by METHOD VERIFICATION PROTOCOL FOR ASSAY OF DICLOFENAC SODIUM BP
| Superseded Protocol No. | Nil |
| Effective Date |
Table of contents :
| Sr. No. | Subject | Page No. |
| Protocol Approval | ||
| Objective | ||
| Scope | ||
| Responsibility of validation team | ||
| Product profile | ||
| Methodology | ||
| Validation Parameter | ||
| Incident / Deviation | ||
| Summary | ||
| Annexure | ||
| Revision History |
- PROTOCOL APPROVAL:
Prepared By:
| Functional Area | Name | Designation | Signature/ Date |
| Quality Control |
Reviewed By:
| Functional Area | Name | Designation | Signature/Date |
| Quality Assurance | |||
| Head Quality Control |
Approved By:
| Functional Area | Name | Designation | Signature/Date |
| Head QA |
Authorized By:
| Functional Area | Name | Designation | Signature/Date |
| Head Quality |
- Objective:
The objective of this verification is to verify the assay method of Diclofenac Sodium by considering the following parameters:
The objective of this verification is to provide documentary evidence that analytical methodology used in the analysis of assay of Diclofenac Sodium potentiometrically has reliable results within the predetermined acceptance criteria.
Analytical method verification will be performed by considering the following parameters:
| Parameters | |
| Specificity | yes |
| Method Precision | yes |
| Intermediate Precision | yes |
| Linearity | yes |
Assay method of Diclofenac Sodium is British Pharmacopoeia method, hence verification of method to be performed to check the suitability of the method under actual condition of use. Assay method of Diclofenac Sodium is titrimetric method (Potentiometric).
- Scope :
This protocol is applicable for the verification of assay method of Diclofenac Sodium BP API.
- Responsibility of Validation Team:
| Departments | Responsibilities |
| QC | Preparation & Review of Protocol. |
| Analysis of samples and recording of data. | |
| Compilation and checking of data | |
| Preparation of Summary Report. | |
| To impart training of protocol to concerned department/persons. | |
| QA | Review and approval of protocol. |
| Co-ordination with QC to carryout Verification. | |
| Review of data and summary report. | |
| Head Quality | Authorization of protocol. |
- Product Profile:
| Category ( API) | Analgesic |
| Reason for Verification | Pharmacopoeial Method |
| Active Ingredient | Diclofenac Sodium |
| Method Reference | British Pharmacopoeia |
| Specification Limits | Assay (on the dried basis). 99.0 per cent to 101.0 per cent |
- Methodology:
Assay
Reagents required: following mention chemical reagents in Table 1.0 required in method analysis used for assay of Diclofenac Sodium BP API.
Table 1.0
| Reagents | Grade |
| Anhydrous Acetic Acid | AR Grade |
| 0.1 M Perchloric Acid | AR Grade |
Procedure:
Dissolve 0.250 g in 60 ml of anhydrous acetic acid R. Titrate with 0.1M perchloric acid, determining the end point potentiometrically.
1 ml of 0.1 M perchloric acid is equivalent to 0.03181 g of C14H10Cl2NNaO2.
Calculation:
V x 0.03181 x 100 x M
——————————————————————-
0.1 x W
Where,
V = volume of 0.1 M perchloric acid
M = Molarity of 0.1 M perchloric acid
W = weight of sample in g
Acceptance Criteria:
If the % Assay on dried basis is within the specification limit (Between 99.0 % and 101.0 %), then the material is deemed as passing this test.
Loss on Drying
Procedure:
Weigh a glass-stopper shallow weighing bottle by drying in an oven at 105°C, cool then weigh. Transfer to the bottle the about 1.000 g quantity of the sample of Diclofenac Sodium, cover it and accurately weigh the bottle and the contents. Distribute the sample as evenly as practicable by gentle side wise shaking to a depth not exceeding 10 mm. Dry the sample by placing the loaded bottle in the drying chamber, Hot air oven at 105°C for 3 hours. After drying is completed open the drying chamber close the bottle promptly and allow it to cool to room temperature in a desiccators before weighing. Weigh the bottle and contents. Check the weight and calculate the loss on drying using the given below formula:
Calculation:
Loss in weight (g) x 100
= ————————————————–
Weight of sample (g)
Analytical Parameters to be verified and acceptance criteria given below Table 2.0:
Table 2.0
| Sr. No. | Analytical Performance Parameter | Acceptance Criteria |
| 1. | Specificity | To check the Interference of the 0.1 M perchloric acid. |
| 2. | Method Precision | The assay values should be in between 99.0 % to 101.0 % on dried basis for all six determinations. The % RSD of six assay values should be NMT 2.0 for Diclofenac Sodium. |
| 3. | Intermediate Precision (Ruggedness) | The assay values should be in between 99.0 % to 101.0 % on dried basis for all six determinations of second analyst and its %RSD should be NMT 2.0.The cumulative % RSD of assay values for both the analysts should NMT 2.0. |
| 4. | Linearity | The correlation coefficient shall be not less than 0.999. |
- Validation Parameter
- Specificity
- Method Precision
- Intermediate
- Linearity
- Specificity
Specificity of analytical method is its ability to assess unequivocally the analyte in presence of components that may be expected to be present in 0.1 M Perchloric acid. Prepare blank solution without analyte and titrate as per methodology and check the interference and record the titrant value consume.
- Method Precision
The precision of an analytical method is the degree of agreement among individual test results when the procedure is applied repeatability to multiple samplings of homogenous sample. It is usually expressed as the standard deviation and the relative standard deviation.
Prepare six individual sample of same batch and analyze as per methodology.
Calculate the % assay of Diclofenac Sodium on dried basis and the % RSD of assay for six individual samples.
Acceptance Criteria:
The assay should be between 99.0 per cent and 101.0 per cent on dried basis for all six determinations. The % RSD for assay of six individual samples should be NMT 2.0.
- Intermediate Precision (Ruggedness):
Intermediate Precision (Ruggedness) of an analytical method is the degree of repeatability of test results obtained by the analysis of same samples by different analyst on different days.
Note: The result obtained from method precision will be considered as the results of first analyst for comparison in ruggedness.
Intermediate precision is performed by analyzing a homogenous sample, by second analyst on second day by preparing six individual sample of same batch and analyze as per methodology. Calculate the % assay on dried basis and the % RSD of assay for six individual samples performed by second analyst.
Acceptance Criteria:
The assay should be between 99.0 per cent and 101.5 per cent on dried basis for all six determinations performed by second analyst. The % RSD for assay of six individual samples should be NMT 2.0.
The cumulative % RSD of assay for both the analysts is NMT 2.0.
- Linearity:
The linearity of an analytical procedure is its ability (within a given range) to obtained test results which are directly proportional to the concentration levels shall be prepared. Determine the linearity by preparing and analyze the sample solution in the range of 80% to 120% of concentration level and calculate the correlation coefficient “r”.
Prepare the sample solutions at five concentrations, typically 80%, 90%, 100%, 110% and 120% of target concentration following material testing procedure. Each concentration will be analyzed. Record results on a datasheet. Plot a graph with concentration (x-axis) versus volume used (ml) on y-axis. Calculate the correlation coefficient (r). Record the calculations on the datasheet.
Level 80 %: Take Sample weight 0.200 g in 60ml of ethanol anhydrous acetic acid R and titrate with 0.1 M perchloric acid, determining the end-point potentiometrically.
Level 90 %: Take Sample weight 0.225 g in 60ml of ethanol anhydrous acetic acid R and titrate with 0.1 M perchloric acid, determining the end-point potentiometrically.
Level 100 %: Take Sample weight 0.250 g in 60ml of ethanol anhydrous acetic acid R and titrate with 0.1 M perchloric acid, determining the end-point potentiometrically.
Level 110 %: Take Sample weight 0.275 g in 60ml of ethanol anhydrous acetic acid R and titrate with 0.1 M perchloric acid, determining the end-point potentiometrically.
Level 120 %: Take Sample weight 0.300 g in 60ml of ethanol anhydrous acetic acid R and titrate with 0.1 M perchloric acid, determining the end-point potentiometrically.
Acceptance criteria
The correlation coefficient (r) between observed titrant value and the weight taken shall be not less than 0.999.
- Incident / Deviation:
Any Incident and deviation during validation exercise shall be reported and justified. Raw data of all non-satisfactory results shall be kept along with validation report.
- Validation Summary:
Data obtained through validation exercise shall be analysed with regard to various parameters of validation and their acceptance criteria. A validation report shall be prepared summarizing all data. All results shall be captured in the method verification report.
Annexure (S) :
Analytical data shall be recorded in annexure – (General test data sheet).
Revision History:
| Revision No. | Details of changes | Reason |
| 00 | Nil | New document |
