Determination And Research Of Thiol Compounds In The Lotus Powder And Flammulina Velutipes

Screening on the basis of this study, determine mainly on lotus powder and Flammulina velutipes in intracellular glutathione (GSH) and thiol compounds containing cysteine residues, in order to screen out rich in GSH and thiol compounds of medicinal and edible plant samples. According to reports in the literature, the content of thiol compounds with cysteine residues in human body related to aging and certain diseases are closely linked. Therefore, screening out rich in GSH and thiol compounds of medicinal and edible food, has practical significance for human medical treatment, disease prevention, prolong life. Although, the determination of thiol compounds in biological samples has a lot of different methods, but these methods using different pretreatment process, which results in larger differences between the different measurement methods. This study using high performance liquid chromatography (HPLC) method and spectrum method for three different pretreatment method of obtaining sample extracts were compared, found using cell wall-breaking, derivative into one of the synchronous pretreatment method, the analysis of the measured signal intensity is the largest, avoiding as much as possible the pre-treatment process in cells (-SH) oxidation degeneration due to extracellular oxidative environment, making the results more real time, reliable which reflected the content of biological cells. Meanwhile, this study also used two different methods which determine the content of GSH and total thiol (-SH) in biological samples, the results of the two methods shows that the content of measured sample GSH and total thiols (-SH) is consistent, This fully shows that the reliability of the selected high performance liquid chromatography and spectrometry on the determination of thiol compounds in biological samples. The study also while using lotus powder as a living cell samples discussed different heavy metal ions stress, which response to the action of the GSH and thiol-containing protein in the cells, which provides basic data for the research of thiol compounds in heavy metal detoxification mechanism.High performance liquid chromatographic method for the determining the levels of GSH and total thiol (-SH) in lotus powderUsing of the excessive derivatization reagent DTNB extracts, cell wall-breaking, derivative synchronous pretreatment method as a whole, determining the content of GSH and total thiols (-SH) in the lotus powder for high performance liquid chromatography. The determination results show that the content of GSH and total thiols (-SH) in the lotus powder are 2.0865 μmol/g and 9.5076 μmol/g, but lotus powder samples do not contain similar to cysteine (Cys) as other small molecular thiol compound, the difference of total thiols (-SH) and GSH is 7.4211 μmol/g, which is the content of protein thiol (P-SH) in lotus powder. The method of recovery is 100.95% and 99.91%, the detection limits are 5.18 μmol/L and 6.22 μmol/L.Determining the content of GSH and total thiols (-SH) in Lotus powder with the method of Fluorescence spectroscopy and UV spectroscopyThis study uses derivatization reagent DTNB and the OPA for determining the content of GSH and total thiols (-SH) in lotus powder by UV spectra and fluorescence spectra. The content of GSH and total thiols (-SH) is 1.8007μmol/g and 9.6829mol/g. Comparing with determination results of Spectrometry and HPLC method, which can be found the results are similar and basic consistent, the reliability of the two methods can be proved.Research on the response mechanism of intracellular thiol compounds under different heavy metal stress in lotus powderIn this study, an HPLC method for the determining changes of the content of GSH and thiol protein (P-SH) in lotus powder under the different heavy metal stress, and use the method of ICP-MS for determining the content of heavy metals in lotus powder under different heavy metal stress. Preliminary discussion on the different heavy metal stress response mechanism of intracellular thiol compounds, which provides data reference for biological cells on heavy metal detoxification and accumulation mechanism.Determining the content of GSH and Total thiol in Flammulina velutipesUsing spectrometric and high performance liquid chromatography for determining the content of GSH and Total thiol in Flammulina velutipes, the determination results show that the content of GSH and total thiols (-SH) in Flammulina velutipes is 1.3665 μmol/g and 1.6366 μmol/g by HPLC, the content of GSH and total thiols (-SH) is 1.3429 μmol/g and 1.6296 μmol/g by spectrometry. The spectrometry of recovery is 100.42% and 100.19%, the detection limits are 0.1177μmol/L and 0.2194μmol/L. The HPLC of recovery is 100.92% and 100.06%, the detection limits are 5.18 μmol/L and 6.22 μmol/L, the recovery rate and the detection line of two methods can achieve the requirement of the sensitivity and accuracy.