Wave-mixing circular dichroism detector for chiral liquid chromatography

A circular dichroism (CD) detector based on laser four-wave mixing (FWM) is demonstrated using separate injections of analyte enantiomers onto a standard silica-based microbore high-performance liquid chromatography (HPLC) column. Using the chiral column, a preliminary "detected" mass detection limit of 180 pg is determined inside a laser probe volume of 200 pL, corresponding to a circular dichroism detection limit, DeltaA, of 2.2 x 10(-5) for (-) camphorquinone. Detection sensitivity levels are dramatically improved when our FWM-CD detector is interfaced to a microbore system due to the lower mobile-phase flow rates and the smaller sample concentrations required for the analysis. Using the microbore column, a preliminary circular dichroism detection limit, DeltaA, of 1.6 x 10(-6) and a preliminary concentration detection limit of 4.1 x 10(-4) M are determined for camphorquinone. This corresponds to a "detected" mass detection limit of 33 pg for the chiral compound. Laser wave mixing offers better detection limits than conventional circular dichroism detection methods and, hence, offers many promising applications.