SOP for Performance Check of UV/ Visible Spectrophotometer : Pharmaceutical Guidelines

SOP for Performance Check of UV/ Visible Spectrophotometer

Standard operating procedure to evaluate the performance of U.V.-Visible spectrophotometer using potassium dichromate.


To check the performance of equipment for reliable and accurate results.

2.0  SCOPE

       This procedure is applicable to calibrate U.V./Visible spectrophotometer, installed in quality control department.

3.0  RESPONSIBILITY             

3.1  Doing      : Technical Assistant/Executive
3.2  Checking : Executive /Manager


       Head of the Department


       Frequency  : Once in a three month
5.1  Operate the instrument as per respective S.O.P.


5.2.1  Weigh accurately about 60mg of potassium dichromate previously dried at 130°C for 3 hrs. & dissolve in 0.01N sulphuric acid to produce 1000ml.
5.2.2  Take the absorbance of the above solution at the wavelength mentioned below and calculate A(1%; 1cm) and record the results as per the Annexure-I.

            Sr. No.                         Wavelength                            Limit
               1                                   235nm                                122.9-126.2
               2                                   257nm                                142.4-145.7
               3                                   313nm                                47.0-50.3
               4                                   350nm                                104.9-108.2


5.3.1  Perform baseline correction with reference to air.
5.3.2  Put the holmium oxide filter into the cell compartment and record the absorbance spectrum between 200nm and 600nm.
5.3.3  Check and record the maxima at the wavelengths given below. And record as per the Annexure-I.

            Sr. No.                          Wavelength                            Limit
               1                                   241.15nm                             + 1 nm
               2                                   287.15nm                             + 1 nm
               3                                   361.50nm                             + 3 nm
               4                                   536.30nm                             + 3 nm


5.4.1  Prepare 0.02% v/v solution of toluene in hexane.
5.4.2  Take spectrum and calculate the ratio of absorption at maximum at 269nm to maximum 266nm.
5.4.3  Record the result in Annexure-I. It should not less than 1.5


5.5.1  Prepare 1.2% w/v solution of KCl in distilled water.
5.5.2  Take spectrum between 190nm to 210nm and take the absorption at 200nm.
5.5.3  Record the results in Annexure-I. It should more than 2.0.
5.5.4  The performance of the instrument is satisfactory, if the observed wavelengths are within the above tolerance limit.


       nm= Nanometer
       U.V= Ultraviolet
       KCl =Potassium chloride
       SOP = Standard operating procedure
       w/v = Weight per volume
       %  = Percentage

Ankur Choudhary is India's first professional pharmaceutical blogger, author and founder of Pharmaceutical Guidelines, a widely-read pharmaceutical blog since 2008. Sign-up for the free email updates for your daily dose of pharmaceutical tips.
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