Design of and First Experiments with a Bioluminescence Detector for HPLC

Abstract

As discussed earlier in this book [Brinkman, #F-2; de Bree, #NC(E)-4], HPLC still lacks a universal sensitive and selective detector, and for biomedical and environmental trace-level analyses HPLC is still outclassed by GC (especially capillary with the new chemically bonded phases). The efficacy of enzymes in metabolizing drugs should be exploitable to furnish a selective post-column reactor for HPLC, since mostly the selective enzymatic conversions involve production of NAD, ATP or H₂O₂. Several bioluminescence kits based on light emission initiated by one of these products are now commercially available. Amongst the many bioluminescence procedures found in the literature, most are in the food and clinical chemistry areas and none are HPLC-linked. In the approach now described, both the coenzyme-generating enzymes and those that emit light are immobilized, which entails problems that are now considered along with prospects for the detection approach in general. At the outset our model compound was flesinoxan