SOP for Performance Check of UV/ Visible Spectrophotometer

1.0  OBJECTIVE

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 the quality control department.

3.0  RESPONSIBILITY             

3.1 Doing: Technical Assistant/Executive

3.2 Checking: Executive /Manager

4.0  ACCOUNTABILITY

       Head of the Department

5.0  PROCEDURE

       Frequency: Once in a three month

5.1  Operate the instrument as per respective S.O.P.

5.2  CONTROL OF ABSORBANCE

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

5.2  WAVELENGTH  ACCURACY (HOLMIUM FILTER SPECTRUM)

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  RESOLUTION POWER

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 the maximum at 269nm to maximum 266nm.

5.4.3  Record the result in Annexure-I. It should not less than 1.5

5.5  LIMIT OF STRAYLIGHT

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.

6.0  ABBREVIATIONS

       nm= Nanometer

       U.V= Ultraviolet

       ml=Millilitre

       KCl =Potassium chloride

       SOP = Standard operating procedure

       w/v = Weight per volume

       %  = Percentage

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