method validation for drug substances and drug product _remodified_2014
DESCRIPTION
Method validation is the process of proving that an analytical method is acceptable for its intended purposes. METHOD VALIDATION = ERROR ASSESSMENT Method validation is the process of demonstrating that analytical procedures are suitable for their intended use and that they support the identity, strength, quality, purity and potency of the drug substances and drug products Validation: Prior ConsiderationsSuitability of Instrument Status of Qualification and Calibration Suitability of Materials Status of Reference Standards, Reagents, Placebo Lots Suitability of Analyst Status of Training and Qualification Records Suitability of Documentation Written and approved standard test procedure and proper approved protocol with pre-established acceptance criteria Compendial vs. Non-compendial Methods Compendial methods-Verification Regulatory analytical procedure in USP/NF Non- compendial methods-Validation Alternative analytical procedure proposed by the applicant for use instead of the regulatory analytical procedure Chromatographic Methods Demonstrate Resolution Impurities/Degradants Available Spike with impurities/degradants Show resolution and a lack of interference Impurities/Degradants Not Available Stress SamplesFor assay, Stressed and Unstressed Samples should be compared. Ability of an analytical method to measure the analyte free from interference due to other components. Selectivity describes the ability of an analytical method to differentiate various substances in a sample Original term used in USP Also Preferred by IUPAC and AOAC Also used to characterize chromatographic columns Degree of Bias (Used in USP) The difference in assay results between the two groups the sample containing added impurities, degradation products, related chemical compounds, placebo ingredients Selectivity: For impurity test, impurity profiles should be compared. Temperature (50-60℃) Humidity (70-80%) Acid Hydrolysis (0.1 N HCl) Base Hydrolysis (0.1 N NaOH) Oxidation (3-30%) Light (UV/Vis/Fl) Intent is to create 10 to 30 % Degradation Change in the analytical procedure, drug substance, drug product, the changes, may necessitate revalidation of the analytical procedures. “The degree of revalidation depends on the nature of the change.” “FDA intends to provide guidance in the future on post-approval changes in analytical procedures.” By Visual Inspection of plot of signals vs. analyte concentration By Appropriate statistical methods Linear Regression (y = mx + b) Correlation Coefficient, y-intercept (b), slope (m) Acceptance criteria: Linear regression r2 > 0.999 Requires a minimum of 6 concentration levels Normally derived from Linearity studies. Established by confirming that the method provides acceptable degree of linearity, accuracy, and precision. Specific range dependent upon intended application of the procedure.TRANSCRIPT
Validation of Validation of Analytical Method Analytical Method
for Drug Substances for Drug Substances & Drug Products& Drug Products
Dr. R.Badmanaban., M.Pharm., M.D(A.M).,PhD., Associate Professor, Head of Dept - Pharmacognosy Shri Sarvajanik Pharmacy college - Mehsana-384001
www.sspcmsn.org Email: [email protected]
Definition:
Method validation is the process of proving that ananalytical method is acceptable for its intended purposes.
METHOD VALIDATION = ERROR ASSESSMENT Method validation is the process of demonstrating that analytical
procedures are suitable for their intended use and that they support the
identity, strength, quality, purity and potency of the
drug substances and drug products
Analytical Method Validation
Body Full name Guidance on
Eurachem Focus for Analytical Chemistry in Europe Method validation
CITAC Cooperation of International Traceability in Analytical Chemistry
Proficiency testingQuality Assurance
EA European Cooperation for Accreditation Accreditation
CEN European Committee for Normalization Standardization
IUPAC International Union of Pure & Applied Chem. Method validation
ISO International Standardization Organisation Standardisation
AOAC
ILAC
Association of Official Analytical Chemists
International Laboratory Accreditation Cooperat.
Internal qual. ControlProficiency testingAccreditation
FDA US Food and Drug Administration Method validation
USP United States Pharmacopoeia Method validation
ICH International Conference on Harmonization Method validation
International regulatory bodies and their guidelines on different aspects of QA
Chromatography todayChromatography todayMore than sixty variants of the technique have been developed.HPLC, GC, SFC, and CE are the most frequently used.
HPLC: almost universal wide range of equipment and columns is commercially available well-understood separation mechanisms sensitive, specific, selective, precise and robust, Rugged, accurate. easy to maintain instrumentation flexible in optimizing separations More efficient than some of the separation techniques
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Liquid Chromatography (HPLC) instrumentLiquid Chromatography (HPLC) instrument
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The Chromatographic The Chromatographic Process – Theoretical Process – Theoretical ConsiderationsConsiderations
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Elution in Column ChromatographyElution in Column Chromatography
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MobileMobilephasephase
StationaryStationaryphasephase
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Intermolecular InteractionsIntermolecular Interactions
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ChromatogramChromatogram
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Retention RelationshipsRetention Relationships
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Validation (4M)
ManMachineMaterialMethod
Why Method Validation is Important
?Analytical Method
Validation
Why Method Validation is Important?
1. Develops confidence in using the method & Proof that
Method is suitable for its intended purpose,
The purpose of analytical measurement is to get consistent,
reliable and accurate data.
Incorrect measurement results can lead to tremendous
costs.
2.2. Regulatory requirementRegulatory requirement, Equal importance for those working
in a regulated and in an accredited environment.U.S. FDA, ISO etc.
When to be validated
?
Analytical Method Validation
When to be validated?Partial validation after development of method.Complete validation after manufacturing formula is finalized.
Which methods are to be validated ?Compendia: Pharmacopoeia method Verification of suitability of methodNon compendia methods: Laboratory developed methods.
Pharmacopoeias methods used outside its scope.
Validation: Prior Considerations
Suitability of Instrument Status of Qualification and Calibration
Suitability of Materials Status of Reference Standards, Reagents, Placebo Lots
Suitability of Analyst Status of Training and Qualification
Records Suitability of Documentation
Written and approved standard test procedure and proper approved protocol with pre-established acceptance criteria
Validation Activity Including the Complete Analytical
Procedure
Sampling
Sample Preparation
Analysis
Data Evaluation Reporting
Validation Step
Define the application, purpose and scope of
the method.
Analytes? Concentration?
Develop a analytical method.
Develop a validation protocol.
Qualification of instrument.
Qualify/train operator
Qualification of material.
Perform pre-validation experiments.
Adjust method parameters and/or acceptance criteria if necessary.
Perform full validation experiments.
Develop Procedures for executing the method in routine analysis.
Document validation experiments and results in the validation report.
Validation Step
Verification vs. Validation
Compendial vs. Non-compendial Methods
Compendial methods-Verification
Regulatory analytical procedure in USP/NF
Non- compendial methods-Validation
Alternative analytical procedure proposed by the
applicant for use instead of the regulatory analytical
procedure
MethodValidation
MethodTransfer
MethodDevelopment
Approved
BACKGROUND-LAB METHOD FLOW
ICH/USP Validation Requirements
Precision
Repeatability
Intermediate Precision
Reproducibility
Limit of Detection
Limit of Quantitation
Robustness
Specificity
System
suitability
Linearity
Range
Accuracy
Validation Parameters
ImpuritiesSpecificityLinearity and RangeAccuracy PrecisionRobustnessLOD & LOQs
Dissolution
SpecificityLinearity and RangeAccuracy PrecisionRobustness
Assay / CU
SpecificityLinearity and RangeAccuracy PrecisionRobustness
Specificity/Selectivity
Ability of an analytical method to measure the analyte free from interference due to other components.
SelectivityBias
Specificity: ICH/USPThe ability to measure accurately and specifically the analyte in the presence of components that may be expected to be present in the matrixThe degree of interference
Active IngredientsExcipientsImpurities (synthetic precursors, enantiomers)Degradation ProductsPlacebo Ingredients
Analytical Method Validation
Continue....
. Combination of 2 or more analytical procedures may be required to achieve necessary level of discrimination
. Stability indicating analytical methods should always be specific.
. Analysts should ascertain whether the peaks within a sample chromatogram are pure or consist of more than one compound. Therefore should know how many compounds are in the sample or use procedures to detect peak purity
Analytical Method Validation
Specificity: Impurities Assay
Chromatographic MethodsDemonstrate Resolution
Impurities/Degradants Available Spike with impurities/degradantsShow resolution and a lack of interference
Impurities/Degradants Not AvailableStress SamplesFor assay, Stressed and Unstressed Samples should be compared.
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SelectivityAbility of an analytical method to measure the analyte free from interference due to other components.
Selectivity describes the ability of an analytical method to differentiate various substances in a sample
Original term used in USP Also Preferred by IUPAC and AOACAlso used to characterize chromatographic columns
Degree of Bias (Used in USP)The difference in assay results between the two groups
- the sample containing added impurities, degradation products, related chemical compounds, placebo ingredients
Selectivity: For impurity test, impurity profiles should be compared.
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Analytical Method Validation
Forced Degradation Studies
Temperature (50-60℃) Humidity (70-80%) Acid Hydrolysis (0.1 N HCl) Base Hydrolysis (0.1 N NaOH) Oxidation (3-30%) Light (UV/Vis/Fl)
Intent is to create 10 to 30 % Degradation
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Analytical Method Validation
Linearity
Ability of an assay to elicit a direct and proportional response to changes in analyte concentration.
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Linearity Should be Evaluated
By Visual Inspection of plot of signals vs. analyte concentrationBy Appropriate statistical methods
Linear Regression (y = mx + b)Correlation Coefficient, y-intercept (b), slope (m)
Acceptance criteria: Linear regression r2 > 0.999
Requires a minimum of 6 concentration levels
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RangeThe interval between the upper and lower concentrations of analyte in the sample that have been demonstrate to have a suitable level of precision, accuracy, and linearity.
Range
Normally derived from Linearity studies.
Established by confirming that the method provides acceptable degree of linearity, accuracy, and precision.
Specific range dependent upon intended application of the procedure.
Range
Acceptable range having linearity, accuracy, precision.For Drug Substance & Drug product Assay
80 to 120% of test Concentration
For Content Uniformity Assay70 to 130% of test Concentration
For Dissolution Test Method+/- 20% over entire Specification Range
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Accuracy
Closeness of the test results obtained by the method to the true value.
Accuracy
Should be established across specified range of analytical procedure.Should be assessed using a minimum of 3 concentration levels, each in triplicate (total of 9 determinations)Should be reported as:
Percent recovery of known amount added or
The difference between the mean assay result and the accepted value
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Accuracy Data Set (1 of 3)
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Precision
The closeness of agreement
(degree of scatter) between a
series of measurements obtained
from multiple samplings of the
same homogeneous sample.
Should be investigated using
homogeneous, authentic
samples.
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Accuracy Vs Precision
Inaccurate &imprecise
Inaccurate butprecise
Accurate butimprecise
Precision… Considered at 3 Levels
Repeatability
Intermediate
Precision
Reproducibility
Repeatabilit
y
Express the precision under the same operating conditions over a short interval of time.
Also referred to as Intra-assay precision
Intermediate Precision
Express within-laboratory
variations.
Expressed in terms of
standard deviation, relative
standard deviation
(coefficient of variation) and
confidence interval.
Known as part of
Ruggedness in USP
(Different Analysts, Different
Laboratories, Different
Instruments, Different
Reagents, Different Days)
Depends on the
circumstances under
which the procedure
is intended to be
used.
Repeatability & Intermediate Precision
Day 1 Day 2100.6 99.5100.8 99.9100.1 98.9100.3 99.2100.5 99.7100.4 99.6
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GrandMean = 100.0RSD = 0.59%
Mean = 100.5RSD = 0.24%
Mean = 99.5RSD = 0.36%
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Definition: Ability to reproduce data within the predefined precision
Determination: SD, RSD and confidence interval
Repeatability test at two different labs.
Reproducibility
Reproducibility Study
Lab 1 Lab 2 Lab 3
Day 1 Day 2 Day 1 Day 2 Day 1 Day 2
Analyst1
Analyst2
Analyst1
Analyst 2
Analyst 1
Analyst 2
3 Preps
3 Preps 3 Preps 3 Preps
3 Preps 3 Preps
Lowest amount of analyte in
a sample that can be
detected but not necessarily
quantitated.
Estimated by Signal to Noise
Ratio of 3:1.
Detection Limit (DL)
Lowest amount of
analyte in a sample that
can be quantified with
suitable accuracy and
precision.
Estimated by Signal to
Noise Ratio of 10:1.
Quantitation Limit (QL)
LOD, LOQ and SNR
Limit of Quantitation (LOQ)Limit of Detection (LOD)Signal to Noise Ratio (SNR)
noise
Peak ALOD
Peak BLOQ
Baseline
S = slope of calibration curves = standard deviation of blank readings or
standard deviation of regression line
Validated by assaying samples at DL or QL
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DL DL ==
3.3s3.3sQL =QL =
10s10s
SS SS
LOD and LOQ Estimated by
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Ybl
LOD LOQ
Statistical estimate of LOD & LOQ
LOD = 3.3 Sbl / b LOQ = 10 Sbl / b
Y = b X + a
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Analytical Method Validation
Definition: Capacity to remain unaffected by small & deliberate variations in method parameters
Determination: Comparison results under differing conditions with precision under normal conditions
Variations may include: stability of analytical solution, variation of pH in a mobile phase, different column (lot/supplier), temperature, flow rate.
Robustnes
s
Robustness Variations
All Assays
HPLC Assays
GC Assays
-Sample Prep Manipulation-Extraction Time-Mobile Phase Composition-Different Columns-Temperature-Flow Rate-Different Columns-Temperature-Flow Rate
RuggednessDegree of reproducibility of test results under a variety of conditions
Different Laboratories
Different Analysts
Different InstrumentsDifferent Reagents
Different Days
Etc.
Expressed as %RSD
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Solutes may readily decompose prior to chromatographicinvestigations e.g. during sample preparation, extraction,cleanup, phase transfer or storage of prepared vials(refrigerators or automatic sampler).Method development should investigate the stability of theAnalytes AND standards.
Stability of analytical solution
Solution stability •Stability of the samples being analyzed in a sample
solution. e.g. 1 – 48 hours using a single solution.•should be determined by replicate analysis of the
samplesolution.
The checking of a system, before or during analysis of unknowns, to ensure system performance.
“No sample analysis is acceptable unless the requirements for system suitability have been met.” (USP Chapter 621)
Plate Count, Tailing, ResolutionDetermination of reproducibility (%RSD)
For %RSD < 2.0%, Five replicatesSystem Suitability "Sample“ - A mixture of main components andexpected by-products utilized to determine system suitability“Whenever There is a Significant change in Equipment or ReagentsSystem Suitability Testing Should be Performed” (USP Chapter 621)
SYSTEM SUITABILITY
Confuse of Precision Terms
Repeatabili
ty
Intermediat
e Precision
Reproducibili
ty
Ruggedness
Robustness
Precision Terms
Instrument Precision
Repeatability
Intermediate Precision
Reproducibility
Ruggedness
Robustness
- 6 Standard Injections
- One Analysis (6 preps)
- Two Analyses
- Two different Lab.
- Many Variables
- Intentional Changes
Change in the analytical procedure, drug substance, drug product, the changes, may necessitate revalidation of the analytical procedures.“The degree of revalidation depends on the nature of the change.”“FDA intends to provide guidance in the future on post-approval changes in analytical procedures.”Revalidation should accompanyformulation changes (new samples with new compounds or new matrices)manufacturing batch changesnew analysts with different skills,new instruments with different characteristics,new location with different environmental conditions,new chemicals and/or reference standards andmodification of analytical parameters.
Revalidation
Validation ReportObjective and scope of the method (applicability, type).Summary of methodology.
Type of compounds and matrix.All chemicals, reagents, reference standards, QC samples with purity, grade, their source or detailed instructions on their preparation. Procedures for quality checks of standards and chemicals used.Method parameters.Critical parameters taken from robustness testing.Listing of equipment and its functional and performance requirements, e.g., cell dimensions, baseline noise and column temperature range.Detailed conditions on conduct of experiments, including sample preparation Statistical procedures and representative calculations. Procedures for QC in routine analyses, e.g., system suitability tests. Representative plots, e.g., chromatograms, spectra and calibration curves. Method acceptance limit performance data and expected uncertainty of measurement results.Criteria for revalidation.The person's) who developed and validated the method.References (if any).
Analytical Method Validation
How do we Know the expectations of the FDA?
FDA Form 483FDA Warning LettersPersonal Experiences
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483 Observations
There was inadequate method validation specificity data to demonstrate that each method was capable of distinguishing the active ingredient from its impurities and degradation products.Specificity studies did not include the minimum stress conditions of acid and base hydrolysis, oxidation, thermal degradation and photolysis, degradation schematic for the active ingredient that identifies the major degradation products was not included for each product.
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FDA Warning Letter
On addition to an example of modifying both compendia methods and customer supplied methods, we also observed the use of invalidated in-house methods
A statement indicating that the method has not been validated in the particular formulation was included in the certificate of analysis for…use of this statement does not absolve…from using valid, accurate, and
reproducible methods. (June 2009)67
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General requirementsQualified and calibrated instrumentsDocumented methodsReliable reference standards Qualified analysts Sample integrityChange control (e.g., synthesis,)
Analytical methods should be used within GMP and GLP environments, and must be developed using the protocols and acceptance criteria set out in the ICH guidelines Q2 (R1)
Analytical Method Validation
Related Site
www.fda.govwww.fda.gov/cder/www.waters.comwww.usp.orgwww.ich.orgwww.aoac.orgwww.pharmweb.net
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Thank you