How to use a Spectrophotometer?

Reviewed by Anurag Mishra (Sr. Technical Consultant)

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Spectrophotometry is a commonly used experimental technique that is meant for analysis of different materials with the help of their chromatic properties. The most prominent use of spectrophotometry is in the analysis of the chemicals and the concentration of the solutes in the solutions. The instrument that is used for performing the spectrophotometry is called as a spectrophotometer. The instrument analyzes the amount of light that is transmitted through the specimen and the amount of light that is absorbed by the. the specimen to detect the absorbency of the products. The procedure of using a portable spectrophotometer is as follows:

  1. First of all the spectrophotometer is turned on.
  2. The cuvette or the test tube in which the sample is to be placed must be cleaned. If the disposable test tubes are used, then there is no need for cleaning. When handling cuvette, one need to be careful as they are very costly.
  3. Insert appropriate amount of sample in the cuvette so that the light incident on the cuvette should pass through the sample for accurate readings.
  4. A control solution must be prepared for the purpose of comparison.
  5. Clean the outer surface of the cuvette.
  6. Now choose and set the wavelength of the light that you want to an incident on the sample.
  7. Make sure that the light source is monochromatic for effective testing.
  8. Calibrate the machine with the control solution you prepare.
  9. Now remove the blank and load the cuvette with the test sample to the machine.
  10. Now wait for 10 seconds after loading the sample until the readings of the display are stable.
  11. Once the readings are stable, note down the readings as % transmittance and % absorbance.
  12. Repeat the test procedure with different successive wavelengths of light.
  13. If you have more than one sample to test, then you have to repeat the above-given procedure for every sample.
  14. After the test data has been obtained, it is analyzed to determine the chromatic and optical properties of the specimen.

With the help of this testing procedure of portable spectrophotometer, it becomes very easy to test the chromatic attributes of the test samples and analyze the quality of the specimen.

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