PROTOCOL FOR CLEANING METHOD VALIDATION OF IBUPROFEN 100MG & PARACETAMOL162.5MG SUSPENSION/5ML FOR IBUPROFEN

PROTOCOL FOR CLEANING METHOD VALIDATION OF IBUPROFEN 100MG & PARACETAMOL162.5MG SUSPENSION/5ML FOR IBUPROFEN IN ORAL LIQUID BLOCK 

Superseded Protocol No.	Nil
Effective Date

TABLE OF CONTENTS :

Sr. No.	Subject	Page No.
	Protocol Approval
	Objective
	Scope
	Responsibility
	Product profile
	Methodology
	Specificity
	Precision
	Method Precision
	Intermediate Precision
	Linearity
	Accuracy
	Range
	Limit of Detection (LOD)
	Limit of Quantification (LOQ)
	Stability of Analytical solution
	Robustness
	Incident/Deviation
	Summary
	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

Authorized By:

Functional Area	Name	Designation	Signature/Date
Head Quality

• Objective:

The objective of this validation study is to validate Cleaning method validation of analysis for cleaning samples of Ibuprofen 100mg & Paracetamol 162.5mg Suspension/5ml for Ibuprofen in oral Liquid block by considering thefollowing parameters:

Parameters	Ibuprofen
Specificity
Precision
System Precision
Method Precision
Intermediate Precision (Ruggedness)
Linearity
Accuracy
Range
LOD
LOQ
Stability of Solution
Robustness

Following parameters will be seen during system suitability:

Sr. No.	Parameters	Limit
1.	% RSD for peak absorbance of Ibuprofen from 6 replicate of standard solution	Not more than 2.0

When the above parameters meet the method, the actual experiment shall be started.

• Scope :

This protocol is based on SOP No. QC-021 and applicable for the validation of cleaning method of sample analysis of Ibuprofen in Oral Liquid block.

• Responsibility of Validation Team:

Departments	Responsibilities
QC	Preparation and Review of Validation protocol.
Perform the validation as per approved protocol and recording of data.
Compilation and checking of data.
Preparation and review of Validation report.
To impart training of protocol to concerned department/persons.
QA	Review and approval of Validation protocol.
Co-ordination with QC to carry out Validation.
Review and approval of Validation report.
Head Quality	Authorization of protocol.

• Product Profile:

Category (% API Content)	Analgesic and Antipyretic
Reason for Validation	To Validate the analytical method for Cleaning sample analysis.
Active Ingredient	Ibuprofen
Methodology	In-House
Method Reference	In-House

• Methodology

         Method of analysis:

               Reagents:

• Methanol
• Diluents: Methanol

• Instrument : UV-VIS Spectrophotometer

Standard Solution preparation:

Weigh accurately about 50 mg Ibuprofen working standard in to a 100.0 ml volumetric flask, and add about 50 ml methanol, sonicate to dissolve and dilute to volume with methanol. Further dilute 2.0 ml of this solution to 100.0 ml with methanol.

Determine the maxima in the range of 400 nm to 200 nm by UV-VIS Spectrophotometer using diluents as a blank.

• Parameters to be Validated:
  • Specificity:

 Prepare blank, swab blank and standard solution 10 mcg/ml as per procedure given below:

Procedure:

Weigh accurately about 50 mg of Ibuprofen working standard in to a 100.0 ml volumetric flask, and add about 50 ml methanol, sonicate to dissolve and dilute to volume with methanol. Further dilute 2.0 ml of this solution to 100.0 ml with methanol.

Blank solution: Diluents

Swab Blank: Take 10.0 ml of diluents in a test tube, dip fresh swab stick in test tube and sonicate for 5 minutes.

Procedure: Take the absorbance ofblank, swab blank and standard solution at maxima in the range of 400 nm to 200 nm.

Acceptance criteria:

There should not be any interference from blank, swab blank with the analyte peak.

• System Precision:

System precision shall be evaluated bymeasuring the absorbance of 6 replicate of standard solution and calculate the relative standard deviation.

Procedure:

Weigh accurately about 50 mg of Ibuprofen working standard in to a 100.0 ml volumetric flask, and add about 50 ml methanol, sonicate to dissolve and dilute to volume with methanol. Further dilute 2.0 ml of this solution to 100.0 ml with methanol.

Acceptance Criteria: % RSD of Six replicate should not be more than 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.

Procedure for Swab Technique:

Dissolved about 50.0 mg of Ibuprofen accurately weigh in a 100.0 ml volumetric flask with diluents and mix well. Further dilute 2.0 ml of this solution in a 100.0 ml volumetric flask with diluents (10mcg/ml).

Prepare the Standard stock solutions of Ibuprofen standard spiked the 0.5 ml solution of each into the area of 10 x 10 cm² in Six different SS Plates. Allow the solutions to dry in air. Swab wipe shall be taken and moisten with diluents, swab one SS plate in the area of 10 x 10 cm². Collect the swab in the test tube containing already 25.0 ml of   diluents in the test tube and sonicate for 10 minutes with swirling. Squeeze the swab and discard. Filter through 0.45µ filter. Final concentrations of standard solutions are 10.0 ppm.

• Procedure for Rinse Technique:

Prepare the Standard stock solutions of Ibuprofen standard spiked the 2.0 ml solution of each into a 1000ml SS Container allow the solutions to dry in air.  Rinse the container with 100.0 ml of diluent. Filter through 0.45µ filter. Final concentrations of standard solutions are 10.0 ppm.

• Intermediate Precision:           

Intermediate precision expresses within laboratory variation with different analysts or equipment on different days using same batch of drug product as per finished product testing procedure.

Procedure for Swab Technique:

Dissolved about 50.0 mg of Ibuprofen accurately weigh in a 100.0 ml volumetric flask with diluents and mix well. Further dilute 2.0 ml of this solution in a 100.0 ml volumetric flask with diluents (10mcg/ml).

Prepare the Standard stock solutions of Ibuprofen standard spiked the 0.5 ml solution of each into the area of 10 x 10 cm² in Six different SS Plates. Allow the solutions to dry in air. Swab wipe shall be taken and moisten with diluents, swab one SS plate in the area of 10 x 10 cm². Collect the swab in the test tube containing already 25.0 ml of   diluents in the test tube and sonicate for 10 minutes with swirling. Squeeze the swab and discard. Filter through 0.45µ filter. Final concentrations of standard solutions are 10.0 ppm.

• Procedure for Rinse Technique:

Prepare the Standard stock solutions of Ibuprofen standard spiked the 2.0 ml solution of each into a 1000ml SS Container allow the solutions to dry in air.  Rinse the container with 100.0 ml of diluent. Filter through 0.45µ filter. Final concentrations of standard solutions are 10.0 ppm.

• Linearity:

The linearity of the method shall be established between 6mcg/ml to 14mcg/ml. The sample absorbance shall be plotted against concentration and correlation coefficient shall be calculated.

Minimum five points shall be covered from the range of 6mcg/ml to 14mcg/ml.

Following concentrations shall be prepared and analyzed for linearity: 6 mcg/ml, 8mcg/ml,   10 mcg/ml, 12mcg/ml and 14mcg/ml.

Procedure: Measure the absorbance of sample solution against blank (diluents) in duplicate. Plot the graph of absorbance v/s actual concentration. Calculate correlation coefficient from linearity curve.

  Acceptance Criteria: Correlation coefficient shall not be less than 0.99

• Recovery:
• Procedure: (for swab and rinse technique)

Dissolved about 50.0 mg of Ibuprofen accurately weigh in a 100.0 ml volumetric flask with diluents and mix well. Further dilute 2.0 ml of this solution in a 100.0 ml volumetric flask with diluents (10mcg/ml).

Prepare the recovery stock solutions of Ibuprofen standard spiked the 0.5 ml solution of each into the area of 10 x 10 cm² in three different SS Plates. Allow the solutions to dry in air. Swab wipe shall be taken and moisten with diluents, swab one SS plate in the area of 10 x 10 cm². Collect the swab in the test tube containing already 25.0 ml of   diluents in the test tube and sonicate for 10 minutes with swirling. Squeeze the swab and discard. Filter through 0.45µ filter. Final concentrations of recovery solutions are 8.0 ppm, 10.0 ppm, and 12.0 ppm. 

        Acceptance criteria:

Recovery at each level should not be less than 70% and % RSD for each concentration should not be more than 10.0 %. 

RECOVERY
	Recovery 8 ppm	Recovery 10 ppm	Recovery 12 ppm
Test 1
Test 2
Test 3

• Procedure for Rinse Technique:

Prepare the Standard stock solutions of Ibuprofen standard spiked the 2.0 ml solution of each into a 1000ml SS Container allow the solutions to dry in air.  Rinse the container with 100.0 ml of diluent. Filter through 0.45µ filter. Final concentrations of recovery solutions are 8.0 ppm, 10.0 ppm, and 12.0 ppm.

• Range:

The range of an analytical procedure is the interval between the upper and lower concentration of analyte in the sample for which it has been demonstrated that the analytical procedure has a suitable level of precision, accuracy and linearity. The range is normally expressed in the same units as test results obtained by the analytical method. (e.g. percent, parts per million).

• Limit of Detection (LOD):

Limit of detection is the lowest concentration of analyte in a sample that can be detected but not necessarily quantified under the stated experimental condition. To determine the lowest amount of analyte in a sample which the analytical method can:

Detect but not necessarily quantitate as an exact value (LOD)

                                    LOD (mcg/ml)     =     3.3 x σ

                                                                           S

 Where σ = Standard deviation of the response of swab blank.

              S = Slope of Linearity Curve.

The LOD was determined for Ibuprofen Calculated the concentration at which the signal to noise ratio is about 3:1.

Prepare the LOD Solution using blank, placebo spiked with known concentration at determined LOD level. Calculate % RSD for six replicates of Ibuprofen.

Acceptance criteria:

% RSD for six replicates shall be not more than 30.

• Limit of Quantification (LOQ):

 Limit of Quantification is the lowest concentration of analyte in a sample that can be quantify with   acceptable precision under the stated experimental condition.

 Based on the standard deviation of the response of swab blank and the slope of linearity curve, LOQ shall be calculated by the following equation:

                                    LOQ (mcg/ml)     =       10 x σ

                                                                           S

  Where σ = Standard deviation of the response of swab blank.

                    S = Slope of Linearity Curve.

     Prepare the LOQ Solution and measure the absorbance of sample in six replicates.

Calculate % RSD for six replicates of Ibuprofen.

Acceptance criteria:

% RSD for six replicates shall be not more than 10.

• Stability of Analytical Solution:

It is essential when validating an analytical method to confirm that the analyte has adequate stability of standard solution during analytical measurement stages of the testing.

Prepare the standard solution and take the absorbance of standard solution at different time intervals.

Acceptance criteria:

Cumulative % RSD of standard absorbance shall not be more than 2.0%.

• Robustness

The method should show reliability of an analysis with respect to deliberate variation in method   parameters.

Following deliberate variations should be done in method parameters:

• By changing the wavelength by ±2nm.
•       Incident/Deviation:

Any Incident or Deviation observed during Analytical Method validation should be recoded and reported in Validation Report.

•       Summary/Conclusion:

Final Conclusion should be drawn from analytical cleaning method validation for its use to analyze the Cleaning sample of Ibuprofen 100mg & Paracetamol 162.5mg Suspension/5ml for Ibuprofen in oral Liquid block by UV.

Summary of validation report should be prepared and accordingly STP to be updated.

1. Revision History:

Revision No.	Details of changes	Reason
00	Nil	New