Superseded Protocol No. Nil
Effective Date  

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
  Protocol Approval
  Responsibility of Validation Team
  Product Profile
  Revision History
  • Objective:

The objective of this verification is to verify the dissolution method of Ofloxacin in Ofloxacin Tablets by considering thefollowing parameters:

  • Specificity
  • Precision
  • System Precision
  • Method Precision
  • Intermediate Precision (Ruggedness)
  • Linearity and Range
  • Stability of Analytical Solutions
  • Scope :

This protocol is based on SOP and applicable for the verification of dissolution method of Ofloxacin in Ofloxacin tablet.

  • 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 Verification.
Review of data and summary report.
Head Quality Authorization of protocol.
  • Product Profile:
Category (% API Content) Antibacterial
Reason for Verification 1st Verification
Active Ingredient Ofloxacin USP
Strength Each Film Coated tablet contains: Ofloxacin USP    200 mg
Methodology In House
Method Reference In House
Specification Limits (At release) NLT 75.0 % ( D) of Ofloxacin is dissolved in 30 minutes
(Shelf Life) 36 Months
  •    Methodology:

Dissolution Parameters:

Dissolution Medium:  900ml of 0.1M Hydrochloric Acid                    

Apparatus: I (Paddle)

Speed          :  50 rpm

Time            :  30 minutes

Preparation of 0.1 M Hydrochloric Acid

Take 51 ml concentrated Hydrochloric Acid carefully and dissolve in 6 ltr water.

Standard Preparation:

Take about 20 mg of Ofloxacin WS / RS in 100.0 ml volumetric flask and dissolve with 0.1M Hydrochloric Acid, mix well. Further dilute 2.0 ml of this solution to 100.0 ml with 0.1M Hydrochloric Acid.


Place the 900 ml of the dissolution medium, free from dissolved air, into the six vessels of the apparatus. Assemble the apparatus and warm the dissolution medium to 36.5°C to 37.5ºC.Place one tablet in to the each vessel taking care to exclude air bubbles from the surface allow the tablet to sink to the bottom of the vessel prior to the rotation of the paddle. Operate the apparatus immediately at the speed of rotation 50 rpm for 30 minutes. After 30 minutes withdraw 20 ml of the sample and filter, further dilute 2.0 ml of filter sample to 100.0 ml with 0.1 M Hydrochloric Acid. Measure the absorbance of sample, at the maximum at about 294 nm. Calculate the content of Ofloxacin in the medium from the absorbance obtained from a solution of known concentration of Ofloxacin RS in the same medium.


      Abs         Ws         2         900       100          100             P       

 ——— x ——— x——–x ——— x——-x————–x———-  = %            

      As          100         100          1           2           200           100      


Abs: Absorbance of Ofloxacin obtained in the sample solution.

As: Absorbance of the Ofloxacin obtained in the standard solution.

Ws: Weight taken for Standard preparation in mg.

P= Potency of Ofloxacin working standard.      

Requirement: As per current version of approved specification.

  • 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 placebo.

Test Procedure:

Prepare and scan the Blank (diluents), Placebo, Standard solution & test solution.


Print chromatograms.

Acceptance criteria

The interference from blank and placebo should not be more than 2 % of standard absorbance.

  • Precision:System Precision:

The system precision is the closeness of agreement between the responses of detector. It is usually expressed as the standard deviation (SD) or the relative standard deviation (RSD).

Prepare and take the absorbance of Blank and Standard solution as per finished product testing procedure and record the absorbance of main peak in the standard solution. Calculate the % RSD of response for six replicate absorbance.

   Acceptance criteria

   The relative standard deviation for absorbance is not more than 2%.

  • Method Precision (Repeatability):

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 finish product testing procedure.


Record the % dissolution of Ofloxacin in Ofloxacin Tablet on the datasheet. Calculate the mean, standard deviation, and % RSD of six samples.

Acceptance criteria

 % RSD should not more be more than 5.

  • Intermediate Precision ( Ruggedness ):

Intermediate precision expresses within laboratory variation with different analysts on equipment within the same laboratory using same batch of drug product as specified under precision.

Test Procedure:

The analysis of the same batch will be done in six replicate analyses by using different analyst, on different day. The mean, standard deviation and relative standard deviation will be calculated.


Calculate the mean, standard deviation, and % RSD for the operators and instruments and record on data sheet.

Acceptance criteria

The relative standard (RSD) of six analyses should not be more than 5% and overall relative standard deviation shall be not more than 5% when compared with method precision results.

  • Linearity and range:

    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 inject the standard solution in the range of 80% to 120% of concentration level and calculate the correlation coefficient “r”.

Test Procedure:

Prepare the standard solutions at five concentrations, typically 80, 90, 100, 110 and 120% of target concentration following finished product testing procedure. Duplicate replicates at each concentration will be analyzed.


Record results on a datasheet. Calculate the mean, standard deviation, and Relative Standard Deviation (RSD) for each concentration. Plot concentration (x-axis) versus mean response (y-axis) for each concentration. Calculate the correlation coefficient (r). Record these calculations on the datasheet.

Acceptance criteria

The correlation coefficient (r) should not be less than 0.999

  • Solution stability:

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

    Test Procedure:

    Prepare the standard solution and perform the dissolution on one tablet as per finish product testing procedure and analyze the standard solution and sample solution at the different time intervals and calculate the difference for the assay result.

Acceptance criteria

    The difference in results should not be more than 2 from initial results.

  •   Revision History:
Revision No.   Details of changes Reason
00   New Document Nil

Bhanu Pratap Singh


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