Visible/ultraviolet absorption spectroscopy, laboratory

Analysis is an integral part of research, clinical, and industrial laboratory methodology. The determination of the components of a substance or the sample in question can be qualitative, quantitative, or both. Techniques that are available to the analyst for such determinations are abundant. In absorption spectroscopy, the molecular absorption properties of the analyte are measured with laboratory instruments that function as detectors. Those that provide absorbance readings over the ultraviolet-visible (UV-vis) light spectrum are commonly used in high-performance liquid chromatography (HPLC). The above method is sufficiently sensitive for quantitative analysis and it has a broader application than other modes of detection. 

Molecular spectroscopy based on ultraviolet, visible, and infrared radiation is widely used for the identification and detennination of many inorganic, organic, and biochemical species. Molecular ultraviolet/visible absorption spectroscopy is used primcirily for quantitative analysis and is probably more extensively applied in chemical and clinical laboratories throughout the world than any other single method. Infrared absorption spectroscopy is a poweiful too for determining the structure of both inorganic and organic compounds. In addition, it now plays an important role in quantitative analysis, particularly in the area of environmental pollution. 

Wide applicability. Enormous numbers of inorganic, organic, and biochemical species absorb ultraviolet or visible radiation and are thus amenable to direct quantitative determination. Many nonabsorbing species can also be determined after chemical conversion to absorbing derivatives. It has been estimated that more than 90% of the analyses performed in clinical laboratories are based on ultraviolet and visible absorption spectroscopy. 

The ultraviolet-visible method is useful for the study of electronic transitions in molecules and atoms. Although various forms of ultraviolet-visible spectroscopy can be used to study a myriad of important chemical and physical properties, we will be most concerned with its use in quantitative analysis. It is probably the single most frequently used analytical method, with the possible exception of the analytical balance. For example, a single clinical analysis laboratory in a major hospital may perform a million chemical analyses a year, primarily on serum and urine, and about 707o of these tests are done by ultraviolet-visible absorption spectroscopy. Atomic absorption and emission spectroscopy (Chaps. 10 and 11) is used primarily to analyze for metallic elements in a variety of matrices—serum, natural waters, tissues, and so forth. 

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