Reference standards in Pharmaceutical Industries


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This presentation is brief introduction about reference standards that are using in pharmaceutical industries for calibration of different instruments, methods and pharmaceutical chemicals...

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Reference standards in Pharmaceutical Industries

  1. 1. SOURCES, PREPARATION, CHARACTERIZATION AND STORAGE OF REFERENCE STANDARDS Presented By, Bhavana Vedantam, I/II M.Pharmacy, Dept. Of Pharmaceutical Analysis.
  2. 2. CONTENTS Introduction Sources Uses Preparation Characterization COA, Handling, Maintenance, Records Storage Dept. Of Pharmaceutical Analysis 2
  3. 3. INTRODUCTION Reference Standards (RSs) are the chemical substances or drug substances with high purity. These are not intended to use as drugs. Reference Standards (RS) plays vital role in all phases of Drug Development process. RS serves as basis for evaluation of both Process and Product performance & serves as bench marks for drug Safety. Dept. Of Pharmaceutical Analysis 3
  4. 4. Sources of Reference Materials Primary Sources United States Pharmacopeia(USP) European Pharmacopoeia (EP) Japanese Pharmacopoeia (JP) Swiss Pharmacopoeia (SP) French Pharmacopoeia (FP) Mexican Pharmacopoeia (MP) In India, National Institute for Standards and Testing (NIST), Central Drug Laboratory, Calcutta. Dept. Of Pharmaceutical Analysis Secondary Sources Custom Manufactures Contract Manufacturers Chemical Suppliers 4
  5. 5. Contd… RSs are generally obtained and prepared from United States of Pharmacopeia-National Formulary (USP-NF) USP RSs collection consists of more than 3,000 items ranging from drug substances, related impurities, residual solvents, biologics, excipients, botanicals, polymers and melting point standards. In India, Central Drug Laboratory is responsible for making available IP RSs. NIST provide a wide variety of standard reference materials (SRMs) for validating and calibrating analytical methods Dept. Of Pharmaceutical Analysis 5
  6. 6. Definition As per USP-NF USP Reference Standards are highly-characterized physical specimens used to ensure the identity, strength, quality, and purity of medicines (drugs, biologics, and excipients), dietary supplements, and food ingredients. Dept. Of Pharmaceutical Analysis Other organizations also involved in establishment of RSs, like Japanese & European pharmacopeias, WHO, Reference Materials Committee of ISO (REMCO), etc. 6
  7. 7. Other Definitions: FDA: Specifically prepared drug substance batch and an authentic material of the highest purity used for structural elucidation and benchmark for working standards ICH (Q6A): A reference standard from new drug substances intended for assays, its impurities should be adequately identified and/or controlled and purity should be measured by a quantitative procedure Dept. Of Pharmaceutical Analysis 7
  8. 8. Uses of Reference Standards (RSs) • Reference standards are used to support measurements concerned with chemical composition, biological, clinical, physical and miscellaneous areas. • Substances characterized for chemical purity and/or trace impurities. • Standard solutions and gas mixtures, often prepared gravimetrically from pure substances and used for calibration of gases. • Physico-chemical reference materials characterized for properties such as melting point, viscosity. • Reference objects or artefacts characterized for functional properties such as taste, odour, octane number, and hardness. Dept. Of Pharmaceutical Analysis 8
  9. 9. Contd… Qualitative analysis of materials by comparison of RS & chemical substance in aspects • IR- Functional Group Conformation • Melting point & HPLC- Purity identification • U.V.- identification, etc, Quantitative analysis • HPLC • Limit tests Specific Uses • Method validation • Calibration of Instruments Dept. Of Pharmaceutical Analysis 9
  10. 10. Qualitative & Quantitative Analysis Includes following aspects, 1) Identification 2) Purity analysis 3) Assay of sample. Results are compared with RS & sample should meet standards of RS. These can be performed by using spectroscopic & chromatographic methods. Dept. Of Pharmaceutical Analysis 10
  11. 11. Calibration Of Instruments When an instrument is giving irrelevant results then it can be calibrated by using RSs. If results are close to acceptance criteria then referred as instrument is working properly. Some examples are given below, Instrument RS Used for Calibration UV-Vis Spectrophotometer Potassium dichromate IR Spectrophotometer Poly Styrene film Karl Fischer Titrimeter Di Sodium Tartrate Flame Photometer Potassium dichromate Gas Chromatography 1.0% v/v Toluene solution in n-Hexane Dept. Of Pharmaceutical Analysis 11
  12. 12. Method validation & Uncertainty of Method  Generally in a method validation estimation of Bias is most critical aspect .  RSs will have limited values of Uncertainty.  Uncertainty associated with RS should be NMT 1/3rd of sample measurement. Dept. Of Pharmaceutical Analysis 12
  13. 13. Classification Analytical Reference Standards (ARS)/ Primary Standard Working / Secondary Standard (WS) Authentic Materials (AM) Dept. Of Pharmaceutical Analysis 13
  14. 14. Analytical Reference Standards (ARS) Definition: A material or substance with well established properties, used for the calibration of apparatus the assessment of a measurement method or for assigning values to materials. USES:  Mainly for Research & Drug development purpose  To characterize WS Dept. Of Pharmaceutical Analysis 14
  15. 15. Working Standard (WS) Definition: A drug substance of established quality and purity as shown by comparison to the reference standards material and used as reference working substance for routine quality control. USES:  Mainly for Laboratory purpose. Dept. Of Pharmaceutical Analysis 15
  16. 16. Comparison Among ARS, WS & AM CATEGORY ARS WS AM Synonym Primary (1˚) / Gold standard Secondary(2˚) standard Authentic Visual Reference Purity NLT 99.5% NLT 95% NLT 80% Quality Highly Characterized Not so as ARS Not so as WS Characterization Acceptable without comparison Requires comparison with 1˚ standard Purpose For industrial purpose in  Drug development process  R&D  Calibration of apparatus, methods and materials  Calibration of 2˚ standard  For Laboratory purpose & in QC As visual images to compare certain test articles to meet requirements Not Used As drug or cosmetic for consumption For research purpose Not for chemical analysis Availability Synthesized independently & available in very less quantity. Procures from Bulk manufacturer or prepared in laboratory itself Available in small quantity Cost Very expensive Low cost Low cost Dept. Of Pharmaceutical Analysis 16
  17. 17. Types of Reference Standards: Dept. Of Pharmaceutical Analysis Reference Solution Compounds which are difficult to handle like very hygroscopic or moisture sensitive, a stable solution of drug substance can be manufactured and filled into tight glass ampule for once use only with a detail of storage condition and shelf life Related Substance include the related impurity, By-products, degradation products and used to prove the system suitability, qualitative or quantitative analysis System Suitability is a mixture of the compound with a small amount of the impurity is required; mixtures of impurities or a mixture of impurities and the compound as reference standard. SST test based on the unadjusted RRT, RT and the resolution. 17
  18. 18. Classification According To ISO & USP Certified Reference Material (CRM) • A reference material whose property values are certified by a technically valid procedure accompanied by or traceable to a certificate or other documentation which is issued by a certifying body. Dept. Of Pharmaceutical Analysis Reference Material (RM) • It is a Working level Reference Material used for the calibration of an apparatus, the assessment of a measurement method, or for assigning values to materials. 18
  19. 19. Reference Standards ARS Preparation: Synthesized independently by the Industry for their use. Preparation of ARS from WS: INCASE OF:  Analytical laboratories facing difficulty to procure ARS from Official sources  For daily analysis CONDITIONS:  Prepared ARS from WS should meet properties of RS. Dept. Of Pharmaceutical Analysis 19
  20. 20. Contd.. Bulk material procured from manufacturer will be characterized & purified (i.e., WS) From above 5-50 grams collected and stored in large glass ampoules Analytical tests will be done according to pharmacopoeial requirements Standardize the content by 3 analysts, if results are close Transfer Standardized WS into amber colored ampoules of capacity 50-100 mg and store properly Dept. Of Pharmaceutical Analysis 20
  21. 21. WS Preparation Bulk material should satisfy following tests • • • • Evidence of chemical structure Assay Purity Accelerated stability testing, etc. Then material can referred as Working standard Dept. Of Pharmaceutical Analysis 21
  22. 22. Characterization Of Reference Standards Qualification of RSs is a critical parameter in which a thorough characterization is necessary in aspects of 1) Identity 2) Strength 3) Quality 4) Purity Dept. Of Pharmaceutical Analysis 22
  23. 23. Basic Requirements For Qualification  Should understand Physico chemical Profile of RS  ARS should have good chemical stability on wide range of storage  Requirements for characterization should be cross-checked to avoid Analytical errors.  The validity (i.e., accuracy, precision, sensitivity, specificity) of the analytical methods used for purity determination should be demonstrated  Highly pure (i.e., ≥99.5%)  In total the degradation and by-products should not be more then 0.5%  The levels of all impurities found (including those not chemically identified, isolated, and characterized) should be provided as a summary tabulation  Content of enantiomer in case of chiral compound is not taken into account as byand degradation products by normalized area percent Dept. Of Pharmaceutical Analysis products 23
  24. 24. • Visual inspection • Optical microscopy Proof of structure Physical Description Steps Involved In Characterization • Elemental analysis • U.V. spectroscopy • I.R. Spectroscopy • Mass Spectroscopy • NMR Spectroscopy • Raman Spectroscopy Dept. Of Pharmaceutical Analysis 24
  25. 25. Contd… Purity • Loss On Drying (LOD) • Karl Fisher Titration • Thermo Gravimetric Analysis (TGA) • Differential Scanning Calorimetry (DSC) • Residue On Ignition (ROI) • Thin Layer Chromatography (TLC) • High Performance Liquid Chromatography (HPLC) • Gas Chromatography (GC) Assay • Titration • Phase - Solubility Analysis Dept. Of Pharmaceutical Analysis 25
  26. 26. Proof Of Structure: Mass spectroscopy Elemental analysis by ICP (inductively-coupled plasma) detects up to 24 metals, measuring less than 5μ in size IR spectroscopy Dept. Of Pharmaceutical Analysis NMR spectroscopy 26
  27. 27. Purity Determination Karl Fischer Titration Determines free content & water of hydration trapped into the crystal • Caution must be taken while doing KF Titration. No external moisture should interrupt. LOD Determines amount of volatile matter • The % of material lost on drying should correlate to the total amount of solvent and moisture obtained by other measurements, when determining mass balance accountability for reference standard material Measures the change in the mass of sample as the temperature is TGA changed • Gives information regarding moisture and solvent levels, an indication if the material is a hydrate or solvate, and the rate of reaction Measures the difference in energy (heat flow) between reference DSC and sample. • Used to accurately measure the melting point and purity of the reference material. Dept. Of Pharmaceutical Analysis 27
  28. 28. Contd… ROI TLC GC HPLC Dept. Of Pharmaceutical Analysis • Gives information on presence of inorganic impurities • Method involves charring, digestion and ignition of RS • ROI can be coupled with spectroscopic methods for identification & quantification of inorganic impurities. • TLC coupled with Densitometer will measure relative amounts of separated components & impurities • Determines presence of Organic Solvents as impurities • Impurities, relative substances detection 28
  29. 29. Assay Of RS Titration • Titration values determines counter-ions & impurities present in RS. • Less selectivity compared to Chromatographic techniques Phase - Solubility Analysis • Absolute method that provides a totally independent assessment of purity and identity of impurities. • Phase-solubility analysis is applicable to all species of compounds that are crystalline solids and that form stable solutions. Dept. Of Pharmaceutical Analysis 29
  30. 30. Working Factor And Drug Content Calculation Contents for Reference standard can be calculated as follow100- (%Loss on drying + %Total related substance + % Residue on ignition*) or 100- %Water + %Residual solvent +% Total related substance + % Residue on ignition*) Working factor for Reference standard can be calculated as follow(100- Volatile correction#+ % Total related substance + % Residues on ignition*)/ 100 Working factor and content are correlated by a factor of 100, e.g. Drug content= 100xWF. If WF is 0.994 then content= 99.4%. But working factor should not be used in the documentation part of Reference Standard. Dept. Of Pharmaceutical Analysis *This is project specific correction and depends again on the information available in the specification #Depends on the product specification, require to consider WC or residual solvents or LOD If Value of WC+Residual solvents or LOD or DP and By products is <0.1% , can be disregarded 30
  31. 31. Stability, Expiry & Retest Expiration and Retest Date: • For any new chemical entity an initial expiry or retest date decided as 6 months if stock material stored at -20⁰C • Availability of stability data for 3 months at 40⁰C/75%RH demonstrates the retest period of 24 months if stored at -20⁰C • Expiry date is periodically extended on the basis of retest performed and stability proved • Reference standard may have a maximum 5 years expiry period Dept. Of Pharmaceutical Analysis 31
  32. 32. Prerequisite for analytical • Essential for bulk drug testing, should be safeguarded by proper storage • No purification required if purity >99.0% for RS and for Impurities >95.0% • Supplied with a CoA including complete identification testing • Stability data indicates the storage conditions • Information of its hygroscopicity and solid-state properties, e.g. amorphous, crystalline, polymorphic form etc.. • A material safety data sheet • A list of potential impurities (if an active substance) with response factors Dept. Of Pharmaceutical Analysis 32
  33. 33. Certificate Of Analysis Information needed for COA: • Name and unique batch number • Manufacturer • Manufacturing date • Retest date • Expiry date • Transport instructions • Storage condition • Content of reference substance • Note on calculation of the contents • Special remark on handling requirement if any • All the required tests with their specifications and results Dept. Of Pharmaceutical Analysis 33
  34. 34. Usage and Maintenance Handling during analysis: Use and maintenance: • Must be equilibrated to room temperature for 30 minutes before use • Protect from light and very tight packaging • For NCEs packaging recommended in amber color bottle with tight sealed cap • Moisture sensitive and hygroscopic reference substance are packed into once use packaging material only and discarded after the use if any remaining quantity available Dept. Of Pharmaceutical Analysis 34
  35. 35. ..HANDLING • Hands should be thoroughly washed before handling. • Transfer should be done with clean stainless steel spatula. The spatula used should be cleaned after use. • While weighing clean spatula has to be used. • Containers should be immediately closed as soon as the sample is taken out. • Containers has to be labelled. Dept. Of Pharmaceutical Analysis 35
  36. 36. STORAGE Qualified Reference standard should be stored in Sealed Ampoules of Single or multiple dose at proper storage conditions. Should be protected from Light Moisture Heat Type of Container Used for Moisture resistant Hygroscopic Drugs Amber colored Light Sensitive Drugs Heat proof Thermo Labile Drugs Dept. Of Pharmaceutical Analysis 36
  37. 37. Records • Only authorized personnel of the laboratory should have an access to reference standards. • Codes should be given to reference standards for easy identification and handling. • A log book should be maintained in order to keep the record of the reference drugs used. • The personnel handling reference standards should take utmost care while maintaining these records. • The log book should have compulsorily have the details like date, name of standard taken, purpose for which it is issued, amount issued, amount remaining, signature. Dept. Of Pharmaceutical Analysis 37
  38. 38. Conclusion RS is an Important Analytical Tool in industry & in QC labs. Result of any analytical procedure or any instrument and drug safety depends on RS. Dept. Of Pharmaceutical Analysis 38
  39. 39. Referen ces Hand Book Of Isolation And Characterization Of Impurities In Pharmaceuticals, By Satinder Ahuja, Elsevier Publications, Volume 5, Pg No: 15, 119-143. alog_LR_04_17_09.pdf 39 Dept. Of Pharmaceutical Analysis
  40. 40. Dept. Of Pharmaceutical Analysis 40