Protocol No. & Rev. No Effective date Reason for revision
    New Document  

Table of contents :

Sr. No. Subject Page No.
  Protocol Approval
  Product Profile
  Summary/Final conclusion/Recommendation
  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 protocol is to perform the additional testing to describe and specify the scientific and technical handling of dissolution data. This study shall be to compare the initial results of dissolution. This study shall be performs only on principal media.

  • Scope :

This protocol is applicable for the dissolution profiling of Cetirizine dihydrochloride tablets.

  • 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 Validation.
Review of data and summary report.
Head Quality Authorization of protocol.
  • Product Profile:
Category  Antihistamine
Reason for Validation First Validation
Active Ingredient Cetirizine Dihydrochloride
Strength Each coated tablet contains: Cetirizine Dihydrochloride               10 mg
Method Reference In House
Specification Limits Not less than 80% (Q) of the labeled amount.
  • Methodology:

 Chemical, reagents and filters:

Table 1.0: Chemical, reagents and filters

Sr. No Material /Chemicals/Filters Grade
1.0 Potassium dihydrogen phosphate (KH2PO4) AR Grade
2.0 Ortho phosphoric acid HPLC Grade
3.0 Water HPLC Grade or Milli Q water
4.0 Methanol HPLC Grade
5.0 Acetonitrile HPLC Grade
6.0 0.45 µm membrane filter Millipore or Equivalent
7.0 0.45 µm Nylon filter Millipore or Equivalent

Dissolution Parameters:

Apparatus                                      :           Paddle (II)

Dissolution Medium                      :           Purified Water

Dissolution Medium volume         :           900 ml

Speed                                            :           50 rpm

Temperature                                 :           37°C ± 0.5 °C

Sampling Time                              :           5, 10, 15, 20, 30, 45 minutes.

Chromatographic conditions (By HPLC):

Instrument                         : High Performance Liquid Chromatography (UV Visible/DAD Detector)

Column                             : ACE 5C18, 150 mm X 4.6 mm, 5 μm

Flow rate                           : 1.5 ml/min

Detector                            : 230 nm  

Column Temperature       : 40ºC

Injection Volume               : 20 µl

Run time                           : 6.0 minutes (Cetirizine RT Approximately 3.0 min)

Program                            : Isocratic Method

Mobile Phase                    : Mixture of Phosphate Buffer: Methanol: (350:650 V/V)

Diluent                               : Water: Methanol: (50:50) filtered through 0.45μ filter

Preparation of Phosphate Buffer:

Weigh accurately about 6.8 g of Potassium dihydrogen phosphate (KH2PO4) in 1000 ml of water. Adjust pH with dilute ortho-phosphoric acid to 2.6 ± 0.05.

Standard Preparation:

Weigh accurately about 56 mg of Cetirizine dihydrochloride working standard, transfer into a 100 mL volumetric flask. Add 60ml of diluent and sonicate till dissolve and dilute to volume with diluent.

Pipette out 10.0 ml of this solution and transfer into 50 ml of volumetric flask and make up the volume with dissolution media and mix. Further pipette 5.0 ml of this solution and transfer into 50 ml of volumetric flask, make up the volume with dissolution media and mix.

Sample Preparation:

Perform the test on six tablets. Place the 1 tablet in each dissolution vessels containing 900.0 ml of dissolution medium, previously maintained specified temperature (37°C ± 0.5 °C) and run the instrument for 45.0 minutes. After specified time interval mention, withdraw specified volume of aliquot from each vessel and filter through 0.45 µm Nylon Syringe filter, discard first 2-3 ml of the filtrate.

 Evaluation of System Suitability:

  1. Area % RSD             :   Should not be more than 2.0 % for peak area due to Cetirizine dihydrochloride from 6 replicate injections of standard solution.
  2. Tailing factor           :   Should not be more than 2.0 for peak due to Cetirizine dihydrochloride.
  3. Theoretical plates        :      Should not be less than 2000 for peak due to Cetirizine dihydrochloride.


Equilibrate the HPLC Column with mobile phase. Inject as per the following sequence or as per requirement.

Sequence of Injections:

Table 2.0: Sequence of Injection of dissolution

Preparation No. of Injections Action
Blank Solution 1 Record the blank Chromatograms
Standard solution 1 6 Check for system suitability Criteria (i), (ii) and (iii)
Sample solution 1 to 6 1 Inject the individual solution, Maximum 6 in a set.
Standard solution 1 (Bracketing standard) 1 %, Area RSD of total 7 injections ≤ 2.0% (6 Injection of standard solution  + 1 injection of bracketing standard)

Calculation for Dissolution (% labeled amount):

Correction factor is used to calculate results at different time interval

                                                                         AT          WS        10       5         900       P        

% Released Content   =        ——- x ——- x —— x —— x —— x —— x 100

                                                            AS       100      50       50        LC      100


AS    =       Average peak area due to Cetirizine Dihydrochloride obtained from six replicate

                  injections of standard solution.

AT    =       Peak area due to Cetirizine Dihydrochloride obtained from sample Solution

WS   =       Weight of Cetirizine Dihydrochloride RS/WS taken in mg.

LC    =       Label claim of Cetirizine Dihydrochloride in mg/tablet (10).

P       =       Potency of Cetirizine Dihydrochloride RS/WS in %, on as basis.

As per method of analysis, record the area on testing data sheet and calculate the % released, mean, standard deviation and % relative standard deviation. Obtained results will be report in tabulated manner as given below.

Dissolution profiling performed on three different batches.

  • Incident /Deviation:

Any incident or deviation observed during dissolution profiling should be recorded and reported.

  • Summary/ Conclusion / Recommendation:

Final conclusion should be drawn from analytical dissolution profiling.

  • Abbreviation

MSP               :           Miscellaneous study protocol

SD                  :           Standard deviation

HPLC             :           High performance liquid chromatography

RT                   :           Retention Time

ml                   :           Milliliter

mg                  :           Milligram

min.                 :           Minutes

QA                   :           Quality Assurance

QC                  :           Quality Control           

%                     :           Percentage

            ºC                    :            Degree centigrade

            hrs                   :           Hours

            µm                   :           Micrometer

            µl                     :           Microlitre

            BP                   :           British Pharmacopoeia

            RSD                :           Relative standard deviation

            NLT                 :           Not less than

NMT                :           Not more than

Vol                    :           Volume

AS                     :           Standard Area

AT                     :           Test Area

Bhanu Pratap Singh


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