Chemiluminescence detection for high-performance liquid chromatography of biomedical samples.
During recent years, much progress has been made in the development of high-performance liquid chromatographic (HPLC) detection systems based on chemiluminescence (CL). CL is now one of the most sensitive detection methods in HPLC. For many compounds detection limits in the femtogram to picogram range have been obtained. Several on-line post-column reactions have been used for chemical excitation of the analytes. Some theoretical aspects of CL detection are presented and special attention is devoted to the coupling of CL to flow systems. The influence of the kinetics of the reaction on the sensitivity of the detection system is stressed. The mechanisms and detection systems of the peroxyoxalate, luminol and lucigenin CL reaction are described. A few examples of the use of bioluminescence for HPLC detection are given, and some less common CL reactions used in flow systems are also mentioned. Many biomedical and related applications are shown. Possibilities and limitations of the various reactions and detection systems are evaluated.
De Jong
G J
GJ
Department of Analytical Chemistry, Free University, Amsterdam, The Netherlands.
Kwakman
P J
PJ
D002624:Chemistry, Clinical / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D008163:Luminescent Measurements; D013050:Spectrometry, Fluorescence