ANALYTICAL METHOD VERIFICATION PROTOCOL OF ASSAY OF NAPROXEN 500 MG TABLETS

ANALYTICAL METHOD VERIFICATION PROTOCOL OF ASSAY OF NAPROXEN 500 MG TABLETS

Superseded Protocol No.	Nil
Effective Date

Table of contents :

Sr. No.	Subject	Page No.
	Protocol Approval
	Objective
	Scope
	Responsibility
	Product profile
	Methodology
	Verification parameters
	Incident/Deviation
	Summary/Final conclusion/Recommendation
	Abbreviation
	Revision History

1. 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

• Objective:

The objective of this protocol is to verify the suitability of Naproxen 500 mg tablet. by considering the following parameters:

Parameters	Naproxen
Specificity	yes
Precision
Method Precision	yes
Intermediate Precision	yes

• Scope :

This protocol is applicable for the verification of assay of Naproxen 500 mg tablet.

• Responsibility of Validation Team:

Departments	Responsibilities
QC	Preparation & Review of Protocol.
Analysis of samples and recording of data.
Compilation and Review of data
Preparation of Summary Report.
To impart training of protocol to concerned department/persons.
QA	Review of protocol.
Co-ordination with QC to carryout Verification.
Review of data and summary report.
Head QA	Approval of Protocol

• Product Profile:

Category	Nonsteroidal Anti-inflammatory Drug (NSAID)
Reason for Verification	First Verification
Active Ingredient	Naproxen 500 mgTablet
Method Reference	European Pharmacopoeia
Specification Limits	Assay Each uncoated tablet contains Naproxen: 475.00 – 525.00 mg 95.0 – 105.0 %

• Methodology:

Assay by UV-Visible Spectrophotometer

Chemical, reagents and filters:

Table 1.0: Chemical, reagents and filters

Sr. No	Material /Chemicals/Filters	Grade
	Methanol	HPLC Grade
	0.45 µ membrane filter	Milli Pore or Equivalent

Sample solution:

Transfer a quantity of tablet powder (from 20 tablets) equivalent to 50 mg of Naproxen into 100 ml volumetric flask, shaken for 30 minutes with 70 ml of methanol; and dilute to volume with Methanol and filter through 0.45 µ membrane filter. Dilute 10 ml of the filtrate to 50 ml with methanol.

Standard solution:

Weigh accurately about 50 mg of Naproxen working standard and transfer into 100 ml volumetric flask, add about 50 ml Methanol, dissolve and make up to volume with Methanol and filter through 0.45 µ membrane filter. Dilute 10 ml of the filtrate to 50 ml with methanol.

Procedure:

Measure the absorbance of the sample solution and standard solution at the maximum of 331 nm against methanol.

Calculation:

                                              At x Ds X P

Content of Naproxen     = ————————- x N

(C₁₄H₁₄O₃) (mg/ tablet)         As X Dt X 100  

Where;

At: Absorbance of sample solution

As: Average absorbance of standard solution.

Dt: Dilution factor for sample solution (Weight / dilution)

Ds: Dilution factor for standard solution (Weight / dilution)

P: Percent purity of Naproxen working standard (on as basis)

N: Average weight of tablet in mg

                                  Content of Naproxen (mg / tablet)

% label amount = ——————————————————- x 100

                                                      500

 Requirement: As per current version of approved specification.

• Verification parameters:

The following parameters to be perform for the verification activity.

Specificity

Method Precision

Intermediate

• 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 blank and placebo.

Specificity of test method should be established by separate titration of blank solution (diluent), Placebo, sample solution and spiked sample solution. Sample solution shall be prepared as per method of analysis given in section 6.0.

Blank Solution: Methanol

Standard solution:

Weigh accurately about 50 mg of Naproxen working standard and transfer into 100 ml volumetric flask, add about 50 ml Methanol, dissolve and make up to volume with Methanol and filter through 0.45 µ membrane filter. Dilute 10 ml of the filtrate to 50 ml with methanol.

Sample solution:

Transfer a quantity of tablet powder (from 20 tablets) equivalent to 50 mg of Naproxen into 100 ml volumetric flask, shaken for 30 minutes with 70 ml of methanol; and dilute to volume with Methanol and filter through 0.45 µ membrane filter. Dilute 10 ml of the filtrate to 50 ml with methanol.

Note: Standard weight can be reduce to achieved the final concentration same.

Placebo solution: Transfer a quantity of placebo powder of 25.6 mg into 100 ml volumetric flask, shaken for 30 minutes with 70 ml of methanol; and dilute to volume with Methanol and filter through 0.45 µ membrane filter. Dilute 10 ml of the filtrate to 50 ml with methanol.

Spike solution: Transfer a quantity of placebo powder of 25.6 mg + 50 mg of Naproxen API into 100 ml volumetric flask, shaken for 30 minutes with 70 ml of methanol; and dilute to volume with Methanol and filter through 0.45 µ membrane filter. Dilute 10 ml of the filtrate to 50 ml with methanol.       

Procedure:

Take the absorbance of placebo, standard, test and spike test solution using methanol as a blank in a UV – Visible spectrophotometer. Scan the samples from 200 nm to 400 nm.

Table 4.0: Specificity data

Sr. No	Sample	Maximum Absorbance at 331 nm
	Blank
	Placebo solution
	Standard solution
	Test Solution
	Spike test solution

Acceptance Criteria:

1. There should be no interference of the methanol and placebo at the wavelength of 331 nm.
2. The Interference of blank and placebo should be not more than 2.0%.
   1. 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 Naproxen for each preparation and the % RSD of assay for six individual samples.

Acceptance Criteria: The assay should be between 99.0 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 99.0 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.

• Incident /Deviation:

Any incident or deviation observed during analytical method verification should be recorded and investigation as per SOP.

• Summary/ Conclusion / Recommendation:

Final conclusion should be drawn from analytical method verification for its use to analyze the assay test of Naproxen Tablet. by Autotitrator.

Summary of verification report shall be prepare and accordingly conclusion and recommendation to be given.

• Abbreviation

ASS                 :           Assay

            VERP              :           Verification Protocol

            SD                   :           Standard deviation

            ml                    :           Milliliter

            mg                   :           Milligram

            min.                 :           Minutes

            QA                   :           Quality Assurance

            QC                  :           Quality Control

            %                     :           Percentage

            ºC                    :            Degree centigrade

            EP/Ph.Eur.      :           Europeian Pharmacopoeia

            RSD                :           Relative standard deviation

            NLT                 :           Not less than

NMT                :           Not more than

            g                      :           Gram

V                      :           Volume

M                     :           Molarity

• Revision History:

Revision No.	Details of changes	Reason
00	Nil	New Document