by METHOD VERIFICATION PROTOCOL FOR ASSAY OF CHLORPHENAMINE MALEATE Ph. Eur.
| Superseded Protocol No. | Nil |
| Effective Date |
Table of contents :
| Sr. No. | Subject | Page No. |
| Protocol Approval | ||
| Objective | ||
| Scope | ||
| Responsibility of validation team | ||
| Product profile | ||
| Methodology | ||
| Verification Parameter | ||
| Incident / Deviation | ||
| Summary | ||
| Abbreviation | ||
| 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 verification is to verify the assay method of Chlorphenamine Maleate by considering the following parameters:
The objective of this verification is to provide documentary evidence that analytical methodology used in the analysis of assay of Chlorphenamine Maleate potentiometrically (non aqueous) has reliable results within the predetermined acceptance criteria.
Analytical method verification will be performed by considering the following parameters:
| Parameters | Chlorphenamine Maleate |
| Specificity | |
| Method Precision | |
| Intermediate Precision | |
| Linearity |
Assay method of Chlorphenamine Maleate is European Pharmacopoeia method, hence verification of method to be performed to check the suitability of the method under actual condition of use. Assay method of Chlorphenamine Maleate is titrimetric method (Potentiometrically non aqueous).
- Scope :
This protocol is applicable for the verification of assay method of Chlorphenamine Maleate.
- 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. | |
| To impart training of protocol to concerned department/persons. | |
| QA | Review and approval of protocol. |
| Co-ordination with QC to carryout Verification. | |
| Review of data and summary report. | |
| Head Quality | Authorization of protocol. |
- Product Profile:
| Category ( API) | antihistamine |
| Reason for Verification | Pharmacopoeial Method |
| Active Ingredient | Chlorphenamine Maleate |
| Method Reference | European Pharmacopoeia / RC007U |
| Specification Limits | Assay (on the dried basis). 98.0 per cent to 101.0 per cent |
- Methodology:
Assay
Reagents required: following mention chemical reagents in Table 1.0 required in method analysis used for assay of Chlorphenamine Maleate Ph. Eur.
Table 1.0: Reagent
| Sr. No | Reagents | Grade |
| 1. | Anhydrous Acetic Acid | AR Grade |
| 2. | 0.1 M Perchloric Acid | AR Grade |
Procedure:
Dissolve 0.150 g in 25.0 ml of anhydrous acetic acid R. Titrate with 0.1 M perchloric acid, determining the end point potentiometrically.
1 ml of 0.1 M perchloric acid is equivalent to 0.01954 g of C20H23ClN2O4 (Chlorphenamine Maleate)
Calculation:
V x 0.01954 x 100 x M
——————————————————————-
0.1 x W
Where,
V = volume of 0.1 M perchloric acid
M = Molarity of 0.1 M perchloric acid
W = weight of sample in g
Acceptance Criteria:
If the % Assay on dried basis is within the specification limit (Between 98.0 % and 101.0 %), then the material is deemed as passing this test.
Loss on Drying
Procedure:
Weigh a glass-stopper shallow weighing bottle that has been dried previously dried at 105°C ± 2°C, cool to room temperature by keeping in desiccator then weigh. Transfer to the bottle the 1.000 g quantity of the sample, cover it and accurately weigh the bottle and the contents. Dry the substance at 105°C ± 2°C for 4 hours. After drying is completed open the drying chamber close the bottle promptly and allow it to cool to room temperature in a desiccator before weighing. Weigh the bottle and contents. Dry the substance to constant mass and calculate the loss on drying using the given below formula:
Calculation:
Loss in weight (g) x 100
=—————————————————–
Weight of sample (g)
Requirement: As per current version of approved specification.
Analytical Parameters to be verified and acceptance criteria given below Table 2.0:
Table 2.0
| Sr. No. | Analytical Performance Parameter | Acceptance Criteria |
| 1. | Specificity | To check the Interference of the 0.1 M perchloric acid. |
| 2. | Method Precision | The assay values should be in between 98.0 % to 101.0 % on dried basis for all six determinations.The % RSD of six assay values should be NMT 2.0 for Chlorphenamine Maleate. |
| 3. | Intermediate Precision (Ruggedness) | The assay values should be in between 98.0 % to 101.0 % on dried basis for all six determinations of second analyst and its %RSD should be NMT 2.0.The cumulative % RSD of assay values for both the analysts should NMT 2.0. |
| 4. | Linearity | The correlation coefficient shall be not less than 0.999. |
- Verification Parameter
The following parameters to be perform for the Verification activity.
- Specificity
- Method Precision
- Intermediate
- Linearity
- 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 0.1 M Perchloric acid. Prepare blank solution without analyte and titrate as per methodology and check the interference and record the titrant value consume.
- 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 six individual sample of same batch and analyze as per methodology.
Calculate the % assay of Chlorphenamine Maleate on dried basis and the % RSD of assay for six individual samples.
Acceptance Criteria:
The assay should be between 98.0 per cent and 101.0 per cent on dried basis for all six determinations. The % RSD for assay of six individual samples should be NMT 2.0.
- Intermediate Precision (Ruggedness):
Intermediate Precision (Ruggedness) of an analytical method is the degree of repeatability of test results obtained by the analysis of same samples by different analyst on different days.
Note: The result obtained from method precision will be considered as the results of first analyst for comparison in ruggedness.
Intermediate precision is performed by analyzing a homogenous sample, by second analyst on second day by preparing six individual sample of same batch and analyze as per methodology. Calculate the % assay on dried basis and the % RSD of assay for six individual samples performed by second analyst.
Acceptance Criteria:
The assay should be between 98.0 per cent and 101.0 per cent on dried basis for all six determinations performed by second analyst. The % RSD for assay of six individual samples should be NMT 2.0.
The cumulative % RSD of assay for both the analysts is NMT 2.0.
- 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 shall be prepared. Determine the linearity by preparing and analyze the sample solution in the range of 80% to 120% of concentration level and calculate the correlation coefficient “r”.
Prepare the sample solutions at five concentrations, typically 80%, 90%, 100%, 110% and 120% of target concentration following material testing procedure. Each concentration will be analyzed. Record results on a datasheet. Plot a graph with concentration (x-axis) versus volume used (ml) on y-axis. Calculate the correlation coefficient (r). Record the calculations on the datasheet.
Level 80 %: Take sample weight of 0.120 g and dissolve in 25.0 ml of anhydrous acetic acid R and titrate with 0.1 M perchloric acid, determining the end-point potentiometrically.
Level 90 %: Take sample weight of 0.135 g and dissolve in 25.0 ml of anhydrous acetic acid R and titrate with 0.1 M perchloric acid, determining the end-point potentiometrically.
Level 100 %: Take sample weight of 0.150 g and dissolve in 25.0 ml of anhydrous acetic acid R and titrate with 0.1 M perchloric acid, determining the end-point potentiometrically.
Level 110 %: Take sample weight of 0.165 g and dissolve in 25.0 ml of anhydrous acetic acid R and titrate with 0.1 M perchloric acid, determining the end-point potentiometrically.
Level 120 %: Take sample weight 0.180 g and dissolve in 25.0 ml of anhydrous acetic acid R and titrate with 0.1 M perchloric acid, determining the end-point potentiometrically.
Acceptance criteria
The correlation coefficient (r) between observed titrant value and the weight taken shall be not less than 0.999.
- Incident / Deviation:
Any Incident and deviation during verification exercise shall be reported and justified. Raw data of all non-satisfactory results shall be kept along with verification report.
- Verification Summary:
Data obtained through verification exercise shall be analysed with regard to various parameters and their acceptance criteria. A verification report shall be prepared summarizing all data. All results shall be captured in the method verification report.
Abbreviation
ASS : Assay
VER : Verification
P : Protocol
SD : Standard deviation
mL : Milliliter
mm : Milimeter
mg : Milligram
g : Gram
min. : Minutes
QA : Quality Assurance
QC : Quality Control
% : Percentage
ºC : Degree centigrade
Corr. coeff.(r) : Correlation coefficient
hrs : Hours
µm : Micrometer
EP : European Pharmacopoeia
RSD : Relative standard deviation
NLT : Not less than
NMT : Not more than
V : Volume
M : Molarity
W : Weight
Revision History:
| Revision No. | Details of changes | Reason |
| 00 | Nil | New document |
