Development of the National System of Medicine Reference Substances in Ukraine

1. Development of the National System of Medicine Reference Substances in Ukraine Leontiev Dmytro Deputy director (science) Group Manager “Validation and Reference Substances” Gryzodoub Oleander Director Ukrainian Scientific Pharmacopoeial Centre for Quality of Medicines (Kharkov ) +38(057)719-06-01 e-mail ogf@phukr.kharkov.ua +38(067 )954-91-74 leontievd@yahoo.com

2. The problem of the Pharmaceutical RS support in Ukraine • The pharmaceutical RS system accepted in the USSRdiffered substantially from that one in the countries in which the Good Manufacturing Practice (GMP) has been applied. • The Pharmaceutical RS in Ukraine were absent after the exit from the USSR • Ukraine, in the pharmacy area, works towards harmonization with the European Community (Regulation of the Ministry of Ukraine No. 244, 19.03.1997). • This requires elaboration of the pharmaceutical RS system meeting the GMP requirements and the leadingPharmacopoeias practice.

3. The Main Types of the Pharmaceutical RS RS for the medicines quality testing: • Official RS (Pharmacopoeia RS). They are the official standards to be used in cases of arbitration • Working RS. May be used for routine analysis, providedthey arecalibrated against the Pharmacopoeia standard RS for the quality assurance of the test results: • Test Samples for the Program of Proficiency Testing (PPT) • Within-lab RS for the quality assurance of the test results • Samples for the analytical procedures validation (for the “direct” methods – titration, refractometry and so on) • RS for the analytical equipment qualification

4. The Main Tasks of the Ukrainian Pharmacopoeia RS System Creation • Elaboration of the theoretical base of the pharmaceutical reference substance attestation • Elaboration of the requirements for the max acceptable uncertainty for the main pharmaceuticals assays and tests (it determinates the requirements to the Pharmaceutical RS characterization) • Elaboration of the Pharmacopoeia RS characterization procedures • Elaboration of the Pharmacopoeia RS documentation system, quality system, including production, labeling, storage and distribution • Establishment and distribution of the Ukrainian Pharmacopoeia RS

5. Elaboration of the Theoretical Base of the Pharmaceutical RS Attestation INSIGNIFICANCE PRINCIPLE The confidence interval (2 )issignificant at P% level (i.e., insignificant at (100 – Р)% level), as compared to a confidence interval (1), when the summarized confidence interval increases not more than by Р%, i.e., the following condition is met: (1) For Р=5% (confidence level accepted in analytical chemistry):

6. Elaboration of criteria for the RS certified value In accordance with the insignificance principle, the following inequality for the certified value uncertainty (RS) should be met: (3)

7. Assay. Approach for the Substances The inspecting approach (some of pharmaceutical substances) The maximal permissible uncertainty of the assay (MAX) is equal to excess of the upper content limit over 100 % (B): (4) Hereinafter the relative confidence interval for the confidence of 95 % is meant by the uncertainty

8. Assay. Approach for the Dosage Forms The confirming approach (all dosage forms) For the symmetrical content limits (В, %) and probability of 95%, taking into account an insignificance principle: (5) MAX -MAXIMAL permitted uncertainty of analytical results (assay), %

9. Test “Uniformity of content” • If we have a positive result one time, it doesn't mean that we must obtain a positive result next time for the same analyzed sample • We can calculate RSDguar for the positive test result for probability ≥0.95 % by use of product rule

10. Test “Uniformity of content” 2 Let р – fraction, for which content must be 85-115% from nominal Stage  I. • The following ratio should be met:р10 0.95; then р  0.995. • The probability of 0.995 corresponds to a of student's factor t(99.5%, 9) = 3.69. • RSDguar(10) = 15/3.69 = 4.07%. Stage II. • The following ratio should be met:р20 0.95; then р  0.9974. • The probability of 0.9974 corresponds to a of student's factor t(99.74%, 29) 3.31, • RSDguar(30)= 15/3.31 = 4.53%. The confidence interval for the two-side student's distribution (t95%,9 =2.262, t95%,29 = 2.045): • StageІ: 4.07* 2.262 = 9.21%. • StageІІ: 4.53*2.045 = 9.26% (e.g. the same as for Stage I).

11. “Uniformity of content” and “Dissolution” Tests Test“Uniformity of content” In accordance with the insignificance principle:  MAX = 0.329.2=3.0% (6) Test “Dissolution”. The same criteria

12. The objective difficulties of the PHARMACEUTICAL RS SYSTEM CREATION IN UKRAINE 1. Information on the pharmaceutical RS attestation (first of all, the theoretical principles) is a restricted one 2. The approaches of other Pharmacopoeias applied now (e.g., a wide application of the inter-laboratory experiment) can’t be transferred automatically into the conditions of Ukraine 3. There was a lack of the clearly defined criteria to the attestation of the pharmaceutical RS. Only the general recommendations on the RS attestation, that did not take into account the specific character of pharmaceutical RS, were available 4. The attestation procedures, as well as documentation and use of the pharmaceutical RS, differ substantially from those ones accepted for the RS used in other areas

13. Elaboration of RS attestation procedures - Assignation of the certified value • Certified value for RS of the Pharmacopoeia of Ukraine must be determined by two methods: by a direct method XAtt (titration, generally) and by subtraction of the obtained content of impurities from 100 % • For the content obtained by the least accurate method (usually, a direct method), the uncertainty of the certified valueAtt is estimated • For the results obtained, the following ratios should be met: (7) (8)

14. Study of the RS uniformityProblems: • In the former USSR/Ukraine the recommendations on uniformity control were available only for bulk material • The requirements to uniformity must be coordinated with МАХ • The former approaches presumed the separation of variation concerned with the uniformity (by ANOVA). It resulted in unacceptably large minimal number of the samples to be analyzed (up to 90!) • The pharmaceutical RS have to meet the very stringent metrological requirements. Estimation of variation concerned with the uniformity may be impossible in practice (i.e., the variation obtained will be caused not by uniformity, but by the errors of the sample preparation). • So, for the pharmaceutical RS other approaches should be used ·

15. Theprocedure for pharmaceutical RS uniformity study • The uniformity is studied for the packaged RS • The maximal permissible non-uniformity is expressed as a confidence interval for an individual content of the certified substance in RS unit (Unit), which must not exceedRS: (9) • For the uniformity, only the practical insignificance is estimated. The uniformity characteristic (Unif) includes both the analysis uncertainty and the variation concerned properly with uniformity. When this summary characteristic is acceptable, the requirements to uniformity are met

16. RS expiry date establishment • Based on the stability under the storage conditions (preferable way, but expiry date must not exceed ½ from that studied experimentally) • Based on the experimental results of accelerated aging data (there is the risk of incorrect data interpretation) • Proposed approach: Based on a priory information about stability of the RS candidate (pharmaceutical substance typically)

17. RS expiry date establishment 2 • Decision based on a priory information – the inequality: (10) - MAX permitted content for the sum of impurities (%), in compliance with specification for the pharmaceutical substance (RS candidate) • Inequality is completed. Expiry date for RS is equal to expiry date for the attestation material • Inequality is not completed. Expiry date for RS must be established by independent way.

18. Elaboration andEstablishment of theUkrainian Pharmacopoeia RS System The following processes, specific for the RS certification have been elaborated and documented : • Choice of the RS candidate • Preparation procedures • Achievement of required degree of homogeneity • Assessment of the stability • Procedures guaranteeing RS characterization • Realization of measurements traceability • Preparation of supporting documents • Establishment of the Pharmacopoeia RS • Arrangements for ensuring adequate storage facilities • Arrangements for the suitable identification, labeling, packaging and delivery procedures • quality monitoring program

19. Elaboration andEstablishment of theUkrainian Pharmacopoeia RS System 2 Elaborated and implemented documents are as follows : • Quality manual (1st level) • Standards of the Pharmacopoeial Committee (ІІ level) • SOPs (ІІІ level) • Other documents – reports, protocols,  logs and electronic databases, etc. (IV level) At the time being the RS Department of Pharmacopoeial Center is certified in Ukraine as a measuring laboratory

20. Progress of the Ukrainian Pharmacopoeia RS system functioning • About 500 various Ph RS were certified • This is more than WHO RS nomenclature • This is about the same as the British Pharmacopoeia RS nomenclature • US pharmacopoeia RS nomenclature is about 2600 • Now the Ukrainian PharmacopoeiaRS are used in most CIS countries by pharmaceutical manufacturers,State Inspections laboratories, and in pre-registration research

21. Progress of the Ukrainian Pharmacopoeia RS system functioning 2 Priority of RS certification: • For Assay (chemical tests, microbiological and biological ones), for the Identification and another tests • For Impurities control (impurities mainly synthesized by the Ukraine Pharmacopoeial Center) • As herbal markers, produced by the Ukraine Pharmacopoeial Center from herbal raw material • As standardized herbal extracts (this permits the use of one summary extract in substitution of several individual herbal markers) • Instead of RS, specific for former USSR (in a majority, of herbal origin) • System suitability check • Other use (turbidity standard, for example, for the turbidity of the bacterial suspensions visual estimation ) • RS for the Quality Assurance of the test results

22. Elaboration of Pharmaceutical WRS system (Local RS) In the USSR, a fundamentally different system of pharmaceutical RS was adopted : • in the presence of Pharmacopoeia RS it is not permitted the use of any other status RS • Laboratory certified all the other local RS as primary (mainly based on the titration results) This required the development of a system of certification of local RS by calibration againstthe official pharmaceutical RS The principles developed for the Pharmacopoeia RS certification were applied successfully to the WRS certification

23. Elaboration ofthe Pharmaceutical WRS system (Local RS) 2 The specificity of the WRS certification • Certification of WRS is fundamentally different from the drugs quality control. Requirements to the WRS are 3.2 times as hard as MAX (see insignificance principle). • WRS certification is fundamentally different from the primary RS Certification. Certified value for the SSF is usually set as 100% - Imp. The content of impurities are usually quite low, so the accuracy of their content determination has a little impact on the RS. • In contrast to the FSO, WRS are calibrated against the Pharmacopoeia RS under the conditions used for the analysis of the medicine. The main methods of WRS certification are chromatography and spectrophotometry. It is much more difficult to achieve the necessary uncertainty of WRS for these methods.

24. Elaboration of the Pharmaceutical WRS system (Local RS) 3 The procedures for certification of WRS and documentation system have been developed. Special analytical techniques for certification shall be applied as specified below: • Volumetric operations performed by gravimetric method (if necessary) • Analyze not less than 2 parallel samples • Sample weight, dilutions and measurements number are optimized to meet WRS • The systematic monitoring of the results quality are used: • Check the differences between the analysis results for the two parallel samples (must not exceed the maximum permissible uncertainty) • Extreme value checking • Check homogeneity of variance for all of the samples.Check the drift The system of Local RS, meeting the requirements of GMP, has been implemented in several largest pharmaceutical enterprises in Ukraine

25. Elaboration of Test Samples for Program of Proficiency Testing Execution of PPT programs requires the use of special Test Samples (TS). TS should be considered as a remedy, which is analyzed by the laboratory. TS in many respects is similar to the RS - for them it is necessary to assign the certified value, study the uniformity and stability as well. The new approach is used (based on the insignificancy principle and MAX): • the uncertainty of certified TS values (TS) should be insignificant compared toMAX: (11) • the result of the participant (Xi) is satisfactory, if Xi deviates from the certified values of TS (XAtt), no more than a MAX: (12)

26. Elaboration of Test Samples for Program of Proficiency Testing 2 Pharmacopoeial Center since 2003 is among the organizers of the Ukrainian PPT Program «Pharma-Test» (testing is conducted annually - in 2012 the 9th round is held). At the end of 2007 the Pharmacopoeial Center received the status of the official coordinator of PPT Programs. Laboratoryof quality control of medicines of Ukraine, CIS countries, and also the countries of the European community (1164 labs in all) participated in different rounds of PPT.

27. Elaboration of Test Samples for Program of Proficiency Testing 3

28. Elaboration of Test Samples for Program of Proficiency Testing 3

29. Elaboration of other RS to ensure the quality of the analysis results The same principles used for the development of other RS: • RS for validation/verification of test procedures (“direct” methods) • RS for analytical equipment qualification (pharmaceutical tests) • RS for in-laboratory quality assurance

30. CONCLUSIONS At the present time, Ukraine is the only country in the CIS, in which all the pharmaceutical RS presented by national Ukrainian Pharmaceutical RS are widely used in the CIS countries

31. Thank you for your attention