Chemical quality control analysis method validation technical guidelines |
I.
Overview
To ensure drug safety,
effectiveness, quality control is the basic principles of drug development and
evaluation should be followed, Among
them, the quality control of medicines is the basis and premise to ensure the
safety and effectiveness of medicines. To achieve control The purpose of
quality control, the need for multi-angle, multi-level control of drug quality,
that is to drugs
Conduct multiple project tests to
thoroughly examine drug quality. In general, each test item is optional
Different analysis methods, in order to make the test results accurate and
reliable, we must analyze the method used Scientific, accurate and feasible
verification to fully demonstrate that the analysis method is in line with the
test project Purpose and requirements, which is commonly referred to as
validation of the method. The purpose of method validation is to determine
whether the analytical method used is scientific, reasonable and effective
The intrinsic quality of
pharmaceuticals. In essence, the method validation is based on the requirements
of the test items, pre- First set a certain content of the verification, and
through the design of a reasonable test to verify the analysis method used Can
meet the requirements of test items. Method validation plays an important role
in the establishment of analytical methods and has become a quality research
and quality The component of volume control. Only validated analytical methods
can be used to control the quality of medicines This method of verification is
the basis for the development of quality standards. Method validation is an
important part of the drug research process.
This guideline focuses on the
essentials of method validation, combining the requirements of analytical
method validation with the desired To the purpose of combining systematic andregular
elaboration, focusing on how to scientifically and reasonably Design of line
demonstration scheme. The guiding principle mainly includes the general
principles of method verification. The method verification involves three main
parties
Surface, method of verification of
the specific content of the method of verification and other content. This
principle together with other relevant technical guidelines constitute a more
complete quality control guidelines. With the continuous improvement of R &
D level of new drugs in our country, the understanding of method validation
will continue to deepen
Guiding principles will be gradually
improved and revised. Due to the particularities of biological products and
Chinese medicines, this principle is mainly applicable to chemical products.
Second, the general principles of method validation
In principle, the method of analysis
used in each test item requires method validation.
The content of the method validation
should be based on the requirements of the test items, combined with the
characteristics of the analysis method used determine. The same analysis method
used for different test items will have different validation requirements. For
example, adopted HPLC for the identification of reagents and impurities
Quantitative test methods should be different requirements of the test The
former focuses on the verification of the specialization, while the latter
focuses on the verification of the specificity, accuracy and reliability
Limits Third, the method validation
involves the three main aspects
(A) the need to verify the test items
Test items is to control the quality
of medicines, to ensure safe and effective set of test items. according to The
purpose of the test items and the verification requirements of the different
requirements of the guidelines will need to verify the test The project is
divided into identification, impurity inspection (limit test, quantitative
test), quantitative determination (content determination, Dissolution, release,
etc.), other specific test items such as four categories. The purpose of
identification is to determine that the analyte is the target compound, but not
other substances, to identify The analytical method requires a strong
specificity. Impurity inspection is mainly used to control the impurities other
than the main component, such as organic impurities, inorganic impurities.
Impurity inspection can be divided into
two kinds of limit test and quantitative test. Method of analysis for limit
testing Verification focuses on specificity and detection limits. Validation of
analytical methods used for quantitative tests emphasizes specificity and
precision
Accuracy and limit of
quantification. Quantitative determination of content, including determination
of the dissolution of the preparation, etc., due to the accuracy of such
projects
Sexual requirements are high, so the
analytical methods used require a certain degree of specificity, accuracy and
linearity. Other specific test items include particle size distribution,
optical rotation, molecular weight distribution, etc., as these seized Test
items and identification of identification, impurity inspection, quantitative
determination of different, for these projects
Analysis method validation should
have different requirements.
(B)
analysis methods
The method of analysis referred to
in this guideline is a test that has been established and established to
accomplish each of the above test items Methods, including analytical methods
and principles, instruments and equipment parameters, reagents, system
suitability tests,
Preparation of test solution,
reference solution preparation, determination, calculation and test results
report. Test methods can use chemical analysis methods and instrumental
analysis methods. These methods have their own characteristics, The same test
method can be used for different test items, but the verification content may
not be the same.
(C)
Verify the content
Verification includes method
specificity, linearity, range, accuracy, precision, limit of detection,
Quantitation, durability and system
suitability. Fourth, the method of verification of the specific content
(A)
exclusive
Specificity refers to the other
components (such as impurities, degradants, excipients, etc.) may exist, the
use of The analytical method can
correctly identify and detect the characteristics of the analyte. In general,
identification, impurity testing, content determination methods should examine
its specificity. As used
Method is not enough, should be used
to supplement a number of ways.
1, identification reaction
Identification tests should confirm that the analyte meets its characteristics.
Specificity tests require that proof be possible with possible
Presence of substances or structural
similar compounds to distinguish, to be confirmed with the analyte for the test
was positive reaction, Negative controls that do not contain the component under
test are negatively responsive, structurally similar, or related compounds in
the component
Should be negative reaction.
2, impurity inspection
As a purity check, the analytical
method used should ensure that impurities in the analyte can be detected
Quantities, such as related substances, heavy metals, organic solvents and so
on. Therefore, impurity inspection requirements analysis method has one Set the
specificity.
In the case of impurities available,
can be added to the test sample a certain amount of impurities that impurities
And coexisting substances can be separated and detected, and with appropriate
accuracy and precision. In the case of impurities or degradation products can
not be obtained, the specificity can be with another proven Reasonable but
separation or detection of different principles, or with a strong ability to
distinguish between methods to compare the results
set. Or the test sample with strong
light irradiation, high temperature, high humidity, acid, alkali hydrolysis and
oxidation method
Destruction (preparation should
consider the impact of excipients), comparing the number and amount of
impurities detected before and after damage. necessary Diode array detection
and mass spectrometry can be used for chromatographic peak purity check.
3, content determination
The purpose of the assay is to
obtain accurate results of the analyte content or potency in the test sample.
In the case of impurities available, for the determination of the main
component can be added to the test sample impurities Or excipients, examining
the determination of whether the interference, and without impurities and
accessories for the test compared to the test Set the result.
4.In the case of impurities or
degradation products can not be obtained, you can use another validated or
Pharmacopoeia Methods to compare the results of two methods compared.
Destructive testing can also be used (bright light Shooting, high temperature,
high humidity, acid, alkali hydrolysis and oxidation) to obtain a sample
containing impurities or degradation products, The two methods were used to
determine the content and compare the results. When necessary, the peak purity
of the chromatographic examination, It is proved that the chromatographic peak
of the content determination component does not contain other components.
(B)
Linear
Linear means within the scope of the
design of the test, the test results and the concentration of the analyte in
the test (Volume) is directly related to the degree of linearity.
Linearity is the basis for
quantitative determination, involving quantitative determination of the
project, such as impurity quantitative test and containing The quantity
determination needs to verify the linearity. The linear relationship should be
determined within the design measurement range. A stock solution can be used
for precision dilution, or Respectively, accurately weighed sample preparation
of a series of measured concentrations of substances were measured, prepared at
least 5 thick degree. Take the measured response signal as a function of the
analyte concentration to see if it is linear and use Least squares method for
linear regression.
When necessary, the response signal
can be mathematically converted, and then linear regression calculation, and explain
the basis.
(C)
range
Range refers to the ability to
achieve a certain degree of accuracy, precision and linearity, the test method
is suitable for the test Sample range of concentration or amount of analyte
level limit.
The range is the specified value,
the scope of the verification and the test method should be determined before
the start of the pilot study. Can be used to meet the requirements of the
preparation of different concentrations of raw materials, in accordance with
the appropriate measurement methods experimenting. The range is usually
expressed in the same units (eg percent concentration) as the test results of
the analytical method. Involving
And to the quantitative
determination of the test items need to verify the scope, such as content
determination, content uniformity Degree, dissolution or release, impurity
quantitative test.
The range should be based on dosage
forms and / or test items to determine the requirements.
1, content determination
The range should be 80% to 100% or
more of the test concentration.
2, formulation content uniformity
The range should be 70% to 130% of
the test concentration. Depending on the dosage form characteristics, such as
aerosols, sprays, When necessary, the scope may be appropriately relaxed.
3, dissolution or release For
dissolution, the range should be ± 20% of the
limit, and if the limit is specified, the lower limit -20% to + 20% of the
upper limit.
For the release, if the limit is in
the range of 20% to 24 hours after 1 hour 90%, the verification range should be
0 ~ 110%.
4, impurities
Determination of impurities, the
scope should be based on preliminary results of the test, to develop the
required limit of ± 20%. If the determination of content and impurity testing
simultaneously measured by area normalization method, the linear range should
be miscellaneous -20% of the specified limit to + 20% of the limit (or upper
limit).
(D)
accuracy
Accuracy means the approximate value
of the result measured by this method and the true value or recognized
reference value degree. Sometimes called reality.
A certain degree of accuracy is a
necessary condition for quantitative determination and thus involves the
detection of quantitative determination
Head are required to verify the accuracy,
such as content determination, impurity quantitative test.
Accuracy should be established
within the scope of the provisions for the preparation of the general recovery
rate test to test certificate. Test design to be considered within the
provisions of the preparation of three different concentrations of the sample,
the determination of 3
Times, that is, measured 9 times,
reported the recovery rate of known addition (%) or the average of the measured
results and true The difference between the real value and its credible limit.
1, content determination
The raw material medicine can use
the known purity reference substance or the qualified raw medicine to carry on
the determination, or use this The results of the method are compared with the
results of another method that has established the accuracy. The formulation
can be determined using a mixture of components containing a known amount of
the analyte. If you can not get the preparation Of the total composition can be
measured by adding a known amount of the test substance to the preparation, if
necessary, A method has been established to compare the accuracy of the
results.
2, impurity quantitative test
Quantitative testing of impurities
can be measured by adding a known amount of impurities to the drug substance or
preparation. in case
Can not get impurities, the method
can be used to determine the results compared with another mature method, such
as Pharmacopoeia Method or proven method. If you can not measure the relative
response of impurities, online measurement of impurities related data, such as
mining
UV detector with diode array
detector, when the impurity spectrum and the main component of the spectrum is
similar, You can use the API response factor approximation to calculate the
impurity content (self-control method). And should be clear single
The total amount of impurities and
impurities corresponds to the weight ratio (%) or area ratio (%) of the main
component.
(E)
precision
Precision refers to the provisions of
the test conditions, the same homogeneous test samples, after repeated sampling
A series of tests to determine the degree of closeness (dispersion).
Precision generally deviations,
standard deviations or relative standard deviation. Use standard deviation or
relative standard
When the quasi-deviation is
expressed, the sampling times should be at least 6 times.
Precision can be investigated from
three levels: repeatability, intermediate precision, reproducibility.
1, repeatability
Repeatability refers to the same
operating conditions, in a relatively short time interval, by the same analyst
The results of the determination of
the precision of the staff. Repeatability determination can be within the
specified range, at least 9 times the measurement results were evaluated, as
prepared 3 different concentrations of the sample, each measured 3 times, or
100% of the concentration level, measured with at least 6 The results of the
evaluation.
2, intermediate precision It means the same intermediate precision laboratory, due to internal conditions of the laboratory changes, such as time, Analysts,
instruments and equipment, the determination of the precision of the results. Variations
in the validation design are typically dates, analysts, equipment.
3, reproducibility
Refers to the different laboratories
between different analysts to determine the precision of the results. When
analytical methods will be adopted by statutory standards, reproducibility
tests should be carried out.
(F)
detection limit
Limit of detection refers to the
minimum amount of analyte in the sample can be detected, but not necessarily
accurate Determine the amount. The significance of the verification index is to
examine whether the method has a sensitive detection capability. So mixed
Quality limit test, to prove that
the method has a sufficiently low detection limit to ensure that the detection
of impurities to be controlled.
1, intuitive method
Intuitive evaluation can be used for
non-instrumental analysis methods as well as instrumental analysis methods.
The detection limit is determined by
analyzing a series of test specimens of known concentration and using Can
accurately and reliably detect the minimum or minimum concentration of the
analyte to establish.
2, signal to noise ratio method
For the analysis of the baseline
noise can be displayed that the known low-concentration samples detected signal
and The noise signal is compared to calculate the lowest detectable
concentration or amount. The general signal to noise ratio of 3: 1
The corresponding concentration or
the amount of equipment injected determines the limit of detection. Other
methods are based on the slope of the working curve and the standard deviation
of the response method Wait. Regardless of the method used, a certain number of
samples are applied at concentrations near or equal to the detection Limits,
analysis is performed to reliably determine the limit of detection.
(7)
Quantitative limit
The limit of quantification refers
to the minimum amount of analyte in a sample that can be quantitatively
determined, and the result of the measurement Should have a certain accuracy
and precision. Quantitative limit reflects the analytical method has the
ability to sensitive quantitative detection. Impurity quantitative test, The
limit of quantitation of the method needs to be investigated to ensure that
impurities with very low contents can be accurately detected.
Common signal to noise ratio to
determine the quantitative limit. The general signal to noise ratio of 10: 1
corresponding concentration or Note Enter the amount of equipment to be
determined.
1, intuitive method
Intuitive evaluation can be used for
non-instrumental analysis methods as well as instrumental analysis methods. The
limit of quantitation is generally determined by analyzing a series of samples
containing analytes of known concentration
Accuracy and precision are required
to meet the requirements of the circumstances, to determine the minimum amount
of measurable substance can be quantified.
2, signal to noise ratio method
For the analysis of the baseline
noise can be displayed that the known low-concentration samples detected signal
and The noise signal is compared to calculate the lowest detectable
concentration or amount. The general signal to noise ratio of 10: 1. Other
methods are based on the slope of the working curve and the standard deviation
of the response method Wait. Regardless of the method used, a certain number of
samples are applied at concentrations close to or equal to the quantitation Limits
for analysis to reliably determine the limit of quantitation.
(H)
Durability
When small changes in durability means the measurement conditions occur,
the result of the measurement is not affected tolerance level. Durability
mainly investigates the anti-interference ability of the method itself for
variable test factors. Start researching When analyzing methods, consider their
durability. If the test conditions demanding, it is recommended in the method
In the statement. Typical variables
include: the composition, flow rate, and pH of the mobile phase in liquid
chromatography, not With the same brand or different batch number of the same
column, such as column temperature. Gas chromatography and carrier gas flow
rate,
Different brands or lot numbers,
stationary phase, support, column temperature, inlet and detector temperature. After
testing, should indicate small changes in line with the system suitability test
requirements to ensure that the method effective.
(IX)
System suitability test
For some instrument analysis
methods, when performing method validation, it is necessary to integrate the
analytical equipment, electronics
The instrument is evaluated as a complete
system together with experimental procedures, test samples and the like. System
suitability It is an indicator of the entire system to be assessed. System
suitability test parameters set to betested Depending on the type of card
method.
Chromatography methods for
analytical equipment, electronic instruments rely on a high degree, so all the
chromatographic methods This indicator should be validated and the system
suitability as part of the analytical method. Experience Card parameters and
methods refer to the relevant provisions of Chinese Pharmacopoeia. Fifth, the
method re-verification In some cases, such as the change of raw material drug
synthesis process, preparation prescription changes, analytical methods
Health changes, etc., are necessary
to once again analyze the analytical methods in full or in part to ensure that
the points Method is reliable, this process is called method re-verification.
Revalidation principle: According to
the degree of change corresponding revalidation.
When the API synthesis process
changes, the introduction of new impurities, impurity inspection methods and The
specificity of the content determination method needs to be verified to prove
that the relevant material inspection method can Newly introduced impurities
are to be detected and the newly introduced
impurities should be free from interference
with the determination of the content of the main components. When the
formulation changes the composition of prescriptions, accessories change, may
affect the identification of the specificity, dissolution
Outcome and determination of the
accuracy of the content, so the need for identification, determination of
content verification method. When the original
Changes in origin of feedstocks may
affect the specificity and accuracy of impurity testing and determination
Therefore, it is necessary to
re-verify the impurity inspection method and content determination method. When
the quality standard of a project analysis method partial change, such as the
use of high-performance liquid color When spectrum content is measured, the
detection wavelength changes, you need to re-test limits, specificity, Accuracy,
precision, linearity and other content verification, proof of the rationality
of the revised analysis method can be Line.
Similarly, there have been national
standards of drug quality research, based on the declaration of raw materials
synthesis process, Preparation of prescriptions and other accessories generally
can not guarantee the consistency of the drugs already on the market, the need
for quality standards The middle part of the project to re-validate the method.
Method validation is the process of
perfection of analytical methods and should be re-tested based on actual
changes
Card to ensure that the analytical
methods used to control the intrinsic quality of the drug.
Sixth, the method validation
evaluation For method validation, the following aspects deserve attention.
(A)
General considerations on methodological validation and evaluation
In general, method validation should
focus on the purpose and general principles of validation, method validation
content The choice and test design should be systematic and reasonable, and the
verification process should be rigorously regulated. Not all methods of
analysis for each test item require verification of all content, but at the
same time Note that the verification should be sufficient to justify the
analytical method used. Such as impurity limits test
Generally need to verify the
specificity and detection limit, and for precision, linearity, limit of
quantification involved The measurement of the project, you generally do not
need to verify.
(B)
method of verification of the integrity and systematic
Method validation content
interrelated, is a whole. Therefore, no matter from the perspective of research
or development Evaluation point of view, method validation are focused on the
integrity and systematic. For example, to identify the specificities needed for
a project, it is generally not likely that an analytical method Fully identify
the analyte, this time using two or more analytical methods can enhance the
identification of the entire project
Body specificity. There is also a
greater correlation between method validation content that can be complemented
with each other. Such as bulk drugs Determination of content using volume
analysis, due to the method itself, the specificity of slightly worse, but if
in the Impurity detection using a more specific chromatography, it is generally
considered the detection method also has Strong specificity.
In short, due to the complexity of
the actual situation, it is not advocated dogmaily during the verification of
the method Line method validation. In addition, more and more new methods are
constantly being used in quality control for these
How to verify the method requires
specific analysis of the specific circumstances, and can not copy the
guidelines. Seven, reference
1. FDA.Guidance for Industry:
analytical procedures and methods
validation, chemistry,
manufacturing, and controls documentation (Draft), 2000.8.
2. ICH Q2A.Test on Validation of
Analytical Procedures 
3. ICH Q2B.Validation of Analytical
Procedures: Methodology
4. Chinese Pharmacopoeia 2000
edition two appendices. Drug quality standard analysis method validation
edition two appendices. Drug quality standard analysis method validation
Eight, author
Study Group on Technical Guiding
Principles for Validation of Chemical Quality Control Analysis Methods
