Workshop on Quality Assurance and GMP of Multisource HIV /AIDS medicines

1. János Pogány, pharmacist, PhD, consultant to WHO Shanghai, 01 March 2005 E-mail: pogany@axelero.hu Workshop on Quality Assurance and GMP of Multisource HIV/AIDSmedicines QUALIFICATION and VALIDATION II. Dr. Pogány - WHO, Shanghai

2. GMP - 4.11 Analytical methods, computers and cleaning procedures • „It is of critical importance that particular attention is paid to the validation ofanalytical test methods, automated systems and cleaning procedures.” • Validation of analytical procedures used in the examination of pharmaceutical materials (WHO Expert Committee on Specifications for Pharmaceutical Preparations. 32nd Report. Geneva, WHO, 1992 (WHO Technical Report Series, No. 823). • Text on Validation of Analytical Procedures Q2A (1994) Validation of Analytical Procedures: Methodology Q2B (1996) ICH Harmonized Tripartite Guidelines Dr. Pogány - WHO, Shanghai

3. Characteristics of analytical procedures (1) • Accuracy (also termedtrueness) • Precision • Repeatability • intermediate precision (within-laboratory variation) • reproducibility (inter-laboratory variation) • Robustness, ruggedness Dr. Pogány - WHO, Shanghai

4. Characteristics of analytical procedures (2) • Linearity • Range • Specificity (selectivity) • Sensitivity (versus robustness) • Limit of detection • Limit of quantitation Dr. Pogány - WHO, Shanghai

5. Accuracy and precision Inaccurate and imprecise Accurate Accurate and precise Precise Dr. Pogány - WHO, Shanghai

6. Classes of analytical tests • „The objective of validation of an analytical procedure is to demonstrate that it is suitable for its intented purpose.” • Class A: To establish identity • Class B: To detect (Bd) and quantitate (Bq)impurities • Class C: To determine quantitatively the concentration, or assay • Class D: To assess characteristics • Other classes not covered in the guides Dr. Pogány - WHO, Shanghai

7. Criteria for analytical classes Dr. Pogány - WHO, Shanghai

8. General requirements • Qualified and calibrated instruments • Documented methods • Reliable reference standards • Qualified analysts • Sample integrity • Change control (e.g., synthesis, FPP composition) Dr. Pogány - WHO, Shanghai

9. HPLC Method Development and Validation for Pharmaceutical Analysis by Ghulam A. Shabir Pharmaceutical Technology Europe, 1 March 2004

10. Prequalification requirements • Analytical method validation is required by WHO for the prequalification of product dossiers. Non-compendial ARV APIs and FPPs were/are tested with methods developed by the manufacturer. • Analytical methods should be used within GMP and GLP environments, and must be developed using the protocols andacceptance criteria set out in the ICH guidelines Q2A and Q2B. Dr. Pogány - WHO, Shanghai

11. HPLC system Dr. Pogány - WHO, Shanghai

12. Linearity and range Dr. Pogány - WHO, Shanghai

13. ICH requirements • Concentration range 0.025–0.15 mg/mL (25–150% of the theoretical concentration in the test preparation, n=3) • Regression equation was found by plotting the peak area (y) versus the analyte concentration (x) expressed in mg/mL: y = 3007.2x + 4250.1 (r2 = 1.000). • The regression coefficient demonstrates the excellent relationship between peak area and concentration of analyte. • The analyte response is linear across 80-120% of the target progesterone concentration. Dr. Pogány - WHO, Shanghai

14. Accuracy The data show that the recovery of analyte in spiked samplesmet the evaluation criterion for accuracy (100 ± 2.0% across 80–120% of target concentrations). Dr. Pogány - WHO, Shanghai

15. Specificity Dr. Pogány - WHO, Shanghai

16. Specificity • An example of specificity criterion for an assay method is that the analyte peak will have baseline chromatographic resolution of at least 2.0 from all other sample components. • In this study, a weight of sample placebo equivalent to the amount present in a sample solution preparation was injected to demonstrate the absence of interference with progesterone elution. Former slide demonstrates specificity. Dr. Pogány - WHO, Shanghai

17. The repeatability precision obtained by one analyst in one laboratory was 1.25% RSD for the analyte and, therefore, meets the evaluation criterionof RSD ≤2%. Repeatability Dr. Pogány - WHO, Shanghai

18. Intermediate precision Dr. Pogány - WHO, Shanghai

19. Limit of detection • The limit of detection (LOD) is defined as the lowest concentration of an analyte in a sample that can be detected, not quantified. It is expressed as a concentration at a specified signal:noise ratio (SNR), usuallybetween 3 and 2:1. • In this study, the LOD was determined to be 10ng/mL with a signal:noise ratio of 2.9:1. Dr. Pogány - WHO, Shanghai

20. Limit of quantitation • The limit of quantitation (LOQ) is defined as the lowest concentration of an analyte in a sample that can be determined with acceptable precision and accuracy under the stated operational conditions of the method. The ICH has recommended a signal:noise ratio (SNR) of10:1. • The LOQ was 20 ng/mL with a signal:noise ratio of 10.2. The RSDfor six injections of the LOQ solution was ≤2%. Dr. Pogány - WHO, Shanghai

21. LOD, LOQ and SNT • Limit of Quantitation (LOQ) • Limit of Detection (LOD) • Signal to Noise Ratio (SNR) Peak BLOQ Peak ALOD noise Baseline Dr. Pogány - WHO, Shanghai

22. Analytical solution stability Standard and sample solutions stored in a capped volumetric flask on a lab bench under normal lighting conditions for 24 h were shown to be stable with no significant change in progesterone concentration during this period. Dr. Pogány - WHO, Shanghai

23. Conclusion of validation study • The relation between concentration and peak area is linear in the range from 25 to 150%. R=1. • Accuracy shows a mean with a RSD of 0.59 over the the range from 50 to 150%. • The method is specific in the given formulation. • Repeatability and intermediate precision are well within the±2% RSD criterion. • LOD is 10 ng/mLand LOQ is 20 ng/mL. • The analytical solution is stable for 24 hours. Dr. Pogány - WHO, Shanghai

24. János Pogány, pharmacist, Ph.D. Shanghai, 03 March 2005 E-mail: pogany@axelero.hu Workshop on Quality Assurance and GMP of Multisource HIV/AIDSmedicines CLEANING VALIDATION Dr. Pogány - WHO, Shanghai

25. Subjects for Discussion • Regulatory background • General considerations • Cleaning validation guideline – Canada • An illustrative approach to the cleaning validation of antiretroviral (ARV) active pharmaceutical ingredient(s) [API(s] • A case study from literature • Conclusions Dr. Pogány - WHO, Shanghai

26. WHO GMP • 4.11 It is of critical importance that particular attention is paid to the validation of...cleaning procedures. • 16.11 Contamination of ... a product by another material or product must be avoided. This risk of accidental cross contamination arises from ... products in process, from residues on equipment. Among the most hazardous contaminants are highly sensitizing materials ...and highly active materials. Dr. Pogány - WHO, Shanghai

27. WHO GMP • 16.15 Before any processing operation is started, steps should be taken to ensure that the work area and equipment are clean. • 16.18 Time limits for storage of equipment after cleaning and before use should be stated and based on data. Dr. Pogány - WHO, Shanghai

28. Why do we validate cleaning processes? • The cleaning processis an integral part of the pharmaceutical manufacturing process. • Industry should view cleaning of equipment as the first manufacturing step.(It will have effect on the safety, efficacy and quality of the batch to be manufactured.) • A cleaning process must be chosen based on products (e.g., ARVs, solid dosage forms), objectives, resources, and limitations within each manufacturing company. Dr. Pogány - WHO, Shanghai

29. Pharmaceutical Process Validation: Second Edition, Revised and Expanded, edited by Ira R. Berry and Robert A. Nash, Marcel Dekker, Inc., New York – Basel – Hong Kong (1993). GENERAL CONSIDERATIONS

30. Potential Contaminants • Chemical contamination • Product residues • Decomposition residues • Cleaning or disinfecting agent residues • Microbiological contamination • Bacteria, moulds, pyrogens • Unintended materials • Airborne (particulate) matter • Lubricants, ancillary material (e.g. pieces of brushes) Dr. Pogány - WHO, Shanghai

31. Manual Cleaning Procedures • Equipment disassembly (if required) • Prewash and inspection (most visible material removed) • Wash (cleaning agent,temperature, multiple steps until visually clean) • Initial rinses (rinse water, temperature) • Final rinse (minimum dissolved solids, microorganisms) • Reassembly (if required) Dr. Pogány - WHO, Shanghai

32. Automated Cleaning Procedures • Clean-in-place (CIP) systems (dishwasher-type equipment) • portable (tank and pump assemblies on wheels) • stationery, cabinet-type • Control system qualification (reproducibility, water temperature control) • Sampling (sampling port, pause capability) • Material supply (hard-plumbed supply lines, volume and dispensing controls, potential impact of long storage periods) Dr. Pogány - WHO, Shanghai

33. Documentation and Traceability • Equipment identification • Equipment use, maintenance and cleaning records • Labeling • Cleaning equipment maintenance and calibration • Utilities(water for injection (WFI), purified water, steam and compressed air systems) qualified and validated. • Standard Operating Procedure(s) [SOP(s)] • Personnel training Dr. Pogány - WHO, Shanghai

34. Cleaning Materials and Tools • Solvents(source and quality controlled) • Cleaning agents(acids, bases, surfactants, etc., qualified type and brand QC controlled) • Ancillary utilities (steam and compressed air qualified) • Scrubbing agents (compression of placebo tablets to clean punches and dies) • Cleaning tools(standard sets of brushes, rags, sponges) • Equipment(thermometers, CIP systems consisting of tanks, metering pumps, heat exchangers, etc. maintaned and kept in calibrated status) Dr. Pogány - WHO, Shanghai

35. Frequency of Cleaning • Cleaning between batches of the same product(abbreviated procedures) • Cleaning between batches of different products • Cleaning after maintenance • Cleaning after accidental contamination Dr. Pogány - WHO, Shanghai

36. Cleaning Validation Guidelines,Health Products and Food Branch Inspectorate, Canadahttp://www.hc-sc.gc.ca/hpfb-dgpsa/inspectorate/clean_val_gui_entire_e.html PRESENTATION IS LIMITED TO SOLID PHARMACEUTICAL DOSAGE FORMS

37. Validation of cleaning processes • Equipment cleaning validation may be performed concurrently with actual production steps during process development and clinical manufacturing. Validation programs should be continued through full-scale commercial production. • All pertinent parameters should be checked to ensure the process, as it will ultimately be run is validated. Therefore, if critical temperatures are needed to effect cleaning, then these should be verified. Any chemical agents added should be verified for type as well as quantity. Volumes of wash and rinse fluids, and velocity measurements for cleaning fluids should be measured as appropriate. Dr. Pogány - WHO, Shanghai

38. Validation of cleaning processes • Validation of cleaning processes should be based on a worst-case scenario including: • challenge of the cleaning process to show that the challenge soil can be recovered in sufficient quantity or demonstrate log removal to ensure that the cleaning process is indeed removing the soil to the required level, and • the use of stress cleaning parameters such as overloading of contaminants, overdrying of equipment surfaces, minimal concentration of cleaning agents and/or minimum contact time of detergents. Dr. Pogány - WHO, Shanghai

39. Validation of cleaning processes • At least three (3) consecutive applications of the cleaning procedure should be performed and shown to be successful in order to prove that the method is validated. Dr. Pogány - WHO, Shanghai

40. Approach for setting limits • Product specific cleaning validation for all products; • Grouping into product families and choosing a worst case product; • Grouping into risk categories (e.g., very soluble products, similar potency, highly toxic products or difficult to detect); • Setting limits on not allowing more than a certain fraction of carryover; • Different safety factors for different dosage forms. Dr. Pogány - WHO, Shanghai

41. Carry-over of product residues • NMT 0.1% of the normal therapeutic dose of any product to appear in the maximum daily dose of the following product (may not be acceptable for parenterals). • NMT 10 ppm of any product to appear in another product(may not be acceptable for parenterals). • No quantity of residue to be visible on the equipment after cleaning procedures are performed. (Spiking studies should determine the concentration at which most active ingredients are visible.) Dr. Pogány - WHO, Shanghai

42. Carry-over of product residues • Residues levels that do not interfere with subsequent manufacturing processes. • For certain allergenic ingredients, penicillins, cephalosporins or potent steroids and cytotoxics, the limits should be below the limit of detection by best available analytical methods. In practice this may mean that dedicated plants are used for these products. • Acceptable limits should be defined for detergent residues after cleaning(there is no normal therapeutic dose, thus e.g. the limit of detection of the most toxic component). Dr. Pogány - WHO, Shanghai

43. Analytical methods • The analytical methods used to detect residuals or contaminants should be specific and be validated before the cleaning validation study is carried out. • The specificity and sensitivity of the analytical methods should be determined. • The analytical method and the percent recovery of contaminants should be challenged in combination with the sampling method(s). Dr. Pogány - WHO, Shanghai

44. Sampling and related issues • Direct surface sampling (swab method) • Indirect sampling (use of rinse solutions) • Indirect testing such as monitoring conductivity may be of some value • In terms of cross-contamination, the main concern is residue left on the internal product-contact surfaces of the manufacturing equipment. Dr. Pogány - WHO, Shanghai

45. An Illustrative Approach to Cleaning Validation ANTIRETROVIRAL FPP(s)

46. Cleaning validation (master) plan • Validation plan is based on risk analysis. • Cleaning of individual pieces of the manufacturing and packaging equipment is validated with products selected as the worst case. • The three regulatory consecutive batches can be extended to include potentially the last batches of one or more campaign productions • Water solubility, toxicity and risk analysis data of all ARV APIs. Dr. Pogány - WHO, Shanghai

47. WATER SOLUBILITY LOW MODERATE HIGH T O X I C I T Y HIGH High High Moderate MODERATE High Moderate Moderate LOW Moderate Moderate Low Risk Analysis RISK FACTORS Dr. Pogány - WHO, Shanghai

48. LD50 (rat or /mouse/) Category < 200 mg/kg High 200 – 2000 mg/kg Moderate > 2000 mg/kg Low Illustrative Indicators for Toxicity Composite toxicity indicators may take into account high activity, hypersensitizing indicators, etc. Dr. Pogány - WHO, Shanghai

49. Descriptive Term for Solubility (Ph.Eur.) Category Very soluble (< 1 ml/g) Freely soluble (1 – 10 ml/g) Soluble (10 – 30 ml/g) Highsolubility (<30 ml/g) Moderately soluble Sparingly soluble (30 – 100 ml/g) Slightly soluble (100 – 1000 ml/g) Moderatesolubility (30 – 1000 ml/g) Very slightly soluble (1000 – 10000 ml/g) Practically insoluble(> 1000 ml/g) Relatively insoluble Insoluble Lowsolubility (> 1000 ml/g) Illustrative Categories for Solubility Dr. Pogány - WHO, Shanghai

50. API TOXICITY INDICATOR SOLUBILITY IN WATER Abacavir Moderate High 77 mg/mL at 25oC Efavirenz Low Low Indinavir sulfate Low High 100 mg/ml Lamivudine Low High Nevirapine Low Low 90 μg/ml at 25°C Ritonavir Low Low Saquinavir Low Low Stavudine Low High Zidovudine Low High 20 mg/mL at 25oC Illustrative Risk Analysis of ARV APIs Dr. Pogány - WHO, Shanghai