v
What Is
Analytical Method Validation?
Ø Analytical method validation establishes documented evidence that the procedure adopted for
a test is fit for the intended purpose in terms of quality, reliability and consistency of results.
Ø
Method validation is the
process of documenting / proving that an analytical method provides analytical
data acceptable for the intended use.
Ø
A pharmaceutical drug product must meet all its
specifications throughout its entire shelf-life.
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Validation
Parameter–
As per USP / ICH –
1. Accuracy:
“Closeness of test results obtained by the method to true value”. i.e. measure the exactness of analytical method. It is expressed as percent recovery by the assay of known amount of analyte in the linearity range.
The accuracy of an analytical procedure expresses the closeness of
agreement between the value which is accepted either as a conventional true
values or accepted reference value and the value found.
Recommended data: Assessed by 9 determination over a minimum of 3 concentration levels covering a
specified range.
2. Precision:
The precision of an analytical procedure expresses the closeness of agreement (degree of scatter) between a series of measurements obtained from multiple sampling of the same homogeneous sample under the prescribed conditions.
Precision include:
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Repeatability
Ø
Intermediate Precision
Ø Reproducibility
v Repeatability: Repeatability expresses the precision under the same operating condition over a short interval of time.
v Intermediate Precision: Intermediate precision expresses variations within laboratories, such as different days, different analysts, different equipment's, etc.
v Reproducibility: Reproducibility expresses the precision between laboratories.
3. Limit of Detection (LOD)
It is the lowest amount of analyte in a sample which can be detected but
not necessarily quantitated.
4. Limit of Quantitation (LOQ)
It is the lowest amount of analyte in a sample which can be quantitatively determined with suitable precision and accuracy.
v
Determination of Limit of Detection (LOD) and
Limit of Quantitation (LOQ)
v Signal of noise
The LOD is the minimum sample concentration at which the substance signal can be reliably detected
when compared to the baseline noise of a blank run. A signal-to-noise between 2:1 and 3:1 is generally
considered acceptable for estimating the detection limit.
5. Specificity
Specificity is the ability to assess unequivocally the analyte in presence of components which may be expected to be present.
6. Linearity
The ability of the method to obtain test result that are directly proportional to concentration within a given range.
Method: Dilution of stock solution / separate weighing
Ø Minimum 5 concentration are used
7. Range
The range (the interval between the upper and lower levels) of an
analytical method has been demonstrated to determine precision, accuracy, and
linearity using the set method. This range is defined by the concentration
range in which the Linearity test was completed.
Specific range dependent upon
intended application of the procedure
Ø Assay:
80 to 120 % test concentration.
Ø Content
Uniformity: 70 to 130 % test concentration.
Ø Dissolution
: 20 % to 120 %
Ø Impurities
reporting level : 120 % of specification limit ( with respect to test
concentration of API)
8. Ruggedness
The ruggedness of an analytical method is the degree of reproducibility
of test results obtained by the analysis of the same sample under a variety of
conditions, such as different laboratories, different analysts, different
equipment's, different days, etc.
i.
Source
ii.
Concentration of stability of solution
iii.
Heating rate
iv.
Column Temperature
v.
Humidity
9. Robustness
The robustness of an analytical procedure is a measure of its capacity to
remain unaffected by small, but deliberate variation in method parameters and
provides an indication of its reliability during normal uses.
Determination: The evaluation of robustness should be considered during
the development phase and depend on the type of procedure under study.
