by DISSOLUTION METHOD VALIDATION PROTOCOL FOR CITALOPRAM TABLETS (BY UV)
| Superseded Protocol No. | Nil |
| Effective Date |
Table of contents :
| Sr. No. | Subject | Page No. |
| Protocol Approval | ||
| Objective | ||
| Scope | ||
| Responsibility | ||
| Product profile | ||
| Methodology | ||
| Specificity | ||
| Precision | ||
| Linearity | ||
| Accuracy | ||
| Range | ||
| Stability of Analytical solution | ||
| Robustness | ||
| Filter paper Selection Study | ||
| Incident/Deviation | ||
| Summary/Conclusion | ||
| 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 provide documentary evidence that analytical methodology used in the determination of Dissolution method content in Citalopram Tablet (10mg, 20mg and 40mg) will yield consistent and reliable results within the predetermined acceptance criteria.
Analytical method validation will be performed by considering the following parameters:
| Parameters | 10mg | 20mg | 40mg | |
| Specificity | yes | yes | yes | |
| Precision | ||||
| System Precision | yes | – | yes | |
| Method Precision | yes | yes | yes | |
| Intermediate Precision(Ruggedness) | yes | yes | yes | |
| Linearity | yes | yes | yes | |
| Accuracy | yes | yes | yes | |
| Range | yes | yes | yes | |
| Stability in Analytical Solutions | – | – | yes | |
| Robustness | – | – | yes | |
| Filter Paper Selection Study | – | – | yes | |
- Scope :
This protocol is based on SOP and applicable for the validation of Dissolution method of Citalopram in Citalopram Hydrobromide tablet.
- Responsibility of Validation Team:
| Departments | Responsibilities |
| QC | Preparation and Review of Validation protocol. |
| Analysis of samples and recording of data. | |
| Compilation and checking of data | |
| Preparation and review of Validation report. | |
| To impart training of protocol to concerned department/persons. | |
| QA | Review and approval of Validation protocol. |
| Co-ordination with QC to carryout Validation. | |
| Review and approval of Validation report. | |
| Head Quality | Authorization of protocol. |
- Product Profile:
| Category | Antidepressant |
| Reason for Validation | First Validation |
| Active Ingredient | Citalopram |
| Strength | Each tablet contains: Citalopram (as Hydrobromide Ph.Eur) 10mg, 20mg &40mg |
| Methodology | In-House |
| Method Reference | In-House |
| Specification Limits | NLT80% |
| Shelf Life | NLT80% |
- Methodology :
Method of analysis of Dissolution:
Reagents:
Citalopram Hydrobromide (Working Standard)
0.1N hydrochloric acid
Membrane filter (pore size 0.45 µm)
Dissolution
Determination by UV-VIS spectrophotometer at λ= 238 nm, using dissolution apparatus and 0.1N hydrochloric acid as the dissolution medium.
Test conditions:
Apparatus : With paddles
Dissolution medium : 0.1 N Hydrochloric Acid
Agitation speed : 75 rpm
Mean volume per vessel : 900 ml
Temperature of the medium: 37 ± 0.5ºC
Duration of test : 30 minutes
Sampling time point : 30 minutes
For the preparation of 10 liters Dissolution Medium or 0.1NHCl Solution:
Dilute 85 ml of 37% hydrochloric acid to 10 liters in purified water.
Blank: Dissolution medium.
Procedure:
Transfer 900 ml 0.1N Hydrochloric acid to each of the vessels and warm to 37ºC ± 0.5ºC.
Prepare a Citalopram calibration curve with 7 different concentration points between 0.0067 mg/ml and 0.044 mg/ml, using Citalopram Hydrobromide WS (with the knowledge that 1 mg of Citalopram base is equivalent to 1.249 mg of Citalopram Hydrobromide).
Linearity solutions
Stock linearity solution 1:
Accurately weigh 85.0 mg of Citalopram Hydrobromide WS and dilute to 50.0 ml with dissolution medium.
Stock linearity solution 2:
Accurately weigh 69.0 mg of Citalopram Hydrobromide WS and dilute to 50.0 ml with dissolution medium.
Linearity solution 1:
(Citalopram base concentration: 0.0068 mg/ml). Dilute 5.0 ml of linearity stock solution 1 to 100.00 ml with dissolution medium. Dilute 1.0 ml of this solution to 10.0 ml.
Linearity solution 2:
(Citalopram base concentration: 0.0088 mg/ml). Dilute 2.0 ml of linearity stock solution 2 to 25.0 ml with dissolution medium. Dilute 5.0 ml of this solution to 50.0 ml.
Linearity solution 3:
(Citalopram base concentration: 0.011 mg/ml). Dilute 1.0 ml of linearity stock solution 2 to 100.0 ml with dissolution medium.
Linearity solution 4:
(Citalopram base concentration: 0.0176 mg/ml). Dilute 5.0 ml of linearity stock solution 2 to 25.0 ml with dissolution medium. Dilute 4.0 ml of this solution to 50.0 ml with the dissolution medium.
Linearity solution 5:
(Citalopram base concentration: 0.027 mg/ml). Dilute 2.0 ml of linearity stock solution 1 to 100.0 ml with dissolution medium.
Linearity solution 6:
(Citalopram base concentration: 0.034 mg/ml). Dilute 5.0 ml of linearity stock solution 1 to 100.0 ml with dissolution medium. Dilute 10.0 ml of this solution to 20.0 ml with the dissolution medium.
Linearity solution 7:
(Citalopram base concentration: 0.0442 mg/ml). Dilute 4.0 ml of linearity stock solution 2 to 100.0 ml with dissolution medium.
Procedure:
Place one film coated tablet from the batch to be tested in each of the vessels.
All standard and test solutions are previously filtered through 0.45 µm filter membrane.
Determine the UV-VIS absorbance of the Standard Linearity solution and samples, filtered into 0.5 cm cells, at wavelength 238 nm and calculate the concentration of Citalopram base dissolved in the samples by extrapolation in the calibration curve.
Calculations:
From the extrapolated concentration, the percentage of solution is calculated using the following formula:
900 x C x 100
% dissolved Citalopram = ————————-
S
In which:
C: concentration, in mg/ml, of Citalopram base in the test solution
S: strength of Citalopram per tablet
- 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 the placebo.
Test Procedure:
Prepare the solution of standard, Blank, Placebo and test solution as per Methodology .
Scan the each solution at wavelength 200nm-400nm.
Acceptance criteria:
No any interference should be observed in the blank and placebo at maxima of standard solution.
If interference observed from blank and placebo should not be more than 2 % of standard absorbance.
- Precision:
- System Precision:
The system precision is the closeness of agreement between the responses of detector. It is usually expressed as the standard deviation (S D) or the relative standard deviation (RSD).
Test Procedure:
Prepare the standard solution as per methodology and determine the correlation coefficient of linearity solution and take absorbance at about 238 nm of linearity level-1 and linearity level-7 to determine the entire range of linearity solution in six times for system precision. Calculate the mean, standard deviation, and % RSD of absorbance Citalopram and record on data sheet.
Acceptance criteria
% RSD should be not more than 2%.
- 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.
Test Procedure:
Prepare six samples of single batch and analyze as per the Methodology procedure and determine the content of six samples. Calculate the mean, standard deviation, and % RSD of Dissolution of Citalopram in Citalopram Tablet and record on data sheet.
Acceptance criteria:
% RSD of the six samples should be not more than 5%.
- Intermediate Precision ( Ruggedness )
Intermediate precision expresses within laboratory variation with different analysts on different days using same batch of drug product as per methodology procedure.
Test Procedure:
Prepare six samples of single batch and analyze as per methodology procedure by two different analysts on different days. Calculate the mean, standard deviation, and % RSD of Dissolution of Citalopram in Citalopram Tablet and record on data sheet.
Acceptance criteria:
% RSD of the six samples should be not more than 5%. Determine the cumulative %RSD of 12 samples preparation should be not more than 5%.
- Linearity:
Its ability (within a given range) to obtain test results which are directly proportional to the concentration (amount) of analyte in the sample. For establishment of linearity minimum six concentration levels should be prepared.
Test Procedure:
Prepare the standard solutions at different concentrations, typically 70 %, 90 %, 100 %,
110 %, 120% and 130 % of target concentration following Methodology procedure. Calculate the correlation coefficient (r). Record these calculations on the datasheet
Acceptance criteria
The correlation coefficient (r) should be not less than 0.99.
Accuracy:
The accuracy of an analytical procedure express the closeness of agreement between the value which accepted either as a conventional true value or an accepted reference value and the value found. The accuracy should be established across the specified range of the analytical procedure.
Test Procedure:
Prepare the sample solution by spiking the Citalopram (Drug substances) to the placebo at about 75%, 100% of 10mg 100%of 20mg 100%and 125% of 40mg test concentration in triplicate in each level and analyze as per methodology procedure and calculate the % overall average recovery.
Acceptance criteria
The % recovery of the drug at each spiking level should be between 95 % and 105 %.The overall average recovery should be between 95% to 105% within %RSD of not more than 5%.
Range:
The range of an analytical procedure is the interval between the upper and lower concentration of analyte in the sample (including these concentrations) for which it has been demonstrated that the analytical procedure has a suitable level of precision, accuracy and linearity. The range is normally expressed in the same units as test results (e.g. Parts per million) obtained by the analytical method.
Stability of Analytical Solutions:
It is essential when validating an analytical method to confirm that the analyte has adequate stability in both the standard and sample solution during analytical measurement stages of the testing.
Test Procedure:
Prepare standard solution and perform the dissolution on the six tablet of higher Strength per finish product testing procedure. Analyze the standard solution and sample solution at the different time intervals and calculate the % Difference for the result.
Acceptance criteria
The difference in the results should be not more than 5 %.
Robustness:
The method should show reliability of an analysis with respect to deliberate variations in method parameters.
- By changing the RPM by ± 5.
- Concentration dissolution Medium ± 10%.
Test Procedure:
Prepare the standard solution and perform the dissolution on the tablets by deliberate variations made in the method for each condition as mentioned in protocol and analyze. Calculate the result and compare with method precision results.
Acceptance criteria
Calculate and compare the % dissolution of above experiment with method precision results. Overall% RSD should be not more than 5% with of the method precision data for individual experiments.
Filter Paper Selection Study :
Test Procedure:
Prepare sample solution as per the methodology procedure. A portion of sample solution should be centrifuge and other portion of sample solution should be filtered with 0.45µ PVDF filter and 0.45µ Nylon filter.
Acceptance criteria:
Calculate the % drug release difference for each sample and calculate the % difference between centrifuges vs. filtered sample. The difference in the results should be not more than 5%.
Incident/Deviation:
Any Incident or Deviation should be observed during Analytical Method validation should be recoded and reported in Validation Report.
Summary/Conclusion:
Final Conclusion should be drawn regarding analysis method validation for its use to analyze the assay test in Citalopram tablets by UV.
Summary of validation report should be prepared and accordingly STP to be updated.
Revision History:
| Revision No. | Details of changes | Reason for change |
| 00 | Nil | New Document |
