method validation in the drug development process-김현성
TRANSCRIPT
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Analytical Method Validation in The Drug
Development Process
Kim HyunSung Ph.D.Berna Biotech Korea
QC Chemistry lab
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Contents
Introduction The importance of Analytical method in drug development
Drug development and GMP
Requirements of analytical method in drug development
Validation
What is validation? Analytical method validation
Analytical performance characteristics
Definitions
How to calculate characteristics?
Summary of validation protocol & report
When do you perform revalidation?
Conclusion
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Pharmaceutical?
A pharmaceutical product manufactured by biotechnologymethods (involving live organisms; bioprocessing)
Introduction
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Scope of Pharmaceuticals
Introduction
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The Importance of Analytical Methodology in
the Drug Development Process
During the drug development, the evaluation of1) Determining optimum crystalline and salt form of the drug
substance, testing support to process development and
cleaning validation
2) The dosage forms bioavailability
3) Determining optimum formulation
4) The setting of specifications for the drug substance,
intermediates, and drug product
5) Identification, quantitation, and qualification of impurities and
degradants
6) The shelf-life of the product (stability)
7) Product safety
8) Support to preclinical and clinical studies (safety and efficacy)
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The Importance of Analytical Methods in the
Drug Development Process
The chemistry, manufacturing, and controls section of a US
regulatory filing is required to contain the following directly
related to analytical testing:
1) Method validation
2) Physical description and characterization of the drug
substance and drug product-proof of the chemical structure
3) In-process controls
4) Characterization of reference standards
5) Specifications-description of analytical methods
6) Evaluation of container/closure system for storage7) Justification of drug product development
8) References to compendial test methods for inactive ingredients
9) Bioequivalence
10) Pharmaceutical development report
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The Importance of Analytical Methodology in the Drug
Development Process
Physicochemical Characterization (1)
1) UV/visible or mass spectrometry
2) Amino acid analysis
3) Amino acid or Nucleic acid sequencing
4) Carbohydrate analysis and, if appropriate, sequencing
5) Peptide mapping
6) Determination of disulfide linkage
7) SDS-PAGE
8) Isoelectric focusing (1D or 2D)
9) HPLC, GC, LC, or thin layer chromatography
10) Nuclear magnetic resonance spectroscopy
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The Importance of Analytical Methodology in the Drug
Development Process
Physicochemical Characterization (2)
11) Assay to detect related proteins including deamidated, oxidated,
processed, and aggregated forms
12) other variants, such as amino acid substitutions and
adducts/derivatives
13) other process contaminants such as sulfhydryl reagetns, urea,
residual host proteins, residual DNA, and endotoxin
3 Stability indicating method
.
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The Importance of Analytical Methodology in the Drug
Development Process
Biological Characterization
1) Specific identity testing such as Western blot analysis or ELISA
2) Cytometric analysis
3) Neutrovirulence testing, if appropriate
4) Serotyping
5) Electrophretic typing
6) Inactivation studies : Irreversibility
7) Neutralization assays
8) Titration
9) in-vivo assay
10) in-vitro assay (ELISA)
11) Specific toxicity
12) Abnormal toxicity
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The Importance of Analytical Methodology in
the Drug Development Process
After the registration of drug product to regulatory authority, the
test methods are used in the pharmaceutical quality control
laboratory to do the following.
1) Identify the drug substance and product
2) Quantitate the pharmaceutical active ingredient
3) Determine level of purity
4) Guarantee the overall quality of the product
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Q 1 : Quality ?
Ans.) 1) Lot (, )
2)
Why?
lot , ,
.
Question & Answer
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Q 2 : Quality ?
Ans.)
1) (Safety)
2) (Stability)3) (Efficacy, Potency, Strength)
Q 3 : Quality ?
Ans.)
1)
2)
3)
Question & Answer
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Stakeholders in Quality Analytical Testing
During Drug Development
Analytical
test
Processdevelopment
/Characteriza
tion/Controls
Support to
Clinical
Trials
Product
development
Toxicology
Pharmacokinet
ics
Regulatory
Filings
Stability /
Release
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Drug Product Development and GMP
Full characterization
Full GMP
21 CFR 210, 211
21 CFR 610
Phase III
Phase I
Phase II
Pre-clinical
QA &QC, Clinical Monitoring Program
Validated MethodPre-qualified
method
(Standard screening
methods)
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Q 4 : ?
Ans.) 2
Why? 2
.
Question & Answer
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Q 5 : 2 fix
?
Ans.)
.
Question & Answer
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Q 6 : Process development Product qualityupgrade ?
Ans.) lot comparability test ()
1) :
2) : ,
Q 7 : Process development Product qualityupgrade ?
Q 8 : ?
Question & Answer
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Requirements of Analytical methods During
the Drug Development Process
Depending on the stage of development, these analytical methods arestandard screening methods at the start of the development
process, which over time are gradually upgraded to thoroughly
validated methods for NDA application.
The filed methods must be simple, robust, and reliable- that is, easy to
use and perform without deviations when appropriately applied in aqualified laboratory.
Note
1. Screening methods are typically not optimized for speed and robustness.
2. The filed methods referred to as VTR2AP methods
(validated, transferable, robust, rapid, accurate, and precise)
James M. Miller and Jonathan B. Crowther, Analytical Chemistry in a GMP
envirinment: A practical guide, John Wiely & Sons,New York (2000)
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Laboratory Analytical Methods Flow During
the Drug Development Process
MethodValidation
Method
Transfer
MethodDevelopment
Approved
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Q 9 : ?
Ans.)
. (R&D or QC)
r R&D
QC .
Question & Answer
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Whats validation?
The FDA defines the term as :
Established documented evidence which provides a high degree of
assurance that a specific process will consistently produce a product
meeting its pre-determined specifications and quality attributes.
General Principles of Validation (1987)
ICH guideline defines the term as :
A documented program that provides a high degree of assurance
that a specific process, method, or system will consistently produce
a result meeting pre-determined acceptance criteria.
Q7A-GMP for active phamarceutical ingredients (2000)
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Q 10 : Quality Specification ?
Ans.) Specification Quality .
Specification Product Quality .
Product Quality .
Question & Answer
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Acceptance criteria
Acceptance criteria means
The product specifications and acceptance / rejection criteria,
such as acceptable quality and unacceptable quality level, with
an associated sampling plan, that are necessary for making a
decision to accept or reject a lot or batch (or any other
convenient subgroups of manufactured units)
CFR Part210.3 Definitions (2000)
ICH guideline define the term as :
Numerical limits, ranges, or other suitable measures for acceptance
of test results.
Q7A-GMP for active phamarceutical ingredients (2000)
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Analytical Method Validation
Initial Method Validation Guidance Issued in 1987
Guideline for submitting samples and analytical data for methods
validation. Food and Drug Administration, February 1987. US
Government Printing Office:1990-281-794:20818.
Updated in August 2000 (Draft Guidance!)
Analytical Procedures and Method Validation. Fed. Reg. 65(169), 52,776-
52,777, 30 August 2000
CFR Part 211.165 Testing and release for distribution (e)
The accuracy, sensitivity, specificity, and reproducibility of test
methods employed by the firm shall be established and documented.
ICH guideline Q7
GMP guide for active pharmaceutical ingredients
ICH guideline Q2(R1)
Validation of analytical Procedure : Text and Methodology
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ICH & USP Method Validation
Analytical Performance Characteristics
Validation parameter ICH USP
Accuracy O O
Precision
Repeatibility O O
Interm. precision O -
Reproducibility O -
Specificity of Selectivity O O
Detection limit O O
Quantitation limit O O
Linearity O O
Range O O
Robustness O O
Ruggedness - O
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Definitions
Specificity:The ability to access unequivocally the analyte in the presence of
components which may be expected to be present. Typically these
might include impurities, degradants, matrix, etc.
Linearity:
Its ability to obtain test results which are directly proportional to the
concentration (mount) of analyte in sample within given range.
Range:The Interval between the upper and lower concentration (amounts) of
analyte in the sample ( including these concentrations) for which ithas been demonstrated that the analytical procedure has a suitable
level of precision, accuracy and linearity.
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Definitions
Precision:
The closeness of agreement (degree of scatter) between a series ofmeasurements obtained from multiple sampling of the same homogeneous
sample under the prescribed conditions.
Accuracy:
The closeness of agreement between the value which is accepted either as a
conventional true value or an accepted reference value and the value found.
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Definitions
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Definitions
Limit of detection:
The lowest amount of analyte in a sample which can be detected but
necessarily quantitated as an exact value.
Limit of quantitation:
The lowest amount of analyte in a sample which can be quantitatively
determined with suitable precision and accuracy.Ruggedness:
The degree of reproducibility obtained under a variety of conditions, such as
different laboratory, different analysts, different instruments, environmental
conditions, operators and materials. (USP)
Robustness:
A measure of its capacity to remain unaffected by small but deliverate
variations in method parameters and provides an indication of its reliability
during normal use. (ICH)
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Comparison of ICH & USP
Analytical Performance Characteristics
Characteristic Refer. ManEquip.
or Cond.Material
Inter-
lab
Intra-
lab.
Precision ICH same same same - O
Interm.Precision
ICH different different same - O
Reproducibility ICH different different different O -
Robustness ICH same
same /
different different - O
Ruggedness USP different different different O -
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1. How to measure the Specificity?
Method : Impurity challenge and Recovery test Impurity : Hydrolyte of HBG by hydrogen peroxide
Challenge : half volume of hydrolyte
Calculation
Measured con. of challenge
Recovery yield = ------------------------------------------- X 100
Calculated con. of challenge
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2. How to measure the Linearity?
The correlation coefficient(r2 or r), slope of regression line and
residual sum of squares should be submitted.
A minimum of5 concentrations is recommended.
Used the standards (5 con.) and samples (4 con.)
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3. How to set the Range?
The specified range is normally derived from linearity studiesand depends on the intended application.
Acceptable degree of linearity, accuracy and precision.
The following minimum specified ranges should be
considered.
Forassay of active substance or a finished product,80~120% of concentration.
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4. How to calculate the Accuracy?
Accuracy should be assessed on samples spiked with knownamounts.
Minimum 9 determinations over a minimum 3 concentration
level covering the specified range(e.g. 3 concentrations/3
replicate)
Should be reported as percent recovery by the assay of
known added amount of analyte in the sample or as thedifference between the mean and the accepted true value
together with the confidence intervals.
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Q 11 : accuracy Spike test
?
Ans.) accuracy
.
.
280nm :
Lowry
. Bradford Lowry
.
Question & Answer
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Q 12 : accuracy specificity Spike test
?
Ans.) challenge .
accuracy challenge
specificity challenge.
Question & Answer
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5. How to calculate the Precision?
Repeatability Minimum 9 determinations covering the specified range(e.g.
3 concentrations/3 replicate) or
A minimum of6 determinations at 100% of the test
concentration.
Intermediate precision Depends on the circumstance under which the procedure is
intended to be used.
Establish the effects of random events on the precision.
Typical deviations to be studied include individually days,
analyst, equipment.
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5. How to calculate the Precision?
Reproducibility
Assessed by means of an inter-laboratory trial.
Should be considered in case of standardization of an
analytical procedure.
This data is not part of the marketing authorization dossier
Recommended data
Standard deviation, relative standard deviation (coefficient ofvariation) and confidence interval should be reported for each
type of precision investigated.
t-test analysis
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6. Detection limit
Based on visual evaluationMostly for non-instrumental methods
Based on signal-to-noise
Analytical procedures which exhibit baseline noise
Compare measured signals from samples with known low
concentration of analyte with those of blank samples.
A sinal-to-noise ratio 3 or 2:1 is acceptable.
Based on the Standard Deviation of the Response
and the Slope
The quantitation limit (QL) may be expressed as:
DL = 3.3/S
where = the standard deviation of blankS = the slope of the calibration curve
The slope S may be estimated from the calibration curve of
the standard.
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7. Quantitation limit
Based on visual evaluationThe quantitation limit is generally determined by the
analysis of samples with known concentrations of analyteand by establishing the minimum level at which theanalyte can be quantified with acceptable accuracy andprecision.
Based on signal-to-noiseA typical signal-to-noise ratio is 10:1.
Based on the Standard Deviation of the Responseand the SlopeThe quantitation limit (QL) may be expressed as:
QL =10/S
where = the standard deviation of blankS = the slope of the calibration curve
The slope S may be estimated from the calibration curve ofthe standard.
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8. How to measure the Robustness?
The evaluation of robustness should be considered duringthe development phase and depends on the type of
procedure under study. It should show the reliability of an
analysis with respect to deliberate variations in method
parameters.
Lowry : reaction time (30min vs. 40 min)
Measurement : confidential variance, standard
deviation, T-test
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Summary of Method Validation Protocol
Parameter Sample Testing No SpecificationAcceptance
Criteria
AccuracyLow, Medium, High &
spiked L, M, H 6 Spiked recovery 80~120%
Repeatability 20, 50, 70, 90ug/mL 6 CV% a
Linearity5 Std &
20, 50, 70, 90ug/mL6
Correlation
coefficient>0.99
SpecificityHBG bulk treated with
H2O25 Recovery 80~120%
Range Low, Medium, High 5 Accuracy, Precision &Linearity pass
LOQ Blank 6 =(10*SD)/slope Calculated value
Robustness Medium 6 (2 person) t-test P> a
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Q 13 : Protocol parameter LOD ?
Ans.) :
.
.
:
parameter .
Question & Answer
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Q 14 : ?
Ans.) , (SD)
2SD, 3SD % .
200ug/mL, SD 13.3ug/mL 2SD
.
Acceptance criteria = [200 (2*13.3)] / 200 *100 = 86.7% (LL)
= [200 + (2*13.3)] / 200 *100 = 113.3% (UL)
* 3SD : Bioassay
Question & Answer
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Summary of Method Validation Report
Parameter SpecificationAcceptance
CriteriaResult
Accuracy Spiked recovery 80~120% ave. 96.02%
Repeatability CV% a(0.05) L 0.768,M 0.293, H 0.293
LinearityCorrelation
coefficient>0.99 >0.995
Specificity Recovery 80~120% ave. 106.18%
RangeAccuracy, Precision &
Linearity pass 25~110ug/ml
LOQ =(10*SD)/slope Calculated value 2.11ug/ml
Robustness t-test P> a 0.652 > 0.05
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Q 15 : validation protocol
?
Ans.)
1) (, , ) review
2)
3) :
4)
- acceptance
criteria ( .)
Why? worst case
acceptance criteria worst case
acceptance criteria .
Question & Answer
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Q 16 : ?
Ans.) : System suitability test
1) Negative control (Blank)
2) Positive control (Standard material)
3) Reference (Normal product or Clinical product)
4) Calibration curve : linearity regression coefficient (r or r2)
Question & Answer
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Q 17 : When do you perform revalidation?
Ans.)
Changes in the synthesis of the drug substance
Changes in the composition of the finished product Changes in the analytical procedure
ICHQ2(R1) Validation of analytical procedures: Text and
Methodology (2005)
In other case, some companies perform the method validation regular
interval.
Question & Answer
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FY 99QS/GMP Deficiencies of Domestic Inspections(483s Issued)
20
22
23
23
25
29
31
36
36
40
42
44
0 5 10 15 20 25 30 35 40 45
Doc.Control
Processing Proc
Design/Dev Plans
Nonconform
Audits
Dev.Spec.
DHRCor/Pre Action
Design Changes
Validation
Inspection/Test Proc.
Complaints
Est. FY99 Total QS/GMP EIs with 483s issued=656
DeficiencyArea
Sample Size = 293
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Q 18 : 483 ?
Ans.) Warning letter inspection GMP
Question & Answer
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Q 18 : 483 ? ()
Question & Answer
The inspection revealed:
1. Analytical results were reported to [redacted] stating that a sample met
specifications when either out-of-specifications (OOS) results were
obtained on the sample analysis or on the quality control samples used
to determine the validity of the analytical results. These OOS results were
not investigated/documented properly to assure results reported to
[redacted] were accurate and valid. [21 CFR 211.165(a)]
2. Inadequate method validation in that OOS findings were discarded
without investigating the cause of the OOS results and analytical data
was selectively reported to support the validation. [21 CFR 211.165(e)]
3. Use of reference standards and reagent solutions for extended periods
of time without data in the analytical records supporting time of use. [21
CFR 211.194(c)]
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Q 19 : 21 CFR 211.165(e)?
Question & Answer
e) The accuracy, sensitivity, specificity, and reproducibility of
test methods employed by the firm shall be established and
documented. Such validation and documentation may be
accomplished in accordance with CFR 211.194(a)(2)
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Conclusion
GMP is Good Manufacturing Practice.
GMP is based on documentation.
Time is money.
Sometimes, Time is more valuable than money
in especially drug development process.