250_OG_LPG 4 Low Pressure Gradient_3 uy_vis_4 mat WVL 265 nm D 0 0 C 0 0 J E A 1004 S aioe ets ar ge Figure 19 Theoretical broken line and real gradients STD_GRAD standard sequence for gradient pumps 5 9 2 Performing the Checks Except for the TSP P2000 Pump the following solvent compositions in B are mixed 0 1 50 99 and 100 The solvent composition in B for the TSP P2000 Pump is as follows 0 50 and 100 This is because the pump does not support gradient programs with more than 9 steps For the described arrangement and non changing solvent composition the ripple is indicated by the signal noise To qu
126. pler DownSpeed Sampler SampleHeight Sampler WashSpeed Sampler DispSpeed Sampler DrawSpeed Sampler SyringeDelay UV Autozero Detector specific settings Inject UV_VIS_1 AcqOn UV_VIS_1 AcqOff End Dionex P580 Automatic 1 00 272 0 50 On Dionex UVD 3408S 03 5 600 Dionex ASI 100 Normal 10 0 10 0 0 50 50 00 50 00 25 00 5 HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 Page 77 of 95 OQ and PQ Operating Instructions 7 6 Sample Temperature Accuracy Autosampler temperature accuracy PGM Version February 02 2007 HPLC System Pump specific settings 7 Pump Dionex DGP 3600A Pump Sampler specific settings Sampler Autosampler Ul1tiMate 3000 De De CECcLlor Sample cector specific settings UltiMate 3000 Dionex WPS 3000 T Pulledloop Dionex VWD 3400 UV_VIS Detector Water Vial uncapped UltiMate 3000 Pump specific settings Log Log Log Sampler specific settings Log Log Log Log Log Column Oven specific settings Log Log UV Detector specific settings Log Log Fluorescence detector specific RI Detector specific settings ELS Detector specific settings Pump ModelNo Pump ModelVariant Pump SerialNo Sampler Sampler Sampler Sampler Sample
127. printing 3 9 Repeating Checks It may be necessary to repeat one or several checks In this case refer to section 6 This section provides possible causes for the failure According to GLP you have to repeat all checks following the one that failed The reason is that almost all checks require that the previous check be passed successfully Example If the UV detector linearity check fails the results regarding the linearity of the injection volume are questionable because the detector linearity is a prerequisite for checking the injection volume Page 44 of 95 HPLC_OQ_PQ_E_Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions 4 Special Test Procedures for Individual Modules 4 1 Introduction This section describes test procedures that fundamentally differ from the procedures described in section 3 These procedures refer to a certain instrument they cannot be used for any other instrument In addition all test sequences must be run one after the other as the tests require different system configurations The test procedures described in sections 3 and 5 serve as a basis this section focuses on the differences in particular Whenever the test steps are identical you are referred to sections 3 and 5 The sequence templates for these tests are available in the SPECIAL_HPLC_TEMPLATES directory see Figure 6 Start the OQ PQ Setup from this directory as described in section 3 5 2 4 2 Dionex VWD 3x00
128. quences STD_GRAD MICRO GRAD or LONG _ GRAD sequences or STD_GRAD_P680DGP_Left or LONG_GRAD_P680DGP_Left sequences 14 Solvent composition for ternary high pressure gradient pumps accuracy precision and ripple between channels C and B Tern_Grad_C_B sequence 15 Temperature accuracy of the column compartments for automatic data acquisition Column Oven sequence 16 Resetting the solvent flow rate to 0 05 ml min Stop sequence o gt oO ND i Tip When you use a manual injection valve make sure that no air is injected with the samples Besides inject five times the sample loop volume that is inject at least 50 ul HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 Page 43 of 95 OQ and PQ Operating Instructions 3 7 Check Time If the column compartment and the RI detector are not included in the check the entire check takes approximately 3 5 hours Depending on the check more time may be required e 2 more hours when checking Dionex P680 pumps UltiMate 3000 pumps with mixing chamber extension or an UltiMate 3000 LPG 3400M B pump LONG_GRAD sequence instead of the STD_GRAD or MICRO_GRAD sequence e 2 more hours when checking Dionex P680 DGP or UltiMate 3000 DGP pumps standard configuration e 4more hours when checking Dionex P680 DGP pumps UltiMate 3000 DGP micro pumps or pumps with mixing chamber extension e 3 more hours when checking the column thermostat e 1 5 more hours when checking the R
129. r ModelNo SerialNo ModelVariant Temperature LoopVolume ColumnOven ModelNo ColumnOven SerialNo UV ModelNo UV SerialNo settings Page 78 of 95 HPLC_OQ_PQ_E_Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions Sampler AcquireExclusiveAccess Pump Pressure LowerLimit 0 bar Pump Pressure UpperLimit 300 bar Virtual channel settings VirtualChannels 01 SamplingStep 1 00 TemperatureOVEN Type Fixed TemperatureOVEN Formula Formula Temperature 1 Sampler settings Sampler TempCtrl On Sampler Temperature Nominal 10 0 C Sampler ReadyTempDelta 0251 7 e 0 000 Message Make sure that th thermometer is located correctly in the vial refer to the manual for details Message Caution To avoid damage to the system do not perform sampler commands while the test is running Wait Sampler Ready Wait Sampler TemperatureReady Sampler Inject TemperatureOven AcqgOn 30 000 Message The test is now complete Please remove th thermometer log Sampler Temperature Value TemperatureOven AcgOff Sampler ReleaseExclusiveAccess End HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 Page 79 of 95 OQ and PQ Operating Instructions 7 7 Solvent Composition of a Gradient Pump Accuracy Precision and Ripple Standard Gradient Low Pressure Gr
130. r Ten injections are analyzed Each of them contains 5 ul of the same standard The relative standard deviation of the peak areas indicates the precision of the injection volume Five injections of the same standard r2 99 99 r2 99 90 sample are analyzed The injection Rsp RSD lt volumes are different 5 ul to 80 ul lt 1 0 1 0 The injection volume and the peak area are represented in a graph The regression coefficient of the resulting line and the deviation from it indicate the linearity After a highly concentrated test sample has been injected a blind sample is injected The blind sample contains only solvent The peak area measured in this chromatogram indicates the carry over Sampler module of the Waters Alliance 2690 Separation Module Precision of Injection Volume Linearity of Injection Volume Ten injections are analyzed Each of them contains 5 ul of the same standard The relative standard deviation of the peak areas indicates the precision of the injection volume Five injections of the same standard r2 99 90 r2 99 90 sample are analyzed The injection Rsp lt RSD lt volumes are different 5 ul to 80 ul 4 9 1 0 The injection volume and the peak area are represented in a graph The regression coefficient of the resulting line and the deviation from it indicate the linearity Page 16 of 95 HPLC_OQ_PQ_E_Manual doc Version 6 8 dated Oktober 2
131. r specific settings Settings specific for the Dionex UVD 340S Detector Diodearray UVD 3405S r Samples Caffeine 10 ug ml 60 ug ml 140 ug ml 220 ug ml 300 ug ml r Pump specific settings Log Pum Log Pum Log Pum Sampler specific settings Log Sam Column Oven specific settings UV Detector specific settings Log Sam Log UV p ModelNo p ModelVariant p SerialNo pler ModelNo pler SerialNo Log ColumnOven ModelNo ModelNo Fluorescence detector specific settings Log Emission ModelNo Pressure LowerLimit 10 Pressure UpperLimit 300 SA Equate Water Pump specific settings Settings specific for the Dionex P580 SA Type Automatic Flow 1 000 Page 72 of 95 HPLC_OQ PQ _E Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions UV_VIS_1 Wavelength UV_VIS_1 Step UV_VIS_1 Average Detector specific settings EEL ELD Se 3DF ngs specific for the Step UV V Bandwidth UV RefWavelength _V I VI UV RefBandwidth Sampler specif Setti er E ic settings ngs specific for the Samp Sampl Sampl Sampl Sampl Sampl Sampl Sampl 0 000 UV Auto InjectMode UpSpeed DownSpeed SampleHeight WashSpeed DispSpeed DrawSpeed SyringeDelay Zero Dete
132. racy Temperature Accuracy Temperature Accuracy Temperature Accuracy Temperature Accuracy A 3 step temperature gradient is programmed An externally calibrated thermometer is used to measure the temperature that is reached If technically possible the data are automatically read by Chromeleon Else enter the values manually during the measurement A 4 step temperature gradient is programmed An externally calibrated thermometer is used to measure the temperature that is reached If technically possible the data are automatically read by Chromeleon Else enter the values manually during the measurement A 3 step temperature gradient is programmed An externally calibrated thermometer is used to measure the temperature that is reached If technically possible the data are automatically read by Chromeleon Else enter the values manually during the measurement A 3 step temperature gradient is programmed An externally calibrated thermometer is used to measure the temperature that is reached If technically possible the data are automatically read by Chromeleon Else enter the values manually during the measurement A 3 step temperature gradient is programmed An externally calibrated thermometer is used to measure the temperature that is reached If technically possible the data are automatically read by Chromeleon Else enter the values manually during the measurement 1 C Measured at Measure
133. rating Instructions UV_VIS_1 Step 4 0 UV_VIS_1 Average On UvV_VIS_1 Wavelength 265 Detector specific settings Settings specific for the Dionex UVD 340S 3DFIELD Step 0 UV_VIS_1 Bandwidth 1 VIS_1 RefWavelength 600 VIS_1 RefBandwidth 1 0 000 UV Autozero Detector specific settings nject UV_VIS_1 AcqOn Flow 2 000 SB 0 0 SC 0 0 6D 0 0 1 000 B 0 0 6B 1 0 4 000 B 1 0 SB 50 0 8 000 B 50 0 6B 99 0 12 000 B 99 0 SB 100 0 15 000 B 100 0 SB 0 0 24 000 UV VIS_1 AcqOff Flow 2 000 SB 0 0 eC 0 0 6D 0 0 a D Q HPLC_OQ_PQ_E_Manual doc Version 6 8 dated Oktober 2008 Page 81 of 95 OQ and PQ Operating Instructions 7 8 Temperature Accuracy of a Column Compartment Automatic Temperature Measurement Temperature Accuracy of Column Oven PGM Version 09 02 2000 This program file is used to determine the temperature accuracy of the column The temperature is measured with an external ther mometer The values are recorded automatically with as analog Signals HPLC System Settings specific for the Dionex TCC 3200 Column Thermostat TCC 3200 UltiMate 3000 Pump specific settings Log Pump ModelNo Log Pump ModelVariant Log Pump SerialNo Sampler specific settings L
134. rnal thermometer Device Integrator Driver as an analog channel named TemperatureOVEN gt Option C Manual data acquisition In the Chromeleon Server Configuration program install the STH_manual device The driver is available under Generic on the Manufacturers list Verify that the driver is in Demo Mode In addition install a virtual channel device name Virtual Channels_01 signal name TemperatureOVEN During the chromatographic run you can then enter the temperature indicated on the external thermometer on the OQ_PQ_STH_manual control panel Option C does not support qualification of the Agilent Shimadzu and Waters column compartments a Tip Sequence templates created with Chromeleon lt 6 50 can be used in Chromeleon 6 50 only after you have deleted the STH_manual connect line from the COLUMN_OVEN program file If you created new sequence templates from the Chromeleon 6 50 or later master templates you do not need to adapt the program file manually i Tip Note that option C qualifying the column compartment with manual data acquisition is not supported for Agilent Shimadzu and Waters instruments HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 Page 33 of 95 OQ and PQ Operating Instructions 3 4 Preparations 3 4 1 Preparing the HPLC System To prepare the HPLC system for OQ or PQ follow the steps below Perform all steps for the instruments in the system observing the correct order UV Det
135. s the precision of the injection volume Five injections of the same standard r2 99 90 r2 99 90 sample are analyzed The injection Rsp lt RSD lt volumes are different 5 ul to 80 ul 4 5 15 The injection volume and the peak area are represented in a graph The regression coefficient of the resulting line and the deviation from it indicate the linearity After a highly concentrated test lt 0 1 sample has been injected a blind sample is injected The blind sample contains only solvent The peak area measured in this chromatogram indicates the carry over Ten injections are analyzed Each of them contains 5 ul of the same standard The relative standard deviation of the peak areas indicates the precision of the injection volume HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 Page 17 of 95 OQ and PQ Operating Instructions OQ PQ Shimadzu LC 2010 SIL 10HTA Linearity of r2 99 90 r2 99 90 Injection Volume Five injections of the same standard sample are analyzed The injection volumes are different 5 ul to 80 ul SIL 10HTC SIL 10Advp Cont d Carry Over 5 ul to 50 ul for the SIL 10ADvp respectively The injection volume and the peak area are represented in a graph The regression coefficient of the resulting line and the deviation from it indicate the linearity After a highly concentrated test sample has been injected a blind sample is inj
136. s solvent for channel B water with 0 1 Vol acetone is used The ripple is determined relative to the absorption of solvent B for each step of the above gradient runs Ten injections are analyzed When the Dionex GINA 50 or GINA 160 autosampler is used six injections are analyzed All of them contain the same standard sample The relative standard deviation and the standard deviation of the retention times indicate the flow precision The greater value is the valid limit A step gradient of two channels is programmed and measured three times The deviation between the measured and theoretical signal heights indicates the accuracy with which the pump forms the gradient The deviations between the three gradient runs indicate the gradient precision For channel A water is used as solvent for channel B water with 0 1 Vol acetone is used The ripple is determined relative to the absorption of solvent B for each step of the above gradient runs RSD s 1 5 or SDs 0 04 min RSD lt 2 0 or SD lt 0 06 min STD lt STD lt 1 0 2 0 i RSD lt RSD lt 0 075 or 0 15 or SDs SDs 0 02 min 0 04 min STD lt STD lt 0 5 0 5 lt 0 5 lt 0 5 1 OQ limits with optimum measuring conditions recommended PQ limits 2 To determine the gradient accuracy and the gradient precision for the TSP P2000 pump the solvent composition must be as follows 0 50 and 100 of solvent B This is b
137. seeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeaeeeeeneaeeeseeeaaeeeeeeaaees 61 5 15 Baseline Noise of the ELS Detector ccccccceceesseeeeeceee eee eeeeeeeeeeseeeeeseeeeaneeseeeeeeseseeeaneeeenees 62 DEVS A TMC ORY ooi eR aA AAAA APEE T AA TEAR AOA teen eatin Saeed 62 5 15 2 Performing and Evaluating the Check ceecceeceeeeeeeeeeeeeeeeeeeeeeeeeeneaeeeseneaeeeseeaaeeeeeeaaees 62 6 Tro bleshooting crsip a r p Ea eee eee eee 63 6 1 G n ral NOE Sa e rAr arara arrra a aaa aAa cles eed a a aaa aaa Kara clade chvetewenss OOS Paa ana peN anain ceesiies 63 6 2 Failure of Individual Checks cccccceceeeeeeeee cnet ee eeeeeeeeeeeeeeeee sees eeaeeeseeeee sens eaneeseeeeeeseneeneneeeeeees 63 6 2 1 UV Detector ciice cava iar eh a eee eine a eee 63 6 2 2 AutoSampler mairei eats eee een ee el Ree 64 6 2 3 PUM oases 2d Pete a area ra rah a ts ioe teag macs So veche ae pet les tn dee A Nes eataent 64 6 2 4 RF2000 Fluorescence Detector ccccccceceeeeceeceeeeeeeceeceeaeeeeeeesesecaecaeeeeeeesetennieaeeeeeeeeees 64 6 2 5 RID ete colar niona Stier eee Set ae es ee ee eS th yale 65 6 2 6 ELS Detector reee Paavat Nesta cBee ane aise eae A tates rE tele Gut tea denna aea a a At 65 7 PGM Files siie oina aaa aiaa a iaaa eaaa tnt a a a aap aana E aaan aTa aaa aan iR 67 7 1 Wavelength Accuracy of the Photodiode Detector sssnusssenunnunnunnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnn 67 7 2 Baseline Noise Drift and Lamp Intensity of the
138. suring and ambient conditions are constant Verify that there are no gas bubbles in the flow cell In addition to the absolute value of the baseline noise the signal height to noise ratio is important The signal height mainly depends on the condition of the lamp and the flow cell A contaminated flow cell may result in a higher fluorescence signal HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 Page 59 of 95 OQ and PQ Operating Instructions 5 11 2 Performing the Check The Fluorescence sequence is used to determine the noise and the signal height When testing the Dionex RF2000 make sure that the detectors ZWAVE parameter is set to 1 gt 3 3 Water is pumped through the flow cell at a flow rate of 1ml min The excitation wavelength is 350 nm the emission wavelength is 394 nm To determine the noise the measuring signal is split into thirty intervals of thirty seconds each For each interval Chromeleon calculates a regression line based on the method of least squares The noise value is the distance between two parallel lines and the regression line through the lowest and highest values For the calculated values the thirty interval values are averaged 5 12 Wavelength Accuracy of the Fluorescence Detector 5 12 1 Performing the Check The Fluorescence sequence is used to determine the wavelength accuracy of the emission spectrum When testing the Dionex RF 2000 detector make sure that the ZWAVE parameter is set t
139. t 3DFELD MeOH UVD2 12 Detector noise drift and lamp intensity UV VIS 1 i mau WVL 254 nm 0 30 4 0 20 4 f 0 30 VA 0 40 WL 0 50 4 mann 0 60 4 Mii 0 76 T T T T T T ma 2 34 3 00 4 00 5 00 6 00 7 00 8 00 9 00 10 00 11 00 11 63 Figure 15 Lamp drift Directly after the lamp has been turned on bottom chromatogram and after it has been burning for six hours top chromatogram The lamp intensity decreases while the lamp is burning Besides the lamp ages when it is turned on and off very often 5 1 2 Performing and Evaluating the Checks The checks for noise drift and lamp intensity are included in the UV_Noise_Drift sequence For those checks water is pumped through the cell at a flow rate of 1 ml min The UV signal is recorded at 254 nm If the lamp intensity can be determined it is read directly from Chromeleon wavelength 254 nm However for the Dionex VWD 3100 and VWD 3400RS detectors the lamp intensity is determined at a wavelength of 230 nm The results are no absolute measured physical quantities such as the luminous density or luminous flux That is why deviations of 5 from lamp to lamp and from detector to detector are quite normal Therefore to evaluate the data compare them to previously determined values HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 Page 49 of 95 OQ and PQ Operating Instructions To calculate noise the measuring signal is split i
140. the following devices all devices of the Agilent 1100 1200 series all devices of the Dionex UltiMate 3000 series all P680 and P580 pumps the Dionex ASI 100 autosampler the Dionex PDA 100 AD25 UVD 340U and UVD 170U detectors and the supported Shimadzu devices For all other devices enter the serial number in column K from line 141 on The fields have a yellow background Delete the value in the related input field by clicking Clear Values on the Edit menu This removes the Chromeleon variable from the cell and clears the audit xxx entry for the cell on the status bar When qualifying instruments that are not listed in the table below enter the model name in column H fields with a yellow background deleting the existing audit xxx entry as before From line 159 on enter the limits in the column with the related model name Enable writing protection on the Edit menu clicking Layout Mode SAVE the report To do so click Save Report Definition on the Workspace menu To check the linearity of the UV detector adapt the amounts in the QNT file of the sequence to the actual amounts of the used standards 3 5 4 Device Names The PGM files of the installed sequences apply to the following devices device names and channel names as determined in the Chromeleon Server Configuration program Device Supported Model Name Entire system Waters Alliance 2690 Separation Module HPLC_System Pumps Dionex HPG 3200RS UltiMate 3000 Di
141. tion Valve Valco Two Position Valve Valve UV detectors Dionex DAD 3000 RS UltiMate 3000 Dionex MWD 3000 RS UltiMate 3000 Dionex PDA 3000 UltiMate 3000 Dionex VWD 3100 UltiMate 3000 Dionex VWD 3400RS UltiMate 3000 Dionex PDA 100 Dionex AD25 Dionex UVD 340U Dionex UVD 170U Dionex UVD 340S Dionex UVD 170S Dionex UVD 160S Dionex UVD 320S Agilent 1100 1200 series G1315A Agilent 1100 1200 series G1315B Agilent 1100 1200 series G1315C Agilent 1100 1200 series G1315D Agilent 1100 1200 series G1314A Agilent 1100 1200 series G1314B Agilent 1100 1200 series G1314C Agilent 1100 1200 series G1365A Agilent 1100 1200 series G1365B Agilent 1100 1200 series G1365C Agilent 1100 1200 series G1365D Waters PDA996 Diode Array Detector Waters PDA2996 Diode Array Detector Waters 2487 Dual Lambda Absorbance Detector TSP UV1000 Single Lambda Detector TSP UV2000 Dual Lambda Detector TSP UV3000 digital and analog data acquisition TSP UV6000 PDA Shimadzu LC 2010 SPD Shimadzu SPD 10Avp Shimadzu SPD 10AVvp UV UV channel names UV VIS 1 UV VIS 2 UV_VIS 3 HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 Page 41 of 95 OQ and PQ Operating Instructions Device Supported Model Name Samplers Dionex ACC 3000 T UltiMate 3000 Dionex WPS 3000 T RS UltiMate 3000 Dionex WPS 3000 T SL UltiMate 3000 Dionex WPS 3000 T PL UltiMate 3000 Dionex WPS 3000TBPL A
142. tor Operator s Name Operator s Jobtitle Execution Date Apr 14 04 Next Qualification Oct 04 e Limits and Test Results Observed max Deviation Result of all Steps Step Accuracy Test passed Step Ripple Test passed Reviewer s signature Date Operator s signature Date Chromeleon c DIONEX 2008 PQ_OQ_Report_6_8 PUMP_GRADIENT Version 6 80 SR5 Build 2413 137116 Printed 27 10 2008 3 15 PM Seq DQ_PQ_OQ_FOQ PQ_OQ HPLC DEMO_for_Manual HPLC_OQ_DEMO_RUNS OQ_STD_GRAD Page 2 of 3 Smp STD Gradient_3 Runtime 14 Apr 2004 11 02 57 e Chromatogram of STD Gradient_3 OQ_STD_GRAD 4 STD Gradient_3 UV_VIS_1 JmAU WVL 265 nm Flow ml min Reviewer s signature Date Operator s signature Date Chromeleon c DIONEX 2008 PQ_OQ_Report_6_8 PUMP_GRADIENT Version 6 80 SR5 Build 2413 137116 Printed 27 10 2008 3 15 PM Seq DQ_PQ_OQ_FOQ PQ_OQ HPLC DEMO_for_Manual HPLC_OQ_DEMO_RUNS OQ_STD_GRAD Page 3 of 3 Smp STD Gradient_3 Runtime 14 Apr 2004 11 02 57 Data of STD Gradient_3 Expected Calculated Abs Critical Calculated Value Value Deviation Deviation Result Observed Value mAU 0 000 1 163 49 826 98 976 100 000 e Ripple of STD Gradient_3 Z Ripple Calculated Critical Step mAU Ripple Ripple 1 00 0 012 0 006 0 500 50 00 0 023 0 011 0 500 99 00 0 017 0 008 0 500 Reviewer s signature Date Operator s signature D
143. uctions OQ PQ UVD 3208 Pure water is pumped through the 0 05 mAU_ 0 10 mAU UVD 160S flow cell The flow rate is 1 ml min 1 0 mAU h 2 0 mAU h UVD 320 Wavelength 254 nm UVD 160 Lamp Intensity gt 400000 gt 200000 analytical flow 16u volume counts s counts s cell Wavelength Pyrene is injected using methanol 2 nm 3 nm Accuracy as solvent The flow rate is 1 ml min The characteristic maximum of pyrene is determined at 239 nm and compared to its theoretical value Linearity Five caffeine solutions are injected r2 99 9 r299 9 in different concentrations RSD lt 5 RSD lt 5 Concentration and peak area are up to up to represented in a graph The 1 5 AU 1 5 AU correlation coefficient of the resulting line and the deviations from it indicate the linearity Agilent 1100 1200 Pure water is pumped through the 5 0 mAU h 5 0 mAU h G1314A Lamp Intensity flow cell The flow rate is 1 ml min Not Not G1314B Wavelength 254 nm checked checked G1314C G1315A Linearity Five caffeine solutions are injected r2 99 90 r2 99 90 G1315B in different concentrations RSD lt 5 RSD lt 5 G1315C Concentration and peak area are up to up to G1315D represented in a graph The 1 5 AU 1 5 AU G1365A regression coefficient of the G1365B resulting line and the deviations G1365C from it indicate the linearity G1365D Agilent 1100 1200 Baseline Noise Pure water is pumped through the 0 05 mAU 0 05 mAU
144. utosampler commands during the test Moving the needle arm or carousel may damage the thermometer or autosampler 4 3 4 Duration The test takes approximately 45 minutes HPLC_OQ_PQ_E Manual doc Version 6 8 dated Oktober 2008 Page 47 of 95 OQ and PQ Operating Instructions Page 48 of 95 HPLC_OQ_PQ_E_Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions 5 Procedures 5 1 Baseline Noise Drift and Lamp Intensity of the UV Detector 5 1 1 Theory Drift and baseline noise are important factors for UV detectors Increased baseline noise considerably reduces the sensitivity as it is not possible to distinguish between low level signals and noise With increased drift it is more difficult to integrate the signals correctly because the less stable the baseline is the more inaccurate is integration The baseline noise of the detector mainly depends on the lamp There is a considerable increase in noise if an old lamp with poor light intensity is used This is also true when the flow cell is dirty In addition make sure that the measuring and ambient conditions are constant and that the flow cell is free from gas bubbles To measure the drift of a UV detector also make sure that the measuring and ambient conditions are constant In addition it is very important that the lamp has been burning for several hours In the detector environment avoid drafts and direct sunlight 0 41 Rauschen Drif
145. ver solution conc 2000 ug ml 5 6 Linearity of the Injection Volume 5 6 1 Theory The linearity of the injection volume and its precision depend on the quality of the syringe and the syringe volume that has been adjusted to the injection volume Besides the quality of the autosampler mechanics also affects the result Select the concentration of the standard which is injected in different volumes in such a way that the detector works in the linear range for all injections usually between 10 mAU and 1000 mAU Page 54 of 95 HPLC_OQ_PQ_E_Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions 5 6 2 Performing the Check With the Sampler_Lin_CO sequence a caffeine standard solvent water at a flow rate of 1 ml min wavelength 272 nm is injected five times The autosampler type determines the injection volume and the standard see table The peak area and injection volume are represented in a graph and the regression line is determined The correlation coefficient and the standard deviation of this line indicate the linearity Autosampler Standard used Injection volume Others Standard 2 5 10 20 40 and 80 ul Dionex Gina 50 Standard 2 10 20 40 60 and 80 ul Dionex Gina 160 Shimadzu SIL 10ADvp Standard 2 5 10 20 40 and 50 ul Dionex ACC 3000 T Sample loop volume 20 ul Standard 3 1 3 5 7 and 10 ul Sample loop volume 50 ul Standard 3 5 10 15
146. wing definitions apply 2 1 1 Operational Qualification OQ The purpose of Operational Qualification is to prove and document that an analytical system functions according to its operating specification while the specific environmental conditions are taken into account In his specification the supplier must therefore define exactly the conditions that must be observed With varying conditions e g different ambient temperatures higher limits must be used Usually Operational Qualification is only performed directly after a new device has been installed 2 1 2 Performance Qualification PQ The purpose of Performance Qualification is to prove and document that an analytical system functions according to a specification that is suitable for the system s routine operation As a system is subject to wear when being operated it may happen that the supplier s specification is no longer met This means The same procedures are used but the tolerances used for Performance Qualification are less restrictive than those used for Operational Qualification are Performance Qualification is usually performed after repair or regular system service procedures have been performed Using the same procedures for OQ and PQ simplifies the handling 2 1 3 System Suitability Check SSC also System Suitability Test SST The purpose of SSC is to prove and document that the necessary limits are met for a specific measuring application The specific condit
147. with 0 1 Vol acetone is used For each step of the above gradient runs the ripple is determined relative to the absorption of solvent B RSD lt RSD lt 0 05 or 0 1 or SDs SDs 0 01 min 0 02 min HPG HPG lt 0 2 lt 0 5 LPG DGP LPG DGP analytical analytical 1 0 lt 2 0 LPG DGP LPG DGP micro micro lt 2 0 lt 2 0 STD lt STD lt 0 5 0 5 lt 0 5 lt 0 5 Page 18 of 95 HPLC_OQ_PQ_E_Manual doc Version 6 8 dated Oktober 2008 OQ and PQ Operating Instructions Instrument Parameter Limits OQ Description PQ M 480 with analytical pump heads Flow Precision M 480 with analytical pump heads low pressure gradient Gradient Accuracy Ripple Gradient Accuracy M480 and M300 High pressure gradients with analytical pump heads Gradient Not checked Not checked Precision N RSD s 0 1 or SD lt 0 02 min RSD lt 0 1 or SD lt 0 02 min Ten injections are analyzed When the Dionex GINA 50 or GINA 160 autosampler is used six injections are analyzed All of them contain the same standard sample The relative standard deviation and the standard deviation of the retention times indicate the flow precision The greater value is the valid limit Two different step gradients of two channels are programmed The deviation between the measured and theoretical signal heights indicates the accuracy with which th
148. xceeds the linearity range of the detector a reference sample with a considerably lower concentration is also injected 5 5 2 Performing the Check The carry over by the autosampler is measured with samples 6 to 9 of the Sampler_Lin_CO sequence solvent water at a flow rate of 0 3 ml min wavelength 272 nm Samples 6 and 9 contain water same vial Sample 7 contains a solution of caffeine in water the concentration is 10 ug ml standard 2 reference sample Sample 8 contains a solution of caffeine in water the concentration is 2000 ug ml standard 7 The carry over CO in is calculated as follows Ar Oyster corr PROD yater CarryOver Area y ater CO P PEA Conc 2000mg ml C HighConcentratedSample Areare ference x C Re ference Are Gy ater CarryOver T AT el yater x CRe ference Arrear ference C Conc2000mg ml AVC pater corr Area of the caffeine peak in the water sample sample 9 sample 6 AT Q conc 2000mg ml Peak area of the highly concentrated caffeine sample sample 8 ATC So CarrvOver Peak area of the water injection sample 9 solvent and caffeine peaks after the carry over sample sample 8 AT ea yy top Peak area of the water injection sample 6 solvent peak before the carry over sample sample 8 AT eare ference Peak area of the reference sample sample 7 Cre frene Caffeine concentration of the reference solution conc 10 ug ml C Cone 2000mg ml Caffeine concentration of the carry o

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