GB 5009.295-2023 English PDF

GB 5009.295-2023: National food safety standard - General rules for verification of chemical analysis methods
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GB 5009.295-2023
GB
NATIONAL STANDARD OF THE
PEOPLE’S REPUBLIC OF CHINA
National Food Safety Standard -- General Principles for
Validation of Chemical Analysis Methods
ISSUED ON. SEPTEMBER 06, 2023
IMPLEMENTED ON. SEPTEMBER 06, 2024
Issued by. National Health Commission of the People's Republic of China;
State Administration for Market Regulation.
Table of Contents
1 Scope... 3
2 Terms and definitions... 3
2.1 Method validation... 3
2.2 Qualitative method... 3
2.3 Quantitative method... 3
2.4 Concentration paid attention to... 3
2.5 Specificity... 3
2.6 Detection limit... 4
2.7 Limit of quantification... 4
2.8 Measuring range... 4
2.9 Accuracy... 4
2.10 Repeatability... 4
2.11 Reproducibility... 4
2.12 Stability... 4
2.13 Robustness... 4
2.14 Intermediate precision... 5
2.15 Linear range... 5
2.16 Detection probability... 5
3 General requirements for validation of chemical analysis methods... 5
3.1 Selection of performance parameters... 5
3.2 Selection of representative foods... 6
3.3 Validation sample requirements... 6
3.4 Data processing requirements... 7
4 Validation of performance parameters of chemical analysis methods... 7
4.1 Validation of method performance parameters... 7
4.1.1 Specificity... 7
4.1.2 Detection limit... 8
4.1.3 Limit of quantitation... 9
4.1.4 Measuring range... 9
4.1.5 Accuracy... 10
4.1.6 Precision... 11
4.1.7 Stability... 12
4.1.8 Robustness... 13
4.2 Validation of method equivalence... 13
4.2.1 Validation requirements... 13
4.2.2 Validation method... 13
Annex A Classification of food samples for validation of chemical analysis methods
... 15
Annex B Robustness test of the method... 18
National Food Safety Standard -- General Principles for
Validation of Chemical Analysis Methods
1 Scope
This Standard specifies the general requirements for the validation of national food
safety standard -- chemical analysis methods.
This Standard is applicable to the validation process during the development and
revision of national food safety standard -- chemical analysis methods.
2 Terms and definitions
2.1 Method validation
Test activities that provide objective and valid evidence to confirm that the performance
parameters of an analytical method meet the intended use of the method.
2.2 Qualitative method
An analytical method that identifies the presence or absence of a target analyte in a
sample based on its specific properties.
2.3 Quantitative method
An analytical method that determines the concentration or amount of target analytes
under conditions that meet specified precision and accuracy.
2.4 Concentration paid attention to
The concentration that is decisive for determining whether the target analyte in the
sample meets the requirements of regulations or standards.
2.5 Specificity
The ability of an analytical method to distinguish between target analytes and
interfering substances.
2.14 Intermediate precision
In the same laboratory, over a long period of time (pay attention to small changes in test
conditions, such as temperature, humidity, light, etc.), the precision of independent
measurement results obtained by using the same method to test the same sample.
2.15 Linear range
Under a certain level of significance, a linear calculation model is used to define the
relationship between instrument response and concentration (or amount), which
conforms to the interval between the lowest concentration (or amount) and the highest
concentration (or amount) of the calculation model.
2.16 Detection probability
The probability that an analytical method will detect the target analyte at a specified
concentration and confidence level.
3 General requirements for validation of chemical analysis
methods
3.1 Selection of performance parameters
3.1.1 For qualitative methods, intra-laboratory and inter-laboratory validation
parameters shall include method specificity and detection limit.
3.1.2 For quantitative methods, laboratory validation parameters shall include method
specificity, detection limit, quantitation limit, measurement range, accuracy, and
repeatability. Interlaboratory validation parameters shall include the detection limit,
quantification limit, determination range, correctness and reproducibility of the method.
3.1.3 When the analysis method is sensitive to small changes in test conditions such as
temperature, humidity, and lighting, the robustness of the method shall be examined,
and the intermediate precision index shall be calculated.
3.1.4 When there are two or more methods with the same applicable scope for the same
target analyte, the equivalence of the different methods needs to be validated.
3.1.5 When literature data on the stability of a reference material are lacking, its stability
shall be validated.
3.1.6 Selection of intra-laboratory and inter-laboratory validation parameters is shown
use of the method.
b) For a sample matrix, at least 3 concentration levels are required, including the
lowest concentration, the middle concentration (generally the concentration of
concern) and the highest concentration of the method's determination range. Their
uniformity and stability are tested.
3.3.4 When selecting a standard addition sample for validation, the validation sample
can be prepared as follows.
a) The sample matrix shall be selected to be representative and meet the intended
use of the method.
b) For a sample matrix, at least 3 concentration addition levels are required,
including the lowest concentration, the middle concentration (generally the
concentration of concern) and the highest concentration of the method
determination range.
3.4 Data processing requirements
All laboratory validation data need to be subjected to outlier testing and statistical
analysis. The analysis method can use statistical methods such as Grubbs test. After
eliminating outliers through technical analysis, the number of laboratories and the
amount of validation data shall meet statistical requirements.
4 Validation of performance parameters of chemical analysis
methods
4.1 Validation of method performance parameters
4.1.1 Specificity
4.1.1.1 Validation requirements
The ability of the analytical method to differentiate between the target analyte and other
substances shall be validated. There shall be no interference factors (signals,
interference peaks, etc.) that lead to errors in the identification of the target analyte and
affect the accuracy of quantitation.
4.1.1.2 Validation method
Analyze representative samples that match the scope of the method. Check whether
there are any interferences (signals, peaks, etc.) that lead to incorrect identification of
the target analytes and/or affect accurate quantitation. A certain concentration of
substances that may interfere with the identification and/or quantification of the target
analyte is added to a representative sample and then analyzed. Check whether these
interfering substances can be effectively identified.
4.1.2 Detection limit
4.1.2.1 Validation requirements
The detection probability of the target analyte at the detection limit level shall be no
less than 95% (at the 95% confidence level). The detection limit needs to be
independently validated for each representative sample matrix.
4.1.2.2 Validation method
4.1.2.2.1 Estimation method
Methods for estimating detection limits include.
a) Signal-to-noise ratio method (applicable to spectroscopy, chromatography, and
mass spectrometry). Add target analytes to a blank sample matrix. Use the added
concentration when the signal-to-noise ratio is 3 as the estimated detection limit.
b) Blank standard deviation method (method suitable for non-zero response values
of blank samples). The detection limit is determined by analyzing a large number
of sample blanks. The number of independent tests shall be no less than 10 (n
≥10). Calculate the standard deviation of the sample blank response. The
concentration corresponding to the average of the sample blank response values
plus 3 times the standard deviation shall be the estimated detection limit.
c) Calibration equation estimation method (applicable to instrument method). If
sample data at or near the detection limit cannot be obtained, use 3S/b (S is the
standard deviation of the calibration curve, b is the slope of the calibration curve)
as the estimated detection limit.
d) Gradual dilution method (applicable to all methods). The target analyte is serially
diluted at certain concentration intervals. Test according to the method to be
validated. The concentration at which the target analyte can be detected shall be
the estimated detection limit.
4.1.2.2.2 Determination method
Select at least 20 parallel samples of blank sample matrix. Target analytes at estimated
detection limit concentrations are added separately. When the detection probability of
the target analyte is not less than 95%, it is determined as the detection limit. When
blank samples are unavailable, isotope-labeled compounds or structural analogs can be
used for validation.
4.1.3 Limit of quantitation
4.1.3.1 Validation requirements
The quantitation limit shall be validated using certified reference materials/standard
samples, quality control samples or standard spiked samples of the same concentration.
Its accuracy and precision shall meet method and regulatory requirements. The limit of
quantification plus 3 times the standard deviation of the sample at the concentration of
interest shall be less than the concentration of interest. For target analytes with limited
values, the limit of quantitation shall be 0.5 times the limit value or less.
4.1.3.2 Validation method
4.1.3.2.1 Estimation method
Methods for estimating quantitation limits include.
a) Add target analytes to blank sample matrix. Use the added concentration when the
signal-to-noise ratio is 10 as the estimated quantitation limit.
b) Three times the detection limit is used as the estimated quantification limit.
c) The concentration corresponding to the average of the sample blank response
values plus 10 times the standard deviation is used as the estimated quantification
limit.
4.1.3.2.2 Determination method
For each representative sample matrix, independent testing is performed using certified
reference materials/standard samples, quality control samples or standard spiked
samples at the quantitation limit concentration level. Test at least 6 replicate samples.
Perform statistical analysis of data. If the accuracy and precision of the measurement
results at this concentration meet the requirements of Table 2 to Table 4, this
concentration can be used as the quantitative limit of the method.
4.1.4 Measuring range
The measurement range of the method shall cover the concentration of interest of the
method. Its validation requirements are as follows.
a) The correctness and precision of at least the lowest and highest concentrations of
the method's measurement range need to be validated. For target analytes with
limited limit values, the accuracy and precision of the limit value concentration
shall also be validated.
b) If the measurement range of the method is linear and the standard curve method
Depending on the properties of the standard solution or sample solution, choose
different storage conditions and time intervals. Use the same analysis method and
analysis conditions to measure the target analyte in the standard solution or sample
solution.
4.1.7.3 Evaluation of stability validation results
4.1.7.3.1 For analytical target substances with only two time point detection results, the
stability can be evaluated using formula (1).
Where,
ρi - the measured concentration of the target analyte at time point i;
ρ0 - the initial measured concentration of target analyte;
UE - the expanded uncertainty of the instrument method (excluding sample pretreatment
steps) at ρ0 concentration, UE≤15%.
4.1.7.3.2 For analytical target substances with multiple time point test results, the
stability can be evaluated using the t test or F test.
4.1.8 Robustness
Robustness testing is a testing procedure that examines the impact of environmental or
other conditional variables on an analytical method. A robust analysis method shall not
be subject to significant changes caused by small changes in environmental factors, that
is, it shall be insensitive to changes in environmental factors such as temperature,
humidity, and air pressure, as well as to other factors such as the amount of reagent
added and reaction time. The robustness test of the method is shown in Annex B.
4.2 Validation of method equivalence
4.2.1 Validation requirements
In the analytical method standards, there shall be no significant difference in the test
results of different analytical methods with the same measurement range.
4.2.2 Validation method
4.2.2.1 Select at least 3 representative samples. For each representative sample, at least
6 standard substances/standard samples or standard spiked samples with concentrations
GB 5009.295-2023
GB
NATIONAL STANDARD OF THE
PEOPLE’S REPUBLIC OF CHINA
National Food Safety Standard -- General Principles for
Validation of Chemical Analysis Methods
ISSUED ON. SEPTEMBER 06, 2023
IMPLEMENTED ON. SEPTEMBER 06, 2024
Issued by. National Health Commission of the People's Republic of China;
State Administration for Market Regulation.
Table of Contents
1 Scope... 3
2 Terms and definitions... 3
2.1 Method validation... 3
2.2 Qualitative method... 3
2.3 Quantitative method... 3
2.4 Concentration paid attention to... 3
2.5 Specificity... 3
2.6 Detection limit... 4
2.7 Limit of quantification... 4
2.8 Measuring range... 4
2.9 Accuracy... 4
2.10 Repeatability... 4
2.11 Reproducibility... 4
2.12 Stability... 4
2.13 Robustness... 4
2.14 Intermediate precision... 5
2.15 Linear range... 5
2.16 Detection probability.....