Accuracy in the chromatographic determination of extreme enantiomeric ratios: A critical reflection

The chromatographic quantitation of very low amounts of an enantiomer in the presence of its antipode can be an extraordinary challenge. If resolution of the peaks is not complete even at extreme mass ratios an integrator will yield inaccurate results due to geometric effects. A given resolution can be adequate for peaks of similar size but result in severe overlap if one of the signals is markedly smaller. If tailing occurs, which is more the rule than the exception, the problem is especially severe for last eluted small peaks. Additional obstacles are detector nonlinearity and other sources of unsatisfactory calibration curves, overloading phenomena, and the possible lack of standards of highest optical purity. These problems have been studied by computer simulations and the liquid chromatographic separation of (R,S)-phenylethyl naphthoic acid amide on a chiral stationary phase. (C) 1995 Wiley-Liss, Inc.