UV SpectroPhotometer - Operation & Calibration

UV VISIBLE SPECTROPHOTOMETER
SOP FOR OPERATION

1.0. PURPOSE:
To provide a procedure for operating the UV – VIS Spectrophotometer.

2.0. SCOPE:
This covers the procedure for operating UV–VIS Spectrophotometer.
MODEL : UV-1700 with PC software
MAKE : Shimadzu.

3.0. RESPONSIBILITY:
3.1. QC-Chemist
3.2. QC-Officer

4.0. ACCOUNTABILITY:
Manager-Quality Control

5.0. DEFINITION:
A device which produces a spectrum of ultraviolet light and is provided with a calibrated scale for measurement of wavelength.

6.0. PROCEDURE:


6.1. For PHOTOMETRIC
6.1.1. Switch on the instrument and computer
6.1.2. Check the connection cable to the PC is properly connected or not.
6.1.3. Select the icon of UV probe and click double
6.1.4. Press PC control (F4) in the instrument.
6.1.5. Select the icon Photometry.
6.1.6. Click the CONNECT icon.
6.1.7. Place the blank solution in sample compartment
6.1.8. Click the method icon(M).The page will be opened enter the wavelength & add the parameter, select calibration and enter raw data and
          close the window and method should be saved in method folder( which is suitable for name of the sample).
6.1.9. Keep blank and perform AUTOZERO
6.1.10. To read the samples keep the test solution and enter the sample ID in sample table and click the Read unknown icon.(If the method
           is already exists open the method file place the blank and make AUTOZERO).
6.1.11. To take print out click on REPORT icon and go to file open , select “ B & B PHOTOMETRIC DISSOLUTION” .Change the necessary
           information and save the report in data folder as XYZ. Where X is name of the sample or code, Y is batch No & Z is date, month &
           data.
6.1.12. Then preview the report and give the print out command.
6.2. FOR SPECTRUM
6.2.1. Switch on the instrument and computer.
6.2.2. Check the connection cable to the PC is properly connected or not.
6.2.3. Select the icon of UV probe and click double.
6.2.4. Press PC control (F4) in the instrument.
6.2.5. Select the icon Spectrum and click.
6.2.6. Place the blank solution in sample compartment
6.2.7. Click the method icon(M).The page will be opened, enter the wavelength range & scan at medium speed, and enter the necessary
          information which are relevant then click and the method should be saved as name of the sample and do the base line correction. (If
          the method is already exists open the method file , place the blank and do the base line correction).
6.2.8. To read the sample spectrum place the sample solution in sample compartment and click the START icon.
6.2.9. After spectrum scanning completion new data set will appear on screen automatically. Enter the file Name :dzu\uvprobe\data\xyz
         where X is sample name or code, Y is B.No , Z is date and Month and data, data storage to be same enter the analyst name and
         enter the analyst name and comment if required, click ok and should be saved
6.2.10. After completion of scanning go to operation click on peak pick, pick table will appear on monitor.
6.2.11. To take print out click on REPORT icon and go to file open select “ B& B SPECTRUM PEAK PICK”
6.2.12. Change the necessary information allotted space and save the report
6.2.13. Then preview the report and then give print out command.

7.0. REFERENCES:
Instrument Manual

8.0. ENCLOSURES:
Nil

9.0. ABBREVIATIONS:
Nil


UV VISIBLW SPECTROPHOTOMETER

SOP FOR CALIBRATION

1.0.CONTROL OF ABSORBANCES

PROCEDURE:
REPARATION OF POTASSIUM DICROMATE SOLUTION: Dry K2Cr2O7 at 130oC for 2 hours. Weigh accurately about the quantity of sample equal to 60mg into 1000ml volumetric flask. Add 0.005m Sulphuric acid to dissolve and dilute with the same solvent.

Find out the absorbances of (A 1% 1cm) above said solution at wavelengths of a 235nm, 257nm, 313nm and 350nm. Calculate the (1% 1cm) value using the following expression.

Absorbance X 1 X 1000 / Weight of K2Cr2O7 in g X 100
Check the value for each wavelength with the tolerances given four wavelengths.


PREPARATION OF REAGENT:

(i) Dilute 3ml of concentrated sulphuric acid and make up 100ml with distilled water.
(ii) Dilute 20ml to 2000ml with distilled water.

2. LIMIT OF STRAYLIGHT:
The absorbance of a 1.2-% w/v solution of potassium chloride at a path-length of 1cm should be greater than 2.0 at about 200nm when compared with water as reference liquid.

1.2% POTASSIUM CHLORIDE SOLUTION:
Take 1.2 g of Potassium chloride previously dried at 105oC for two hours into the 100ml volumetric flask. Add water to dissolve and make up to volume with water.

3. RESOLUTION POWER:
Record the spectrum of a 0.02% v/v solution of Toluene in Hexane UV. The ratio of the absorbance at the maximum at about 269nm to that at the minimum at about 266nm is not less than 1.5 unless other wise specified in the monograph.
0.02% v/v

TOLUENE:
Take 0.02ml of Toluene into the 100ml volumetric flask. Add Hexane to volume.