by METHOD VERIFICATION PROTOCOL FOR DETERMINATION OF RESIDUALSOLVENT IN SILDENAFIL CITRATE
| Superseded Protocol No. | Nil |
| Effective Date |
- 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 |
Table of contents:
| Sr. No. | Subject | Page No. |
| Objective | ||
| Scope | ||
| Responsibility of Validation Team | ||
| Product Profile | ||
| Methodology | ||
| Specificity | ||
| System Precision | ||
| Intermediate Precision (Ruggedness) | ||
| Accuracy | ||
| Robustness | ||
| Method Verification | ||
| Revision History |
- Objective:
- The objective of this validation is to validate the method validation of residual solvent in Sildenafil Citrate by considering the following parameters:
- Specificity
- Precision
- System Precision
- Intermediate Precision (Ruggedness)
- Accuracy
- Robustness
Following parameters will be seen during system suitability:
| Sr. No. | Parameters | Limit |
| 1. | % RSD for peak areas of each solvent obtain by 6 replicate injections of standard solution. | Not more than 15.0 |
When the above parameters meet the method, the actual experiment shall be started.
- Scope :
This protocol is based on SOP. and applicable for the method validation of Residual solvent in Sildenafil Citrate.
- 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. | |
| 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 | Erectile Dysfunction |
| Reason for Validation | First Verification Validation |
| Methodology | In-House |
| Method Reference | In-House |
| Specification Limits (ppm) | Acetone : NMT 5000 ppm Methylene Dichloride : NMT 600 ppm Methanol : NMT 3000ppm Ethyl Acetate : NMT 5000ppm Toluene : NMT 890ppm |
- Methodology:
Residual solvents by GC-HS (Agilent)
Instrument: A gas chromatograph system is equipped with flame ionization detector and
integrator.
GC Conditions
Column : OPTIMA-624 or equivalent
Length : 30.0 meters
Film thickness : 3.00 µm
ID : 0.53 mm
Carrier gas : Nitrogen
Flow rate : 4.0 ml/min
Injector port temperature : 180°C
Detector port temperature : 260°C
Split ratio : 1:12
Injection volume : 1000 µl
Sample Run time : 21.7
Column temperature : Start at 40°C, hold for 8minutes, then raise at a rate
35ºC/min to 240°C and hold for 8 minutes.
Detector : FID
Nitrogen (Makeup flow) :10ml/min
Hydrogen Flow : 30ml/min
Air Flow : 300ml/min
Head Space Condition:
Vial Equilibration Temperature : 95ºC
Vial Equilibration Time : 30min
Transfer line Temperature : 105ºC
GC Cycle Time : 55min
Pressurization Time : 30Seconds
Preparation of stock solution:
- Take 100ml volumetric flask Containing 25ml DMSO Transfer 5000 mg of Acetone, 5000mg of Ethyl acetate, 3000 mg of Methanol, 600 mg of Methylene Dichloride and 890mg of Toluene Make up to volume with DMSO.
Preparation of standard solution:
- Transfer 1.0ml of stock solution in to a 100 ml volumetric flask containing 20 ml of DMSO Make up to the volume with DMSO.
Preparation of sample solution:
- Transfer about 500 mg of test sample into a 20 ml GC-Head Space Vial Add 5ml of DMSO Seal the vial immediately.
Preparation of Blank solution:
- Transfer 5ml of DMSO in 20 ml GC-Head Space Vial Seal the vial immediately.
System suitability:
- Inject standard solution six times into the chromatographic system.
- Record the chromatograms obtained with standard solution and measure the peak responses.
- The relative standard deviation of the individual peak responses from six replicate injections should be Not More than 15.0 %.
Procedure:
- Inject DMSO as blank solution and record the chromatographic system.
- Inject standard solution six times into the chromatographic system.
- Record the chromatograms obtained with standard solution and measure the peak responses.
- Inject test sample solution two times into the chromatographic system.
- Record the chromatogram obtained with sample solution and measures the peak responses.
- Identify the peaks in the chromatograms of the sample preparation, based on the retention time of corresponding peaks present in the standard preparation and disregard the DMSO peak in the Standard and Test Solution.
- Calculate and note down the peak responses of individual components from the sample and standard preparations.
- Apply the blank correction in case of any interference from the solvent used as diluent.
- Calculate the amount of residual solvents present in the sample by the formula given below.
Calculation:
AT DS
————- x ————x P x10000
AS D
Where,
AT = Peak area of each solvent observed in test sample chromatogram.
AS = Average area of each solvent observed in standard chromatogram.
Ws = Weight of the respective standard in standard solution in mg test.
Wt = Weight of the test sample taken.
P = Potency of respective standard.
- Specificity:
Specificity of test method shall be established by injecting all solvents to be examined simultaneously. Prepare blank solution, standard solutions individually, mixes standard and spiked test sample as per method of analysis. Retention time observed for each solvent shall be recorded
Acceptance criteria:
Retention times of each solvent should match in, individual, mixed and spiked solution and there should no interference at the retention time of the solvents.
- System Precision:
Prepare and inject separately Blank and Standard solution at target level. Calculate the % RSD of response for six replicate injections.
Analysis sequence
Blank – 1 injection
Standard solution – 6 injections
Acceptance criteria
The relative standard deviation for peak area counts is not more than 15.0%.
- Intermediate Precision (Ruggedness):
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.
Documentation:
Calculate the mean, standard deviation, and % RSD for the operators and instruments and record on data sheet.
Acceptance criteria:
% RSD of content of each solvent in six sample preparations should be NMT 10.0
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:
Accuracy can be accessed by spiking the standard solution of each solvent into test sample at levels 50%, 100% and 150% with respect to target levels. Analyze the solutions in three sample preparation at each level. Calculate the % RSD for recovery obtained at each level separately and overall %RSD.
Acceptance criteria
The mean recovery at each level shall be between 85% to 115 %.
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 flow rate by ±5%
- By changing the Oven temperature by ±2°C
Test Procedure:
Inject blank, six replicate injections of standard solution and two injection of spiked sample Solution.
Acceptance criteria
The system suitability and specificity should meet.
- Method Verification .
This method is validating using of Perkin Elmer GC-HS.
- Revision History:
| Revision No. | Details of changes | Reason |
| 00 | New Document | Nil |
