I've tried many solvents and precipitation agents but still don't know which one would serve the best, bearing in mind that glutathione is a labile compound and changes its concentration rapidly.
Hello, why without derivatisation? do you mean extraction before derivatization?
Hi! I mean an extraction technique from plant to determine glutathione level but as a non-derivatised substance. There are many publications concerning glutathione determination with derivatisation but I want to extract it and derermine it, without changing its structure. I already have a method on my LC-MS/MS for glutathione so now I need a plant extract with it. Do you know what I mean? Is it possible? Thanks:-)
Hi, I don't have data on the stability of GSH in plant extracts but in tissues and biological fluids post-extraction derivatization is a must because the GSH is oxidized into GSSG quite fast. Derivatization with e.g. NEM (N-ethyl-maleimide) can be very straightforward and you could also use LCMS/MS for measuring GS-NEM
Hi, thank you very much for your answer. I'll try to find the best method.
Try this one.
Plant Cell Physiol. 2011 Nov;52(11):2006-15. doi: 10.1093/pcp/pcr133. Epub 2011 Sep 30.
Detection and quantification of S-nitrosoglutathione (GSNO) in pepper (Capsicum annuum L.) plant organs by LC-ES/MS.
Airaki M1, Sánchez-Moreno L, Leterrier M, Barroso JB, Palma JM, Corpas FJ.
Author information
1Departamento de Bioquímica, Biología Celular y Sistemas Molecular de Plantas, Estación Experimental del Zaidín, CSIC, Apartado 419, E-18080 Granada, Spain.
Abstract
Glutathione (GSH) is one of the major, soluble, low molecular weight antioxidants, as well as the major non-protein thiol in plant cells. However, the relevance of this molecule could be even greater considering that it can react with nitric oxide (NO) to generate S-nitrosoglutathione (GSNO) which is considered to function as a mobile reservoir of NO bioactivity in plants. Although this NO-derived molecule has an increased physiological and phytopathological relevance in plants cells, its identification and quantification in plant tissues have not be reported so far. Using liquid chromatography-electrospray/mass spectrometry (LC-ES/MS), a method was set up to detect and quantify simultaneously GSNO as well reduced and oxidized glutathione (GSH and GSSG, respectively) in different pepper plant organs including roots, stems and leaves, and in Arabidopsis leaves. The analysis of NO and GSNO reductase (GSNOR) activity in these pepper organs showed that the content of GSNO was directly related to the content of NO in each organ and oppositely related to the GSNOR activity. This approach opens up new analytical possibilities to understand the relevance of GSNO in plant cells under physiological and stress conditions.
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