Identification of selenium-containing glutathione S-conjugates in a yeast extract by two-dimensional liquid chromatography with inductively coupled plasma MS and nanoelectrospray MS/MS detection

被引:76
作者
Lindemann, T [1 ]
Hintelmann, H [1 ]
机构
[1] Trent Univ, Water Qual Ctr, Peterborough, ON K9J 7B8, Canada
关键词
D O I
10.1021/ac020196g
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
An approach for the identification of unknown selenium-containing biomolecules was developed, enabling the identification of selenodiglutathione (GS-Se-SG) and the mixed selenotrisulfide of glutathione and cysteinylglycine (GS-Se-SCG) in aqueous yeast extracts. The method consists of two-dimensional liquid chromatography, inductively coupled plasma mass spectrometry (ICPMS) and nanoelectrospray tandem mass spectrometry. Analytes were separated by size-exclusion chromatography followed by preconcentration and separation on a porous graphitic carbon HPLC column. The HPLC effluent was monitored for selenium by ICPMS, and two selenium-containing fractions were isolated and analyzed by nanoelectrospray MS. The nanoelectrospray technique has a low sample consumption of similar to80 nL/min, enabling a preconcentration of the sample to a few microliters. Mass spectra of the two fractions showed the characteristic Se isotopic pattern centered at m/z 693.1 and 564.0 for the [M + H] Se-80 ions. MS/MS spectra of adjacent parent ions confirmed the presence of Se. The two selenium species were identified as GS-Se-SG and GS-Se-SCG by collision induced dissociation (CID). The accurately measured masses of the most abundant 691 and 693 u parent ions are in good agreement (differences = 3 ppm) with the theoretical masses. To our knowledge, this is the first identification of GS-Se-SG and GS-Se-SCG in biological matrixes by MS/MS.
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页码:4602 / 4610
页数:9
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