Derivatization of alcohols using acridone-9-N-acetyl-benzene-disulfonate as a condensation agent and its application for the determination of volatile alcohols in human plasma by liquid chromatography with fluorescence detection Original Research Article

A sensitive liquid chromatographic (LC) method for the determination of alcohols including the volatile alcohols from spiked plasma, using the fluorescent condensation agent, acridone-9-N-acetyl-benzene-disulfonate (AABS), has been developed. A mixture of alcohols and triethylamine in dichloromethane or chloroform is treated with AABS to give a quantitative yield of esters. Emission maxima for the derivatized alcohols are at 435 nm (λex 404 nm). The labeled derivatives are very stable, no remarkable decomposition is observed after heating in 15% acetonitrile solution (pH 6.5) at 40°C for 24 h. The method, in conjunction with a multi-gradient program, offers a baseline resolution of the common alcohol derivatives on a reversed-phase C18 column. Studies on derivatization conditions indicate that alcohols react very fast with AABS in the presence of triethylamine in dichloromethane or chloroform to give the corresponding fluorescent derivatives. The established method for the analysis of alcohol derivatives is more convenient and more efficient than previous methods which require the prior conversion of the carboxylic acids to the acyl chlorides. The application for the analysis of volatile alcohols in spiked human plasma is also investigated. The LC system shows good selectivity and reproducibility for volatile alcohols from spiked plasma. The relative standard deviations (n=5) for 50 pmol of each alcohol are <4%. The detection limits are at the fmol level. The method is also suitable for the determination of other alcohols in various matrices.