Analysis of microbial components using LC-IR.

Characterization of bacteria is currently an important research area in the medical, military, food, and agricultural sciences. In recent years, FT-IR has found an application as a microbiological detection method and as a general research tool. When coupled with a liquid chromatographic system, a new facet of research has evolved. By utilizing the separation ability of typical liquid chromatography systems, matrix elimination is possible, therefore allowing for clean spectra of cellular components. Information about the compositional makeup of various bacteria enhances the overall understanding of biology at the cellular level, provides a quantification of the chemistry of cellular processes, and can be used as a general identification tool. Both whole cells and lysed Escherichia coli cells were investigated in the present study. The cellular components consisting of proteins, glycoproteins, phospholipids, fatty amides and acids, and genomic materials were separated, isolated, and identified by FT-IR.

Perspectives on analyses of nucleic acid constituents: the basis of genomics.

The recent mapping of the human genome was a tremendous achievement made possible to a large degree by the development of analytical methods for sequencing purine and pyrimidine bases in nucleic acids. In the last 3 decades, the number of analyses of nucleic acids and their constituents by HPLC and capillary electrophoresis (CE) has exploded. These techniques have been used not only for genomics, but also for the determination of free nucleotides, nucleosides and their bases in body fluids and tissues. Although a large number of HPLC and CE papers have been published on nucleic acid constituent applications, relatively little has been written on the mechanisms of the separations. However, to optimize analytical conditions knowledgeably and rapidly, it is important to know why and how these separations occur and the factors that affect them. The HPLC methods for the analysis of nucleic acid constituents and the information available on some of the mechanisms of separation of nucleotides, nucleosides and their bases, as well as the analysis of these compounds by CE and the factors that affect these separations are discussed.