by PROTOCOL FOR VALIDATION OF METHOD OF ANALYSIS FOR CLEANING SAMPLE OF CITALOPRAM HYDROBROMIDE TABLET
| Superseded Protocol No. | Nil |
| Effective Date |
Table of contents :
| Sr. No. | Subject |
| Protocol Approval | |
| Objective | |
| Scope | |
| Responsibility of validation team | |
| Product profile | |
| Methodology | |
| 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 validation is to validate method of analysis for cleaning samples of Citalopram Hydrobromide tablet by considering the following parameters:
- Specificity
- System Precision
- Linearity
- Limit of Detection ( LOD )
- Limit of Quantification ( LOQ )
- Recovery Study
- Stability of Solution
- Robustness
- Scope :
This protocol is applicable for the validation of method of analysis of cleaning samples of Citalopram Hydrobromide tablet.
- Responsibility of Validation Team:
| Departments | Responsibilities |
| QC | Preparation, Review and execution of Protocol. |
| Analysis of samples and recording of data. | |
| Compilation and checking of data | |
| Preparation of Summary Report. | |
| QA | Review and approval of protocol. |
| Co-ordination with QC to carryout Validation. | |
| Review of data and summary report. | |
| Head Quality | Authorization of protocol. |
- Product Profile:
| Category | Antidepressant |
| Reason for Validation | To Validate the analytical method for Cleaning sample analysis. |
| Active Ingredient | Citalopram Hydrobromide |
| Methodology | In-House |
| Method Reference | In-House |
- Methodology
Method of analysis:
Reagents:
- Hydrochloric Acid
- Diluents: 0.1 M Hydrochloric Acid
- Instrument : UV-VIS Spectrophotometer
Standard Solution preparation:
Weigh accurately about 50 mg of Citalopram Hydro bromide working standard in to a 100.0 ml volumetric flask, and add about 50 ml 0.1 M Hydrochloric Acid, sonicate to dissolve and dilute to volume with diluents. Further dilute 2.0 ml of this solution to 100.0 ml with diluents (10mcg/ml).
Determine the maxima in the range of 400 nm to 200 nm by UV-VIS Spectrophotometer using diluents as a blank.
- Specificity:
Prepare blank, swab blank and standard solution 10 mcg/ml as per procedure given in protocol.
Blank solution: Diluents
Swab Blank: Take 10.0 ml of diluents in a test tube, dip fresh swab stick in test tube and sonicate for 5 minutes.
Procedure: Take the absorbance ofblank, swab blank and standard solution at maxima in the range of 400 nm to 200 nm.
Acceptance criteria:
The interference from blank and swab blank shall not be more than 2% of Standard absorbance.
- System Precision:
System precision shall be evaluated bymeasuring the absorbance of 6 replicate of standard solution and calculate the relative standard deviation.
Procedure:
Prepare the standard solution (10mcg/ml) as per procedure given in protocol. Take the absorbance ofblank and standard solution in six replicate at maxima in the range of 400 nm to 200 nm.
Acceptance Criteria: % RSD of Six replicate absorbance of standard solution should not be more than 2.0%.
- Linearity:
The linearity of the method shall be established between LOQ level to 200 % of concentration level. The sample absorbance shall be plotted against concentration and correlation coefficient shall be calculated.
Minimum five points shall be covered from the range of LOQ level to 200 % of concentration level.
Following concentrations shall be prepared and analyzed for linearity: LOQ level, 5mcg/ml, 10 mcg/ml, 15mcg/ml and 20mcg/ml.
Procedure: Measure the absorbance of sample solution in duplicate. Plot the graph of absorbance vs. actual concentration. Calculate the correlation coefficient and slope from linearity curve.
Acceptance Criteria: Correlation coefficient shall not be less than 0.99.
- Limit of Detection (LOD):
Limit of detection is the lowest concentration of analyte in a sample that can be detected but not necessarily quantified under the stated experimental condition.
Based on the standard deviation of the response of swab blank and the slope of linearity curve, LOD shall be calculated by the following equation:
LOD (mcg/ml) = 3.3 x σ
S
Where σ = Standard deviation of the response of swab blank.
S = Slope of Linearity Curve.
- Verification of Limit of Detection (LOD):
Prepare the LOD Sample and measure the absorbance of LOD sample in six replicates. Calculate % RSD for six replicates of Citalopram Hydro bromide.
Acceptance criteria:
% RSD for six replicates shall be not more than 30.
- Limit of Quantification (LOQ):
Limit of Quantification is the lowest concentration of analyte in a sample that can be quantify with acceptable precision under the stated experimental condition.
Based on the standard deviation of the response of swab blank and the slope of linearity curve, LOQ shall be calculated by the following equation:
LOQ (mcg/ml) = 10 x σ
S
Where σ = Standard deviation of the response of swab blank.
S = Slope of Linearity Curve.
- Verification of Limit of Quantification (LOQ):
Prepare the LOQ Solution and measure the absorbance of sample in six replicates. Calculate % RSD for six replicates of Citalopram Hydro bromide.
Acceptance criteria:
% RSD for six replicates shall be not more than 10.
- Recovery:
Standard Solution preparation:
Weigh accurately about 50 mg of Citalopram Hydro bromide working standard in to a 100.0 ml volumetric flask, and add about 50 ml 0.1 M Hydrochloric Acid, sonicate to dissolve and dilute to volume with diluents. Further dilute 2.0 ml of this solution to 100.0 ml with diluent.
Recovery Stock Solution:
Weigh accurately about 50 mg of Citalopram Hydro bromide working standard in to a 100.0 ml volumetric flask, and add about 50 ml diluents, sonicate to dissolve and dilute to volume with diluents.
- Spiked the 0.8 ml, 2.0 ml and 3.0 ml of recovery stock solution into the area of 10 x 10 cm2 in three different SS Plates for each concentration.
- Allow the solutions to dry in air.
- Take the Swab wipe and moisten with diluents, swab one SS plate in the area of 10 x 10 cm2. Collect the swab in the test tube containing already 10.0 ml of diluents in the test tube and sonicate for 10 minutes with swirling. Squeeze the swab and discard and transfer into a 100.0 ml volumetric flask and make up volume to 100.0 ml with the diluents. Filter through 0.45µ filter. Final concentrations of recovery solutions are 4 ppm, 10 ppm, and 15 ppm.
- Similarly do the recovery on 10× 10 cm2 Acrylic plates, PVC and Conveying belt.
Calculation:
Test Abs. Std. Weight 2.0 100.0 100.0 Potency
_________ X ____________ X _________ X _________ X __________ X _________ x 100
Std Abs. 100.0 100.0 Test Weight Spiked Vol. 100
Acceptance criteria:
Recovery at each level should not be less than 70% and % RSD for each concentration should not be more than 10.0 %.
- Stability of Solution:
It is essential when validating an analytical method to confirm that the analyte has adequate stability of standard solution during analytical measurement stages of the testing.
Prepare the standard solution and take the absorbance of standard solution at different time intervals.
Acceptance criteria:
Cumulative % RSD of standard absorbance shall not be more than 2.
- Robustness:
The method shall show reliability of an analysis with respect to deliberate variation in method parameters.
Following deliberate variations shall be done in method parameters:
- By changing the wavelength by ±2 nm.
Test Procedure:
Prepare the standard solution as per the method mentioned in protocol and take the absorbance of solution at different wavelength by changing the wavelength by ±2 nm with respect to System Precision data.
Acceptance criteria
Overall % RSD shall be not more than 2 with system precision data.
- Revision History:
| Revision No. | Details of changes | Reason |
| 00 | Nil | New |
