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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
Department of Pharmacy Health & Well-Being, Faculty of Applied Sciences, University of Sunderland, Sciences Complex, Sunderland, UK
Department of Pharmacy Health & Well-Being, Faculty of Applied Sciences, University of Sunderland, Sciences Complex, Sunderland, UK
Department of Pharmacy Health & Well-Being, Faculty of Applied Sciences, University of Sunderland, Sciences Complex, Sunderland, UK
Cambridge Cancer Research Fund, Cambridge, UK
- 1 Department of Pharmacy Health & Well-Being, Faculty of Applied Sciences, University of Sunderland, Sciences Complex, Sunderland, UK
- 2 Department of Pharmacy Health & Well-Being, Faculty of Applied Sciences, University of Sunderland, Sciences Complex, Sunderland, UK
- 3 Department of Pharmacy Health & Well-Being, Faculty of Applied Sciences, University of Sunderland, Sciences Complex, Sunderland, UK
- 4 Cambridge Cancer Research Fund, Cambridge, UK
American Journal of Analytical Chemistry·Volume 08 (2017)·Pages 1–18·Published 4 January 2017·DOI10.4236/ajac.2017.81001
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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.
KeywordsProstate CancerLow Molecular Weight ThiolsThiol AnalysisUltra-High Performance Liquid ChromatographyNovel ThiolLC-MSNMREllman’s Reagent
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