The basic working principle of a spectrophotometer is based on the selectivity of matter for the absorption of light (the wavelength of light). Different substances have their own absorption bands, so when the spectrum after light dispersion passes through a solution, Light of certain wavelengths is absorbed by the solution. At a certain wavelength, the concentration of the substance in the solution has a proportional relationship with the degree of attenuation of the light energy, that is, according to Beer's law.

Construction of spectrophotometer

The spectrophotometer allows a measurement wavelength range of 360 to 800 nm, and its structure is relatively simple, and the sensitivity and precision of the measurement are high. Therefore, the application is more extensive. The instrument structure of the 721 spectrophotometer is shown in the figure below.

The continuous radiation from the light source is incident on the condenser lens, and after convergence, it is rotated through the plane mirror by 90° and reflected to the entrance slit. It is thus incident into the monochromator, and the slit is located on the focal plane of the spherical collimating objective lens. When the incident light is reflected by the collimating objective lens, a pair of parallel light is incident on the prism. After the light enters the prism, the dispersion is performed. After the dispersion, the light coming back, after being reflected by the collimator lens, will gather on the light exit slit, and then enter the cuvette through the condensing mirror. Part of the light is absorbed, and the transmitted light enters the photocell, generating corresponding photocurrent. After zooming in, read it on the microampere meter.

How to use the spectrophotometer

1 Turn on the power first, turn on the power switch 1, the indicator light is on, open the cuvette black box cover 8, and warm up for 20 minutes.

2 Wavelength selection knob 6, select the desired wavelength of monochromatic light, and use sensitivity knob 2 to select the desired sensitive file.

3 Put in the cuvette, rotate the zero knob 5 to zero, close the cuvette cover, push the cuvette lever 3, so that the reference cuvette is in the blank correction position, so that the photocell sees light, the rotation transmittance Adjust knob 4 so that the micro-amplifier 9 pointer is exactly 100%. The measurement can be performed by adjusting the zero position and the 100% position several times in the above manner.

Precautions for use and maintenance of the spectrophotometer:

1 The time for continuous use of the instrument should not exceed 2 hours, preferably 0.5 hours after the interval, and then continue to use.

2 After each use, cuvettes should be washed with deionized water, inverted and dried, and stored in a cuvette box. In daily use, care should be taken to protect the translucent surface of the cuvette from damage or scratches so as not to affect the light transmittance.

3 The instrument should not be damp. In daily use, always pay attention to whether the moisture-proof silicone on the monochromator (at the bottom of the instrument) is discolored. If the color of the silica gel has turned red, it should be taken out or dried immediately.

4 Care should be taken when handling or moving the instrument.

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