1. 1 Preparation of rh-Insulin National Reference standard National Institute of Biologicals Noida E-mail: info@nib.gov.in Meeting of the Manufacturer’s March 20, 2009

2. 2 WHY the need ╩ There is no National Reference Standard available in our country on rh-Insulin with any metrological trace ability as per ISO guidelines – 17511. ╩ The certificate of Analysis also varies from one Manufacturer to another as there are no guidelines and Recommendations laid down for National Reference standard of rh-Insulin incorporated in “ Indian Pharmacopoeia ” ╩ International biological standards are the “ Primary standards ” against which Secondary standards are calibrated. Secondary standards may be - a) National standard, b) Regional standard, c) Working Reference material. rh-Insulin Reference Standard

3. 3 International conventional reference measurement procedure by International Scientific organization, WHO a International conventional Calibrator b Manufacturer’s Working calibrator d Manufacturer’s Product calibrator f Manufacturer’s selected measurement procedure c Manufacturer’s standing measurement procedure e METROLOGICAL TRACEABILITYMETROLOGICAL TRACEABILITY International scientific organization: WHO, NIBSC, USP End User’s routine measurement procedure g Routine Sample h RESULT i Manufacturer Or end user End user CALIBRATION HIERARCHY & METROLOGICAL TRACEABILITY MATERIALCALIBRATION VALUE ASSIGNMENTPROCEDUREIMPLEMENTATION Source ISO 17511-2003, First edition, 2003-08-15

4. 4 Memorandum of Understanding ╩ All the information of API-bulk material supplied during the study will be treated as confidential and proprietary to the manufacturer ’ s. ╩ A MoU will be developed between a) NIB, b) Industry and c) IPC ╩ All persons involved herein by signing the agreement confirms that there is no financial or vested commercial interest.

5. 5 Collaborators in the Study Collaborating labNameStudy Investigator Pharmacopoeia Indian Pharmacopoeia Commission To Propose Government 1.National Institute of Biologicals-NCL 2. Office of DCG(I)- NRA 3. CDSCO-zone wise -do- Industry rh-Insulin Manufacturer’s: 1. Biocon Bangalore 2. Wockhardt Aurangabad 3. MJ Biopharm Navi Mumbai 4. Cipla Goa 5. Eli-Lily Gurgaon 6. Aventis Mumbai 7. Novo Nordisk Denmark & Bangalore -do- Other Standard setting organization 1. European Pharmacopoeia 2. Japanese Pharmacopoeia 3. US Pharmacopoeia 4. WHO 5. NIST-USA 6. REMCO ( Ref material committee of ISO ) To maintain continuous contacts Academia To be ProposedTo Propose

6. 6 Steps in establishing National Reference standard 1. Incorporation of human Insulin National Reference Standard in IP-Monograph for Human Insulin 2. Provide collaborators with set of necessary documents 3. Establish – 3.1 Inter laboratory ” collaborative study for selecting a Candidate RS material 3.2 coded material to be distributed maintaining confidentiality 33 Proficiency testing program based on NABL 162 guidelines

7. 7 1. Quantitative uses in assays of rh-insulin bulk & formulation, either by: ╔ Biological methods ╔ Physiochemical method-HPLC 2. Quantitative uses in Limit tests by Physiochemical method-HPLC 3. Qualitative uses in Identification tests, system suitability tests, chromatographic peak markers 4. Method development, evaluation of method performance Intended Use

8. 8 Goals of Collaborative study Selection of rh-insulin Candidate material Obtain API-Bulk material from a major manufacturer of rh-insulin Detailed rh-insulin testing Protocol to include elements of : ╩ Type of tests ╩ No of tests ╩ No of collaborators ╩ Elements of validation ╩ No. of replicates ╩ References to the procedure to be used. ╩ Data recording forms for result compilation Material is Calibrated and Characterized, with regular use of CRM Statistical control in design and Analysis of Results Assign Property values and Uncertainties

9. 9 Assignment of Property Values and Their Uncertainties ╔ Investigation of statistical outliers and/or use of robust statistic ╔ Methods used to assign uncertainties to property values ╔ Acquire time-zero information for future continued-suitability for use of rh-insulin Reference standard ╔ 1 Unit of Human insulin =0.0347 mg by definition (28.8 IU/mg). ╔ Value is assigned to rh-insulin bulk in IP. ╔ Need to Work Out : Assignment of Property Value to serve as Standard for Assay of 40 IU and 100 IU Insulin preparations.

10. 10 Calibration test as per requirements of IP-2007 & USP Characterization test on Candidate Reference material as per requirements of IP- 2007 & USP Calibration test as per requirements of IP-2007 & USP Characterization test on Candidate Reference material as per requirements of IP- 2007 & USP Calibration Potency content RP- HPLC Characterization Identification RP-HPLC Peptide map Purity % related compounds RP-HPLC % insulin aggregates SE-HPLC Characterization Identification RP-HPLC Peptide map Purity % related compounds RP-HPLC % insulin aggregates SE-HPLC Lead Role of Collaborators

11. 11 Work Progress in the study 1. NIB has received rh-insulin working Ref material from Four manufacturer's 2. Material received : i. powder and liquid form ii. 4 vials or ampoules from each iii. Certificate information from 3 manufacturer 3. NIB has initiated Calibration using: i. Certified Ref material-NIBSC & USP ii. Method – Potency, USP, EP as per Manufacturer CoA iii. Method – Potency -IP not given in Manufacturer ’ s CoA

12. 12 S. No. Logical order for presentation Information requiredInformation given by producer Information given by highest metrological trace ability 1. The identity of the producer body and identification of the reference material Name and address of certifying body √√ 2. Title of the document √√ 3. Name of the material √√ 4. Reference material code and batch number √√ 5. A description of the material and its intended use Description of the material, origin and history No√ 6. Intended use No√ 7. Instructions on the correct use of the material No√ 8. Safety √ 9. Level of homogeneity Partial√ 10. The certified values, their Traceability and the period of validity of the certificate Certified values and their uncertainty intervals at a stated confidence level Partial√ 11. Traceability √√ 12. Values obtained by individual laboratories or methods √√ 13. Uncertified values √√ 14. Date of certification or assignment of property values √√ 15. Stability No√ 16. Other information Further information √√ 17. Legal considerations Partial√ 18. Signatures and names of certifying officer(s) or officer(s) signing an analysis certificate √√ & Leaflet CERTIFICATE INFORMATION from rh-insulin Reference Material Producers

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14. 14 Recommending a Strategy 1 Time Lines Attached schedule to be agreed upon 2 Label Text & LeafletDraft to be reviewed & approved 3 rh-Insulin Candidate materialAcceptance from 2-3 manufacturer’s Material f or supply, filling & packing 4 Testing for Homogeneity Batch of Candidate Ref material a) intermediate checks b) checks in final packed form 5 Authorization of Property value to Reference Standard By Technical competent body for Reference Standard 6 Distribution of designated Ref.Standard Recommendation of Expert committee 7 Commercial cost of Ref Standard per vial or Ampoule Cost to be proposed

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