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Ultraviolet visible spectrophotometers (often referred to as a UV Vis spectrophotometer or just

a UV spectrophotometer) quantify the optical properties of samples in the ultraviolet and visible
wavelength ranges of light (typically 190 to 900/1100 nm). Specifically, UV Vis spectrophotometers
determine how much light of a given wavelength passes through a sample, and how much is absorbed.
UV / Vis spectrophotometer uses visible light and ultraviolet to analyze the chemical structure of
substance. A spectrophotometer is a special type of spectrometer, which is used to measure the
intensity of light, and the intensity is proportional to the wavelength. When ultraviolet project to various
organic compounds, these compounds will absorb it. So, you can use UV / Vis spectrophotometer to
measure the absorption of a compound by the result, and have its molecular structure, as well as the
related information.

The UV VIS Spectrophotometer has different parts namely: sample compartment, where the
sample will be placed; wavelength control knob, to change the wavelength; zero control knob,
transmittance/ absorbance control, and mode select button. Before getting started the researchers
should prepare the following samples: blank solution, the solution needed in aqueous form except the
molecule needed to measure; sample solution, the molecule/ sample being measured. Select
“Transmittance” before making adjustments by pressing the Mode Select Button.

In the calibration process, the researchers should set the machine first, by doing so, use the
Wavelength Control Knob to select the desired wavelength. Then, use the Zero Control Knob to set the
Transmittance to 0.0%. Next, open the Sample Compartment and put the blank solution in, but making
sure to wipe off the bottom of the cuvettes to ensure accurate readings. The cuvettes have a vertical
line and match it up with the mark at the front edge of the Sample Compartment. Then, use the
Transmittance/ Absorbance Control Knob to set the Transmittance to 100.0%.

When the calibration process is completed take the Blank Solution cuvette out and put the
Sample Solution in the Sample Compartment. Make sure the vertical line match up the sample
compartment. When removing the cuvette in the Sample Compartment, the Transmittance should go
back to 0.00%. The researchers can take multiple readings at the same wavelength. However, when
needed to change the wavelength make sure to recalibrate the machine before proceeding. To
recalibrate, first, change the wavelength then empty the sample compartment and set the
Transmittance to 0.00% then proceed with the rest of the process as mentioned earlier.


(Sept 28, 2019)

(Sept 28, 2019)