American Journal of Analytical Chemistry

Volume 8, Issue 1 (January 2017)

ISSN Print: 2156-8251   ISSN Online: 2156-8278

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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

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DOI: 10.4236/ajac.2017.81001    1,749 Downloads   3,073 Views  Citations

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.

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Childs, S. , Haroune, N. , Williams, L. and Gronow, M. (2017) 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. American Journal of Analytical Chemistry, 8, 1-18. doi: 10.4236/ajac.2017.81001.

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