REFERENCE STANDARD Smt R D Gardi B pharmacy

REFERENCE STANDARD Smt. R. D. Gardi B. pharmacy college M. Pharm (sem-I) Quality Assurance Prepared By: paras shah Guided By: Mr. Ketan Dadhania

DEFINATION Reference substances are authentic specimens that have been approved by the designated agency of the country as suitable for use as comparison standards in the test and assays specified in the respective official pharmacopoeia of the country.

For your knowledge Some of these reference standards have been identified as: - Ph. Eur: CRS (Chemical Reference Standards) - BP: BPCRS (British Pharmacopoeia) - Intern. Pharm. : WHO international CRS - USP: USP reference Standard At USP, sights are set on the milestone of reaching 3000 RS, to meet by the end of 2012.

Types Primary Reference Standard of active compound: Secondary Standard (Working Standard /In-House reference material) Related Compound Standard Compendial Standard Internal standard Other Standards: Former USP and NF Reference Standards FCC Reference Standards Authentic Substances (AS)

Sources of reference-standard materials Mainly two types of sources 1. Compendial (primary) 2. Noncompendial (secondary

Preparation The primary reference standard of the Current lot of drug substances procured from the corresponding Pharmacopoeial commission. Primary standards used for Standardization, calibration, system suitability etc. are procured from the approved vendors. Their purity is considered as per the label claim by the vendors. The working reference standard of drug substances are prepared by taking the released commercial batch of the drug substance, which is further purified in Research and Development Department and analyzed by in-house Quality control Department against the primary reference standard of the drug substance. The reference standards of non pharmacopoeial product are prepared by taking the highly pure chemical substances synthesized by in house Research and Development department which is identified, fully characterized analyzed before making use for routine analysis.

Preparation The impurity standards are procured from the concerned pharmacopoeial commission and are used for the routine analysis. If official standards for impurities are not available, in house R&D department prepares the impurity standards. They re fully identified and characterized before use for routine analysis. Standards for the solvents are prepared by taking the previously approved lots of the solvents and record thereof is maintained.

Characterization characterization requirement measurement technique 1. Physical properties NMR, MS, FTIR, UV-visible, functional group analysis, crystallinity Proof of structure Physical description Appearance melting point, pka, optical rotation X-ray diffraction Physical properties

Characterization 2. purity Related compound Chiral purity Inorganic impurities Solvents HPLC purity HPLC, ZE chiral purity Residue on ignition, ICP , AAS ICP, GS Water Karl Fischer titration Ion chromatography Counter ions Verification organic impurities TLC 3. Assay Verification Titration, Assay by difference, C, H, N analysis Mass balance calculation

Usage of reference standards: The following are the areas wherein the reference substances are used: A. infrared Spectrophotometry. B. ultraviolet absorption spectrophotometry. C. quantitative methods based on the development of a color and the measurement of its intensity, whether by instrumental or visual comparison. D. chromatographic separation. E. quantitative methods (including automated methods) based on other separative techniques that depend upon partition of the material to be determined between solvent phases, where the precise efficiency of the extraction procedure might depend upon ambient conditions. F. assay methods based on measurement of optical rotations.

Handling and Distribution of Reference Substances

Packaging and Storage ØSuitable packaging: Light protection: brown glass Humidity protection: tight closing containers Inacceptable is parafilm One dose Standards (HPLCvials) O 2 protection: N 2 Atmosphere, tight closing container ØSuitable container material Residual solvent: glass ØSuitable place Thermolability: fridge, deep freezer

Handling Adapt to room temperature prior use Planned operations enable the technicians to ensure short periods for opening and handling the standards Hygroscopic substances Photo lability Some standards have to be dried prior use - possible time consuming error source Contaminations during use One standard - one spattle - one weighing glass

Documentation Information to be supplied with Reference Substances Labels on chemical reference substances should give the following information: a. name of the substance; b. type of reference substance (e. g. International Chemical Reference Substance, or National Chemical Reference Substances, or Authentic Specimen); c. name and address of the issuing authority; d. approximate quantity of material in the container; and e. batch or control number

The following information should be given, as necessary, either on the labels or in associated documents: i. recommended storage conditions ii. Intended use of the reference substances iii. Directions for use iv. Information about the composition of the reference substances v. a disclaimer of responsibility when reference substances are misused, or stored under inappropriate conditions, or used for other purpose than those intended by the Issuing Authority. Reference Materials Calibrated Against International Chemical Reference Substances: It is advisable to establish the reference substances that have been co-related with International Reference Substance.

Establishment of expiry dates: Unopened CRS vials are fit for use as single use only Expiry dates for in-house standards Based on stability data of the API Taking into account the following: Hygroscopic properties Photo lability Thermo lability Based on scientific literature Based on supplier information Based on own experience – analytical data, stability program

Certificate Of Analysis PREDNISONE CERTIFIED REFERENCE MATERIAL LOT #: P 500042 CATALOG #: PHR-1042 NOMINAL WT: 1 g CERTIFICATE VALIDITY DATE: 9 September 2009 VERSION: 500042. 3 Note: Certificates may be updated due to Compendial Lot changes or the availability of new data. Check our website at: www. RTPharma. com for the most current version. EXPIRATION: 12 Months from Receipt (Proper Storage and Handling Required). STORAGE: Store at Room Temperature, keep container tightly closed. Attachment of a 20 mm aluminum crimp seal recommended for unused portions. CHEMICAL FORMULA: C 21 H 26 O 5 MW: 358. 4 PHYSICAL DESCRIPTION: White powder in amber vial CAS #: 53 -03 -2 HAZARDS: Read MSDS before using. Always handle as if potentially hazardous using prudent laboratory techniques. INSTRUCTIONS FOR USE: Do not dry. The internal pressure of the container may be slightly different from the atmospheric pressure at the user’s location. Open slowly and carefully to avoid dispersion of the material.

Certificate Of Analysis In general all standards should have a certificate of analysis generated either from a qualified supplier or through analysis of in house data. A typical certificate of analysis of working reference standard should bear the following information: Name of product: WRS No: Date of preparation: Calibrated Against: Quantity: Control No: Use before: Reference Standard Lot No:

Reference Standard Control Strategy

Ø Ø Regulations Governing RS Operations Food and Drug Administration (FDA) – CFR Title 21 – Food and Drugs GMP, 211. 194(c), 211. 160(b) (1) and 299. 5(c) – ICH Guidelines Q 7, Q 6 B – Various FDA Guidance Documents – FDA regulatory observations – United States Pharmacopeia European Agency for the Evaluation of Medicinal Products (EMEA) – EU GMP 32, Annex 18 (transcription of ICH Q 7) – EU Quality Guideline 32 (regulatory submission requirements) – ICH Guidelines Q 7, Q 6 B European Pharmacopoeia Japan Ministry of Health Labor and Welfare (MHLW) – Japan Pharmacopoeia Technical Information (JPTI) 1995, section 2 – ICH Guidelines Q 7, Q 6 B – Japan Pharmacopoeia

Reference Standards for Biologics Global measurement standards Global written standards More than 250 WHO measurement Standards are available; define the IU of biological activity Support scientific evaluation of emerging issues

Underlying principle A “biological” analyte is considered by WHO as one “…of biological origin, which cannot be characterized adequately by chemical and/or physical means alone…” This is a practical definition, relating the structural complexity of the material being standardized to the current utility of analytical methods. Approaches to the calibration and value assignment of global biological reference standards Global biological reference materials are established via one of two approaches: 1 Those which support a measured quantity, and are assigned a value in SI units (eg mg, mol) 2 Those which are used to quantify an effect (e. g. biological activity) and are assigned a value in arbitrary units (e. g. WHO International Units)

Approaches for value-assignment can change with time – example of growth hormone Somatropin (recombinant growth hormone) Growth hormone has undergone the transition from a biological to a nonbiological Assay method 1985 191 amino-acid 22 k. D Non- glycosylated Biosynthetic Weight gain or tibial width increase In hypophysectomised rats 2005 Size exclusion -HPLC

Reference Standards for Biologics Reference Standard must be representative of product including manufacturing pathway. Storage stability is critical. Protocol with more extensive testing to ensure quality that is not dependent on quality of a previous reference standard. Identity and purity often requires several different assays (ELISA, Western Blot, PCR, bioassay, etc. ). Actual structure may not be confirmed, but may need to be inferred from biological activity. Bioassays need to be expressed in units of activity relative to a reference standard using e. g. WHO international units where applicable Warning Letters: RS Extracts.

Examples of reference standard USP Warfarin RS - This is the acid form of Warfarin. Dry portion in vaccum over phosphorus pentoxide for 4 hour before using. Keep container tightly closed. Protect from light. USP Urea RS - Do not dry. USP Salicylic acid tablet RS (Dissolution calibrator, Nondisintegrating) - The label states the nominal weight of each tablet. Use only whole tablet-extra tablets are provided. Remove any surface dust with a soft brush before using. Keep container tightly closed. USP Ibuprofen RS - Do not dry. Keep container tightly closed. USP Anhydrous lactose RS- Dry portion at 80 0 C. For 2 hour before using. Keep container tightly closed.

REFERENCES USP/NF 2007 www. who. int/biologicals http: //bit. ly/QOS-Expl www. pharmatech. com http: //www. fda. gov/cder/guidance/2396 dft. htm.

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