SOP for operation , cleaning and maintenance of UV-VIS Spectrophotometer


1.0 Scope : This procedure is applicable for the operation, cleaning and maintenance of UV-VIS spectrophotometer installed in Quality Control laboratory of pharmaceutical industries.

2.0 Responsiblities :

QC officer : Responsible for the operation and maintenance of the instrument.

QC Head/ Manager : Responsible for accountability and effective implementation of the standard procedure for operation of instrument.

3.0 Definition-Spectrophotometer : It is a device which measures the absorbance of sample at a fixed wavelength or in a wide scanning range of wavelength and produces spectrum when the beam of monochromatic light passes through sample.

4.0 Procedure  :

4.1Before operation ensure that the instrument is clean and calibrated.

Turn ON the main power of the instrument and computer connected with the instrument.

4.3   After turning ON the power, spectrophotometer will starts the initialization to checks the functions of all               parameter . Initialization will completed about after 5 minutes.

4.4  Wait for 15 minutes for the proper stabilization and illumination of UV lamp before operation.

Do not interrupt the initialization procedure it is important as it performs the cheks for functioning of all instrumental parameters each parameter will be highlighted which functions normally as the initialization proceeds and if any parameters is not functioning normally and the initialization will stop automatically.

4.6   Spectrophotometer can also be operated by it is functions keys provided on the instrument which displays         their response on the LCD screen of the instrument.

4.7  UV Probe software is commonly installed to operate the UV VIS spectrophotometer.

4.8   To run the software double click on the UV Probe software icon displayed on the desktop of the computer.         Login the main window by entering the individual login ID and password.

4.9   The two modes of measurements were:

  • Photometric mode
  • Spectrum mode

4.10   Photometric mode : It is used to measure the absorbance transmittance (T%) at affixed wavelength.

4.11   To connect spectrophotometer with software click on connect after connecting a green color indicator will           be highlighted on the computer

4.12  Select [Window] →[Photometric], it will be display UV Probe (system Administrator)- [Photometric].

4.13  Create the method enter the wavelength & add the parameter, select calibration and enter raw data and            save the method.

4.14  For blank correction, take two clean cuvettes, after proper rinsing keep the blank solution prior to the                measurement in the both cuvette and wipe them clean from the outside using tissue paper.

4.15  Place one cuvette in the reference cell holder and the other in the sample cell holder. Close the sample            compartment.

4.16  Press ‘AUTO ZERO’ key. In case of multi wavelength press ‘Baseline key.

4.17  After Blank correction, keep the sample in sample compartment. Click on ‘Read Unk.’ to measure the               absorbance.

4.18  After completion of measurement remove the cuvettes and wash them with purified water followed by               acetone, so that the cuvettes get dried easily.

4.19  Click Report button, it will displayed report template of respective test.

4.20  Press the “PRINT” key to take the result print.

4.21 Spectrum mode : Used to measure the absorbance of sample in wide range of selected wavelength it will scan the sample in selected range and produces the spectra.

4.22  Select [Window] → [Spectrum], it will be display UV Probe (system Administrator) – [Spectrum].

4.23  To create the measurement file select method enter the wavelength range from start to end select scanning rate at medium speed then save the method file at desired path (If method is already exist then open the method and do the baseline correction and scan the sample).

4.24 For baseline correction take two clean cuvettes, After proper rinsing, keep the blank solution prior to the measurement in the both cuvettes and wipe them clean from the outside using tissue paper.

4.25 Place the cuvette in the sample cell holder. Close the sample compartment.

4.26 Click on baseline button, the “Baseline Parameters” flie gets opened, select the wavelength range (nm) start to end. And click on ‘OK’ to measure the baseline.

4.27 Keep the standard or sample in sample compartment. Click on Start button for scanning, it will display the [New Data Set] , Save the data with respective name and path.

4.28 After scanning, Select [Operation] → [Peak Pick] to the data at each wavelength of the spectrum can be read.

4.29 Select Report button, it will displayed report template of respective test.

4.30 Press the “PRINT” key to take the result print.

5.0 Preacautions :

5.1 Do not open sample compartment during initialization.

5.2 Do not open sample compartment during baseline correction. Auto zero and sampling measuring mode.

5.3 Before placing the cuvettes in the sample holder clean the outer surface of cuvettes with tissue paper.

5.4 Do not hold the cuvettes from its transparent optical surface.

5.5 Operate the instrument after proper initialization.

6.0 Maintenance :

6.1 Press the keys and handle the instrument carefully.

6.2 When instrument is not in use switch off the instrument from mains.

6.3 Check and replace the silica gel placed inside the instrument once in week or whenever required.

6.4 Avoid any spillage of solution nearer to the instrument or if it happened the clean the area immediately.

6.5 Instrument should be serviced by service engineer as per AMC or as and when required.

6.6 Calibrate the instrument as per its calibration frequency or after any major maintenance.

7.0 Cleaning :

7.1 Clean the cuvettes after every analysis, wash the cuvettes several time with purified water and then rinse with methanol or acetone and wipe with tissue paper, dry and store in the box.

7.2 Clean the Outer surface of instrument with isopropyl alcohol and then dry with lint free cloth.

8.0 Abberviations :

UV-VIS : Ultra violet visible

QC : Quality Control

LCD : Liquid crystal display

AMC : Annual maintenance contract


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