amv protocol for chloramphenicol 250 capsule

ASSAY TESTBYUV SPECTROPHOTOMETRY

analytical method validation protocol for chloramphenicol 250 capsule

DOCUMENT NO. :
PRODUCT NAME :	CHLORAMPHENICOL 250 CAPSULE
ISSUE DATE :
SUPERSEDES :	Nil

TABLE OF CONTENT

Analytical Method Protocol Approval Sheet

S.No.	Subject	Page no.
1	Introduction
2	Objective
3	Scope
4	Responsibilities
5	Procedure
5.1	Selection Of Analytical Performance Parameters
5.2	Description of Analytical Method
6.0	Description of Analytical performance parameter
6.1	Specificity Study ( Placebo Interference)
6.2	Accuracy Study
6.3	Precision Study
6.3.1	System Precision Study (Repeatability )
6.3.2	Intermediate Precision Study (Reproducibility)/Ruggedness
6.4	Linearity & Range study
6.5	Robustness Study
7.0	Acceptance Criteria
8.0	Re-Validation Criteria
9.0	Documentation
10.0	Summary Report And Conclusion
11.0	Conclusion

PROTOCOL PREPARATION AND APPROVAL

The approval of this AMVP (CHLORAMPHENICOL 250 CAPSULE) will be joint responsibility

of the following functional areas. Any modification in this document shall be authorized and documented.

Prepared By

Name

Designation/Department

Signature

Date

Checked By

Name

Designation/Department

Signature

Date

Reviewed By

Name

Designation/Department

Signature

Date

Approved By

Name

Designation/Department

Signature

Date

2.0 INTRODUCTION:
Analytical method validation is performed to validate the suitability ofanalytical method. This protocol describes the validation study of theanalytical method, which is to be used for the determination of “Chloramphenicol”content in 250 Capsule.

3.0 OBJECTIVE
To generate the documentary evidence for analytical method validation,for determination of Chloramphenicolcontent in 250 Capsule., which is carried out byUVSpectrophotometer technique. The performance characteristics of themethod should meet the requirements for the intended analytical applicationof content determination at finished product stage.

Analytical Performance Characteristics or parameters that will beconsidered in the method validations are as follows:

1. Specificity.
2. Accuracy / Recovery.
3. Precision: – Method Precision.
Intermediate Precision
4. Linearity and Range.
5. Stability of Solution

4.0 SCOPE

This protocol is applicable to validate the analytical method forDeterminationof Chloramphenicol” content in Chloramphenicol 250 Capsule.
RESPONSIBILITY

No	Activity	Responsibility
1	Writing Of Protocol	Officer Quality Control
2	Review And Checking Of Protocol	Manager Quality Assurance
3	Approval Of Protocol	Manager Quality Assurance
4	Validation Study	Chemist/ Officer Quality Control
5	Evaluation And Approval Of Results	Manager Quality Assurance

4.0 PROCEDURE

4.1 SELECTION OF ANALYTICAL PERFORMANCE PARAMETERS

As per cGMP guidelines the test methods, which are used for assessing the quality of pharmaceutical products with established specifications, must be validated for analytical performance parameters viz; Limit ofdetection,Limit of quantitation, Precision, Accuracy, Linearity & range and Ruggedness.

However, if the analytical method employed is in the IP, BP., USP /NF, Association of official analytical chemists, Book of methods or inother recognized standard references, or is detailed in an Approved NewDrug Application (ANDA) and the preferred method is not modified, isnot required to be validated for accuracy and reliability parameters butmerely verify their suitability under actual conditions of use.i.e. For itssuitability under actual conditions of use,(Placebo interference),Precision, Accuracy, Linearity & Range, Ruggedness.

Listed below in Table 1 are analytical parameters required to bevalidated depending upon the test and pharmacopoeial status.

TABLE 1. Analytical parameters required for validation

TEST	PHARMACOPOEIAL		NON – PHARMACOPOEIAL
TEST	Bulk Drug	Drug Product	Bulk Drug	Drug Product
Assay	Precision Linearity Range Ruggedness	Specificity AccuracyPrecisionLinearityRuggedness	Specificity Accuracy Precision Linearity Range Ruggedness	Specificity Accuracy Precision Linearity Ruggedness
Related Substances Impurity Profile	Precision Accuracy Linearity Range LOD LOQ Ruggedness	Specificity Accuracy Precision Linearity LOD LOQ Ruggedness	Accuracy Precision Linearity Range LOD LOQ Ruggedness	Specificity Accuracy Precision Linearity LOD LOQ Ruggedness

• LOD – Limit of Detection
• LOQ – Limit of Quantitation
• NA – Not Applicable

Analytical method validation study forChloramphenicol” content in Chloramphenicol 250 Capsuleshall be carried out forthe following analytical performanceparameters viz, accuracy, precision, specificity, linearity, range, andRuggedness study.

5.2 DESCRIPTION OF ANALYTICAL METHOD
The detailed analytical method for the determination of Chloramphenicol” content in 250 Capsuleis described below:
5.2.1 Apparatus

* UV Spectrophotometer (Make:- )
* Analytical balance (Make: – )
5.2.2 Glassware
*Volumetric flask 200 ml,100ml.
*Volumetric Pipette 10 ml
* Glass Beaker
* Glass Funnel
5.2.3 Reagents
Water

5.2.4 Procedure:

Assay

TEST DILUTION:
Weigh accurately a quantity of the mixed contents of 20 capsules equivalent to about 0.2 g of Chloramphenicol, dissolve in 800 ml of water, warming if necessary to effect solution and add sufficient water to produce 1000.0 ml. Dilute 10.0 ml of this solution to 100.0 ml with water and measure the absorbance of the resulting solution at the maximum at about 278 nm, . Calculate the content of C11H12Cl2N2O5, taking 297 as the value of A (1%, 1 cm) at the maximum about 278 nm.
Limits : not less than 92.5% w/w and not more than 107.5% w/w of the stated amount of Chloramphenicol C11H12Cl2N2O5.

Calculation

Test Abs. * 10 * 1000 * 100

———- x ——–x ——–x——x ——Av. Wt.

Spe. Abs * 1000 * Wt. Test *10

Acceptance LIMIT: Estimation of Chloramphenicol , 92.5 – 107.5 %

6.0 DESCRIPTION OF ANALYTICAL PERFORMANCE PARAMETERS

6.1 SPECIFICITY STUDY(PLACEBOINTEREFERENCE)

Specificity of an analytical method is its ability to measure accuratelyand specificallythe analyze in the presence of components viz. knownand unknown impurities, degradation products or placebo ingredients thatmay be expected to be present in the sample matrix.
Placebo study of analytical method use for drug product is checkingthe positive or negative interference due to placebo on final resultsof analytical method. Placebo study of the Chloramphenicol” content in CAFCOL 250 Capsule shall be established by analyzing the placebo as perassay method and calculating the % interference of the same.

6.2 ACCURACY STUDY

The accuracy of an analytical method is the closeness of test results obtained by that method to the true value. Accuracy may often be expressed as percent recovery by the assay of known, added amount of Chloramphenicol.

The accuracy of the method is determined by adding known amount of Chloramphenicol to cover both, above and below (80%, 100% & 120%) the normal level of the analyte concentration.

The accuracy of the assay method of Chloramphenicol” content in Chloramphenicol 250 Capsule shall be determined by adding known amount of Chloramphenicolin placebo to cover both above and below ( 80% , 100% & 120% ) the normal levels
(label claim)expected in the samples.

6.3 PRECISION STUDY (Repeatability)

The precision of an analytical method is the degree of repeatability of the results in a series of experiments run during a single session by single operator with identical reagents andequipments.

Precision of the Chloramphenicol” content in Chloramphenicol 250 Capsule shall be stablishedby separately weighing and analyzing the samples from a homogeneous sample for six times and by calculating % RSD.

The precision of the analytical method is established by performing two types of precision study.i. e. System precision study& Intermediate precision study.

6.3.1 SYSTEM PRECISION STUDY (REPEATABILITY)

This study is to confirm the degree of repeatability of the results in a series of experiments during a single session by single operator with identical reagents, glassware’s and equipment’s. Hence the reliability of standard analyte, reagents as well as equipment is evaluated by this study.

6.3.2 INTERMEDIATE PRECISION STUDY (REPRODUCIBILITY) / RUGGEDNESS

This study is to confirm the degree of repeatability of the results in a series of experiments within the same laboratory but different days, analysts, equipment’s/reagents.

6.4 LINEARITY &RANGE STUDY

The linearity of an analytical method is its ability to elicit test results directly proportional to the concentration of the analyte in samples within given range.

Linearity of Chloramphenicol” content in 250 Capsule shall be studied by taking minimum of five concentration of test sample in the range of drug substance from 40 % to 140 % content Linearity curve shall be plotted for absorbance against concentration. Correlation coefficient is calculated using above plot.

6.5STABILITY OF SOLUTION:

To demonstrate Stability of solution study, test solution of the Chloramphenicol” content in 250 Capsules shall be checked for stability of test solution over 24hrs and comprised the test results of sample containing impurities or degrading product

7.0 ACCEPTANCE CRITERIA

S.No	PERFORMANCE PARAMETERS	ACCEPTANCE LIMITS
1	Specificity study (Placebo interference)	NMT 1%
2	Accuracy study (in terms of % recovery)	98 % to 102%
3	Precision study 1. Method ( Repeatability) 2. Intermediate	%RSD = NMT 2%
4	Linearity study	Correlation Coefficient ( r = ³ 0.999)
5	Stability of solution	Variance ± 1 % of each other test condition

REVALIDATION CRITERIA

Analytical method once validated need to be revalidated when, Change control procedure follows for any of the below listed reason.

1. Some major or minor changes made in the validated method.
2. Change of product manufacturing process.
3. Change in product formulation. i.e. excipients.
4. Change in manufacturing site.

9.0 DOCUMENTATION (PRODUCT SPECIFIC)

After completion of activity of analytical method validation, detailed reports including result findings shall be prepared as per the above outlined guidelines which shall cover the details of analytical method, selection of analytical performance parameters, acceptance criteria and complete details about the validation procedure and plan.

10.0 SUMMARY REPORT

Summary of the results of completed analytical method validation study shall be tabulated.

11.0 CONCLUSION

After completion of analytical method validation study, the final report, summary and conclusion shall be reviewed and approved by Head Quality Assurance.

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