Transcript Guidelines - Good laboratory practice

Pharmaceutical Development with Focus
on Paediatric formulations
WHO/FIP Training Workshop
Hyatt Regency Hotel
Sahar Airport Road
Andheri East, Mumbai, India
28 April 2008 – 2 May 2008
1|
Birgit Schmauser | April 2008
Analytical Method Development
Presented by:
Birgit Schmauser, PhD
Federal Institute for Drugs
and Medical Devices (BfArM)
[email protected]
2|
Birgit Schmauser | April 2008
Analytical Method Development
In this presentation:
 Standards in developing analytical methods for
– Originator and multisource generic FPPs
• Specifications
• Stability
 Parallel development of analytical methods for
cleaning validation
3|
Birgit Schmauser | April 2008
Analytical Method Development
 Originator, First-time Generic and Multisource Generic
Originator
First-time Generic Multisource Generic
Originator´s
specifications
Information from
regulatory agencies
(publicly available) &
literature data
Pharmacopoeias
FPP quality Originator´s
standards specifications
Information from
regulatory agencies
(publicly available) &
literature data
Pharmacopoeias
Analytical
methods
Derive identity, potency, Verify identity, potency,
purity of API and FPP
purity of API and FPP by
by in house methods
pharmacopoeial methods
and in-house methods
API quality
standards
4|
Establish identity,
potency, purity of
API and FPP by
in-house methods
Birgit Schmauser | April 2008
Analytical Method Development
 HPLC-method to assay potency and purity – risk assessment
Originator
First-time Generic
Multisource Generic
Characterise all
impurities/degradants
Calculate Response factors
(qualification by clinical
use)
Derive impurities/degradants
from Originator
Characterize „in-house“
impurities/degradants
Calculate response factors
Verify impurities from
Pharmacopoeia
Characterise „in-house“
impurities/degradants
(Response factors)
Establish reference
materials
Extract (& reproduce) reference Use pharmacopoeial
materials
reference materials
Adapt to routine use
Adapt/modify to/for routine use „Implement“ for routine use
Selectively screen/detect
any impurity or degradant
Establish potency
Identify
impurities/degradants
5|
Birgit Schmauser | April 2008
Analytical Method Development
Interchangeability (IC) of multisource generic FPPs
(Essential similarity with Innovator FPP)
Pharmaceutical + Bioequivalence
Equivalence
IC = PE + BE
6|
Birgit Schmauser | April 2008
Analytical Method Development
 Pharmaceutical equivalence
– FPPs meet the same or comparable standards by
use of equivalent analytical methods
• Same API (chemical and physical equivalence)
• Same dosage form and route of administration
• Same strength
• Comparable labeling
– Equivalence in pharmaceutical development
– Equivalence in stability
– Equivalence in manufacture (WHO-GMP)
7|
Birgit Schmauser | April 2008
Analytical Method Development
 Prequalification requirements
– Validation of analytical methods is a prerequisite for
prequalification of product dossiers
• Non-compendial APIs and FPPs are tested with methods developed by the
manufacturer
• For compendial APIs and FPPs the „applicability“ of pharmacopoeial methods to
particular products must be demonstrated (verification)
– Analytical methods must be developed and validated
according to TRS 823, Annex 5, Validation of analytical
procedures used in the examination of pharmaceutical
materials ; ICH Q2 (R1)
• To be used within GLP and GMP environments
8|
Birgit Schmauser | April 2008
Analytical Method Development
Use of analytical methods - generics
CLINICAL
PHARMACEUTICAL
METHODS
At initial phase of pharmaceutical development
To determine
bioavailability in
healthy volunteers
To develop a stable and
reproducible formulation for
the manufacture of
bioequivalence, dissolution,
stability and pilot-scale
validation batches
To understand the profile of related
substances and to study stability
To start measuring the impact of
key product and manufacturing
process parameters on consistent
FPP quality
At advanced phase of pharmaceutical development
To prove
bioequivalence after
critical variations to
the prequalified
dossier
9|
To optimise, scale-up and
transfer a stable and
controlled manufacturing
process for the
prequalification product
Birgit Schmauser | April 2008
To be robust, transferable, accurate
and precise for specification
setting, stability assessment and
QC release of prequalified product
batches
Analytical Method Development
 Prerequisites for analytical method validation
– Six “M”s
Machine
Man
Methods
calibrated
qualified
robust
skilled
Reference
standards
Quality
Material
10 |
qualified
Milieu
Birgit Schmauser | April 2008
documented
suitable
Quality of the
analytical method
Vibrations
Temperature
characterised
Irradiations
Humidity
Time
Analysts´
support
Supplies
Management
Analytical Method Development
 Method development life cycle
Planning
Method development
Development and Validation Policy
Initital Method Development
Pre-Validation Evaluation
Objectives/Requirements of Method
Development
Plan –
Project
Information Gathering
Resource Gathering
Validation Experiments
Method Transfer
Experiments
Method Optimization
Robustness
System Suitability
Customer Evaluation
Testing
Filed Method in Use
From: Analytical Chemistry in a GMP Environment. Edited by J.M. Miller and J.B. Crowther, ISBN 0-471-31431-5, Wiley & Sons Inc.
11 |
Birgit Schmauser | April 2008
Periodically
Monitoring/Review
of Methods
in Control Labs
Analytical Method Development
 Validation should verify the suitability of an
analytical method for its intended purpose
 Validation should be founded on method
development performed beforehand that
suggest the suitability and robustness of the
method
 Validation may be performed in different ways
(individual purpose) according to common
standards
12 |
Birgit Schmauser | April 2008
 Validation protocol
– Method principle / objective
– Listing of responsibilities
• Laboratories involved and their role in the validation
– Method categorization
– List of reagents (including test lots) and standards
– Test procedures to evaluate each validation parameter and proposed
acceptance criteria
– Plan or procedure when acceptance criteria are not met
– Requirements for the final report
 The validation process cannot proceed until the protocol and
all parties involved approve the acceptance criteria
13 |
Birgit Schmauser | April 2008
Analytical Method Development
 Innovator versus Generics
Innovator
Generics
R & D on API
+
-
Preclinical trials
+
-
Clinical trials phase I and II
Method validation
summary
-
Clinical trials phase III
Method validation
completed
-
Post marketing phase IV
Validated methods
-
Entering of Generics;
Pharmaceutical development,
Comparability with Innovator
Validated methods
Validated methods:
GMP and GLP
14 |
Birgit Schmauser | April 2008
Analytical Method Development
 Validation Characteristics
Identification
Impurities
Assay
quantitative
limit
Accuracy
-
+
-
+
Precision
-
+
-
+
Specificity
+
+
+
+
Detection Limit
-
-
+
-
Quantitation Limit
-
+
-
-
Linearity
-
+
-
+
Range
-
+
-
+
Robustness
+
+
+
+
15 |
Birgit Schmauser | April 2008
Analytical Method Development
 Accuracy and precision
Accurate &
precise
16 |
Birgit Schmauser | April 2008
Accurate &
imprecise
Inaccurate &
precise
Inaccurate & imprecise
Analytical Method Development
 Precision
– Expresses the closeness of agreement between a series of
measurements obtained from multiple sampling of the same
homogenous sample
– Is usually expressed as the standard deviation (S), variance
(S2) or coefficient of variation (RSD) of a series of
measurements
– Precision may be considered at three levels
• Repeatability (intra-assay precision)
• Intermediate Precision (variability within a laboratory)
• Reproducibility (precision between laboratories)
17 |
Birgit Schmauser | April 2008
Analytical Method Development
 Normal distribution, probability function [P(x)]
and confidence interval [CI]
xn
P
s
0.6826895
2s 0.9544997
3s 0.9973002
4s 0.9999366
5s 0.9999994
 An interval of ± 3 s
covers 99.73% of values
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Birgit Schmauser | April 2008
Number of times each value occurs
– Probability (P), that measurements from a normal distribution fall within [µ-xn, µ+xn]
for xn = ns is described by the “erf-function” (µ = mean):
3s
2s
s Values s
2s
3s
Analytical Method Development
 Normal distribution, probability function [P(x)] and confidence
interval [CI]
– Probability-P Confidence interval [CI]
centered around the mean [µ]
in units of sigma [s] described by
“inverse erf-function”:
– A CI of 95% includes values
± 1.95 s around the mean
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Birgit Schmauser | April 2008
P
xp
0.800
1.28155s
0.900
1.64485s
0.950
1.95996s
0.995
2.57583s
0.999
3.29053s
Analytical Method Development
 Relationship of variability, probability and reliability of data
– High variability of data (large s) generate large confidence intervals and
thus lower the reliability of the mean
– Low variability of data (small s) generate small confidence intervals and
thus increase the reliability of the mean
20 |
Birgit Schmauser | April 2008
Analytical Method Development
 Repeatability
– Six replicate sample preparation steps from a homogenously prepared tablet
mixture (nominal value of API 150 mg)
Injection Peak area Assay
1
173865
147.10 mg/98.06%
2
174926
148.00 mg/98.66%
3
172933
146.32 mg/97.54%
4
175011
148.08 mg/98.72%
5
179557
151.95 mg/101.30%
6
176425
149.28 mg/99.52%
Mean
175453
148.45 mg/98.96%
SD (s)
2329
1.98 mg/1.32%
RSD
1.32%
1.32%
21 |
Birgit Schmauser | April 2008
Mean ± 3 SD =
Confidence interval of 99.73%
98.96 ± 3x1.32% = 95% - 102.92%
Analytical Method Development
 Intermediate precision
– Expresses within-laboratories variations (different days, different analysts,
different equipment etc.)
Injection
1
2
3
4
5
6
Mean
SD (s)
RSD
22 |
Peak area
analyst 1
173865
174926
172933
175011
179557
176425
175453
2329
1.32%
Birgit Schmauser | April 2008
Peak area
analyst 2
175656
175878
176004
176344
175332
174959
175695
495
0.28%
Peak area
analyst 3
177965
178556
177342
178011
179466
179688
178504
918
0.51%
Mean ± 3 SD: (177252  100%)
Analyst 1: 98.96% ± 3 x 1.32%
Analyst 2: 99.12% ± 3 x 0.28
Analyst 3: 100.70% ± 3 x 0.51
Average of 3 analysts ± 3SD:
95% - 102.23%
Analytical Method Development
 Reproducibility
– Expresses the precision between laboratories
• Collaborative studies, usually applied to standardisation of
methodology
– Transfer of technology
– Compendial methods
23 |
Birgit Schmauser | April 2008
Analytical Method Development
 Accuracy
– Expresses 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
• Sometimes referred to as „TRUENESS“
mean
24 |
Birgit Schmauser | April 2008
true
Analytical Method Development
To find out whether a method is accurate:
 Drug substance (assay)
– Application of the method to an analyte of known purity (e.g. reference
substance)
– Comparison of the results of one method with those of a second wellcharacterised method (accuracy known)
 Drug product (assay)
– Application of the method to synthetic mixtures of the drug product component
to which known quantities of the analyte have been added
• Drug product may exceptionally be used as matrix
 Drug substance/Drug product (Impurities)
– Application of the method to samples spiked with known amounts of impurities
25 |
Birgit Schmauser | April 2008
Analytical Method Development
 Accuracy: Application of the method to synthetic mixtures of the
drug product components
to which known quantities
of the analyte
have been added
 Recovery reduced
by ~10 – 15%
From: Analytical Method Validation and Instrument Performance Verification, Edited by Chung Chow Chan,Herman Lam, Y.C.
Lee and Xue-Ming Zhang, ISBN 0-471-25953-5, Wiley & Sons
26 |
Birgit Schmauser | April 2008
Analytical Method Development
 When to expect Accuracy problems
– Insufficient selectivity of the method
• Impurity peaks are not resolved and account for assay value
– Recovery is < 100%
• Irreversible adsorption of analyte to surfaces of the system
– Incorrect assay value of a reference standard
• Due to decomposition of reference standard
– Incorrect assay value due to change in matrix
• Analytical laboratory still uses the preceding matrix as standard
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Birgit Schmauser | April 2008
Analytical Method Development
 Specificity
– Is the ability to assess unequivocally the analyte in the presence of components
which may be expected to be present (impurities, degradants, matrix…)
Identity testing
– To ensure the identity of an analyte
Purity testing
– To ensure accurate statement on the content of impurities of an analyte
Assay
– To allow an accurate statement on the content of an analyte in a sample
28 |
Birgit Schmauser | April 2008
Analytical Method Development
 Specificity: Overlay chromatogram of an impurity solution with a
sample solution
From: Analytical Method Validation and Instrument Performance Verification, Edited by Chung Chow Chan,Herman Lam, Y.C.
Lee and Xue-Ming Zhang, ISBN 0-471-25953-5, Wiley & Sons
29 |
Birgit Schmauser | April 2008
Analytical Method Development
Specificity and stability
 Stress stability testing to ensure the stability indicating potential of an
analytical method
– Apply diverse stress factors to the API
– Apply diverse stress factors to the FPP
Stress conditions: e.g. Supplement 2 of Generic Guideline; TRS 929, Annex 5
Assure that the API can be assessed specifically in the presence of
known and unknown (generated by stress) impurities
Assure that known impurities/degradants can be specifically assessed
in the presence of further degradants
By peak purity assessment and (overlay of) chromatograms
30 |
Birgit Schmauser | April 2008
Analytical Method Development
 Stress stability studies versus forced degradation studies
Stress
parameter
Forced degradation
Stress stability
(5 – 15% decomposition)
Acid
0.2 ml 1N HCl / 5 ml API-solution /
3h, 6h, 12h, 24h…7d (RT & 60°C)
pH ± 2 (2 weeks)
Base
0.2 ml 1N NaOH / 5 ml API-solution /
3h, 6h, 12h, 24h…7d (RT & 60°C)
pH ± 10 (2 weeks)
H2O2 /
Oxygen
0.2 ml 5% or 35% H2O2 / 5 ml APIsolution (RT, to 7d & 60°C, 3h)
1 g/ml oxygen bubbled through (8 hours)
0.1 – 2% H2O2 (24 hours)
Heat
60°C / 5 ml solution (3h, 6h…7d)
-
Heat
105° C / solid API (1d and 7d)
60°C (4 weeks)
UV or Light
365 nm or white fluorescent light /
solid API (1d and 7d)
Humidity
-
31 |
Birgit Schmauser | April 2008
50°C / 80% RH (4 weeks)
Analytical Method Development
 Limit of Detection (LOD, DL)
– The LOD of an analytical procedure is the lowest amount of analyte in sample
which can be detected but not necessarily quantitated as an exact value
 Determination is usually based on
– Signal to noise ratio (~3:1) (baseline noise)
or
– Standard deviation of response (s) and Slope (S)
• 3.3 s/S
32 |
Birgit Schmauser | April 2008
Analytical Method Development
 Limit of Quantitation (LOQ, QL)
– The LOQ is the lowest amount of analyte in a sample which can
be quantitatively determined with suitable precision and accuracy
• The quantitation limit is used particularly for the determination of
impurities and/or degradation products
 Determination is usually based on
– Signal to noise ratio (~10:1) (baseline noise)
or
– Standard deviation of response (s) and Slope (S)
• 10 s/S
33 |
Birgit Schmauser | April 2008
Analytical Method Development
LOD, LOQ and Signal to Noise Ratio (SNR)
LOQ
Signal to Noise = 10:1
Signal to Noise = 3:1
LOD
Noise
34 |
Birgit Schmauser | April 2008
Analytical Method Development
 LOQ
– Quantitation by SNR is accepted
– Quantitation by Standard deviation of response (s) and Slope (S)
(10 s/S) is more adequate as it involves the response of the actual
analyte
– Best to calculate in the region close to y-intercept
35 |
Birgit Schmauser | April 2008
Analytical Method Development
 LOQ and impurities
– In determination of impurities in APIs and FPPs the LOQ should
be determined in the presence of API
• LOQ should be NMT reporting level
• LOQ should be given relative to the test concentration of API
– Specificity of impurity determination should always be
demonstrated in the presence of API at API specification levels
• Spiking of test concentration (API/FPP) with impurities at levels of their
specification range
36 |
Birgit Schmauser | April 2008
Analytical Method Development
 Spiking
– API test concentration (normalised)
• 0.1 mg/ml (100%)
– Impurity spiking concentrations
• 0.001 mg/ml (1%) – specification limit
• 0.0001 mg/ml (0.1%) – limit of quantitation (minimum requirement)
API at test concentrations
API below test concentrations
37 |
Birgit Schmauser | April 2008
Analytical Method Development
 Linearity
of an analytical procedure is its ability (within a given range)
to obtain test results which are directly proportional to the
concentration (amount) of analyte in the sample
– If there is a linear relationship test results should be
evaluated by appropriate statistical methods
•
•
•
•
•
38 |
Correlation coefficient (r)
Y-intercept
Slope of regression line
Residual sum of squares
PLOT OF THE DATA
Birgit Schmauser | April 2008
Analytical Method Development
 Usual acceptance criteria for a linear calibration
curve
– r > 0.999; y-intercept a < 0 to 5% of target concentration
RSD (wrt calibration curve) < 1.5-2%
r > 0.997
r < 0.997
From: Analytical Method Validation and Instrument Performance Verification, Edited by Chung Chow Chan,Herman Lam, Y.C. Lee and
Xue-Ming Zhang, ISBN 0-471-25953-5, Wiley & Sons
39 |
Birgit Schmauser | April 2008
Analytical Method Development
 Range
– The range of an analytical procedure is the interval
between the upper and lower concentration (amounts) of
analyte in the sample for which it has been demonstrated
that the analytical procedure has a suitable level of
precision, accuracy and linearity
40 |
Birgit Schmauser | April 2008
Analytical Method Development
 Range
– Assay
• 80 to 120% of test concentration
– Content uniformity
• 70 to 130% of test concentration
– Dissolution
• Q-20% to 120%
– Impurities
• Reporting level – 120% of specification limit (with respect
to test concentration of API)
– Assay & Impurities
• Reporting level to 120% of assay specification
41 |
Birgit Schmauser | April 2008
Analytical Method Development
 Linearity is limited to 150%of shelf life specification of impurities
– Test concentration can be
used to determine impurities
To determine drug substance
(assay) the test concentration
must be diluted
The range is 0 – ~ 150% of
impurity specification
From: Analytical Method Validation and Instrument Performance Verification, Edited by Chung Chow Chan,Herman Lam, Y.C.
Lee and Xue-Ming Zhang, ISBN 0-471-25953-5, Wiley & Sons
42 |
Birgit Schmauser | April 2008
Analytical Method Development
 Robustness
– Robustness of an analytical procedure should show
the reliability of an analysis with respect to deliberate
variations in method parameters
– The evaluation of robustness should be considered
during the development phase
– If measurements are susceptible to variations in
analytical conditions the analytical conditions should
be suitably controlled or a precautionary statement
should be included in the procedure
43 |
Birgit Schmauser | April 2008
Analytical Method Development
 Influence of buffer pH and buffer concentration in mobile phase
on retention times of API and impurities
API
Impurity A
Impurity B
Impurity C
As is
10.46
3.86
7.43
8.26
buffer pH 5.9
10.45
3.94
7.51
8.38
buffer pH 6.9
10.46
3.94
7.49
8.34
Buffer conc. 83%
7.84
3.43
6.16
6.66
Buffer conc. 87%
15.26
4.77
9.61
11.18
 Conclusion: The buffer composition should be maintained in a range of
85 ± 0.5%
– Missing: Acceptance criterion for maximal deviation of retention time should be
defined unless justified
44 |
Birgit Schmauser | April 2008
Analytical Method Development
 System suitability testing
– Based on the concept that equipment, electronics,
analytical operations and samples to be analysed
constitute an integral system that can be evaluated as
such
– Suitability parameters are established for each analytical
procedure individually
• Depend on the type of analytical procedure
45 |
Birgit Schmauser | April 2008
Analytical Method Development
 Method stability
– System suitability over time
• Sample solution stability
– A solution of stavudine is stable for ~ 2 h, then it starts to degrade
to thymine
• Impurity-spiked sample solution stability
A solution containing stavudine spiked with its impurity thymine does not allow to
clearly distinguish between degradation and spike
A solution containing stavudine of a FPP-stability sample solution does not allow
to clearly distinguish between FPP-stability degradation and sample solution
degradation
Should be analysed immediately
46 |
Birgit Schmauser | April 2008
Analytical Method Development
 When to be „surprised“ about validation data:
47 |
– Precision of
impurity determination
System precision
% RSD 0.33 – 2.25
Method precision
% RSD 0.0
– Precision of
API determination
Average peak area
% RSD 0.08
– Method precision of
released API (dissolution)
Average peak area
Birgit Schmauser | April 2008
Acceptance criterion % RSD ≤ 2.0
% RSD 0.4
Acceptance criterion % RSD ≤ 10.0
Analytical Method Development
 Specification range (USL-LSL)
– Process variability (usually ± 2 SD)
– Analytical variability (± 3s)
• ~ NMT 30% of total specification range
 Analytical variability
Process variability
– Reliability of evaluation of major process variables by analytical
procedures depends on analytical variability
– Impurities
• LOQ and specification limit (e.g. qualification limits NMT 0.15%)
– Response factors (LOQ modified by response factor)
48 |
Birgit Schmauser | April 2008
Analytical Method Development
 Methods for cleaning validation
– Method for assay and related substances used in stability studies of API
and FPP
• Specificity (in samples taken from a cleaning assessment)
• Linearity of response (from 50% of the cleaning limit to 10x this concentration; R2 ≥ 0.9900)
• Precision
– Repeatability (RSD ≤ 5%)
– intermediate precision [ruggedness (USP)]
– Reproducibility
• Limits of detection and quantitation
• Accuracy or recovery from rinsate (≥ 80%), swabs (≥ 90%), and process surface (≥ 70%)
• Range (lowest level is at least 2x higher than LOQ)
49 |
Birgit Schmauser | April 2008
Analytical Method Development
Summary
 Analytical procedures play a critical role in pharmaceutical
equivalence and risk assessment/management
– Establishment of product-specific acceptance criteria
– Assessment of stability of APIs and FPPs
 Validation of analytical procedures should demonstrate that
they are suitable for their intended use
 Validation of analytical procedures deserves special attention
during assessment of dossiers for prequalification
50 |
Birgit Schmauser | April 2008
Analytical Method Development
THANK YOU
51 |
Birgit Schmauser | April 2008