Absorbance detection of amino acids by laser wave mixing in microbore liquid chromatography

Nonlinear optical phase conjugation by degenerate four-wave mixing is demonstrated as a sensitive "absorbance" detection method for microbore high-performance liquid chromatography. An argon ion laser operating at the 488-nm line is used as the excitation light source to generate the wave-mixing signal for dabsyl-labeled amino acids. Advantages of the nonlinear laser detection method include: virtually 100% optical signal collection efficiency, generation of the signal in the form of a coherent laser beam, signal measurement against a virtually dark background, reliable detection of small absorbance values, excellent detection sensitivity for both fluorescing and non-fluorescing analytes, relatively simple one-color one-laser optical setup, and low power or energy requirements for continuous-wave or pulsed lasers. Using our one-laser one-color nonlinear laser detector for "absorbance" measurements in liquid chromatography, we report a crude preliminary "injected" detection limit of 780 fmol for glycine.