SOP for Analytical Method Validation

1.0 OBJECTIVE

To establish the procedure that the performance characteristic of the analytical procedure meet the requirement for the intended analytical applications.

2.0 SCOPE

This procedure is applicable to the method of analysis used for analysis of material/ products at [Company name].

3.0 RESPONSIBILITY

• Analytical Development Laboratory – To provide method for analysis Provide assistance for validation Activity Approval of protocol and report

• Quality Assurance (QA) Department – Preparation and approval of protocol Review of report and Approval Certification

• Quality Control (QC) Department – Checking of protocol Perform the method validation Activities Preparation and checking of report.

4.0 PROCEDURE

(A) When analytical method validation Required

(i) Method validation may be necessary for submitting revised analytical procedure or use of an established procedure with a new product or raw material (Refer annexure – I)

(ii) For any new finished product or non-compendial active Pharmaceutical Ingredient (API).

(B) Analytical Method shall be verify or validated for following test.

(I) Identification test for active and coloring agent

(II) Dissolution Test

(III) Related Substance/ Related Compound

(IV) Stability indicating Assay method

(V) Organic Volatile Impurity

(VI) Residue in cleaning Validation

(C) Typical analytical characteristic used in method validation

Typical parameter for validation of analytical method of analysis. Design for providing quantitative result shall include

(I) System Suitability

(II) Accuracy

(III) Precision

(IV) Specificity

(V) Detection Limit

(VI) Quantitation Limit

(VII) Linearity and range

(VIII) Robustness

(IX) Filter paper interference

(X) Solution Stability

(I) System suitability: Prior to the acceptance of any validation study, the suitability of the system should be   determined.

In case of column Chromatography (i.e. HPLC etc) It can be done by evaluating resolution between peaks. In case of UV/Visible or other it can be done by repeatedly taking the response of standard appropriate time as specified in protocol and the RSD should be within the limit specified in protocol. The system suitability data must meet the acceptance criteria prior to the acceptance of validation data with which it is associated.

(II) Accuracy: The accuracy of an analytical procedure is the closeness of the test result obtained by that procedure to the true value.

Accuracy shall be assessed using a minimum of nine determinations over a minimum of three concentration level covering specific range. i.e. three concentration and three replicates of each concentration. (80%, 100%, 120%)

Guideline Table for accuracy recovery records and % recovery refer Annexure – II

(III) Precision: – It is degree of assignment among individual test results when the procedure applied respectively to multiple sampling of homogeneous sample.

It is usually expressed as the standard deviation or relative standard deviation.

The representative shall be assessed using minimum of nine determination covering the specific range i.e. three concentration and three replicate of each concentration or using minimum of six determination of 100% of test concentration.

System Precision:  Nine or six replicate as expressed above for sample & RSD calculated. Target RSD as given in protocol shall meet

Method Precision:  Repeatability & Intermediate precision or ruggedness

(i) Repeatability: Full analysis method carried out for nine or six sample as expressed above for homogenized mixture/ LOT and RSD calculated. RSD should within limit as given in protocol shall meet.

(ii) Intermediate precision or ruggedness: The intermediate precision or ruggedness of analytical method of determination of  content of analyte study by analyzing the sample by two different analysts on two different dates using different instruments as per the method described under repeatability.  % Difference between the two mean assays determine and report

For tabulation and calculation refer Annexure – III as a guideline

(IV) Specificity: Specificity means ability to assess unequivocally (unmistakably) the analyte in presence of components that may be expected to present like impurities, excipients, preservatives and other active etc.

The peak purity study carried out on forcefully degraded samples of active, which can be achieved by acid hydrolysis, alkali hydrolysis and oxidation or other degradation pathway.

Accelerated compatibility study between API and excipients alone and in combination at elevated temperature and photostability chamber against standard.

For the tabulation & calculation refer Annexure IX as a guideline, for detail method for acid hydrolysis alkali hydrolysis and oxidation respective protocol to be followed

(V) Detection Limit (DL): It is lowest amount of analyte in a sample that can be detected.

For non instrumental method the minimum level at which the analyte can be reliably detected by using known concentration of analyte by serially diluting analyte. For instrumental method the same approach may be use.

i.e if 0.1 require to detect as impurity level. It should be demonstrated that the procedure will reliably detect the impurity at that level.

DL data evaluated from the calculation from the calibration curve (Response on Y-axis and Concentration on X-axis) using formula DL= [3.3 * SyX/Slope]

(VI) Quantitation limit (QL): This is lowest level of compound in analyte in sample that can be determined with acceptable precision and accuracy under stated experimental conditions. Quantitation limit is generally determined by using known concentration of analyte by serially diluting analyte. I.e. It is require that an analyte be assayed at the level of 0.1 mg per tablet, it should be demonstrated that the procedure will reliably quantitate the analyte at that level.

QL data evaluated from the calculation from the calibration curve (Response on Y-axis and Concentration on X-axis) using formula QL= [10 * SyX/Slope]

(VII) Linearity and range

Linearity: It is ability to elicit test result that are directly or by well defined mathematical Transformation, proportionate to the concentration of analyte in sample with in given range

For linearity minimum five concentrations to be consider over range of analytical procedure. Linearity shall assess by regressing the mean peak area of each solution against their known concentrations. The graphical data shall be plotted & The correlation coefficient, Y-intercept, slop of the regression line and residual sum of squares shall be reported.

Range: The specified range desired from linearity study and depend on the intended application of the procedure.

For Assay 80 to 120 % of test concentration

For uniformity 70 to 130 % test concentration

For DR  20% over specified Range

For impurity: from 50 % to 120 % of the acceptance criteria or LOQ to 120 % of specified limit in which whichever less i.e. 50% or LOQ or as per specified in protocol.

Plot the linearity curve for response and concentration. For tabulation and calculation refer annexure V as a guideline.

(VIII) Robustness: It is measure of it’s capacity to remain unaffected by small but deliberate variation in procedural parameter that can be but not limited to Flow rate, pH, Concentration of buffer, RPM in case of dissolution, Different make Column. Refer ANNEXURE VI as a guideline.

(IX) Filter Paper interference: The possible interference from different filter papers used during sampling shall be checked. A required parameter value (e.g. % assay, % impurity) obtained from the filtered solution shall be checked against required parameter value of the unfiltered centrifuged solution (or specified in protocol). The absolute difference should be calculated and reported. Refer ANNEXURE VII as a guideline.

(X) Solution Stability: The stability of the standard and sample solutions shall be determined at the time intervals representative for storage (at least 24 hrs with two intermediate time points). Standard solution stability should be done by assessing the standard preparation as per the method at initial time point or 0 hrs then at another intervals including 24 hrs (and so on) and compare against freshly assessed standard. % Difference of reading within the mean of each interval should not be more than % specified in protocol. Sample solution stability will be determined by comparing to its initial value. Absolute difference should be calculated. Refer ANNEXURE VIII as a guideline

5. ANNEXURE (S)

ANNEXURE I: Data elements require for validation 

ANNEXURE II: Guideline table and calculation for accuracy study

ANNEXURE III: Guideline table for precision

ANNEXURE IV: Guideline tables for specificity study

ANNEXURE V: Guideline linearity and range study

ANNEXURE VI: Guideline table for robustness

ANNEXURE VII: Guideline table for filter evaluation study

ANNEXURE VIII: Guideline table for solution stability study of standard

Annexure I: Data Elements require for validation

Category I – Analytical methods for quantitation of major components of bulk drug substances or active ingredients (including preservatives) in finished pharmaceutical products.

2. Table for method precision

Annexure IV: Guideline tables for specificity study

Annexure V: Guideline linearity and range study

Annexure VI: Guideline table for robustness

Annexure VII: Guideline table for filter evaluation study

Annexure VIII: Guideline table for solution stability study of standard