高速液体クロマトグラフィーによる生体中のN-Acetyl-L-cysteineの微量けい光測定法

概要

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A new sensitive fluorometric HPLC method for the determination of a mucolytic agent N-acetyl-L-cysteine (NAC) and its metabolite L-cysteine (CySH) in biological fluids was developed. By the addition of a fluorescent agent, N-(9-acridinyl) maleimide (NAM), to NAC and CySH, stable fluorescence of NAM derivatives were developed at room temperature in a buffer solution, and the derivatives were separated and determined by HPLC. To standard or test solutions containing NAC and CySH were added 0.5 M Na_2CO_3-H_3BO_3-KCl buffer solution (pH 8.8), 5N NaOH and acetone solution of NAM (0.5-2×10^<-6> mol/ml), and the mixture was allowed to stand for 2 h at room temperature. An aliquot of this solution (5-20 μl) was injected into the HPLC equipped with fluorometric detector. The separation of the derivatives were carried out on an aminoalkyl chemically bonded silica by using acetonitrile/0.05 M CH_3COONa as mobile phase system for the analysis of NAC and on an octadecyl chemically bonded silica by using acetonitrile/0.05 M CH_3COONH_4 for the analysis of CySH. The fluorescence intensities of the derivatives were determined at 360 nm (excitation) and 435 nm (emmision). This method is simple, specific and sensitive compared with the gas chromatographic procedure for the determination of NAC and CySH in the plasma and tissues of rats after single oral administration of NAC.
社団法人日本薬学会の論文
1981-11-25