REFERENCE STANDARD

1
REFERENCE STANDARD

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

2
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.

3
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.

4
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)

5
Sources of reference-standard materials

• Mainly two types of sources
• 1.Compendial (primary)
• 2.Noncompendial (secondary

6
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 and analyzed
  before making use for routine analysis.
•  

7
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 RD
  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.
•  

8
Characterization
characterization requirement measurement technique
1. Physical properties
Proof of structure NMR,MS,FTIR,UV-visible, functional group analysis, crystallinity
Physical description Appearance
Physical properties melting point, pka, optical rotation X-ray diffraction
9
Characterization
2.purity
Related compound HPLC purity
Chiral purity HPLC,ZE chiral purity
Inorganic impurities Residue on ignition, ICP ,AAS
Solvents ICP, GS
Water Karl Fischer titration
Counter ions Ion chromatography
Verification organic impurities TLC
3.Assay
Verification Titration, Assay by difference, C,H,N analysis Mass balance calculation
10
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.

11
Handling and Distribution of Reference Substances
12
Packaging and Storage

• Suitable packaging
• Light protection brown glass
• Humidity protection tight closing containers
  Inacceptable is parafilm One dose Standards
  (HPLC-vials)
• O2 protection N2 Atmosphere, tight closing
  container
•  
• Suitable container material
• Residual solvent glass
•  
• Suitable place
• Thermolability fridge, deep freezer
•  

13
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

14
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

15

• 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.

16

• 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

17
Certificate Of Analysis
PREDNISONE CERTIFIED REFERENCE MATERIAL LOT
P500042 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 C21H26O5
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 users location. Open slowly and
carefully to avoid dispersion of the material.
18
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
  Control No
• Date of preparation Use
  before
• Calibrated Against
  Reference Standard Lot No
• Quantity
•  

19
Reference Standard Control Strategy
20
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 Q7, Q6B
• 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 Q7)
• EU Quality Guideline 32 (regulatory submission
  requirements)
• ICH Guidelines Q7, Q6B
• European Pharmacopoeia
• Japan Ministry of Health Labor and Welfare (MHLW)
• Japan Pharmacopoeia Technical Information
  (JPTI) 1995, section 2
• ICH Guidelines Q7, Q6B
• Japan Pharmacopoeia
•  

21
Reference Standards for Biologics
Support scientific evaluation of emerging issues
Global measurement standards
Global written standards
More than 250 WHO measurement Standards are
available define the IU of biological activity
22
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)

23
Approaches for value-assignment can changewith
time example of growth hormone
Somatropin
(recombinant growth hormone)
Somatropin (recombinant growth hormone)
Growth hormone has undergone the transition from
a biological to a nonbiological
Assay method
1985
2005
191 amino-acid 22kD Non- glycosylated Biosynthetic
Weight gain or tibial width increase In
hypophysectomised rats
Size exclusion -HPLC
24
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. 

25
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, Non-disintegrating) - 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 0C.
  For 2 hour before using. Keep container tightly
  closed.
•  

26
REFERENCES

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

27
Thank you for your attention