TITLE:
Investigation of the Low Molecular Weight Thiol Composition in a Metastatic Prostate Cancer Cell Line (LNCaP) by LC-UV-MS and NMR after Labelling with the Ellman Reagent
AUTHORS:
Stephen Childs, Nicolas Haroune, Lee Williams, Michael Gronow
KEYWORDS:
Prostate Cancer, Low Molecular Weight Thiols, Thiol Analysis, Ultra-High Performance Liquid Chromatography, Novel Thiol, LC-MS, NMR, Ellman’s Reagent
JOURNAL NAME:
American Journal of Analytical Chemistry,
Vol.8 No.1,
January
4,
2017
ABSTRACT: The low molecular weight thiols present in the deproteinized extract of a prostate cancer cell line (LNCaP-FGC) were analysed after derivatization with the Ellman reagent (ESSE). The mixed disulphides formed (RSSE) were fractionated, characterized and quantified by liquid chromatography on a C-18 column using UV detection. This revealed the presence, in femtomoles per cell, of glutathione (8.30 ± 0.73), cysteine (2.71 ± 0.04) and cysteinylglycine (0.83 ± 0.10), accounting for the bulk of the thiol present. Further analysis of the cell extracts using a novel and sensitive mass spectrometry technique allowed the detection of low level of an additional derivative which was identified as cysteinylglycerate using NMRspectroscopy.