1. Lect. 5+6. 2009-10 Eli Solomon M.Sc 1 VALIDATION PRINCIPLES Principles of qualification and validation which are applicable to the manufacture of medicinal products. Analytical Methodאנליזה מעבדתית Medical Devicesאביזרים ומכשור רפואי (אמ"ר) Cleaning and Sterilization ניקוי ועיקור Process: Development, Analysis, and Control. פיתוח ניתוח ובקרת תהליך

2. Lect. 5+6. 2009-10 Eli Solomon M.Sc 2 Laboratory Method Flow APPROVED Method Development פיתוח Method Validation Method Transfer וולידציה העברה

3. Lect. 5+6. 2009-10 Eli Solomon M.Sc 3 Why Validate Method validation is completed to ensure that an analytical methodology is accurate, reproducible and rugged over specific range that an analyte will be analyzed.Method validation is completed to ensure that an analytical methodology is accurate, reproducible and rugged over specific range that an analyte will be analyzed. Provides assurance of reliabilityאמינותProvides assurance of reliabilityאמינות FDA Compliance התאמה מתועדת לשיטת הבדיקה The process of providing documented evidence that something does what it is indented to do”FDA Compliance התאמה מתועדת לשיטת הבדיקה The process of providing documented evidence that something does what it is indented to do”

4. Lect. 5+6. 2009-10 Eli Solomon M.Sc 4 הגדרת המושג תיקוף Validation (מתוך ת”י 4335 - נספח מתורגם) 3.24 הליך מתועד לצורך השגה, רישום וניתוח של התוצאות, הנדרשות כדי לקבוע שתהליך יניב בעקביות מוצר המתאים למפרטים שנקבעו מראש.

5. Lect. 5+6. 2009-10 Eli Solomon M.Sc 5 Validation Process Validation Instruments Method Validation System Suitability Software

6. Lect. 5+6. 2009-10 Eli Solomon M.Sc 6 USP – Analytical Method Validation Method Validation דיק - PRECISION דיוק - ACCURACY LIMIT OF DETECTION (LOD) LIMIT OF QUANTIFICATION (LOQ) ייחודי - SPECIFICITY LINEARITY AND RANGE "קשיחות" - RUGGEDNESS ROBUSTNESS "חוסן" -

7. Lect. 5+6. 2009-10 Eli Solomon M.Sc 7 The closeness of agreement between the values obtained in an assay. When the method is applied repeatedly to multiple sampling of a homogenous sample Expressed as % SD for a statistically significant number of samples Should be performed at 3 levels. Repeatability (הדירות) Intermediate Precision ( ביניים) Reproducibility (שעתוק) Precision Definition דיק -

8. Lect. 5+6. 2009-10 Eli Solomon M.Sc 8 Precisionדיק - Explanation and Enlargement : (USP analytical Procedure Criterion) Repeatability: (USP analytical Procedure Criterion) Same operating condition, short time interval. Inter-assay Precision Within-lab variations: Different days, analysts, equipment. Multiple determinations of a single sample, controls and reagents analyzed in several runs, same lab.

9. Lect. 5+6. 2009-10 Eli Solomon M.Sc 9 Minimum of 9 determinations covering specified range of procedure (3 levels 3 replica. each), or Minimum of 6 determination at 100% test conc. (expressed as % CV) Reproducibility Precision between labs Collaborative studies, not relevant in manufacturing facility Precisionדיק - Explanation and Enlargement

10. Lect. 5+6. 2009-10 Eli Solomon M.Sc 10 Accuracy Definitionדיוק - The closeness of agreement between the actual values of the drug and the measured value Spiked (known added amount) and Recovery studies are performed to measure accuracy

11. Lect. 5+6. 2009-10 Eli Solomon M.Sc 11 Expressed as % error between the observed value and the true value (assay value / actual value x100%) Accuracy Definitionדיוק – (Cont’)

12. Lect. 5+6. 2009-10 Eli Solomon M.Sc 12 Accuracy Determination (Cont’) Drug Substance Analysis of Reference material Compare results to a second, well characterized method Impurities (Quantification) Analysis of Samples (Drug substances) spiked, with known amounts of impurities

13. Lect. 5+6. 2009-10 Eli Solomon M.Sc 13 ACCURACY vs PRECISION Good Accuracy Poor Precision Good Precision Poor Accuracy Good Precision Good Accuracy The closeness of test results to the true value obtained by the method

14. Lect. 5+6. 2009-10 Eli Solomon M.Sc 14 Limit of Detection (LOD) Definitionגבול זיהוי - Lowest Concentration of Analyte in a Sample that can be detected (not necessarily quantitated) Limit Test: Above or Below a Certain Level – P / F results Expressed as Concentration (%, ppb)

15. Lect. 5+6. 2009-10 Eli Solomon M.Sc 15 Almost never necessary to be determined May be based on standard deviation (STD) of the response and slope DL=(3.3)STD / S STD = Standard Deviation of the response S = Slope of calibration curve Limit of Detection (LOD) Definitionגבול זיהוי - Limit of Detection (LOD) Definitionגבול זיהוי - (Cont’)

16. Lect. 5+6. 2009-10 Eli Solomon M.Sc 16 Limit of Quantification (LOQ) Definition Lowest concentration of analyte in a sample that can be determined with acceptable precision and accuracy under stated operational conditions Expressed in Concentration of Analyte Signal To Noise Ratio (10:1 is typical)

17. Lect. 5+6. 2009-10 Eli Solomon M.Sc 17 Limit of Quantification (LOQ) Explanation and Enlargement QL = (10) STD / S STD = Standard Deviation of the response S = Slope of Calibration curve DL and QL are validated by analyzing a suitable number of samples Method should be documented

18. Lect. 5+6. 2009-10 Eli Solomon M.Sc 18 The ability to measure accurately and specifically the analyte in the presence of components that may be expected to be present in the product The Degree of Interference Active Ingredients ExcipientsImpurities Degradation Products Placebo Ingredients Specificity (Selectivity) Definition

19. Lect. 5+6. 2009-10 Eli Solomon M.Sc 19 Specificity Determination  Qualitative Identification Test Demonstrate ability to select between compounds of closely related structure  Confirm Positive and Negative results Assay Demonstrate that the results are unaffected by spiked impurities or excipients

20. Lect. 5+6. 2009-10 Eli Solomon M.Sc 20 Linearity and Range ליניאריות ותחום Linearity The ability of the method to elicit test results that are directly proportional to concentration within a given range -Expressed as the Variance of the Slope of the Regression Line Range Interval between upper and lower levels of analyte demonstrated by the method

21. Lect. 5+6. 2009-10 Eli Solomon M.Sc 21   Prepare Minimum 5 Concentrations (Preferable 6-8 sample dilutions across the range)   Test each dilution in triplicates for 3 runs. (5x3x3=45) Linearity Evaluation

22. Lect. 5+6. 2009-10 Eli Solomon M.Sc 22 Linearity Evaluation   Record expected values, actual values, and % recovery for each one.   Analyze each set of dilutions for statistical methods: Regression, Correlation Coefficient, y-intercept, Slop, Plot.

23. Lect. 5+6. 2009-10 Eli Solomon M.Sc 23 Range Determination קביעת התחום Minimum Specified Ranged Assay:80 - 120 %   Impurities Test:From QL – 120% of Spec.   Toxic or Potent impurities: Fit with the controlled level.   Content Uniformity: 70 – 130% of Test Concentration.   Dissolution Testing + 20% over Specified Range.

24. Lect. 5+6. 2009-10 Eli Solomon M.Sc 24 Ruggedness "קשיחות" Degree of Reproducibility of Test Results under a Variety of Conditions Different: Laboratories, Analysts, Instruments, Reagents, Days Expressed as % RSD

25. Lect. 5+6. 2009-10 Eli Solomon M.Sc 25 Measure The Capacity to Remain Unaffected by Small Variation in Method Parameters. Indication or Reliability During Normal Use. Robustness "חוסן"

26. Lect. 5+6. 2009-10 Eli Solomon M.Sc 26 Why is Method Validation Considered a Bottleneck צוואר בקבוק - - Every Method validation Process Consists About 80 -100 Analyses - Each of Them Produce About 4 specific Results; These results in a total of about 400 Numbers

27. Lect. 5+6. 2009-10 Eli Solomon M.Sc 27 - All of These Numbers Need to be mathematically Processed: Manual (Calculator, Excel Macro) Dedicated Method Validation Program Why is Method Validation Considered a Bottleneck (Cont’)

28. Lect. 5+6. 2009-10 Eli Solomon M.Sc 28 To Take Advantage of a New Technology - Cost/Benefit Economic Exercise - Instrumentation, Methods Formulation Changes Why Revalidate

29. Lect. 5+6. 2009-10 Eli Solomon M.Sc 29 Why Revalidate (cont’) Manufacturing Batch Changes Changes in Incoming Raw Material Changes in Method

30. Lect. 5+6. 2009-10 Eli Solomon M.Sc 30 CONCLUSION סיכום  Defined Validation and its Importance  Examined The steps of Method Validation  Learned How each Step is Documented and Measured:( System Suitability, USP/ICH) Measured:( System Suitability, USP/ICH)  Protocol and Pitfalls: Revalidation

31. Lect. 5+6. 2009-10 Eli Solomon M.Sc 31 ANALYTICAL METHOD VALIDATION REFERENCE Initial Method Validation Guidance -1987 Guidance for submitting samples and analytical data for methods validation. FDA, February 1987. US Government Printing office: 1990-281-794:20818. Update in August 2000 (Draft Guidance) Analytical Procedures and Method Validation. Fed.Reg.659169), 52,776-52,777, 30 August 2000 All Guidance are on FDA Web site: www.fda.gof/cder/guidance