Determining The Content Of GSH And Total Thiolsin Bee-Collected Pollenand Humulus Scandens

This study with cysteine residues of selective thiol compounds derived function of derivatization reagent, using the improved pretreatment method, respectively, using HPLC method and the spectral analysis method, through the screening of Bee-Collected pollen and Humulus scandens was measured respectively reduced glutathione (GSH) and thiol proteins(SH).The results showed that; Bee-Collected pollen contains rich GSH and thiol proteins, these thiol compounds is an important active ingredient of Bee-Collected pollen. Humulus scandens as it grows fast and has resistance, low temperature resistance, drought resistance of herbaceous plants, the seed also contains rich GSH and thiol proteins, means that the low temperature, drought resistance maybe related to that it contains rich thiol compounds.Humulus scandens seed cells in heavy metal stress, the content of thiol with heavy metal stress response, its resistance to heavy metal stress may be related to the interaction of intracellular thiol compounds.In this research, choose DTNB as the derivatization reagent and use HPLC to determine GSH and total Thiols in Bee-Collected pollen。The content of GSH and total Thiols in Bee-Collected pollen are2.1132-2.2106μmolg-1and4.4288-4.3965μmolg-1the recovery rate are101.84%and100.81%, the detection limit are5.43μmolL-1and4.26μmolL-1.Second, use spectrometry to determine GSH and total Thiols in Bee-Collected pollen The content of total thiols and GSH in bee pollen are4.7687-4.9075μmolg-1and1.9079-1.9098μmolg-1, recovery rate are98.32%and98.32%, the detection limit of respectively are0.39μmolL-1and3.58μmolL-1.Finally, GSH and total Thiols in Humulus scandens are dertermined by using UV-vis Spectrophotometry and fluorescence Spectro-metry with "in-situ" derivatizations. The content of GSH and total Thiols in Humulus scandens are0.1671-0.1720μmolg-1and1.9937-2.2973μmolg-1, the recovery rate are101.35%and100.32%, the detection limit are1.365μmolL-1and0.401μmoL-1.This research adopt the Humulus scandens containing GSH and thiol protein as the biological cell sample, and use spectroscopy and chroma to graphy to study different heavy metal stress response function of thiols compound. Thus discusses the thiol compounds such as GSH interact with Cd, Cu, Fe, Pb, and provide basic experiment basis for discussing biological cells in heavy metal detoxification and accumulation mechanism...