Conclusions and Summary

This article has provided an overview of derivatiz-ation for LC. It is clear that this approach has undergone significant developments over the past few decades, to the point where it is now a mature area of LC science. Numerous books and reviews have appeared in recent years, and the literature continues to grow. Several excellent primers are available on the use of derivatization in LC and other separation areas, such as capillary electrophoresis (CE). Derivat-ization serves several useful functions in LC, but can especially improve chromatographic performance and peak shape for the original analyte, improve detector response to permit trace determinations, and improve quantitation for the original analyte by improving signal-to-noise ratios in complex sample matrices.

Derivatization can also stabilize an otherwise reactive analyte by the formation of a more stable derivative. The formation of multiple derivatives, using either solution or solid-phase (mixed-bed) approaches, has enabled improved qualitative identification of an analyte, as well as confirmation of quantitation by providing two to three different peaks for such purposes, all from the same sample undergoing one or a series of tagging reactions. Automation of derivatization, both pre- and post-column and online or offline, has developed such that it has become virtually a routine part of LC analysis. It is quite common to perform derivatization of amino acids pre-column, offline or online, in order to improve the identification and quantitation of these species, for example in a protein hydrolysate or intravenous solution. Derivatization for trace level detection of many analytes has also become commonplace, particularly when combined with preconcentration as part of the sample preparation-derivatization-LC steps. These tagging approaches permit the trace analysis of many analytes in complex sample matrices that otherwise would not be detectable by direct LC analysis.

Derivatization has thus become very commonplace in much of LC analytical work and applications. A wide variety of suitable reagents are commercially available, providing enhanced detection in several modes (UV, FL, EC and MS). Derivatization approaches are being developed for proteins and pep-tides that would lead to directed fragmentation and/or improved ionization for lowered detection limits in various forms of mass spectrometry or LC-MS. These efforts to develop improved derivatiz-ation reagents for further LC-detector applications will undoubtedly continue for many years to come.

See also: II/Chromatography: Liquid: Detectors: Ultraviolet and Visible Detection. III/Peptides and Proteins: Liquid Chromatography.

Solar Panel Basics

Solar Panel Basics

Global warming is a huge problem which will significantly affect every country in the world. Many people all over the world are trying to do whatever they can to help combat the effects of global warming. One of the ways that people can fight global warming is to reduce their dependence on non-renewable energy sources like oil and petroleum based products.

Get My Free Ebook


Post a comment