方法验证（详细参数,英文）

方法验证（详细参数,英文）
Category I Category III Category IV Definition Validation Procedure Acceptance Criteria Verification
Target of analytical
methods Assay
Impurity
Quantitative
Impurity
Limit Tests
Dissolution Identification
Accuracy Yes Yes**No The nearness of a measured
value to the true value
Spike certain amount and calculate the
percentage recovery (3
concentrations/three replicates, minimum
9 determinations over minimum of three
concentration levels per ICH).
For Assay: 5 Conc. X 3 replicates
98% - 102%
For Impurity: 3 Conc. X 3 replicates
85% - 115%
*
Precision Yes Yes No Yes No Closeness of agreement
between a series
measurements
Repeatability, Intermediate precision, and
reproducibility
See Below for Detail Yes
Repeatability Yes Yes No Yes No Measure of the precision
under
the same operationconditions
over a short interval of time
(with the same equipment)
%RSD (3 concentrations/three replicates,
minimum 9 determinations over minimum of three concentration levels, or minimum of 6 determinations at 100% of the test
concentration per ICH
Assay (Category I) NMT 2%
Impurity (Category II) NMT 15% or
Based on repeatability of instruments
Yes
Intermediate Precision Yes Yes No Yes No
Measure the variation within
the same laboratory, including
Day-to-day variation, analyst
variation, and equipment
variation
SD, %RSD, and Confidence interval
The obtained variability that is in the
same range or less than repeatability
variation.
Yes
Reproducibility Yes Yes No Yes No
Measures the precision
between laboratories (Method
transfer between laboratories)
Means of repeatability data
The means of the two laboratories
are not significant different from each other
Yes
Specificity Yes Yes Yes*Yes
The ability to assess
unequivocally an analyst in the presence of componenets
thatmay be expected to be
present
1. When Impurities are available:
1.1 Assay Method: Comparing test results between Sample with Impurities and sample without impurities.
1.2 Impurities Method: Spike drug substance with the appropriate level of impurities (LOQ);
2. When Impurities are not available, demonstrated specificity by comparing the test results of samples containing impurities to a second well characterized procedure or other validated procedure, should include samples stored under relevant stress conditions:
2.1 Assay method: The two results should be compared;
2.2 Impurities Method: The impurity profiles should be compared and peak purity check (using DAD or MSD) should be performed;
1. When Impurities are available:
1.1 Assay Method: Impurities peaks
need to be resolved from the analyte.
Impurities do not need to resolved
from each other;
1.2 Impurities Method: impurities are
determined with appropriate
accuracy, precision, and seperation
of these impurities individually;
2. When Impurities are not
available
2.1 Assay method: The two results
should be compared;
2.2 Impurities Method: The impurity
profiles should be compared and
peak purity check (using DAD or
MSD) should be performed to shown
that the analyte chromatographic
peak is not attributable to more than
one component
*
Detection Limit (LOD)No No Yes*No The lowest amount of analyte
in a sample that can be
detected, but not necessarily
quantitated, under the stated
experimental conditions.
1. For noninstrumental procedure:
generally determined by the analysis of
samples with known concentration of
analyte;
2. For instrumental procedures: The detection limit is shown to be sufficiently low by the analysis of samples with known concentrations of analyte above and below the required detection level. Usually at least 3-5 times lower than the limit (20% to 33% of the required limit). Prepare several (at least 5, recommend 7) samples at the LOD level and test.
1. Visual evaluation for noninstrumental procedures and instrumental procedures;
2. Signal-to-noise ratio of 3:1 at least;
3. LOD = 3σ/S, where σ is the
standard deviation of multiple blank samples response and the S is the
slope of the calibration curve. Usually
the RSD of LOD level is NMT 10%
for HPLC method
No
Quantitation Limit
(LOQ)No Yes No*No
A characteristic of quantitative
assays for low levels of
compounds in sample
materices, such as impurities
in bulk drug substances and degradation products in
finished pharmaceuticals.
1. For noninstrumental procedure: generally determined by the analysis of samples with known concentration of analyte;
2. For instrumental procedures: The quantitation limit is shown to be sufficiently low by the analysis of samples with known concentrations of analyte above and below the required quantitation level, and the procedure will reliably quantitate the analyte at that level;
Usually at least 1-2 times lower than the limit (50% to 100% of the required limit). Prepare several (at least 5, recommend
7) samples at the LOD level and test.
1. For noninstrumental procedure, the analyte can be determined with acceptable accuracy and precision;
2. Signal-to-noise ratio of 10:1 at
least;
3. LOQ = 10σ/S, where σ is the
standard deviation of multiple blank samples response and the S is the
slope of the calibration curve. Usually
the RSD of LOD level is NMT 5% for HPLC method
No
Linearity Yes Yes No*No
The ability of an analytical
procedure to elicit test results
that are directly or by a well-
defined mathematical transformation, proportional to
the concentration of analyte in
samples within a given range.
In the other way, is the linearity
of the relationship of
concentration and assay measurement.
Should be established across the range of the analytical procedure.
At least 5 concentrations are used to determine the linearity:
1. For assay method: at least cover 80%
to 120% of the test concentration;
2. Impurity method: at least cover from 50% to 120% of the acceptance criterion;
3. For dissolution test: ± 20% over the specified range, for example: 30%@1h, and 90%@24h, so the range will be 10% to 110%;
4. For Metered-dose inhalers content uniformity, a minimum 70% to 130% of the test concentration, or wider;
1. Calculate the regression line by the method of least squares, and the co- relation co-efficient (R, not R square)
is NLT 0.99, and co-efficient R
square is NLT 0.997, the y-intercept should be closed to origin.
2. ICH requires to report Slope, Residual Sum of squares, and y- intercept
No
Range Yes Yes**No
The range of an analytical
procedure is the interval
between the upper and lower
levels of analyte (including
these levels) that have been demonstrated to be
determined with a suitable
level of precision, accuracy,
and linearity.
Should be established across the range of the analytical procedure.
At least 5 concentrations are used to determine the linearity:
1. For assay method: at least cover 80%
to 120% of the test concentration;
2. Impurity method: at least cover from 50% to 120% of the acceptance criterion;
3. For dissolution test: ± 20% over the specified range, for example: 30%@1h, and 90%@24h, so the range will be 10% to 110%;
4. For Metered-dose inhalers content uniformity, a minimum 70% to 130% of the test concentration, or wider;
At each level, multiple trials are performed
to access a suitable level of precision, accuracy, and linearity.
1. Calculate the regression line by the method of least squares, and the co- relation co-efficient (R, not R square)
is NLT 0.99, and co-efficient R
square is NLT 0.997, the y-intercept should be closed to origin.
2. The RSD of multiple trials at each level are the same level or lower the level of variation in Repeatability
No
Robustness (System
suitability determination)*****
The measure of the analytical
method to remain unaffected
by small but deliberate
variations in method
parameters and provides an
indication of its reliability during normal usage
1. Sample preparation: Extration time, pH ± 0.05 unit, solvent composition ±2% absolute, with/without membrane filters, Sample and standard stability, and so on;
2. HPLC Methods: Mobile-phase composition (pH ± 0.05 unit, solvent composition ±2% absolute, and so on), different column used, Temperature (±2°C), Flow rate (± 10%);
3. GC Methods: different column used,
Temperature (± 2°C), Flow rate (± 10%);
Still meet the required system
suitability, such as resolution between
peaks, relative retentions, or tailing
factor, signal-to-noise ratio, and so
on.
*
*
Category II
May be required, depending on the nature of the specific test Validation of Analytic Methods for Pharmaceutical Analysis Other Characteristics。