Principles of Double Beam UV Spectrophotometer

A spectrophotometer is an optical device that implements the property of light rays to measure a particular wavelength. Hence, these instruments are also called analytical optical instruments. The prime use of a double beam UV spectrophotometer is to detect and confirm the presence of chemical species present within the biological specimens grounded on their chemical structures, concentration, etc. A double beam spectrophotometer uses the light rays in their visible wavelength and calculates in the same wavelength of light.

The double beam spectrophotometer emits the photons as light packets. These photons pass through multiple biological specimens where the light beam is absorbed for measuring the presence of biochemical entities. A spectrophotometer calculates the intensities of light at different wavelengths through their precise instrumentation.

What are the principles of a double beam UV spectrophotometer?

A double beam UV spectrophotometer works on the same principle as the single beam spectrophotometer. However, there are some additional phenomena of light rays in the UV spectrophotometers. The working principles of a double beam spectrophotometer are listed below:

1. Dispersion of light rays: You can see the dispersion of light rays in nature just after the rain. The droplets diffract the light rays of the sun by generating a spectrum of color. In this case, the spectrum is termed the rainbow. The same principle applies in the double beam UV spectrophotometer where the diffraction of light happens through the specimens. This refraction of light makes the spectrophotometry process more reproducible. In this equipment, the diffraction angles rely on the wavelength of the main incident light.
2. Absorbance: It is defined as the ratio of incident light to transmitted radiant light rays through a biological sample during the process of spectrophotometry. The cells contained in the samples reflect the light and also absorb the same that gets transferred to the detectors present in the spectrophotometer. When the case is of biological samples that scatter light, absorbance can be defined as the ability of the sample to absorb more and more light passing through it.
3. The visible spectrum scattering of light rays: In a double beam spectrophotometer, when white light passes through the sample and is reflected by a colored object, a certain portion of the wavelengths is absorbed. The remaining light beam will be guessed as the complementary color to the wavelength of light absorbed. It is demonstrated by the color spectrum.

The double beam UV spectrophotometer analyzes different biological specimens for good results. The electromagnetic spectrum is also implemented in the process of making measurements out of samples. Every process relating to dispersion, reflection, and refraction happens in spectrophotometry which further helps in making analytical measurements.