by ANALYTICAL METHOD VALIDATION PROTOCOL FOR DISSOLUTION METHOD OF SILDENAFIL CITRATE TABLETS 50 mg
| Superseded Protocol No. | Nil |
| Effective Date |
Table of contents :
| Sr. No. | Subject |
| 1.0 | Protocol Approval |
| 2.0 | Objective |
| 3.0 | Scope |
| 4.0 | Responsibility |
| 5.0 | Product profile |
| 6.0 | Methodology |
| 7.0 | 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 method validation is to provide documentary evidence that analytical methodology used for Dissolution test of Sildenafil Citrate Tablets 50 mg will yield consistent and reliable results within the predetermined acceptance criteria.
The method validation for Dissolution method will be performed as per Protocol. This protocol describes the methodology which will be used in the validation study for the following parameters;
- Precision
- System Precision
- Method Precision
- Intermediate Precision (Ruggedness)
- Specificity
- Linearity
- Accuracy
- Stability in Analytical Solutions
- Robustness
- Filter Paper Selection Study
- Scope :
The scope of this protocol is based on SOP. and applicable for the validation of Dissolution method of Sildenafil Citrate tablet 50 mg.
- 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 validation report. | |
| To impart training of protocol to concerned department/persons. | |
| QA | Review and approval of protocol. |
| Co-ordination with QC to carryout Validation. | |
| Review of validation data. | |
| Head Quality | Authorization of validation protocol. |
- Product Profile:
| Category | For Erectile Dysfunction |
| Reason for Validation | Ist Validation and regulatory requirement |
| Type of validation | Complete |
| Reference | ICH Q2 (R1) |
| Active Ingredient | Sildenafil Citrate EP |
| Strength | Each Film Coated tablet contains: Sildenafil Citrate EP equivalent to Sildenafil – 50 mg |
| Methodology | In House |
| Method Reference | In House |
| Specification Limits | NLT 75 % |
| Sample details | Sildenafil Citrate tablets 50 mg |
- Test Methodology:
- Method of analysis of Dissolution:
Reagents:
Sildenafil Citrate EP (Working Standard)
Hydrochloric Acid
Placebo,
Instruments:
Dissolution Apparatus
UV- VIS Spectrophotometer
Sample details:
Dissolution medium:
0.01 M Hydrochloric Acid: Mix 8.5 ml of Conc. Hydrochloric Acid with 10000 ml of water.
Apparatus : Apparatus No. 1 (Paddle)
Temperature of Dissolution Medium : 37 ± 0.5°C
Speed : 100 rpm
Volume : 900 ml
Time : 30 minutes
Standard Solution:
Weigh 75 mg of Sildenafil Citrate WS in to a 100.0 ml volumetric flask add 50 ml of dissolution medium sonicate to dissolve and dilute to 100.0 ml with dissolution medium. Further dilute 2.0 ml to 50.0 ml with dissolution medium.
Procedure:
Place the 900 ml of the dissolution medium, free from dissolved air, into the six vessels of the apparatus. Assemble the apparatus and warm the dissolution medium to 36.5°C to 37.5°C. Place one tablet in to the each vessel allows the tablet to sink to the bottom of the vessels prior to the rotation of the paddle. Operate the apparatus immediately at the speed of rotation for 30 minutes.
Test Preparation:
After completion of the dissolution withdraw sample from a zone midway between the surface of the dissolution medium and the top of the rotating blade, not less then 10 mm from the wall of the vessel. Filter through 0.45m Nylon filter or equivalent filter. Discard the first few ml of the filtrate, Dilute 20.0 ml of the filtrate to 50.0 ml with dissolution medium. Calculate the amount of dissolved active ingredient in solution as a percentage of the stated amount.
Measure the absorbance of the resulting solutions of standard preparation and test preparation at the maximum at about 294 nm, taking dissolution medium as a blank. Calculate the content of Sildenafil using the given below formula.
Calculation:
At Ws 2.0 900 50.0 P
———-x ——— x ———- x ——– x ———- x ——– x 0.71064 x 100
As 100.0 50.0 50 20.0 100
Where,
At: Absorbance of test solution
As: Absorbance of Std. Solution
Ws: Weight taken for Standard preparation in mg
P: Potency of Working Standard
- 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).
Standard solution will be scanned six times and record the absorbance as main peak in standard solution.
Test Procedure:
Standard solution shall prepare and take the absorbance at about 294 nm as per test methodology.
Documentation:
Calculate the mean, standard deviation, and % RSD of absorbance of Sildenafil and record on data sheet.
Acceptance criteria
The % RSD for six replicate injections of standard solution should be NMT 2.0.
- 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:
Six sample of single batch (same batch) shall be prepared and analyzed as per test methodology and calculated the % RSD for six samples results.
Documentation:
Calculate the mean, standard deviation, and % RSD of Dissolution of Sildenafil in Sildenafil Citrate Tablet and record on data sheet.
Acceptance criteria
% RSD of the six samples shall be not more than 5%.
- Intermediate Precision ( Ruggedness )
Intermediate precision expresses within laboratory variation with different analysts on different days by using of same batch of drug product as per test methodology.
Test Procedure:
Six sample of same batch shall be prepared and analyzed as per test methodology by different analyst on different days.
Documentation:
To be calculated the mean, standard deviation, and % RSD of Dissolution of Sildenafil in Sildenafil Citrate Tablet and recorded on data sheet.
Acceptance criteria
% RSD of the six analyses shall be not more than 5%. Overall (two analyst results value, 12 units) % RSD shall be not more than 5%.
- 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:
Specificity of test method shall be established by scanning of the blank (diluents), placebo and standard solution.
Documentation:
Print chromatograms.
Acceptance criteria
The interference from blank and placebo shall be not more than 2 % of standard absorbance.
- Linearity :
Its ability (within a given range) to obtain test results which are directly proportional to the concentration levels shall be prepared. For establishment of linearity minimum five concentration levels shall be prepared.
Test Procedure:
The linearity of response for Sildenafil shall be demonstrated by using the standard solutions at at six different concentrations as 25 %, 50 %, 75 %, 100 %, 150 % and 200 % of target concentration (specification) as per test methodology.
Documentation:
To be recorded results on a datasheet and calculated the correlation coefficient (r).
Acceptance criteria
The correlation coefficient (r) shall 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 shall be established across the specified range of the analytical procedure.
Test Procedure:
Known amount of Sildenafil Citrate (Drug substances) working standard shall be spiked with placebo at about 80%, 100% and 120% of test concentration and analyze as per test methodology. Calculate the % overall average recovery.
Documentation:
For each level of percentage recovery shall be calculated and recorded on test data sheet. Calculate the mean percent recovery for all samples. Record results on the datasheet.
Acceptance criteria
The % recovery of the drug at each spiking level shall be between 95 % and 105 % and RSD shall be not more than 5.0 %.
- Filter Paper Selection Study :
Test Procedure:
Prepare sample solution as per the test methodology. A portion of sample solution shall be centrifuged and other portion of sample solution shall be filtered with 0.45m PVDF filter and 0.45m Nylon filter.
Documentation:
Calculate the % drug release difference for each sample and calculate the % difference between centrifuges vs. filtered sample.
Acceptance criteria
The difference in the results shall be not more than 5 %.
- Robustness:
The method shall show reliability of an analysis with respect to deliberate variations in method parameters.
Robustness of the method shall be verified by varying the instrumental conditions such as
By changing the RPM by ± 5.
By changing the wavelength ± 2 nm.
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.
Documentation:
Calculate and compare the % dissolution of above experiment with method precision results.
Acceptance criteria
Overall % RSD shall be not more than 5 with of the method precision data for individual experiments.
- 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 tablet as per finish product testing procedure. Analyze the standard solution and sample solution at the different time intervals and calculate the % Difference for the result.
Documentation:
Calculate the % Dissolution.
Acceptance criteria
The difference in the results shall not be more than 2 %.
- Revision History:
| Revision No. | Details of changes | Reason |
| 00 | Nil | New Document |
