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

A sensitive liquid chromatographic (LC) method for the determination of alc ohols including the volatile alcohols from spiked plasma, using the fluores cent condensation agent, acridone-9-N-acetyl-benzene-disulfonate (AABS), ha s been developed. A mixture of alcohols and triethylamine in dichloromethan e or chloroform is treated with AABS to give a quantitative yield of esters , Emission maxima for the derivatized alcohols are at 435 nm (lambda(ex) 40 4 nm). The labeled derivatives are very stable, no remarkable decomposition is observed after heating in 15% acetonitrile solution (pH 6.5) at 40 degr ees C for 24 h, The method, in conjunction with a multi-gradient program, o ffers a baseline resolution of the common alcohol derivatives on a reversed -phase C-18 column. Studies on derivatization conditions indicate that alco hols react very fast with AABS in the presence of triethylamine in dichloro methane or chloroform to give the corresponding fluorescent derivatives. Th e established method for the analysis of alcohol derivatives is more conven ient and more efficient than previous methods which require the prior conve rsion of the carboxylic acids to the acyl chlorides. The application for th e analysis of volatile alcohols in spiked human plasma is also investigated . The LC system shows good selectivity and reproducibility for volatile alc ohols from spiked plasma. The relative standard deviations (n=5) for 50 pmo l 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. (C) 1999 Elsevier Science B.V. All rights reserved.