Heavy Metal StressResponse Function Of ThiolCompounds In Biological Samples

At present, an extensive literature reported the determination of GSH and other thiol compounds. The determination bases on the derivatized reagent spectro-photometry and separation, but there are different sample pre-treatment processes between different methods, and the results of quantitative have significant differences. In this paper, instant yeasts, as active cell samples, were analyzed using spectroscopy and chromatography to identify the content of GSH and total sulfhydryl. During the experiment, this research used OPA and DTNB as the derivatized reagent and new pretreatment method which breaking the cell and deriving were accomplished simultaneously.Then compare the determinantion results of different pretreatment methods and analysis methods. Finally, establish a accurate quantitative analysis methods for GSH and other thiol compounds which are from biological cells.GSH has the effectiveress of heavy metal poisoning that is detoxification. Until now, a large number of literatures reporting the functional groups which come from GSH, such as thiols, carboxyl, amidogen, etc that can coordinate heavy metal. But it isn’t obvious that GSH in the cell interact with exogenous and poisonous heavy metal stress in detail as well as the detoxification mechanism. This research use the analysis method that has been estabilished to detect the response function of GSH and total thiols when the different heveay metals effect the biological cells. Futher more, disscuss the interact mechanism about exogenous heavy metal and GSH, thiol compounds in the cells. Finally, provide the basic data for thiol compounds such as GSH about heavy detoxification mechanism of the chemical research.Chapter1:IntroductionSummarize the detection of GSH ect thoil compounds in the biological cells and the troubles about heavy metals detoxification mechanism research. In the meantime, put forward the thought of the choice and main research contents.Chapter2:glutathione(GSH) and total thiols(-SH) in instant yeast are detemined by "in-situ" derivatization spectrometry.This study through the experiment confirmed the derivatization reagent DTNB that don’t have selectivity to GSH, Cys and cysteine residues of protein content of sulphur. So can determine total thiols content in the samples. Derivatization reagent OPA can select GSH from thiol compounds and generate fluorescence derivatization. This study use OPA and DTNB as derivatization reagents. GSH and total thiols in yeast are dertermined by using spectrometry with "in-situ" derivatizations. The GSH and total thiols in yeast biological samples are12.0213μmol/L-12.0823μmol/L and28.9859-29.2515μmol/g, the recovery rate are100.65%and99.47%, the detection limit are0.2655μmol/L and0.7649μmol/L.Chapter3:glutathione(GSH) and total thiols(-SH) in yeast detemined by DTNB column derivatization HPLC chromatography.High performance liquid chromatography (HPLC) can be used to detect various thiol compounds in biological samples (cysteine, glutathione, etc.). This study use DTNB as derivatization reagent. GSH and total thiols in yeast are dertermined by using high performance liquid chromatography with "in-situ" derivatizations. The GSH and total thiols in yeast biological samples are12.0569μmol/g and29.2823μmol/g, the recovery rate are99.43%and100.87%, the detection limit are0.2296μmol/L and0.7649μmol/L.Chapter4:The research of the response mechanism that Heavy metal interact with thiol compound in yeast cellsThis research adopt the instant yeast containing GSH and thiol protein as the biological cell sample, and use spectroscopy and chromatography to study different heavy metal stress response function of thiol compounds. Thus discusses the thiol compounds such as GSH interact with Cd, Cu, Cr, Fe, Pb, and provide basic experiment basis for discussing biological cells in heavy metal detoxification and accumulation mechanism.Chapter5:glutathione(GSH) and total thiols(-SH) in Corn silk detemined by "in-situ" derivatization using and spectrometry.GSH and total thiols in Corn silk are determined, choosing o-Phthalaldehyde (OPA) and4,4’-disulfide generation of pyridine(4-DPS) as the derivatization reagent, by using fluorescence spectrometry and UV-Vis spectrophotometry with "in-situ" derivatizations. The content of GSH and total thiols in Corn silk are0.0773-0.0788μmol/g and1.0751-1.0874μmol/g, the recovery rate are99.09%and101.30%, the detection limit are0.0395μmol/L and0.6947μmol/L.