Chemical quality control analysis method validation technical guidelines

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

Eight, author

Study Group on Technical Guiding Principles for Validation of Chemical Quality Control Analysis Methods