Determination Of Thiol Compounds And Responses To The Heavy Metal Stresses In Abutilon Theophfasti Medic

Thiol compounds of the peptide (like Glutathione,GSH) and protein (Prot-SH) contain cysteine residues in biological cells, possess the functions such as antioxidant, promoting amino acid absorption, heavy metal detoxification, cell signal transduction, is the activity of plant physiological activities play an important role in a variety of substances. which is the active substance Play an important role in the various plants physiological activities. To develop a sensitive and convenient analysis method, By determining and screening the plants rich in thiol compounds, investigate the relationship between thiol compounds and its biological function is one of the hot spot of current research.This research use 5,5’-dithio-bis-nitrobenzoic acid(DTNB) can selective derivative react with the thiol of cysteine residues, Using the pretreatment method of simultaneous derivatization during cell rupture, Glutathione and Protein thiol (Prot-SH)in Abutilon Theophfasti Medic were determined by HPLC and spectroscopy, found there are high content of GSH and Prot-SH distribution in Abutilon Theophfasti Medic. Using the analysis method to determine the content change of GSH and Prot-SH in Abutilon Theophfasti Medi under the heavy metal stress of Cd2+、Cr3+、Pb2+2、As3+、Fe3+、Cu2+ Combined with the determination of ICP-MS, Discusses the response function and relationship with heavy metal adsorption quantity of the thiol of GSH and Prot-SH in Abutilon Theophfasti Medic when the different heavy metal stress. At the same time discusses the role and the mechanism of the different thiol compounds in heavy metal detoxification and accumulation. In order to further research the practical application of using herbaceous plant bioremediation of heavy metal pollution to provide important reference data.Chapter 1:IntroductionSummarizes the present research situation of Abutilon Theophfasti Medic and the mechanism of Heavy metal with thiol compounds in cell, And put forward the problem of Thiol compounds in the Abutilon Theophfasti Medic cell in detecting method and the research of its effect on heavy metal accumulation and stress. Summarizes the thought and main research contents of this paperChapter 2:Glutathione(GSH) and Total Thiols(-SH) are detemined by High performance liquid chromatography in Abutilon Theophfasti Medic.Sample were prepared by simultaneous derivatization during cell rupture. Reduced glutathione (GSH) and total thiols in Abutilon Theophfasti Medic were determined by High performance liquid chromatography using 5,5’-dithio-bis-nitrobenzoic acid (DTNB) as derivatization agents. Chromatographic conditions including Sample preparation and chromatographic mobile phase are optimized, and established a determination method of simultaneous determination cells contain cysteine residues of thiol compounds. The determination results show that; The determination of three average are 1.6053μmol/g of GSH and 4.8759μmol/g of total thiols in Abutilon Theophfasti Medic. Abutilon Theophfasti Medic are rich in GSH and the thiol protein. Under optimized conditions, recoveries were 101.58% and 98.47%, the detection limit are 5.18μmol/L and 6.23μmol/L.Chapter 3:Glutathione and Total Thiols are determined by spectrometry in Abutilon Theophfasti Medic.Reduced glutathione (GSH) and total thiol in Abutilon Theophfasti Medic were determined by fluorophotometry and UV-vis spectrophotometry using O-phthaldehde (OPA) and 5,5’-dithio-bis-nitrobenzoic acid (DTNB) as derivatization agents. Sample were prepared by simultaneous derivatization during cell rupture. Measurement results showed that Abutilon Theophfasti Medic are rich in GSH and total thiol,1.4637μmol/g of GSH and 5.12082μmol/g of total thiols were detected in the sample. The detection limits for fluorophotometry and UV-vis spectrophotometry were 0.04698μmol/L and 1.5152μmol/L, and recoveries were 99.69%～100.27% and 99.44%～100.89%.Chapter 4:The response mechanism of Heavy metal interact with thiol compounds in Abutilon Theophfasti Medic.This research use the Abutilon Theophfasti Medic containing thiol compounds as the experimental samples. Using derivatization reagent DTNB research different heavy metal stress response function of thiol compounds by high performance liquid chromatography with "in-situ" derivatizations. Mainly discusses the thiol compounds in Abutilon Theophfasti Medic interact with Cd2+、Cr3+、Pb2+、As3+、Fe3+、Cu2+ of exogenous heavy metal ions in different time. Determine the content of heavy metal ions in Abutilon Theophfasti Medic cells with metal ions stress response. Obtained the relevant data of thiol compounds with the accumulation and coordination of exogenous metal ions in Abutilon Theophfasti Medic cells. Provide basic experiment basis for discussing hyperaccumulation plants in heavy metal detoxification and accumulation mechanism.