Stability_Indicating_HPLC_Method.ppt

PRESENTATION
BY
P.G.SAINI
SR,VICE PRESIDENT
THEMIS LABORATORIES PVT. LTD
THANE
STABILITY INDICATING METHOD
DEVELOPMENT

Information, objective and
definitions related to Stability
testing
DEVELOPMENT

DEFINITIONS
STABILITY
The ability of a pharmaceutical product to retain its chemical, physical,
microbiological and biopharmaceutical properties within specified limits
throughout its shelf-life.
SHELF-LIFE
The period of time during which a drug product, if stored correctly, is
expected to comply with the specification as determined by stability
studies on a number of batches of the product. The shelf-life is used to
establish the expiry date of each batch.
SHELF-LIFE SPECIFICATION
Shelf-life specification means the requirements to be met throughout
the shelf-life of the drug product (should not be confused with "release
specification").
DEVELOPMENT

TYPE OF STUDY
ACCELERATED STABILITY TESTING
Studies designed to increase the rate of chemical degradation
and physical change of a drug or drug product by using
exaggerated storage conditions as part of the formal stability-
testing programme. The results of accelerated testing studies are
not always predictive of physical changes.
REAL-TIME (LONG-TERM) STABILITY STUDIES
Experiments on the physical, chemical, biological,
biopharmaceutical and microbiological characteristics of a drug,
during and beyond the expected shelf-life and storage periods of
samples under the storage conditions expected in the intended
market. The results are used to establish the shelf-life, to
confirm the projected shelf-life, and to recommend storage
conditions.
DEVELOPMENT

Main objectives of stability testing
OBJECTIVES TYPE OF STUDY USE
To select adequate (from the viewpoint Accelerated Development of the product
of stability) formulations and container
-closure systems
To determine shelf-life and storage Accelerated Development of the
Conditions and real-time Stability product and registration
dossier
To substantiate the claimed shelf-life Real-time Registration dossier
To verify that no changes have been Accelerated Quality assurance in
introduced in the formulation or general, including
process that can adversely affect the stability quality control
manufacturing of the product
DEVELOPMENT

Main CLIMATIC ZONES
Four climatic zones can be distinguished for the purpose of
worldwide stability testing, as follows:
CLIMATIC DESCRIPTION DERIVED STORAGE CONDITIONS
ZONE (FOR REAL TIME STUDIES)
Zone I Temperate 21°C 45%RH
Zone II Subtropical, with 25°C 60%RH
possible high humidity.
Zone III Hot/dry. 30°C 35%RH
Zone IV Hot/humid. 30°C 70%RH
DEVELOPMENT

CONDITIONS FOR REAL TIME STABILITY TESTING OF
RELATIVELY STABLE DRUG SUBSTANCE AND ITS
DRUG PRODUCT FOR ZONE I, II AND III
Storage temperature Relative humidity Duration of studies
(°C) (%) (months)
25 ± 2 60 ± 5 3,6,9,12,18,24,36,48 & 60
AND FOR ZONE IV
Storage temperature Relative humidity Duration of studies
(°C) (%) (months)
30 ± 2 65 ± 5 3,6,9,12,18,24,36,48 & 60
DEVELOPMENT

CONDITIONS FOR ACCELERATED STABILITY
TESTING OF RELATIVELY STABLE DRUG
SUBSTANCE AND ITS DRUG PRODUCT
FOR ALL ZONES AND GLOBAL MARKET
Storage temperature Relative humidity Frequency
of studies
(°C) (%) (months)
40 ± 2 75 ± 5 1,2,3 and 6
DEVELOPMENT

SIGNIFICANT CHANGE
Significant change for a drug product is defined as:
1. A 5% change in assay from its initial value; or failure to meet the
acceptance criteria for potency when using biological or immunological
procedures;
2. Any degradation product’s exceeding its acceptance criterion;
3. Failure to meet the acceptance criteria for appearance, physical
attributes, and functionality test (e.g., color, phase separation, re-
suspendibility, caking, hardness, dose delivery per actuation); however,
some changes in physical attributes (e.g., softening of suppositories,
melting of creams) may be expected under accelerated conditions; and,
as appropriate for the dosage form:
4. Failure to meet the acceptance criterion for pH; or
5. Failure to meet the acceptance criteria for dissolution for 12 dosage
units.
DEVELOPMENT

APPROACH
TO
STABILITY INDICATING
METHOD
DEVELOPMENT

REGULATORY STATUS OF STABILITY INDICATING
ASSAY
ICH GUIDELINE
Q1A
New Drug Substance & Product
Testing by Stability –indicating method
stress testing at 10 C increment above accelerated temp & extremes
pH, Oxidative and Photolytic conditions
Q3B
Impurities in New product emphasizes providing documented
evidence suitable for detection & Quantitation.
DEVELOPMENT

REGULATORY STATUS OF STABILITY
INDICATING ASSAY
ICH GUIDELINE
Q6A
Provides guidance on Specifications and requirement on
Stability –indicating assay under UNIVERSAL TESTS /
CRITERIA for both Drug Substance & Drug Product.
Q5C
The same on stability Testing of Biotechnological /
Biological Products
DEVELOPMENT

REGULATORY STATUS OF STABILITY
INDICATING ASSAY
UNFORTUNATELY none of the ICH GUIDELINE
provide exact definition of
Current ICH guideline on GMP for API (Q7A),
under adoption by WHO also mention that the
test procedures used in stability testing should
be Validated and Stability – indicating.
DEVELOPMENT

REVIEW OF LITERATURE ON STABILITY INDICATING
INDICATING ASSAY
A General Review Published as early as 1971 that gave
general principles and discussed the method developed
till date.
Subsequently Review on Stability – indicating methods
by HPLC assay methods reported till 1996.
A compilation of Stability – indicating methods > 500
for various was published by Xu and Trissel.
Recent pubilcation-Book , ‘Drug Stability: Principles and
Practices’ by Carstensen and Rhodes.
DEVELOPMENT

ASSEMENT AND CURRENT REGUTATORY
REQUIREMENTS
Very Few Methods Claimed to be Stability – indicating
fits in to current definition of Stability – indicating assay
in true sense.
Few Studies are truly stability indicating where
drug has been expopsed to all types of stress
conditions and attempts made to separate the
drug from degradation products and themselves.
Also reports on the formulations subjected to stress
studies and also combination products.
DEVELOPMENT

TECHNIQUES EMPLOYED IN STABILITY INDICATING
ASSAY
TITRIMETRIC AND SPECTROPHOTOMETRIC
Methods employed for drug alone in the matrix of
excipients, additives, degradation products, impurites
etc.
CHROMATOGRAPHY
TLC, HPTLC, GC, HPLC;
Hyphenated Techniques
GC-MS; LC-MS; LC-MS-MS; LC-NMR and CE-MS
Miscellaneous:
NMR and CE (Capillary Electrophoresis)
DEVELOPMENT

DEVELOPMENT OF STABILITY INDICATING ASSAY
Step 1. Detailed study of drug structure to under
stand the possible decomposition routes.
Functional groups:- amides,esters,lactams,lactones etc.
that can undergo hydrolysis.
Thiols, thio-ether etc undergo oxidation.
TITRIMETRIC AND SPECTROPHOTOMETRIC
Methods employed for drug alone in the matrix of
excipients, additives, degradation products, impurities
etc.
DEVELOPMENT

DEVELOPMENT OF STABILITY INDICATING
ASSAY
Step 1. Detailed study of drug structure to under
stand the possible decomposition routes.
Functional groups:- amides, esters,lactams,lactones etc.
that can undergo hydrolysis.
Thiols, thioether etc undergo oxidation and compounds
like olefines, aryl halo-derivatives, aryl acetic acids,
aeromatic nitro gr. N-oxides undergoes
photodecomposition.
Most of the new drugs are congener of existing drugs.
For more information look for Analytical Profile of Drug
substances and monograph provided by Connors et al.
DEVELOPMENT

Step 2. Information on Physicochemical Properties
IMPORTANT PARAMETERS
SOLUBILITY: Aqueous, organic and solvents commonly used in
HPLC
LAMBDA Max: Detection of drug and its degradation products.
ABSORPTIVITY: Wavelength maxima & Extinction.
pKa: pH related changes in RT takeplace at pH value within +/- 1.5
units
LogP: Information on drug and identified degradation products
provides valuable information in the separation behavior on a
column stationary phase.
DEVELOPMENT

ASSAY
Step 3. FORCED DECOMPOSITION STUDIES
(STRESS TESTING)
Q1A. ICH GUIDELINE
I) 10º C increments above Accelerated Temp.
II) Humidity 75% or Greater
III) Hyhrolysis across a wide range of pH value.
IV) Oxidation.
V) Photolysis.
DEVELOPMENT

ASSAY
(STRESS TESTING)
I) 10º C increments above Accelerated Temp.
Stability study conditions.
II) Humidity 75% or Greater
Stability study conditions.
DEVELOPMENT

(STRESS TESTING)
III) Hyhrolysis across a wide range of pH value.
Reflux the drug in 0.1N HCL/NaOH for 8 hrs. if no degradation
then reflux in higher strength for longer time and vies-versa.
IV) Oxidation.
Concentration in the range of 3-30% of H2O2
DEVELOPMENT

ASSAY
(STRESS TESTING)
V) Photolysis.
Exposure to combination cool white and Ultera-Violet
fluorescent lamp or Xenon and Metal Halide lamps.
Exposure energy min 1.2 million lux h fluorescent light
and 200
W h/m2 UV. If no degradation increase intensity by 5
times.
DEVELOPMENT

FORCED DECOMPOSITION STUDIES
(STRESS TESTING)
Min 4 samples for Stress Studies for every stress conditions for
conc of 1mg/ml.
Blank Solution at normal condition
Blank solution subjected to stress condition
Zero time samples containing drug stored under normal
conditions
Drug Solution subjected to stress conations
DEVELOPMENT

Step 4. PRELIMINARY STUDIES ON STRESSED SAMPLES.
The first and the simplest choice is Reversed phase
Octadecyl column.(New).
Mobile phase Water/methanol or Water/Acetonitrile. (50:50)
Modifier, previous knowledge on the method to obtain Capacity
Factor 5-10 for drug.
Selection of the Wavelength based on the spectral behaviour,
Injection volume and flow rate suitably adjusted.
Drug and its products can have different extinction values.
If non chromatographic products or low molecular weight
fraction use LC-MS.
DEVELOPMENT

Step 5. FINAL METHOD DEVELOPMENT AND OPTIMASATION
Tabulate the RRT of ALL product formed for each condition.
Components with Close peak’s (RT or RRT) acertain with PDA
or LC-Ms profile.
Mixture of all the Reaction Solution based on the data
indicating different products formed instress study.
Care taken while mixing different ph solutions.
Co-eluting peaks separated and optimised by changing the
Mobile phase ratio, pH, Gradient, Flow Rate, Temperature,
Solvent Type, and the Column and its Type.
Components
DEVELOPMENT

Step 6. IDENTIFICATION AND
CHARACTERATION OF DEGRADATION
PRODUCTS
Identify the Drug Degradation Products .
Arrange for their Standards.
Establish Specificity / Selectivity of the method.
(For Standard product direct procurement)
Known impurities confirmed by Spiking.
DEVELOPMENT

Step 7. VALIDATION OF DEVELOPMENT METHOD
FIRST STEP: In General
ICH Guidelines- Q2A and Q2B, FDA guideline and USP
Establishment of Specificity / Selectivity based on the
knowledge drug degradation carried while Forced
Degradation.
1. Specificity
2. Linearity:- 0-100 %
3. Range 80 – 120 % ; (0 - 20 % for degradation Product)
4. Precision
5. LOD
6. LOQ
7. Robustness
DEVELOPMENT

DEVELOPMENT

Step 7. VALIDATION OF DEVELOPMENT METHOD
SECCOND STEP
The Method Extended to FORMULATIONS:
Emphasis on limited to Prove the pertinence of Established
Validation parameters in the presence of Excipients or other
constituents of Formulation.
Specially the most important parameters are:
1. Specificity
2. Accuracy
3. Precision
4. Robustness
5. Linearity:- 80 to 120 % of Assay Conc.
(50-120% Injection & Unstable drug.)
6. Range
DEVELOPMENT

VALIDATION OF DEVELOPMENT METHOD
CRITICAL ISSUES
SPECIFIC AND SELECTIVE STABILITY INDICATING
METHODS
SPECIFIC STABILITY INDICATING METHOD: A method
that is able to measure the drug substances in the presence all
degradation products, excipients and additives expected to be
present in the formulation.
(Titrimetric, UV)
SELECTIVE STABILITY INDICATING METHODS
A method that can measure the drug(s) and all degradation
products in the presence of excipients and additives expected to
be present in the formulation.
(Chromatographic)
DEVELOPMENT

CRITICAL ISSUES
Is It necessary to follow the Stress testing Route to Develop ?
Well answered in the ICH Guidelines.
“However it may not be necessary to examine specially for
certain degradation products if it has been demonstrateed
that they are not formed under accelerated or long term
storage conditions.”
The so developed method Stress testing can be applied to broad range
of situations and extended from Drug substance to Formulation
or one Formulation to another.
DEVELOPMENT

CRITICAL ISSUES
Can Formulation instead of Drug substance be subjected
to Stress Testing for STABILITY INDICATING METHOD.
ICH Guidelines Q1AR and the ICH’s Common Technical Document
suggest Stress Testing only of drug substance.
For Drug Product Stress Testing is provided in Q1AR, read as
“Studies under taken to assess the effect of sever conditions on the drug
product, such studies include Photo stability Testing and specific testing
on certain products (e.g. metered dose inhalers, Creams, emulsions,
refrigerated aqueous liquid products).
Carry out Stress Studies on Drug Formulations for Formulators is
rational to do so.
DEVELOPMENT

CRITICAL ISSUES
Mass Balance in Development of Stability Indicating
Method.
There are situtation where Mass Balance may be difficult to be
Established.
For the following reasons:
Formation of multiple degradation products involving complex
reaction pathways and drugs excipient interaction products.
 In complete detection due to loss of UV chromophore or lack of
universal detection
 Loss of drug / degradation products as volatiles.
 Diffusive loss into or through containers
 Elution / resolution problems
 Inappropriate or unknown response factors due to lack of standards
 Errors and variabiltiy in the drugs contains assay
DEVELOPMENT

CRITICAL ISSUES
Mass Balance in EMERGING TECHNIQUES
FOR ANALYSIS OF STABILITY SAMPLES
AN INCREASING TREND IN HYPENATED
TECHQUES
GC-MS,LC MS OR LC-MS-MS, CE-MS, LC-NMR ETC
LIMITATION HIGH COST.
DEVELOPMENT

STABILITY INDICATING METHOD DEVELOPMENT
THANK YOU
Prescription Pharma Support
Themis Laboratories Private Limited
THANE
R&D Driven Novel Drug Delivery Company
Reference- Review by Monika & Dr. Saranjit Singh
NIPER

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