Derivatisation methods for trace-level quantification of B-lactam antibiotics using HPLC

The B-lactam antibiotics are a therapeutically and commercially important group of antibacterial agents. Quantification of this group was originally achieved using microbial assays. High performance liquid chromatography (HPLC) as become the method of choice for the analysis of low concentrations of these compounds and their metabolites in complex biological matrices such as body fluids and fermentation broths. Chemical derivatisation allows selective introduction of chromophores, fluorophores, and electrochemically active functionalities into B-lactam molecules This can enhance separation, sensitivity, and selectivity wit the aid of spectrophotometric, spectrofluorometric and electrochemical detectors. Although precolumn and postcolumn derivatisation methods have been refined, the former are more suited to fully automated analytical procedures. DErivatisation methods using HPLC for analysis of selected classes of B-lactam are reviewed. The most recent developments include the use of on-line chemiluminescence and photolytic techniques.