usp chemical medicines & excipients - evolution of validation practices
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Track I, Session I: Chemical
Medicines and Excipients-Evolution
of Validation Practices Wednesday, April 17, 2013 (9:00 a.m. to 11:00 a.m.)
IPC–USP Science & Standards Symposium
Partnering Globally for 21st Century Medicines
Moderator: Milind Joshi, Ph.D. Chair, USP South Asia Stakeholder Forum
Acceptable Analytical Method Variation
Setting System Suitability
Requirements
Todd L. Cecil, Ph.D.
Vice President, Chemical Medicines
USP
Sources
– Instrument Characteristics
– Sample Characteristics
– Method Parameters
– Environmental Affects
– Analyst
– Instrument Settings
– Many others
Method Variation
Systematic Error
– Determinate Error
– Discoverable source
(in theory)
Estimated using Accuracy
Measuring Variation
Random Error
– Indeterminate Error
– Experimental error
Estimated using Precision
Developed in late 1980’s for the Pharmaceutical
Industry
– PhRMA -> USP <1225> -> ICH Q2A -> USP <1225>
Defined “Analytical Performance Characteristics”
– Accuracy
– Precision
– Specificity
– Detection Limit
– Quantification Limit
– Linearity
– Range
Validation to Measure Variability
Trueness
Bias
Intermediate precision
Repeatability
Reproducibility
Ruggedness
Robustness
Depends upon two factors
Acceptable Variability
Expectation
• Analyst Experience
• Instrument knowledge
• Matrix complexity
Application
Test
Procedure
Acceptance criteria
ICH and USP do not describe acceptable limits
Therefore, Acceptable Validation/ Variation is
open to interpretation by:
– Bench Chemist
– Supervisory Chemist
– Regulatory Affairs Professional
– The Regulator
– The Pharmacopeial Professional
– And fights ensue…
Acceptable Validation
Recent publications
– Pharma’s Analytical Target Profile (ATP)
– USP’s Performance-Based Procedures
Upcoming publications
– USP Validation and Verification Expert Panel
– USP Statistics Expert Committee
– USP “Requirements for Compendial Validation
<1200>” (working title)
There is Another Way!
Critical Validation Parameters
– What are the critical features (parameters) of
an acceptable procedure?
Procedure Performance Measures
– How do we measure the critical parameters?
Procedure Performance Acceptance Criteria
– What defines “good enough” for each
performance measure?
Defining Another Way Forward
Measuring the Parameters
Precision – % RSD with sufficient degrees of freedom
Accuracy – Spike Recovery or Comparison to Primary
Standard
Specificity – Resolution or Spike Recovery
Linearity – Slope, Intercept, R2
Range – Precision and accuracy
Limit of Detection – Precision
Limit of Quantification – Precision
Collapsing the Parameters
Precision – Measure of Random Error
Accuracy – Measure of Systematic Error
Specificity – Measure of Systematic Error
Linearity – Measure of Systematic Error
Range – ?
Limit of Detection – Measure of Random Error
Limit of Quantification – Measure of Random Error
Why are we measuring things so many different ways?
Does agreement mean quality? or are we hiding behind tradition?
IF we combine critical components can we gain efficiency?
Extract from <1200>
Category I* Category II*
(Quantitative)
Category II* (Semi-
quantitative)
Analytical
Performance
Characteristics
<1225> <1200> <1225> <1200> <1225> <1200>
Accuracy Y 1 Y 4 ? N
Precision Y 1 Y 5 N 5
Specificity Y 2 Y 2 Y 2
Detection Limit N N N N Y 6
Quantification
Limit N N Y 4 N N
Linearity Y 3 Y 4 N N
Range Y 3 Y 4 ? N 1 Covered in the Precision-Accuracy Study 2 Covered in the Specificity Study 3 Covered in the Range Study 4 Covered in Accuracy Study 5 Covered in Precision Study 6 Covered in the Detectability Study
Precision and Accuracy Study
When properly combined Precision and Accuracy yield a
probability of passing.
Bias-%CV Tradefoff, 98%-102% limits, True Value = 100, Prob'y Passing 0.95
0
0.2
0.4
0.6
0.8
1
1.2
2.001.801.601.401.201.000.800.600.400.200.00
Bias
%C
V
Precision
– % RSD of 6 independent samples at 100%
Accuracy
– Δ from RS label at 100%
– The data obtained for Precision can be used for
Accuracy
Combine with Acceptance Criteria to calculate
probability
Result = NORMDIST(Upper, Mean, SD, TRUE)
-NORMDIST (Lower, Mean, SD, TRUE)
Limit: NLT 0.95
Study Detail
Specificity is a special case of Accuracy.
Interferences considered
Separation Sciences
– Resolution of NLT 1.5
Specificity Study
Non-Chromatographic Procedures are harder
Spiked samples with interferences
Measure the error caused by the addition of an
interference
Limit is linked to the acceptance criteria of the
analyte
– The error caused by all interferences cannot
exceed the allowable bias from the Precision-
Accuracy Study
Specificity Study
Retasked Range
Precision-Accuracy evaluation at 80%, 90%,
100%, 110%, and 120%
Instead of Mean in the calculation, use recovery
value
Recovery Value = [Mean]/[Known]*100%
Limit: Each concentration is NLT 0.95
Range Study
Response vs Concentration
Calibration curve
Technique dependent application
Calculated vs Known Concentration
Slope =1
Intercept =0
Accuracy evaluation
How do you measure linearity?
Slope: not correlated to error
Intercept: not correlated to error
R2: limited correlation to error
Linearity
Linearity
Slope and Intercept
– Overwhelmed by random noise
– Not correlatable to systematic noise
– Adds no additional information
Basis for Linearity is not supported
Range adequate
Limit of Detection
Only applies to “Limit” procedures
S/N of 3
– independent samples at LOD
– “adequate precision and accuracy”
What is adequate?
What is the purpose of the test?
Limit Test
Measure a Standard solution of the impurity at the limit
Measure a Sample solution
– Is the response of the impurity in the Sample < Standard
– Pass
– Is the response of the impurity in the Sample ≥ Standard
– Fail
Is the Δ between pass/fail adequate?
If the limit is 0.1%, then acceptable values are
– 0.14% to 0.05%
– LOD does not assure the measurement
–Detectability does.
Detectability
A new term included in <1200>
Replaces LOD
3 steps
–1: Standard of impurity at limit
–2: Sample spiked with impurity at limit
–3: Standard spiked with impurity at 100%-%RSD* for the impurity
–*can be estimated with Horwitz
If 1=2 and 3<2, then the difference is detectable
Otherwise, procedure is not adequate
What is Horwitz?
Limit of Quantitation
Why do we make this measurement?
–10x S/N . . .
A meaningful quantity in development?
–Yes
Necessary to validate?
–No
Validation presumes
–A known procedure
–A typical value for the analyte
–Known acceptance criteria
You already know the typical range of the analyte…
Use the Range Study
–80%-120% for Assay; 50%-150% for impurities
Summary
USP is challenging validation concepts
Including DOE and QbD through the ATP
Include measurable parameters and clear criteria
Focus on Precision and Accuracy results
Use Specificity to aid understanding of Accuracy
Retask Range
Introduce detectabiltiy
Eliminate LOD, LOQ and Linearity
But Wait, There’s More…
Setting System Suitability Requirements
Validation is measured only once
System suitability is measured on a daily basis
– Traditionally uses instrument dependent measurements
– Resolution
– Tailing
– %RSD
System suitability rarely linked to variance
Use Validation protocol to evaluate Precision and Accuracy
across the days run.
System suitability can then be linked to validation
Specificity should represent necessary minimums, but should
exceed criteria of validation
Precision, Accuracy & Linearity
Harry Yang, Ph.D.
Member, USP Statistics Expert Committee
Validation is a snapshot (at any given time) of
the assay’s performance.
It is confirmation that the assay is fit for its
intended use
Required by regulatory guidelines
Method Validation
History of ICH Guidelines on Method Validation
Other Related Guidelines
Accuracy
Precision
Repeatability
Intermediate precision
Specificity
Limit of detection
Limit of quantitation
Linearity
Range
Common Validation Characteristics
Accuracy
Precision
Repeatability
Intermediate precision
Specificity
Limit of detection
Limit of quantitation
Linearity
Range
Common Validation Characteristics
Closeness between a series of measurements
obtained from multiple sampling of the same
homogeneous sample
Precision
Repeatability: intra-assay precision. Usually
same day, operator, equipment
Intermediate Precision: same laboratory but
different operators, equipment, etc.
Reproducibility: precision between laboratories
Expressed as standard deviation (SD) or relative
standard deviation (RSD)
Precision
X
SDRSD
n
XX
SDn
X
X
n
i
i
n
i
n
,
1
)(
, 1
2
1
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 (ICH Q2(R1))
Accuracy
]var[)(])[( 22 XXE TXT
True value Mean measurement
Precision Bias
Closeness of agreement between the average
value obtained from a large series of test results
and an accepted reference value
Bias
TXBias
µT µX
Accuracy = bias + precision
Validation of Accuracy and Precision
Model
or
or
An Example
Test result Bias Intermediate
precision
Repeatability
Burdick, LeBlond, Sandell, Yang, 2013
TX TX
An Example
Test result Bias Intermediate
precision
Repeatability
Burdick, LeBlond, Sandell, Yang, 2013
TX TX
Test the hypothesis that bias = 0
H0: µ = 0 vs. H1: µ ≠ 0
Reject H0 if (which is the same as p-value < 0.05)
where
Assessment of Bias: Traditional Approach
025.0,1/
ntns
X
025.0,1nt - cutpoint of t-distribution
025.0,12 /
ntns
Y
,1
)(
, 1
2
21
n
YY
sn
Y
Y
n
i
i
n
i
n
P-value < 0.05 is equivalent to that the 95%
confidence interval contains zero, i.e.
Assessment of Bias: Traditional Approach
p-value < 0.05
p-value ≥ 0.05
nstYnstY nn /,/0 025.0,1025.0,1
Penalize more precise assay
Award small sample size
Issue with the Traditional Approach
Huberta et al, 2004
With of the 90% CI is
proportional to assay
precision (s) and
reciprocal
of the squared root of
sample size n.
nstYnstY nn /,/0 025.0,1025.0,1
Bias is deemed acceptable if the 90%
confidence interval of bias is bounded by pre-
specified acceptance limits (e.g., ±15%)
Equivalence Method
Huberta et al, 2004
nstYnstY nn /,/ 025.0,1025.0,1
Comparison Between Significance and Equivalence
Is bias acceptable?
Significance Equivalence
Yes
Yes
Yes
No
Yes
Yes
No
No
Bias is deemed acceptable if the 90%
confidence interval at each concentration level is
contained with in pre-specified range (LAL, UAL)
Equivalence Method
0
UAL
a b c
LAL
0
UAL
a b c
LAL
Plot of Bias vs. True Value
True value
Accessing Conformance to Acceptance Criteria: Precision
Intermediate precision is considered acceptable
if the 95% confidence interval is bounded by a
pre-selected number UAL
< UAL
Burdick, LeBlond, Sandell, Yang, 2013
Total Error Approach
Bias cannot be assessed independent of
precision
Huberta et al, 2004; Hoffman & Kringle, 2007
Measured value = True value + Method Bias + Method error
Total Error Approach
Test result True value Y = -
Accuracy of a method is acceptable if it is very
likely that the difference between every
measurement of a sample and the true value is
inside pre-chosen acceptance limits
Total Error Approach
Huberta et al, 2004
Total Error Approach
Risk = 1 - Probability of meeting acceptance criterion
Huberta et al, 2004
Beta-expectation tolerance interval (Huberta et al, 2004)
With 100β% confidence that bias of a future measurement is bounded by λ
Average (expected) probability for bias of a future observation is no smaller than 100β%
Beta-content tolerance interval (Hoffman & Kringle, 2007)
With 100γ% confidence that bias of 100β% future measurements is bounded by λ
Bayesian analysis (Burdick, LeBlond, Sandell, Yang, 2013)
Conditional on validation data, probability for bias of a future observation is no smaller than 100β%
Methods for Testing H0:
.)|( dataXYP T
Accuracy Profile
Huberta et al, 2004
1 Graybill FA, Wang CM. Confidence intervals on nonnegative linear combinations of variances. J Am Stat Assoc. 1980;75:869–
873.
2. Nijhuis MB, Van den Heuvel ER. Closed-form confidence intervals on measures of precision for an interlaboratory study. J
Biopharmaceutical Stat. 2007;17:123–142.
3. Satterthwaite FE. An approximate distribution of estimates of variance components. Biometric Bull. 1946;2:110–114.
4. Huberta P, Nguyen-Huub JJ, Boulangerc B, et al. Harmonization of strategies for the validation of quantitative analytical
procedures: a SFSTP proposal—part I. J Pharm Biomed Anal. 2004;36:579–586.
5. Huberta P, Nguyen-Huub JJ, Boulangerc B, et al. Harmonization of strategies for the validation of quantitative analytical
procedures: a SFSTP proposal—part II. J Pharm Biomed Anal. 2007;45:70–81.
6. Huberta P, Nguyen-Huub JJ, Boulangerc B, et al. Harmonization of strategies for the validation of quantitative analytical
procedures: a SFSTP proposal—part III. J Pharm Biomed Anal. 2007;45:82–96.
7. Mee RW. b-expectation and b-content tolerance limits for balanced one-way ANOVA random model. Technometrics.
1984;26:251–254.
8. Hahn GJ, Meeker WQ. Statistical Intervals: A Guide for Practitioners. New York:Wiley; 1991:204.
9. Hoffman D, Kringle R. Two-sided tolerance intervals for balanced and unbalanced random effects models. J Biopharm Stat.
2005;15:283–293.
10. Montgomery D. Introduction to Statistical Quality Control. 3rd ed. New York: Wiley; 1996:441.
11. Kushler RH, Hurley P. Confidence bounds for capability indices. J Quality Technol. 1992:24(4):188–195.
12. Wolfinger RD. Tolerance intervals for variance component models using Bayesian simulation. J Quality Technol.
1998;30:18–32.
13. Ntzoufras I. Bayesian Modeling in WinBUGS. New York: Wiley; 2009:308–312.
14. Spiegelhalter D, Thomas A, Best A, and Gilks, W (1996) BUGS 0.5 Examples Volume 1(version i), Example 7, Dyes, pp 24-
26. Available from http://www.mrc-bsu.cam.ac.uk/bugs/documentation/Download/eg05vol1.pdf (accessed November 20,
2012).
15. Burdick R, LeBlond D, Sandell D, Yang H. Statistical methods for validation of method accuracy and precision.
Pharmacopeia Forum, May –June Issue, 39 (3)
.16. USP. USP 36–NF 31, Validation of Compendial Procedures <1225>. Rockville, MD: USP; 2013:983–988.
17. ICH. Validation of analytical procedures: text and methodology Q2(R1). 2005.
http://www.ich.org/fileadmin/Public_Web_Site/ICH_Products/Guidelines/Quality/Q2_R1/Step4/Q2_R1__Guideline.pdf.
Accessed 27 November 2012.
References
Linearity
Response vs concentration linear curve
This is a calibration curve
It provides a means to convert a signal to a desired measured
value
Predicted concentration vs known concentration
This is a surrogate for Accuracy
Slope should be 1 and intercept should be 0
Two Types of Linearity
Todd L. Cecil, Personal communication, 2013
We wish to measure the concentration of an
analyte in a test sample.
Standards = known concentrations of an analyte
To estimate the concentration, we create a
standard curve
Calibration Curve
Standard Curve
Novick and Yang, 2013
Standard Curve
Sample
Novick and Yang, 2013
Evaluate linearity by visual inspection
Test of Linearity - ICH Q2(R1) guideline
Novick and Yang, 2013
Test of Linearity – Pearson Correlation
r = 1 r = -1
r = 0
Determine how close the predicted values to the mean
values at each concentration level
Test of Linearity – Lack of Fit (LOF)
Evidence of lack of fit
Clinical and Laboratory Standards Institute
http://www.clsi.org/source/orders/free/ep6-a.pdf
Compare straight-line to higher-order polynomial
curve fits
Recommendation: Test higher-order coefficients.
The EP6-A Guidelines
Novick and Yang, 2013
The EP6-A Guidelines
Novick and Yang, 2013
Literature
Novick and Yang, 2013
Conduct hypothesis testing with linearity claim
as the null hypothesis
Rely on failing to reject the null hypothesis to
conclude linearity
Penalize precise assay
Award small sample size
Drawbacks of Significance Test
More Literature
Novick and Yang, 2013
Two Practical Approaches
Two one-sided tests (TOST) for calibration error
Estimate bias in concentration due to approximating either
quadratic curve or proportional model using linear line
Bias is expressed as a function of a ratio of two model
parameters. Thus the Fieller’s Theorem can be applied to
obtained 90% confidence interval of the bias
Akaike information criterion (AIC)
Based on the principle of parsimony – the smallest possible
number of parameters for adequate representation of the data
where N – total number of data points, and K – the total number
of estimated regression model parameters
LeBlond, Tan and Yang, (2013a, 2013b)
Estimating Calibration Bias: Linear vs Quadratic
Models:
Bias:
Assumption: Concentration levels used in the experiment are
symmetrically spaced.
LeBlond, Tan and Yang, (2013a, 2013b)
90% Confidence Interval (CI) of Bias
Linearity is accepted if the above 90% CI is contained
Within pre-specified limits.
LeBlond, Tan and Yang, (2013a, 2013b)
Fieller’s exact 90% confidence Interval
Linear Model vs Proportional Model
LeBlond, Tan and Yang, (2013a, 2013b)
Models:
Bias:
90% Confidence Interval of Bias
Linearity is accepted if the above
90% CI is contained
Within pre-specified limits.
LeBlond, Tan and Yang, (2013a, 2013b)
90% CI of ratio of two model parameters:
90% CI of bias in concentration:
An equally-spaced experimental design is not a
necessary condition
Linearity can be tested under general conditions
Test Linearity for More General Experiment Design Conditions
Yang, Novick and LeBlond, 2013; Novick and Yang, 2013
1. USP. USP 36–NF 31, Validation of Compendial Procedures <1225>. Rockville, MD: USP; 2013:983–988.
2. ICH. Validation of analytical procedures: text and methodology Q2(R1). 2005. http://www.ich.org/fileadmin/Public_Web_Site/ICH_Products/Guidelines/Quality/Q2_R1/Step4/Q2_R1__Guideline.pdf. Accessed 27 November 2012.
3. Clinical and Laboratory Standards Institute. EP06-A02 Evaluation of the linearity of quantitative measurement procedures: a statistical approach. 2003. http://www.techstreet.com/standards/clsi/ep06a?product_id=1277866. Accessed 27 November 2012.
4. Anscombe FJ. Graphs in statistical analysis. Am Stat. 1973;27(1):17–21.
5. Van Loco J, Elskens M, Croux C, Beernaert H. Linearity of calibration curves: use and misuse of the correlation coefficient. Accred Qual Assur. 2002;7:281–285.
6. Bruggemann L, Quapp W, Wennrich R. Test for nonlinearity concerning linear calibrated chemical measurements. Accred Qual Assur. 2006;11:625–631.
7. Mandel J. (1964) The Statistical Analysis of Experimental Data. New York: Wiley; 1964.
8. Mark H, Workman J., Chemometrics in Spectroscopy, Linearity in Calibration How to Test for Non-linearity, Spectroscopy 2005;20(9):26–35
9. Liu J, Hsieh E. Evaluation of linearity in assay validation. In: Encyclopedia of Biopharmaceutical Statistics. 2nd ed. London: Informa Healthcare; 2010:467–474
10. Finney DJ. Statistical Method in Biological Assay. 2nd ed. London: Charles Griffin; 1964:27–29.
11. Berger RL, Hsu JC. Bioequivalence trials, intersection-union tests and equivalence confidence sets. Stat Sci. 1996:11(4):283–319.
12. Burnham KP, Anderson DR. Model Selection and Multimodel Inference: A Practical Information–Theoretic Approach. 2nd ed. New York: Springer; 1998:31.
13. Burnham KP, Anderson DR. Multimodel inference: understanding AIC and BIC in model selection. Sociol Meth Res. 2004;33(2):261–304.
14. David LeBlond, Charles Y Tan, Harry Yang (2013), Confirmation of Analytical Method Calibration Linearity, Pharmacopeial Forum 39(5), pp XX – XX.
15. David LeBlond, Charles Y Tan, Harry Yang (2013), Confirmation of Analytical Method Calibration Linearity: Practical Application, Pharmacopeial Forum ??(??), pp ?? – ??.
16. Steve Novick and Harry Yang (2013), Directly Testing the Linearity Assumption for Assay Validation, Accepted for publication in Journal of Chemometrics.
17. Steve Novick and Harry Yang (2013), Directly Testing the Linearity Assumption for Assay Validation, Accepted for publication in Journal of Chemometrics, The 36th Mid-west Biopharmaceutical Statistics Workshop, Muncie, Indiana, May, 2013i
18. Harry Yang, Steve Novick and David LeBlond (2013). Testing linearity under general experimental conditions. In preparation.
References
Lifecycle Management of Analytical
Procedures
Joachim Ermer, Ph.D.
Member, USP Validation and Verification Expert Panel
Adaptation of the lifecycle concept [ ICH Q8] and of modern concepts for process validation to analytical procedures
to holistically align analytical procedure variability with the requirements of the product to be tested
to demonstrate that the analytical procedure meets the predefined criteria over the whole lifecycle
to facilitate continual improvement
Proposal to revision and compile USP General Chapters <1225>, <1226> and <1224> into a single General Information Chapter on Lifecycle Management of Analytical Procedures
Stimuli article to be published in PF 39(5), Sep - Oct 2013
Objectives of Expert Panel Validation & Verification
“systematic approach that begins with predefined objectives and emphasizes product and process understanding and process control, based on sound science and quality risk management” [ICH Q8]
systematic approach that begins with predefined objectives and emphasizes analytical procedure understanding and analytical control, based on sound science and quality risk management”
Quality by Design – Also Relevant for Analytics
Alignement of Process and Analytical Procedure
Quality Target
Product Profile
Prospective summary of the quality
characteristics of a drug product
to ensure quality, safety, efficacy
Analytical Target
Profile
Defines the objective of the test
and quality requirements
for the reportable result
PROCESS ANALYTICAL PROCEDURE
Developed starting 2008 by EFPIA / PhRMA Working
Group “Analytical Design Space”
M. Schweitzer, M. Pohl et al.: QbD Analytics. Implications and
Opportunities of Applying QbD Principles to Analytical
Measurements, Pharmaceutical Technology, Feb. 2010, 2-8
http://pharmtech.findpharma.com/pharmtech/article/articleDetail.
jsp?id=654746
Quality (data) attributes of the reportable result
performance requirements for use
accuracy and measurement uncertainty including precision
Analytical Target Profile (ATP)
Based on the understanding of the target measurement
uncertainty
Maximum allowed uncertainty to maintain acceptable levels of
confidence
Reference point for assessing the fitness of an analytical
procedure
towards predetermined performance requirements
In development phase and during all changes within the lifecycle
linked to the purpose, not to a specific analytical technique.
Analytical Target Profile (ATP)
Any analytical procedure that conforms to the ATP is
acceptable
USP Medicines Compendium, General Chapter <10>
May be also established for existing procedures
including compendial procedures
based on (monograph) specifications, existing knowledge
Analytical Target Profile (ATP)
The procedure must be able to quantify [Analyte]
in presence of X, Y, Z
over a range of A% to B% of the nominal concentration
with an accuracy and uncertainty such that the reportable result falls
within ±1.0% of the true value
with at least a 90% probability
determined with 95% confidence
ATP Example Assay
Aligned with process validation terminology:
Three Stage Approach to Analytical Validation
Stage 1
Procedure Design (Development and Understanding
Stage 2
Procedure Performance Qualification (PPQ)
Stage 3
Continued Procedure Performance Verification
Ris
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es
sm
en
t
Kn
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An
aly
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on
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gy
Changes
According to ATP requirements
Procedure selection, development and understanding
Identification and investigation of potential analytical variables
Risk assessment
Robustness studies (Method Design Space)
Analytical Control Strategy
Knowledge gathering and preparation
Stage 1 – Procedure Design
Confirmation the analytical procedure, operated in the
routine environment is capable of delivering
reproducible data which consistently meet the ATP
Includes analytical transfer
Implementation of compendial procedures
Precision study to finalize the Analytical Control Strategy
e.g. format of the reportable result (number of determinations)
May / should be built on results generated in Stage 1
Iterative character of procedure development/optimisation
Stage 2 - Procedure Performance Qualification (PPQ)
To provide ongoing assurance that the analytical
procedure remains in a state of control throughout its
lifecycle
Routine Monitoring: Ongoing program to collect and
process data that relate to method performance, e.g.
from analysis / replication of samples or standards during batch
analysis
by trending system suitability data
by assessing precision from stability studies
[J. Ermer et al.: J. Pharm. Biomed. Anal. 38/4 (2005) 653-663]
Stage 3 – Continued Procedure Performance Verification
Risk assessment to evaluate
Impact of the respective change
Required actions to demonstrate (continued) appropriate performance
Accordingly, apply
Stage 3 (if within Method Design Space)
Stage 2 (e.g. transfer)
Stage 1 (e.g. outside Method Design Space, new procedure)
Continual Improvements (Changes)
Gregory P. Martin, (Chair) Complectors Consulting
Kimber L. Barnett, Pfizer Inc.
Christopher Burgess, Burgess Analytical Consultancy, Ltd.
Paul D. Curry, Abbvie,
Joachim Ermer, Sanofi-Aventis GmbH
Gyongyi S. Gratzl, Ben Venue Laboratories, Inc.
Elizabeth Kovacs, Apotex, Inc.
David J. LeBlond, Statistical Consultant
Rosario LoBrutto, Teva Pharmaceuticals USA
Anne K. McCasland-Keller, Eli Lilly & Company
Pauline L. McGregor, PMcG Consulting
Phil Nethercote, GlaxoSmithKline
David P. Thomas, Johnson & Johnson Pharmaceutical R&D
M. L. Jane Weitzel, Quality Analysis Consultants
Government Liaison(s): Lucinda F. Buhse, FDA
USP Scientific Liaison(s):
Todd L Cecil, Kenneth Freebern, Walter Hauck, Horacio N. Pappa, Tsion Bililign
2010-2015 V&V Expert Panel
Analytical Method Validations
Current Practices and Industry
Perspective
Rajiv A. Desai, Ph.D.
Dishman Pharmaceuticals and Chemicals Ltd.
Impurity: The procedure must be able to quantify [impurity] relative to [drug]
in the presence of components likely to present in the sample
over the range from reporting threshold to the specification limit.
The accuracy and precision of the procedure must be such that the reportable result falls
within ± X% of the true value for impurity levels from 0.05% to 0.15% with 80% probability with 95% confidence,
and within ± Y% of the true value for impurity levels >0.15%, with 90% probability determined with 95% confidence.
ATP Example Impurity
Purpose of the Analytical measurement is
to get consistent, reliable and accurate data
Origin of Impurities
Impurities in Drug Substances From Equipment and
Packaging material
Earlier stage
material
Side reactions Degradents
Residual solvents
Genotoxic Impurities
Extractables Leachables
Source of Impurities in the Drug Substance and products
General Process for the Synthesis of Drug Substance
Stage 1
Solvent W
A + B C + ( traces of A and B )
Stage 2
Solvent X
C + D E + ( traces of C and D ) + M ( reaction between A and C )
Reagent R
Stage 3
Solvent Y
E + F Crude API + ( traces of E and F ) + traces of D + degradent of E
Metal catalyst
Stage 4
Solvent Z
Crude API Final API + Traces of earlier stage material
Side reactions
Degradents
Solvents
Reagents
Analytical Method Validation Criteria ….
- Suitability of Instrument
- Status of Instrument Qualification and calibration
- Suitability of reference standard , reagent, placebo, etc
- Suitability of documentation, written analytical procedure
approved protocol with pre-established acceptance criteria
USP General Chapter <1224>
Transfer of Analytical Procedures
1. Comparative testing of same lot or standards
2. Co-validation between laboratories
3. Complete or partial validation of Analytical procedures
by receiving laboratory and hence a transfer waiver
Validation of Compendial Procedures
As per cGMP regulations 211.194(a), the test methods
with established specifications, must meet standards of
accuracy and reliability
As per 211.194(a)(2) users are not required to validate
the accuracy and reliability of these methods. But
verify their suitability under actual conditions of use.
USP General Chapter <1225>
Data Elements Required to be Validated
Analytical
Performance
Charecteristics
Category I Category II
Category III Category IV
Quantitative Limit tests
Accuracy Yes Yes * * No
Precision Yes Yes No Yes No
Specificity Yes Yes Yes * Yes
Detection limit No No Yes * No
Quantitation limit No Yes No * No
Linearity Yes Yes No * No
Range Yes Yes * * No
• May be required, depending on the nature of the specific test
Category I : Procedures for Quantitation of major component or Active substance
Category II : Procedures for determining Impurities
Category III : Procedures for determining performance characteristics ( eg., dissolution, drug release, etc )
Category IV : Identification Tests
Rationale for revisiting the compendial method
An appropriate justification for a testing procedure
Elaborating the capability of the proposed method over other
types of determinations.
For revisions, a comparison should be provided for the
limitation of the current method and advantage offered by the
new method.
USP General Chapter <1225>
Verification of Compendial Procedures
Verification for a compendial test procedure is an
assessment of whether the procedure can be used for its
intended purpose, under actual conditions of use for a
specific drug substance or drug product.
User should have the appropriate experience, knowledge
and training to understand and be able to perform the
compendial procedure.
USP General Chapter <1226>
Verification of Compendial Procedures
If the verification of the compendial procedure is not
successful and the USP staff is unable to resolve the
problem, it may be concluded that the procedure may not
be suitable for use
It may be necessary to develop and validate an alternate
procedure. This alternate method can be submitted to
USP , along with appropriate data to support the inclusion
or replacement of the current compendial procedure.
USP General Chapter <1226>
Verification of Compendial Procedures
Method verification should be based on an assessment of
the complexity of both the procedure and the material to
which the procedure is applied
Verification should assess whether the compendial method is
suitable for the drug substance and the drug product matrix.
Taking into account the drug substance synthetic route, the
method of manufacture for the drug product or both.
US General Chapter <1226>
Verification of Compendial Procedures
Drug substance from different suppliers may have different
impurity profile that may not necessarily be addressed by the
compendial method
Excepients in the drug products can vary widely among
manufacturers and may interfere directly or cause formation
of impurities that are not considered by the compendial
procedure.
US General Chapter <1226>
Chromatography
System Suitability is an integral part of chromatography
methods
These are based on the concept that equipment, electronics,
analytical operations and samples analysed constitute an
integral system that can be evaluated as such.
Factors affecting chromatography
US General Chapter <621>
Mobile phase Composition, strength , temperature, pH, flow rate
Column Flow rate, dimention, Temperature, pressure, Stationary phase
Adjustments allowed in HPLC Compendial methods
pH of Mobile phase : ± 0.2 units
Concentration of salts in buffer : within ± 10%
Ratio of components in mobile phase : ± 10%
Wavelength : ± 3 nm Column length : ± 70 %
Flow rate : ± 50% Column Temperature : ± 10 deg C
Injection volume : Can be reduced, but not increased
US General Chapter <621>
Adjustments allowed in GC Compendial methods
Gas carrier flow rate : ± 50 %
Oven temperature : ± 10%
Temperature program : ± 20 %
Column length : ± 70 %
Injection volume and split volume : Can be adjusted, if detection
and repeatability are satisfactory
US General Chapter <621>
Techniques Used for Analysis
Additional testing parameters are now considered along with the
conventional methods
Analytical Instruments moving from Research to Quality Control
NMR ICP XRD
LC-MS GC-MS NIR
Used mainly for low level detections of impurities
Method validation parameters to be selected appropriately along
with sampling and sample preparations
QbD and PAT
Quality by Design (QbD ) is being encouraged by the Regulatory
guidelines, the analysis conducted at every step of the process
needs to be reliable.
Testing methods adopted under the Process Analytical technology
(PAT) should be able to provide real time analysis in the shortest
possible time.
Validation should be definitely done for analytical methods used
under the QbD and PAT environment. No matter what the stage
of the process and not just restricted to final product.
A validated method gives assurance of process control at each
stage, concept of QbD will be further reinforced.
Compendial Method and Non-compendial Method
Compendial Method Verification / Validation
Non-compendial methods Validation
Alternate to Compendial method Validation + Equivalence
Genotoxic Compounds have a potential to damage DNA at
any level of exposure. Its scientifically proved that there
are certain chemical structures which damage the DNA.
The accepted levels of such chemicals is required to be
maintained at a very low to avoid any cause of concern.
Potential Genotoxic Impurity (PGI)
When can a specification of a drug substance exclude a limit of Potential
Genotoxic Impurity ?
1. Is just a theoretical impurity, but not found during manufacturing.
2. Is formed or introduced in intermediate steps and is controlled in the
intermediate stage and does not exceed 30% of the limit derived by TTC or
any defined acceptable limits
3. Is formed or introduced in final synthesis step, it should be included.
4. However, it is possible to apply skip test if the level does not exceed 30% of
the limit. Data of atleast 6 consecutive pilot scale batches or 3 consecutive
production batches would support the justification
Method validation becomes a very important aspect
which ever stage the analysis is performed
Guideline on the limits of genotoxic impurities' (EMEA/CHMP/QWP/251344/2006),
Potential Genotoxic Impurity (PGI)
Threshold of toxicological concern (TTC) values for genotoxic impurities
above 1.5 μg /day will be treated on a case-by-case basis. For short-duration
treatments, the acceptability of higher levels will be in line with the principles
outlined below
For more than one PGI in a drug substance, the TTC limits will be individually
applied, if the impurities are structurally different.
For more than on PGI, but structurally similar, it is expected that the mode of
action would be same, hence a sum of the limits will be accepted.
Duration of
Exposure
Single
dose
≤1 month ≤3
months
≤6
months
≤12
months
Allowable
daily intake
120 μg 60 μg 20 μg 10 μg 5 μg
Potential Genotoxic Impurity (PGI)
Your firm did not validate analytical methods used to test APIs.
The inspection revealed that your firm had not validated the HPLC
method for assay and related substances for finished API for human
use..
Your response states that XX of the APIs manufactured at your
facility, are compendial products. The remaining YY % are non-
compendial APIs had no method validation. You committed to complete
these method validations by (Date) . However, this does not address
product currently on the market, or product that will enter the market
tested with an unvalidated method. Your proposal to verify “key
parameters” for the first API batch produced does not provide the same
level of assurance as method validation.
Regulatory Audit Warning Letter
Inadequate Instrument Qualification and Analytical Method Validation
Improvements to analytical techniques and transfer of methods to at-
or on-line applications emerged as important opportunities to reduce
risk and increase efficiency in today’s modern manufacturing facility.
A pharmaceutical company was cited for not adequately performing
the required steps to support the transition to a new testing approach.
There was no method comparison or equivalency study performed to
show that the “changes were superior to the original approved
method.
The data was used for OOS closure and lot release.
Regulatory Audit Warning Letter
Four Level Control on Analysis and Results
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