Method Transfer Crossing Multiple Borders
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Method Transfer Crossing Multiple Borders Document Transcript

  • 1. 8/2/2010 Method Transfer: Crossing Multiple Borders Xiande (Andy) Wang, Ph.D. Analytical Research and Development Cordis Corporation, A Johnson & Johnson Company, Warren, NJ 07059 IVT Annual Method Validation 2010 Outline1. Lifecycle of Analytical Methods2. Border I: Between Methods – Case study: combination of 3 methods into one3. Border Between Instrumentation/Technique – Case study: validation of an HPLC and UPLC method side by side4. Border Between Groups – Case study: Troubleshooting cross functions during method transfer5. Conclusions 1
  • 2. 8/2/2010Life Cycle of an Analytical method Method Development Method Method transfer validation Life Cycle of Analytical methods Development transfer validation 2
  • 3. 8/2/2010Life Cycle of Analytical Methods - Examples Development transfer validationLife Cycle of Analytical Methods - Examples UPLC HPLC Development HPLC UPLC UPLC HPLC UPLC HPLC UPLC HPLCHPLC UPLC HPLC UPLC UPLC HPLC UPLC HPLC validation transfer 3
  • 4. 8/2/2010 Borders to Cross in Lifecycle of MethodBorder II UPLC HPLC DevelopmentBorder I HPLC UPLC UPLC HPLCBorder III UPLC HPLC UPLC HPLC HPLC UPLC HPLC UPLC UPLC HPLC UPLC HPLC validation transfer Border I: Between Methods UPLC HPLC UPLC HPLC UPLC HPLC 4
  • 5. 8/2/2010 Questions Around Border I• Can the method be used interchangeably?• Methods require the same instrumentation, column, reagents, materials?• Solution (standard, sample, mobile phase) storage and stability• Timing of validation, transfer; effective date, versions• Will methods be run in the same or different lab?• Sample shipping, storage at different environment• Consistency in any other areas How is HPLC Assay Method Developed? Method Objectives– Stability indicating • Peak purity • Resolution of all species • LC-MS compatible • Elution of all species/compounds– Long gradient method– Orthogonal– Robust– Sensitivity 5
  • 6. 8/2/2010 Systematic HPLC Assay Method Development Samples are Stressed samples Representative stressed under are analyzed with stressed samples are different a generic method chosen conditions Method screening is The method is A primary method conducted with optimized is identified selected samplesStressed samples are The primary method is ready analyzed with for further optimization / optimized method validation 11 General guidelines for HPLC Column Selection• Select high-purity silica-based columns• C18 and C8– Hydrophobic, retentive and stable• Phenyl – medium polarity components; unique selectivity for aromatics• Hydrophilic end-capped phases (retentive for water soluble compounds)• Polar-embedded phases (amide, carbamate, ether, sulfonamide) – Less tailing for basic analytes – “Orthogonal” to C8/C18, No phase collapse• Explore selectivity differences between C18, polar-embedded or phenyl bonded phases – Consult column selectivity chart – For low pH Applications, select column resistant to hydrolytic cleavage (e.g., StableBond, X-Bridge C18) – For high-pH application, select columns stable at high pH (e.g., Gemini, X- Bridge, Extend, Luna) 12 M.W. Dong, Modern HPLC for Practicing Scientists, Wiley, 2006, Chap. 3. 6
  • 7. 8/2/2010 Some Popular HPLC Columns• Waters: Symmetry, SunFire, XTerra *, ACQUITY*, X-Bridge*, Atlantis, NovaPak, m-Bondapak, Spherisorb• Agilent: Zorbax StableBond, Eclipse XDB, Extend C18*, Bonus• Phenomenex: Luna*, Prodigy, Synergi*, Gemini*• Supelco: Discovery, Ascentis, Supelcosil• Varian: Inertsil, Polaris*• Thermo: HyPURITY, Hypersil, Prism, Hypersil Gold *• MacMod: ProntoSIL, ACE (Adv. Chrom. Tech.)• YMC: YMCbasic, Pack Pro• Eka Chemicals: Kromasil• GL Sciences: Inertsil• Macherey Nagel: Nucleosil• Merck KGaA: Chromolith (Monolith)• Bischoff: ProntoSIL*• Grace: Vydac, Platinum (Alltech)• Dionex: Acclaim, Acclaim PA, Acclaim PA2*Columns based on high-purity silica are underlined. Hybrid particles are in bold. Phases stable in high pH are italicized and marked with *. 13 M.W. Dong, Modern HPLC for Practicing Scientists, Wiley, 2006, Chap. 3. HPLC Assay Method Development: Column Screening E. F. Hewitt, P. Lukulay, and S. Galushko, J Chromatography A, 1107, 79. 7
  • 8. 8/2/2010 HPLC Column Screen Set: An ExampleOrthogonal Screening – Columns Stationary Phase Column pH Rangea Manufacturer Part Number C18 – Twin Technology Gemini C18, 5 μm, 110A, 4.6 x 150 mm 1-12 Phenomenex 00F-4435-E0 Phenyl with Hexyl (C6) linker, Luna Phenyl-Hexyl, 3 μm, 4.6 x 150 mm 1.5-10 Phenomenex 00F-4256-E0 endcapped C18-20% C loading Discovery HS-C18, 3μm, 4.6 x 150 mm 2-8 Supelco 569252-U C18 – polar embedded, hybrid XTerra RP18, 3.5 μm, 4.6 x 150 mm 1-12 Waters 186000442 particle with Shield Technology C18– silica Sunfire C18, 3.5 μm, 4.6 x 150 mm 2.8 Waters 186002554 Pentafluorophenyl Curosil PFP, 3 μm, 4.6 x 150 mm 2-7.5 Phenomenex 00F-4122-E0aColumns were screened only against mobiles phases within their compatible pH range. Slide courtesy of H. Rasmussen et al 15 HPLC Screening Conditions: An Example Orthogonal Screening Method Description a Time (min) %Water %Acetonitrile % Modifier Flow Rate (ml/min) 0 85 10 5 1.0 40 10 85 5 1.0 45 10 85 5 1.0 45.10 85 10 5 1.0 60 85 10 5 1.0 Injection Volume 5 μL Detection 280 nm; DAD (190 – 400 nm) Column Temperature Ambient o Sample Temperature 5C aModifierstock solutions are prepared at a concentration 20 times higher than the desired mobile phase concentration since mobile phases are prepared at time of use with the HPLC quaternary pump. Modifier Mobile Phase Approximate pH Concentration Trifluoroacetic Acid (TFA) 0.05% 2 Formic Acid 0.1% 2.8 Ammonium Acetate + Acetic Acid 8 mM + 0.1% 4 Ammonium Acetate 8 mM 7 Ammonium Acetate + Ammonium 8 mM + 0.05% 10.2 Hydroxide Ammonium Hydroxide 0.05% 10.8 16 Slide courtesy of H. Rasmussen et al 8
  • 9. 8/2/2010 Summary of HPLC Assay Method Development• There is a systematic approach for assay method development.• Column screening is an effective tool.• It will enhance efficiency by narrowing down the list of columns for screening/selection.• It is critical to seek input from other labs (QA, QC) during method development.• It’s important to look at the big picture across methods. What is Important in Developing HPLC CU Method? Method Objectives– Short– Specific– Isocratic preferred– Robust– Compatible with assay method– Can be used for dissolution testing 9
  • 10. 8/2/2010 Systematic HPLC CU Method Development samples are Specificity, retention, No analyzed with a peak shape etc. generic method acceptable Yes The method is A primary method is Method screening is optimized identified conductedThe primary method is ready forfurther optimization / validation 19 HPLC CU Method Development: Column Screening E. F. Hewitt, P. Lukulay, and S. Galushko, J Chromatography A, 1107, 79. 10
  • 11. 8/2/2010 What is Important in Developing HPLC Method for Dissolution Testing?– Short– Specific– Isocratic preferred– Robust– Compatible with CU method– Sensitive How to Minimize Border I (Between Methods)• Use same materials/chemicals: grade, vendor• Adopt similar standard/sample solution: procedure of preparation, concentration, pH, storage, expiry• Use same column: vendor, stationary phase, dimension, particle size; alternative column• Use same Instrumentation• Be consistent in write-up: same product description, formulation number, etc. 11
  • 12. 8/2/2010 Case Study: Combination of 3 methods into 1 Method Method title number N1 Identification, Assay and Content Uniformity Determination of … by RP-HPLC-UV O1 Identification and Assay of … by RP-HPLC-UV O2 Determination of Content Uniformity of … by RP-HPLC-UV O3 Identification of … by UV Spectroscopy Method O1: Identification and Assay of … by RP-HPLC-UV Parameter Value HPLC Column Agilent, Zorbax Eclipse XDB-C18, 150 mm x 4.6 mm x 3.5 µm 02 .00 Flow Rate 1.0 ± 0.1 mL/minInjection Volume 25 µL Column 01 .05 40 ± 2 °C Temperature Detection 278 nm. Note: If using an Agilent DAD detector, set the bandwidth to 4 01 .00 Wavelength AU nm and the reference wavelength off. A: 80:20% (v/v) 0.02% formic acid:THF Mobile Phases B: 75:20:5:0.02% (v/v/v/v), acetonitrile: THF : water: 00 .05 formic acid Time % Mobile Phase A % Mobile Phase B (minutes) 00 .00 Gradient 0.0 55 45 Program 3 55 45 1 0 2 0 3 0 4 0 5 0 6 0 7 0 8 0 9 0 1 .0 1.0 1.0 1 .0 1 .0 1.0 .0 .0 .0 .0 .0 .0 .0 .0 .0 0 0 1 0 2 0 3 0 4 0 5 0(Linear Gradient 7 42 58 Mu s in te Profile) 13 10 90 15 55 45 12
  • 13. 8/2/2010Method O2: Determination of Content Uniformity of and Dissolution … by RP-HPLC-UV Parameter Value Phenomenex Luna C18 (2), 4.6 x 50 mm, 3 μm or Phenomenex Column Gemini C18, 4.6 x 50 mm, 3 μm HPLC Column Column 35oC ± 2oC Temperature Ambient (20 to 25 °C if Autosampler temperature control is Temperature available) 55:45, 0.02% v/v Formic Mobile Phase Acid:THF Flow Rate 1.2 mL/minute 278 nm Note: If using Agilent PDA Detector detector, set bandwidth to 4 nm and reference wavelength off.Injection Volume 25 μL Run Time 10 minutesMethod O3: Identification of … by UV Spectroscopy 13
  • 14. 8/2/2010 Method N1: Identification, Assay and Content Uniformity Determination of … by RP-HPLC-UV Parameter Value R A P A M Y C IN - 4 .05 5 0.028 HPLC Column Agilent Zorbax Eclipse XDB-C18, 100 mm x 4.6 0.026 mm, 3.5 µm 0.024 0.022 Flow Rate 1.0 mL/min 0.020 3.3 90Injection Volume 20 µL 0.018 B H T - 7 .19 6 0.016 Column o 45 C ± 2°C 0.014 Temperature 0.012 AU 0.010 Assay Identification 3.5 95 0.008 2.1 81 3.0 59 Detection 278 nm 2.5 77 0.006 4.9 79 200 to 400 Wavelength 0.004 Note: If using an Agilent DAD detector, set the bandwidth to 4 nm and nm 0.002 0.000 the reference wavelength off. -0.002 A: (20:80)THF: Formate Buffer -0.004 Mobile Phases B: (75:20:5)ACN:THF:Formate Buffer 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 Minutes % Mobile % Mobile Phase Time (min) Phase A B 279.0 0.50 0.0 47 53 0.45 Gradient 3.0 47 53 0.40 269.0 291.0 Program 5.0 20 80 0.35 (Linear 6.0 2 98 0.30 Gradient) AU 0.25 7.5 2 98 0.20 7.6 47 53 0.15 208.0 10 47 53 0.10 0.05 349.0 0.00 200.00 220.00 240.00 260.00 280.00 300.00 320.00 340.00 360.00 380.00 400.00 nm Benefits of a Combined MethodBenefit DetailsShortened One, instead of three, set of test methods, validationproject timelines protocols, validation reportsEnhanced Run time cut from 18 to 10min;throughput one instrument/set-up for both assay/CU; eliminate requirement of a UV spectrometer.Reduce cost One , instead of two, set of HPLC column is needed Less reference standard, organic solvent Less sample shipment costEliminated When two methods are used, possible bias could comecauses for bias from different instrument set up, different way of peakbetween assay integration, different separation capacity, etc. One methodand CU eliminate all these possible causes. 14
  • 15. 8/2/2010 Benefits of Elimination of Border I• Same columns• Same reagents, vendor, expiry dates• Same materials such as glassware, HPLC vials, etc. Assay/CU/dissolution• No discrepancy (bias) of results due to integration, separation capacity, etc.• Same solution (standard, sample, mobile phase) storage and stability• Transfer at the same time• Same method, same version Assay/CU/dissolution Assay/CU/dissolution• One HPLC system needed• Minimize sample shipping, storage at different environment Summary of Strategy to Minimize/Eliminate Border I• Assay, CU, and dissolution (HPLC) methods have different objectives and hence the strategy for method development may be different.• However, it is important to keep the other method in mind during method development.• Column screening is an effective tool for assay, CU or dissolution methods.• It is effective to narrow down the list of columns for screening/selection.• It is critical to seek input from other labs (QA, QC) during method development.• It’s important to leverage the knowledge, as much as possible, of one method and apply it to another.• It offers a lot of benefit to use similar or the same procedures, instrumentation and materials for all methods, and transfer them at the same time. 15
  • 16. 8/2/2010 Border II: HPLC vs. UPLC UPLC HPLC UPLC HPLC UPLC HPLC UPLC vs. HPLC: Example of Assay MethodRef: L. Pereira, Poster at Pittcon 2007, Chicago, Illinois, February 2007. 16
  • 17. 8/2/2010 UPLC vs. HPLC: Example of CU Method http://www.waters.co m/waters Issues Around Border II (HPLC vs. UPLC)• Availability of instruments at all sites• Different UHPLC systems have different design, not a direct transfer• Training/knowledge and technical challenges on the new instruments• Limited availability and technical challenges of columns• Project history• Cost 17
  • 18. 8/2/2010 How to Minimize Border II (HPLV vs. UPLC) • Use the same stationary phase with different dimension and particle size • Same mobile phase but slight different gradient • Same protocol template • Use the same solutions and validate the methods side by side Case study: Validation of HPLC and UPLC Assay Method Side by Side Chromatograms of the Final Methods RAPAMYCIN - 4.055 0.028 0.026 0.024 0.022 0.020 HPLC 3.390 0.018 BHT - 7.196 0.016 0.014AU 0.012 0.010 UPLC 3.595 0.008 2.181 3.059 2.577 4.979 0.006 0.004 0.002 0.000 -0.002 -0.004 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 Minutes RAPAMYCIN - 2.358 0.012 BHT - 4.202 0.010 0.008 0.006 AU 0.004 0.002 0.000 -0.002 0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00 Minutes 18
  • 19. 8/2/2010Case study: Validation of HPLC and UPLC Assay Method Side by Side Final Method Parameters HPLC UPLC Parameter Value Parameter Value UPLC AgilentZorbax Eclipse XDB-C18, 100 HPLC Column Agilent Zorbax Eclipse XDB-C18, 100 mm x 4.6 Column mm x 3.0 mm, 1.8 µm mm, 3.5 µm Flow Rate 0.7 mL/min Flow Rate 1.0 mL/min Injection 5 µLInjection Volume 20 µL Volume Autosampler 10 oC Column o 45 C ± 2°C Temperature Temperature Column 45 oC ± 2 °C Assay Identification Temperature Sampling Detection 278 nm Assay 200 to 400 Rate Wavelength Note: If using an Agilent DAD 278 nm detector, set the bandwidth to 4 nm and nm Detection Note: If using an Agilent the reference wavelength off. Wavelength DAD detector, set the 20 A: (20:80)THF: Formate Buffer bandwidth to 4 nm and (points/sec) Mobile Phases B: (75:20:5)ACN:THF:Formate Buffer the reference wavelength off. % Mobile % Mobile Phase A: (20:80)THF: Formate Buffer Time (min) Mobile Phase A B B: (75:20:5)ACN:THF:Formate Phases 0.0 47 53 Buffer % Mobile % Mobile Gradient 3.0 47 53 Time (min) Phase A Phase B Program 5.0 20 80 0.0 47 53 Gradient (Linear 2.0 47 53 6.0 2 98 Program 3.5 20 80 Gradient) (Linear 7.5 2 98 Gradient) 4.2 2 98 5.2 2 98 7.6 47 53 5.3 47 53 10 47 53 6.5 47 53 6.5 minutes (Retention time is ~2.5 min Run Time for sirolimus, ~4.3 min for BHT. Seal Wash Acetonitrile Acetonitrile/water: 50/50, 600 µL Weak Wash volumeCase study: Validation of HPLC and UPLC Assay Method Side by Side Accuracy (spiked recovery) Actual Individual % Mean % % ~Level Concentration Recovery Recovery RSD (μg/mL) HPLC 99.9 40% 67.4 99.7 99.9 0.1 99.9 99.5 UPLC 100% 151.7 99.8 99.6 0.2 99.5 100.6 160% 242.7 100.6 100.5 0.2 100.3 Actual Individual % Mean % % ~Level Concentration Recovery Recovery RSD (μg/mL) 100.9 40% 67.4 101.0 101.0 0.1 101.1 99.4 100% 151.7 99.6 99.6 0.2 99.8 99.3 160% 242.7 98.9 99.0 0.2 98.9 19
  • 20. 8/2/2010Validation of HPLC and UPLC CU Method Side by Side: Accuracy (spiked recovery) Actual Individual Mean % ~Level Concentration % % RSD Recovery (μg/mL) Recovery 100.9 HPLC 40% 12.1 101.6 101.2 0.3 101.3 100.4 UPLC 100% 30.3 100.0 100.0 0.4 99.5 100.4 160% 48.5 102.2 101.3 0.9 101.5 Actual Individual Mean % ~Level Concentration % % RSD Recovery (μg/mL) Recovery 100.6 40% 12.1 100.7 100.4 0.4 100.0 98.9 100% 30.3 98.8 98.9 0.1 99.0 99.6 160% 48.5 100.0 99.8 0.2 99.6Validation of HPLC and UPLC Assay Method Side by Side: Intermediate Precision Determination %LC Analyst 1 %LC Analyst 2 1 103.1 103.4 2 101.9 102.6 HPLC 3 101.4 103.0 4 101.6 103.0 5 102.4 103.4 UPLC 6 102.2 104.0 Mean (n=6) 102.1 103.2 %RSD (n=12) 0.8 Determination %LC Analyst 1 %LC Analyst 2 1 103.8 101.3 2 101.8 102.1 3 100.9 102.3 4 100.6 102.2 5 101.8 102.7 6 102.2 102.9 Mean (n=6) 101.9 102.2 %RSD (n=12) 0.5 20
  • 21. 8/2/2010 Validation of HPLC and UPLC CU Method Side by Side: Intermediate PrecisionDetermination %LC Analyst 1 %LC Analyst 2 1 102.0 103.4 2 101.6 103.7 3 101.3 103.4 4 100.4 102.7 HPLC 5 102.6 102.2 6 101.0 101.7 7 101.7 103.6 UPLC 8 100.8 102.6 9 101.6 102.9 10 101.4 102.2Mean (n=10) 101.4 102.7%RSD (n=20) 0.9 Determination %LC Analyst 1 %LC Analyst 2 1 101.5 102.7 2 102.0 104.2 3 101.0 102.3 4 103.4 102.3 5 105.1 102.3 6 101.3 101.7 7 103.1 102.6 8 103.1 102.7 9 102.1 102.7 10 102.9 102.0 Mean (n=10) 102.6 102.6 %RSD (n=20) 0.9Validation of HPLC and UPLC Assay Method Side by Side: Linearity HPLC UPLC R=0.9997 21
  • 22. 8/2/2010Validation of HPLC and UPLC CU Method Side by Side: Linearity HPLC UPLC R=0.9997 R=1.0000Validation of HPLC and UPLC CU Method Side by Side: Method Equivalency Determi- %LC %LC with %LC old %LC with Determi- nation with old new CU new nation CU method method method method 1 98.4 98.7 1 102.0 101.3 2 99.2 99.6 2 101.6 100.6 3 98.7 99.9 3 101.3 100.8 4 100.1 100.3 4 100.4 100.4 5 100.0 99.9 5 102.6 102.2 6 100.4 100.6 6 101.0 100.7 7 100.4 100.2 7 101.7 101.1 8 100.1 100.5 8 100.8 100.6 9 100.7 100.2 9 101.6 101.4 10 99.9 100.4 10 101.4 101.0 Mean 99.8 100.0 Mean 101.4 101.0 (n=10) (n=10) %RSD 0.8 0.6 %RSD 0.6 0.5 (n=10) Absolute Absolute difference 0.4 Difference 0.2 (%) (%) HPLC UPLC 22
  • 23. 8/2/2010Summary of Case Study to Minimize Border II (HPLC vs. UPLC) • Use the same stationary phase with different dimension and particle size • Same mobile phase but slight different gradient • Same protocol template • Use the same solutions and validate the method side by side • The knowledge of both methods were exchanged during the process • HPLC and UPLC method proved to be equivalent and can be used interchangeably Border III. Between Functions Border II UPLC HPLC Development Border I HPLC UPLC UPLC HPLC Border III UPLC HPLC UPLC HPLC HPLC UPLC HPLC UPLC UPLC HPLC UPLC HPLC validation transfer 23
  • 24. 8/2/2010 III-a: Method Development vs. Validation• Often reside in the same functional group• Natural redundancy in terms of robustness, forced degradation, alternative columns Method Development Method Method transfer validation What Is Analytical Method Transfer• Protocol driven study with pre-defined acceptance criteria• Transfer of validated analytical procedures to a new laboratory• Verification of a method’s suitability for its intended use• Demonstration of a laboratory’s proficiency in running a particular method• No official guidelines 24
  • 25. 8/2/2010 Options for Method Transfer• Comparative testing: A set of samples are tested in both labs and resulting data are compared with predetermined acceptance criteria.• Co-validation between two labs: The receiving laboratory is involved in method validation but have to identify which validation parameters are to be generated or challenged by the two labs.• Complete or partial method validation: A repeat of method validation either completely or partially.• Transfer waiver (omission of formal validation): Needs justification as to why method transfer was not needed. For example, lab is already testing the product. Border III-b: Method Transfer and Validation• Method transfers are closely related to validation• Method transfer is more challenging because multiple laboratories and companies are involved – Different approaches to Validation and Transfer – Different expectations of what is an acceptable validation – Different instruments and facilities 25
  • 26. 8/2/2010 Method Transfer and Validation Method transfer Method validation• Can be part of the validation • Protocol driven study with pre-defined acceptance criteria• Protocol driven study with pre-defined acceptance criteria • Validation of analytical procedures in a laboratory• Transfer of validated analytical procedures to a new laboratory • Verification of a method’s suitability for its intended use• Verification of a method’s suitability for its intended use in a new laboratory • Demonstration of a laboratory’s proficiency in running a particular method• Demonstration of a laboratory’s proficiency in running a particular method • http://www.ich.org/LOB/media/MEDIA41 7.pdf;• No official guidelines http://www.fda.gov/cder/guidance/2396d ft.pdf Objectives of Method Transfer• Maintain the validated state of the method and meet all regulatory requirements• Minimize surprises! – Open and responsive communication – Pre-determined expectations – Clearly documented and communicated technical details – Pre-transfer evaluation by experienced technical staff at receiving site – Technical contact available for troubleshooting at transferring site 26
  • 27. 8/2/2010 Typical Method Transfer Steps• Discussions Initiated• Review of Method and Validation• Laboratory Evaluation• Protocol (Transfer or validation) Written• Protocol Approved• Experimental evidence from a transfer study generated• Report (Transfer or validation) Written• Report Approved• Transfer Complete Preparation for a Method Transfer• Method – Details about method – Specific instrument• Method development history report• Training/discussion on the method• Materials – Reference Standard – Samples for Evaluation – Difficult to purchase supplies• Specifications• Technical Contact• Details about product 27
  • 28. 8/2/2010 Prior to Formal Method Transfer• Receiving laboratory should perform the method – Helps to determine where there are differences and gaps in documentation • Lack of detailed test method instructions – Assay Conditions – Calculations – System Suitability – Differences with instrumentation or reagents Prior to Formal Method Transfer• Training of Personnel – Review of relevant SOPs – Observation of test procedure – Performing test procedure• Helpful to include development, qualification and validation reports to recipient laboratory 28
  • 29. 8/2/2010 Method Validation and Transfer• Method transfers are closely related to validation• Method transfer is usually part of validation Method Development Method Method transfer validation Border III-c: Method Transfer and Method Development: Before or During Method Development• Define goals of end method• Dynamic platform of communication• Exchange of knowledge 29
  • 30. 8/2/2010 Border III-c: Method Transfer and Development: After method Transfer• Is the method inadequate by today’s scientific standard or regulatory requirement?• Is sufficient data available to permit simplification of the method?• Does monitoring of laboratory deviation suggest a need for method improvement ?• Do newer method for similar products significantly outperform?• Is the volume of testing justify further method optimization or automation? Border III-c: Method Transfer and Development: During Method Transfer– Concerns?– Observations?– Investigations/troubleshooting: Must involve multiple labs 30
  • 31. 8/2/2010Case Study: Extraneous peaks associated with HPLC vials RAPAMYCIN - 4.133 0.010 0.009 Deactivated glass vials 0.008 0.007 Polypro 0.006 pylene AU 0.005 0.004 3.670 0.003 0.002 0.001 3.108 0.000 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 MinutesCase Study: Extraneous peaks associated with Glass Pipettes Rapamycin - 3.947 0.011 0.010 0.009 0.008 0.007 Unknown4 - 3.496 0.006 Unknown3 - 2.977 AU 0.005 Unknown1 - 1.690 Unknown2 - 2.526 0.004 0.003 0.002 0.001 0.000 -0.001 -0.002 0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 Minutes Above: chromatograms of two different lots of glass pipettes . Plastic transfer pipettes, or no pipettes, were recommended. 31
  • 32. 8/2/2010Case Study: Extraneous peaks associated with Extraction Vials Volu Number Price Price Extraction Vial me Comments per case ( $/case) ($/vial) (mL) 10 10 224.70 22.47 Re usable. VWR Cat# 2100 Nalgene-Teflon 30 10 333.30 33.33 Re usable. VWR Cat# 2100 BD Falcon-Poly 5 500 160.00 0.32 VWR Cat # 60819-706 Propylene, snap cap 14 500 195.30 0.39 VWR Cat # 60819-740 BD Falcon-Poly Propylene, 15 500 220.89 0.44 VWR Cat# 21008-918 screw cap only 2 sizes available VWR Starplex- 5 1500 186.18 0.12 14216-262 Polypropylene only 2 sizes available VWR screw cap 10 1000 135.96 0.14 14216-266 NUNC Poly 15 500 192.00 0.38 Sigma Aldrich Cat# Z7204 propylene snap cap Pre-cleaned* Sigma Aldrich 7 100 92.50 0.93 27341 Supelco pre-cleaned Pre-cleaned* Sigma Aldrich 15 100 102.50 1.03 clear glass 27342 Pre-cleaned* Sigma Aldrich 22 100 116.00 1.16 27343 Larger sizes are available u Supelco Silanized 4 1000 267.00 0.27 request, caps not included. Clear Glass Sigma Aldrich Cat# 27114 Currently being used VWR 8 144 152.49 1.06 66009-984 Kimble-glass Currently being used VWR 16 144 193.44 1.34 66009-986 Treatment to Glass Extraction Vials • Rinse with 0.02% formic acid in acetonitrile • Rinse with acetonitrile • Wash with a regular washing cycle for other glassware • Soak with 0.02% acid (formic, acetic or nitric) in water and rinse with DI water • Soak/sonicate in DI water • Details are critical to ensure accurate comparison cross labs. 32
  • 33. 8/2/2010 Standard Solution in Unwashed Glass Vials 3.50 3.00 2.50Total Impurity Area% 2.00 1.50 1.00 0.50 0.00 0 1 2 3 4 5 6 7 8 Day Standard Solution in Pre-cleaned Glass Vials 5 4.5 4 3.5 2ml Total Impurity Area % 3ml 3 4ml 2.5 5ml 6ml 2 7ml 1.5 9ml 1 10ml 0.5 0 0 1 2 3 4 5 6 7 8 Day 33
  • 34. 8/2/2010 Standard Solution in Silanized Glass VialsStandard Solutions in Glass Vials Soaked and Sonicated in DI Water 34
  • 35. 8/2/2010Standard Solution in Glass Vials Rinsed with MeCN 3 Times 3 2.5 2 Total Impurity area % Direct pour 8ml- std soln-1 1.5 8ml- std soln-2 8ml- std soln-3 1 8ml- std soln-4 8ml- std soln-5 0.5 0 0 1 2 3 4 5 6 7 8 Day Standard Solution in Glass Vials Rinsed with 0.02% Formic Acid in MeCN 3 Times 1 0.9 0.8 0.7 Total Impurity Area% 0.6 Direct pour 8ml- std soln-1 0.5 8ml- std soln-2 0.4 8ml- std soln-3 8ml- std soln-4 0.3 8ml- std soln-5 0.2 0.1 0 0 1 2 3 4 5 6 7 8 Day 35
  • 36. 8/2/2010Standard Solutions in Glass Vials Treated with 0.02% Acid (Nitric, Acetic or formic) Standard Solution in Washed Glass Vials Standard in Washed Extraction Vial 0.1 2 0.08 Total impurity % 3 0.06 4 0.04 5 0.02 6 0 7 0 1 2 3 4 5 6 7 9 Day 10 36
  • 37. 8/2/2010 Standard Solution in BD Falcon Polypropylene Vials 0.16 small-1 0.14 Total Impurity area% 0.12 small-2 0.10 small-3 0.08 small-4 0.06 small-5 0.04 small-6 0.02 0.00 small-7 0 1 2 3 4 5 6 7 small-8 Day small-9Summary to Case Study: Extraneous peaks Associate with Glassware • Pay attention to the glass grade, vendor and treatment. • Glass vials vary within the same lot/box. • Rinse with 0.02% acid (formic, acetic, nitric) acid in water is effective for this method. • Wash the glass vials with acidic detergent is effective. • Use of polypropylene vials eliminates the problem. • It is critical for multiple labs to be involved, to carry our experiments and share data with details. 37
  • 38. 8/2/2010 Method transfer vs. method development• Start from the beginning of the lifecycle (define goals, communicate limitations)• Maintain a dynamic and continuous process• Build strong partnership and co-ownership through regular meetings, visits, design and execute experiments together• Transfer knowledge, not just method Method Transfer vs. validation Vs. Development• Dynamic platform of communication• Design and execute experiments (AMERT, Investigations)• Share ownership Method Development• Exchange of knowledge Method Method transfer validation 38
  • 39. 8/2/2010 Conclusions• Analytical method lifecycle is a dynamic and continuous process• Transfer of knowledge, instead of method, is desired in every stage of the life cycle• Scientists/managers need to zoom in and zoom out to consider needs of other projects, methods or labs.• It enhances efficiency to eliminate borders between methods, techniques and functional groups, as much as possible• Strong partnership and co-ownership is the key to successful methods 39