1. © AL IPA/EDQM/IDMA Symposium 2007, All rights reserved The Ph. Eur. policy on Reference standards Dr Andrea Lodi Deputy Head, Laboratory Department, EDQM, Council of Europe

2. © AL IPA/EDQM/IDMA Symposium 2007, All rights reserved Content of the presentation Definition Role in the monograph How established CRS Database Conclusion Definition Role in the monograph How established CRS Database Conclusion

3. © AL IPA/EDQM/IDMA Symposium 2007, All rights reserved European Pharmacopoeia nearly 2,000 monographs ; >1,900 reference standards

4. © AL IPA/EDQM/IDMA Symposium 2007, All rights reserved Classification of reference standards TYPE :   Chemical Reference Substances   Biological Reference Preparations ESTABLISHMENT :   Primary standards   Secondary standards PURPOSE :   QUALITATIVE (Id, system suitability)   QUANTITATIVE (Assay, external standard)

5. © AL IPA/EDQM/IDMA Symposium 2007, All rights reserved European Pharmacopoeia reference standards Chemical reference substance (CRS) A chemical compound or mixture of compounds which has been established for use as a standard in an identity test, a purity test and/or an assay as prescribed in a monograph. The standard is valid only for the specific test(s) for which it has been established

6. © AL IPA/EDQM/IDMA Symposium 2007, All rights reserved European Pharmacopoeia reference standards A European Pharmacopoeia reference standard is an integral and essential part of the monograph and as such is an official standard that is alone authoritative in case of doubt or dispute.

7. © AL IPA/EDQM/IDMA Symposium 2007, All rights reserved European Pharmacopoeia Reference Substances

8. © AL IPA/EDQM/IDMA Symposium 2007, All rights reserved Extent of analytical testing: according to the CRS use Identification => structure elucidation + compliance Peak ID, system suitability => structure elucidation + fitness for purpose External standard => structure elucidation + purity Assay: assignment based on interlaboratory study involving five laboratories: EDQM Laboratory + Official Laboratories and Experts; the manufacturer is normally invited to participate. Identification => structure elucidation + compliance Peak ID, system suitability => structure elucidation + fitness for purpose External standard => structure elucidation + purity Assay: assignment based on interlaboratory study involving five laboratories: EDQM Laboratory + Official Laboratories and Experts; the manufacturer is normally invited to participate.

9. © AL IPA/EDQM/IDMA Symposium 2007, All rights reserved Requirements for a candidate CRS Pharmaceutical Substance subject of a monograph: The CRS must comply with the requirements of the monograph, no special purity required. Impurity CRS (i.e. not subject of a monograph) Minimum content : qualitative use: 90%; quantitative use: 95%, otherwise value assigned Pharmaceutical Substance subject of a monograph: The CRS must comply with the requirements of the monograph, no special purity required. Impurity CRS (i.e. not subject of a monograph) Minimum content : qualitative use: 90%; quantitative use: 95%, otherwise value assigned

10. © AL IPA/EDQM/IDMA Symposium 2007, All rights reserved CRS for identification Compliance / structure elucidation : The EDQM laboratory carries out all tests of the monograph. The structure of the substance is elucidated by means of NMR, MS and FT-IR data as well as by comparison to literature data. Compliance / structure elucidation : The EDQM laboratory carries out all tests of the monograph. The structure of the substance is elucidated by means of NMR, MS and FT-IR data as well as by comparison to literature data.

11. © AL IPA/EDQM/IDMA Symposium 2007, All rights reserved CRS used in the test for related substances Qualitative use -System suitability -Peak identification Qualitative use -System suitability -Peak identification Quantitative use -Limit test -Quantitative test

12. © AL IPA/EDQM/IDMA Symposium 2007, All rights reserved CRS used in the test for related substances In general an impurity is to be identified, i.e. located in the chromatogram or electropherogram: -when it has an individual limit and/or -when a correction factor must be applied In general an impurity is to be identified, i.e. located in the chromatogram or electropherogram: -when it has an individual limit and/or -when a correction factor must be applied

13. © AL IPA/EDQM/IDMA Symposium 2007, All rights reserved Impurity CRS : qualitative use Individual impurity : preferred option but sometimes difficult to obtain  Mixtures of impurities  Batches containing one or more impurities  Spiked samples Individual impurity : preferred option but sometimes difficult to obtain  Mixtures of impurities  Batches containing one or more impurities  Spiked samples

14. © AL IPA/EDQM/IDMA Symposium 2007, All rights reserved Impurity CRS : qualitative use A representative chromatogram is supplied together with the CRS when indicated in the monograph A representative chromatogram is supplied together with the CRS when indicated in the monograph

15. © AL IPA/EDQM/IDMA Symposium 2007, All rights reserved Rs CF Elution order Example : Torasemide for system suitability CRS

16. © AL IPA/EDQM/IDMA Symposium 2007, All rights reserved foeniculin Example Monograph Anise oil, GC-test for foeniculin

17. © AL IPA/EDQM/IDMA Symposium 2007, All rights reserved Info provided with a CRS Non-official information, intended to facilitate the use of the CRS can be found in the knowledge database (web site). Non-official information, intended to facilitate the use of the CRS can be found in the knowledge database (web site).

18. © AL IPA/EDQM/IDMA Symposium 2007, All rights reserved Impurity CRS : quantitative use Individual impurities, with an assigned value if necessary Batch containing the impurity, with an assigned value Individual impurities, with an assigned value if necessary Batch containing the impurity, with an assigned value

19. © AL IPA/EDQM/IDMA Symposium 2007, All rights reserved Assay CRS Assay methods : LC GC UV spectrophotometry Microbiological assay Bio- or immunoassay Assay methods : LC GC UV spectrophotometry Microbiological assay Bio- or immunoassay

20. © AL IPA/EDQM/IDMA Symposium 2007, All rights reserved Establishment of assay CRS: internal preliminary work Elucidation of the structure of the candidate CRS Verification of compliance with the monograph requirements Additional tests, such as residual solvents, DSC purity, non-aqueous titration. Elucidation of the structure of the candidate CRS Verification of compliance with the monograph requirements Additional tests, such as residual solvents, DSC purity, non-aqueous titration.

21. © AL IPA/EDQM/IDMA Symposium 2007, All rights reserved 1.Protocol (method-specific approach) developed and checked in the EDQM laboratory 2.Samples out to the participants 3.Analysis of results 4.Report with proposed assigned value 5.Approval from the Group of Experts 6.Adoption by the Commission 1.Protocol (method-specific approach) developed and checked in the EDQM laboratory 2.Samples out to the participants 3.Analysis of results 4.Report with proposed assigned value 5.Approval from the Group of Experts 6.Adoption by the Commission Establishment of assay CRS: interlaboratory study

22. © AL IPA/EDQM/IDMA Symposium 2007, All rights reserved Example protocol: CRS for LC-assay Collaborative study: Loss on drying or det. of water (by KF / coulometry) Quantitative determination of the impurities by LC Residual solvents by head-space GC Collaborative study: Loss on drying or det. of water (by KF / coulometry) Quantitative determination of the impurities by LC Residual solvents by head-space GC

23. © AL IPA/EDQM/IDMA Symposium 2007, All rights reserved Repeatability (water determination/ LOD) Selectivity test (LC-GC) Symmetry factor (LC-GC) Repeatability (LC-GC) Repeatability (water determination/ LOD) Selectivity test (LC-GC) Symmetry factor (LC-GC) Repeatability (LC-GC) Establishment of assay CRS: Results acceptance criteria

24. © AL IPA/EDQM/IDMA Symposium 2007, All rights reserved Estimation of the uncertainty of the assigned content Ph. Eur. Commission policy : the uncertainty of the value assigned as a result of an interlaboratory trial should be within a predetermined limit which is calculated in relation with the intended use. Ph. Eur. Commission policy : the uncertainty of the value assigned as a result of an interlaboratory trial should be within a predetermined limit which is calculated in relation with the intended use.

25. © AL IPA/EDQM/IDMA Symposium 2007, All rights reserved Dealing with uncertainty

26. © AL IPA/EDQM/IDMA Symposium 2007, All rights reserved Content If the CRS is supplied as a powder to be weighed, the value assigned is the % (m/m) content on an «as is» basis. NO NEED TO DRY. Example: Benzathine benzylpenicillin CRS 2 If the CRS is supplied as a powder to be weighed, the value assigned is the % (m/m) content on an «as is» basis. NO NEED TO DRY. Example: Benzathine benzylpenicillin CRS 2

27. © AL IPA/EDQM/IDMA Symposium 2007, All rights reserved CRS with assigned content

28. © AL IPA/EDQM/IDMA Symposium 2007, All rights reserved Non-assay CRS

29. © AL IPA/EDQM/IDMA Symposium 2007, All rights reserved Content If the CRS is supplied in a pre-dispensed form such as a freeze-dried substance to be reconstituted, the value assigned is the quantity per vial. Example: Calcitonin (salmon) CRS 5 If the CRS is supplied in a pre-dispensed form such as a freeze-dried substance to be reconstituted, the value assigned is the quantity per vial. Example: Calcitonin (salmon) CRS 5

30. © AL IPA/EDQM/IDMA Symposium 2007, All rights reserved

31. Use of Ph. Eur. CRS Ph. Eur. CRS are to be used in conjunction with the corresponding monograph(s). Ph. Eur. CRS are to be used for the purpose for which they are intended. Any different use is in the responsibility of the user. Storage of opened containers is discouraged and is in any case the responsibility of the user. Ph. Eur. CRS are to be used in conjunction with the corresponding monograph(s). Ph. Eur. CRS are to be used for the purpose for which they are intended. Any different use is in the responsibility of the user. Storage of opened containers is discouraged and is in any case the responsibility of the user.

32. © AL IPA/EDQM/IDMA Symposium 2007, All rights reserved Monitoring programme Once established and adopted, the CRS undergoes periodic testing in order to ensure its continuous fitness for use. The periodicity of testing depends on the use of the CRS and the stability information available. The extent of testing depends on the use of the CRS. In general the focus is on the properties that may change during the life cycle of a CRS, i.e.: appearance, purity by HPLC or GC, DSC, water content. Once established and adopted, the CRS undergoes periodic testing in order to ensure its continuous fitness for use. The periodicity of testing depends on the use of the CRS and the stability information available. The extent of testing depends on the use of the CRS. In general the focus is on the properties that may change during the life cycle of a CRS, i.e.: appearance, purity by HPLC or GC, DSC, water content.

33. © AL IPA/EDQM/IDMA Symposium 2007, All rights reserved REFERENCE STANDARDS CATALOG ON LINE Paper or PDF 3 issues a year

34. © AL IPA/EDQM/IDMA Symposium 2007, All rights reserved Reference standards database Knowledge database Availability Origin Assigned Value Batch Validity Statement Safety Data Sheets Info on reagents and columns Chromatograms

35. © AL IPA/EDQM/IDMA Symposium 2007, All rights reserved

37. Example of a BVS

38. © AL IPA/EDQM/IDMA Symposium 2007, All rights reserved New Chapter of the European Pharmacopoeia 5.12 Pharmaceutical Reference Standards Effective Jan 1st, 2007 ESTABLISHMENT USE PRESENTATION of pharmaceutical reference standards in general & European Pharmacopoeia reference standards in particular. Effective Jan 1st, 2007 ESTABLISHMENT USE PRESENTATION of pharmaceutical reference standards in general & European Pharmacopoeia reference standards in particular.

39. © AL IPA/EDQM/IDMA Symposium 2007, All rights reserved Impurity Reference Standards New opportunities / Development Areas The Challenge: The new Ph. Eur. policy on Impurities requires that most if not all the specified impurities be identified. Our Strategy: - Develop new compounding/dispensing techniques - Hyphenated techniques for establishment - New rapid techniques for monitoring - Keep no. of CRS/monograph as low as possible - Keep price reasonable The Challenge: The new Ph. Eur. policy on Impurities requires that most if not all the specified impurities be identified. Our Strategy: - Develop new compounding/dispensing techniques - Hyphenated techniques for establishment - New rapid techniques for monitoring - Keep no. of CRS/monograph as low as possible - Keep price reasonable

40. © AL IPA/EDQM/IDMA Symposium 2007, All rights reserved Ph. Eur. CRS take home messages They are official, primary standards They constitute an essential part of the monograph They are guaranteed only for the intended purpose The assay value is assigned “as is” Their continuous fitness for purpose is assured by a regular monitoring programme User interface => website The Ph. Eur. policy on reference standard is reflected in general chapter 5.12