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A UV-Vis spectrophotometer is an analytical instrument that measures the amount of ultraviolet (UV) and visible light that is absorbed by a sample. It is a widely used technique in chemistry, biochemistry, and other fields, to identify and quantify compounds in a variety of samples.
Ultraviolet–visible spectroscopy (UV–vis) can distinguish between enantiomers by showing a distinct Cotton effect for each isomer. UV–vis spectroscopy sees only chromophores , so other molecules must be prepared for analysis by chemical addition of a chromophore such as anthracene .
UV-Vis absorption SEC is a recent technique that is continuously evolving. However, many advantages have been observed over other techniques. However, many advantages have been observed over other techniques.
Ultraviolet-visible (UV-vis) spectroscopy involves energy levels that excite electronic transitions. Absorption of UV-vis light excites molecules that are in ground-states to their excited-states. [5] Visible region 400–700 nm spectrophotometry is used extensively in colorimetry science. It is a known fact that it operates best at the range ...
The restriction of the spin selection rule makes it even easier to predict the possible transitions and their relative intensity. Although they are qualitative, Tanabe–Sugano diagrams are very useful tools for analyzing UV-vis spectra: they are used to assign bands and calculate Dq values for ligand field splitting. [13] [14]
In ultraviolet-visible spectroscopy or spectroscopy in general a 1 cm pathlength cuvette is used to measure samples. The cuvette is filled with sample, light is passed through the sample and intensity readings are taken. The slope spectroscopy technique can be applied using the same methods as in absorption spectroscopy.
It is commonly used for analysis of compounds that do not absorb UV-VIS radiation significantly, such as sugars, antiviral drugs, antibiotics, fatty acids, lipids, oils, phospholipids, polymers, surfactants, terpenoids and triglycerides. [3] [4]
In the case of UV-visible spectroscopy, for example, this means that the system must conform to the Beer-Lambert law. In addition, the total concentration of the two binding partners, the pH and ionic strength of the solution must all be maintained at fixed values throughout the experiment.