by ASSAY METHOD VALIDATION PROTOCOL FOR CITALOPRAM TABLETS
| Superseded Protocol No. | Nil |
| Effective Date |
TABLE OF CONTENTS :
| Sr. No. | Subject | Page No. |
| Protocol Approval | ||
| Objective | ||
| Scope | ||
| Responsibility | ||
| Product profile | ||
| Methodology | ||
| Precision | ||
| Linearity | ||
| Accuracy | ||
| Range | ||
| Incident/Deviation | ||
| Summary | ||
| 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 Assay content in Citalopram Hydrobromide (API) will yield consistent and reliable results within the predetermined acceptance criteria.
Analytical method validation will be performed by considering thefollowing parameters:
| Parameters | Citalopram |
| Method Precision | yes |
| Intermediate Precision (Ruggedness) | yes |
| Linearity | yes |
| Accuracy | yes |
| Range | yes |
- Scope :
The scope of this protocol is based on SOP and applicable for the validation of Assay method of Citalopram Hydrobromide.
- Responsibility of Validation Team:
| Departments | Responsibilities |
| QC | Preparation and Review of Validation protocol. |
| Perform the validation as per approved protocol 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 carry out Validation. | |
| Review and approval of Validation report. | |
| Head Quality | Authorization of protocol. |
- Product Profile:
| Category (% API Content) | Antidepressant |
| Reason for Validation | First Validation |
| Active Ingredient | Citalopram |
| Strength | Citalopram (as Hydrobromide Ph. Eur ) |
| Methodology | Non- compendia (In-House) |
| Method Reference | Provided by customer |
| Specification Limits | 99 % -101.5 % |
- Standard testing procedure
Assay of Citalopram Hydrobromide by Potentiometrically
Required reagents
Ethanol 96%
0.1M Hydrochloric Acid
0.1M Sodium Hydroxide (volumetric solution)
Blank titration
Take 50 mL of Ethanol and 0.5ml hydrochloric Acid in a titration vessel.
Titrate the solution with 0.1 M Sodium Hydroxide solution.
Test titration
Take 50 mL of Ethanol and 0.5ml hydrochloric Acid in a titration vessel. Accurately weigh and transfer about 0.300g of test sample to above vessel and dissolve completely. Titrate the above solution with 0.1 M Sodium Hydroxide solution.
Determine the end-point potentiometrically.
Conditions
Instrument : Auto-titrator (Mfg.- Metrohm; Model- Titra+)
Electrode : Epoxy/Glass body comb pH electrode
Stirring rate : 6
Method Parameter
Sample Name : Citalopram Hydrobromide
Titrant Name : 0.1MNaOH
Measurement type : Potentiometric
Initial dose : 100µl
Titration mode : Equilibration
P.D Criteria : 0MV
Min Constant Dose : 10 µl
Stir time : 10 sec
Const1 (Factor) : 40.5300
Calculation : Last end point
Limit:Between 99.0 and 101.5%.
- Precision
- 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 the six samples of same batch of and analyze as per method of analysis. Record the area on data sheet and calculate the % Assay, mean, standard deviation and % relative standard deviation.
Acceptance criteria:
RSD of Assay results in six sample preparations should be not more than 2%.
- Intermediate Precision ( Ruggedness ):
Intermediate precision expresses within laboratory variation with different analysts or equipment or /same on different days using same batch of drug product as per Methodology.
Test Procedure:
The analysis of the same batch will be done in six replicate analyses by using different columns by different analyst, by different system on different day. The mean, standard deviation and relative standard deviation will be calculated.
Acceptance criteria
% RSD should be not more than 2%.
- 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 should be prepared.
Determine the linearity by preparing and inject the standard solution in the range of 70% to 130% of concentration level and calculate the correlation coefficient “r”.
Test Procedure:
Prepare the standard solutions at five concentrations, typically 70, 90, 100, 120 and 130% of target concentration following Methodology. Three individually prepared replicates at each concentration will be analyzed. Calculate the mean, standard deviation, and Relative Standard Deviation (RSD) for each concentration. Plot concentration (x-axis) versus mean response (y-axis) for each concentration. Calculate the correlation coefficient (r). Record these calculations on the datasheet.
Acceptance criteria
The correlation coefficient (r) should be not less than 0.999.
- 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 at about 70, 90, 100, 120 and 130% of test concentration level in triplicate in each level and analyze as per Methodology. Further the % recovery for each preparation was calculated against the average % assay of method precision. Calculate the % RSD for recovery obtained at each level separately and overall %RSD.
Acceptance criteria
The mean recovery at each level should be between 98% to 102 % of the theoretical value and the % RSD not more than 2%.The overall average should be between 98% to 102% with %RSD not more than 2%.
- Range:
The range of an analytical procedure is the interval between the upper and lower concentration of analyte in the sample 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 obtained by the analytical method.
- Incident/Deviation:
Any Incident or Deviation observed during Analytical Method validation should be recoded and reported in Validation Report.
- Summary/Conclusion:
Final Conclusion should be drawn from analytical method validation for its use to analyze the assay test in Citalopram Hydrobromide by Potentiometrically.
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 |
