Selective 3,4-Dihydroxyphenylalanine Analysis in Human Urine as Ethoxycarbonyl-tert-butyldimethylsilyl Derivatives by Gas Chromatography-Mass Spectrometry.

Abstract

A new analytical method for measurement of 3,4-dihydroxyphenylalanine (DOPA) in human urine was developed. DOPA from an aqueous solution was converted into an ethoxycarbonyl (EOC) derivative. A tert-butyldimethylsilyl (TBDMS) reaction under anhydrous conditions was then attempted for analysis by gas chromatography-mass spectrometry in selected ion monitoring mode. A new mass spectral data on DOPA as a tri-EOC/mono-TBDMS derivative was built. This method showed good linearity (r ≥ 0.999), precision (% relative standard deviation = 3.1-9.2), and accuracy (% relative error = −7.2-8.8), with a detection limit of 0.05ng/mL. This selective and accurate method of DOPA analysis will be useful for biochemical monitoring of various neurological disorders including Parkinson’s disease in biological fluids.