SOP for Performance Check of UV - Visible Spectrophotometer (Jasco)

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 (Jasco), installed in the Quality control laboratory

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 S.O.P
5.2 Control of Absorbance
5.2.1 Weigh accurately about 60 mg of potassium dichromate previously dried at 130°C for 3 Hrs. and dissolve in 0.01N sulphuric acid to produce 1000 ml.
5.2.2 Take the absorbance of the above solution at the wavelength mentioned below and calculate A (1 %, 1 cm) and record the results.

Sr.No.        1     2     3     4

Wavelength     235 nm     257 nm     313 nm     350 nm

Limit 122.9 – 126.2 142.4 – 145.7 47.0   -  50.3 104.9 – 108.2

5.3 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 200 nm and 600 nm.
5.3.3 Check and record the maxima at the wavelengths given below, and record.

  Sr. No.                   Wavelength                          Limit

    1                             241.15 nm                          ± 1 nm

    2                             287.15 nm                          ± 1 nm

    3                             361.50 nm                          ± 3 nm

    4                             536.30 nm                          ± 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 maximum at 269 nm to minimum 266 nm.
5.4.3 Record the result.

5.5 LIMIT OF STRAY LIGHT

5.5.1 Prepare 1.2% w/v solution of KCl in distilled water.
5.5.2 Take spectrum between 190 nm to 210 nm and take the absorption at 200.00 nm.
5.5.3 Record the result.
5.5.4 The performance of the instrument is satisfactory , if the observed wavelengths are within the above tolerance limit, otherwise follow the S.O.P.

Related: Principle of Ultraviolet and Visible Absorption Spectrophotometry

6.0 ABBREVIATIONS

6.1 nm = Nanometer
6.2 U.V = Ultraviolet
6.3 ml = Millilitre
6.4 KCl = Potassium chloride
6.5 w/v = Weigh per volume
6.6 % = Percentage

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