Spectrum-effect Analysis On The Anti-type 2diabetes Mellitus Activity And Serum Pharmacochemistry Of Dimocarpus Longan Lour.

Objective:（1）In this paper,the profile-effect of HPLC finger-print at different polar parts of leaves of Dimocarpus longan Lour.with its extracorporal oxidation resistance and inhabitation of activity ofα-glucosidase has been researched.（2）A new strategy of quantitative analysis of multi-components via a single marker（QAMS）to synchronously determine the quantity of 5 components（ethyl gallate（C₁）,astragalin（C₂）,quercetin（C₃）,luteolin（C₄）,kaempferol（C₅））in Dimocarpus longan.by UPLC and HPLC was firstly established.（3）The antioxidant effect and identify the blood components of drug serum were verified to explore the pharmacodynamic material basis of Dimocarpus longan Lour.Methods:（1）DPPH method was adopted to measure the extracorporal antioxidant activity of different polar parts and research the inhabitation function of that on activity ofα-glucosidase through enzyme inhabitation dynamics and fluorescene quenching spectroscopy.the HPLC finger-print of5 polar parts of leaves of Dimocarpus longan Lour.was established and the profile-effect was researched through gray relational analysis grade method and Pearson correlation coefficient method.（2）Quercetin（C₃）was chosen as the internal reference.Calculating relative correction factors(RCF_s,f_s/i)of the other 4 components by two correction methods（multipoint correction and slope correction）to effectuate QAMS.At the same time,the difference between the results measured by the method of QAMS and external standard method was compared to verify the accuracy of QAMS.（3）（1）Firstly EFD was selected as the active fraction,after feeding drug（EFD）to the rats,blood from abdominalis of rats was fetched to prepare drug serum.EFD and it’s drug serum’s scavenging ability and total antioxidant capacity of the O₂^-·and·OH two free radicals were observeed.Then the inhibiting effect generated by H₂O₂^-induced oxidative hemolysis and mouse liver microsomal lipid peroxidation product MDA was researched.（2）UPLC-Q-TOF-MS/MS was used to separate EFD and serum samples of rats after drug administration,and total ion detection and second-order fragment ion scanning were performed in the mode of anion detection,and the prototype components and metabolic components extracted from Dimocarpus longan.in rats blood were preliminarily identified.Results:（1）It shows that different polar parts of leaves of Dimocarpus longan Lour.had an obvious scavenging effect on DPPH free radical,and could obviously inhibit the activity ofα-glucosidas;it was indicated through the profile-effect based on gray relational analysis grade method that Peak No.18,1and 22（Quercetin）had relatively large contribution on the antioxidant function,and those that had relatively large contribution on enzymatic activity inhabitation were Peak No.5,6,8,16 and 7;while Peak No.14（Ethyl gallate）and 18 had relatively strong dependency with oxidant resistance through Pearson correlation coefficient method,and those that had relatively strong dependency with enzymatic activity inhabitation were Peak No.16,8,18,7and 5.（2）The results showed that within the linear range,f_s/iwere all in the good durability under diverse chromatographic conditions（RSD<2.28%）.The quantitative results of 5 components in Dimocarpus longan collect from 10producing area by different chromatographic systems and quantitative methods were significantly correlated（Pearson’s r>97.0%）.（3）（1）Compared with blank group,EFD and drug serum has an obvious scavenging effect on free radicals of O₂^-·and·OH（P<0.05）,and the scavenging rate increases with the increase of sample concentration.Drug serum with different dilution ratios has a significant inhibiting effect on erythrocyte hemolysis in mice（P<0.05）and effectively reduces the content of MDA produced by mouse liver microsome（P<0.05）.（2）A total of 17 drug-derived components were preliminarily identified from the drug serum,among which 1 prototype component was isolated from the extract of Dimocarpus longan.,and the rest were mostly glycoside metabolites of the prototype component.Conclusion:（1）The antioxidant function and inhabitation of 5 different polar parts of leaves of Dimocarpus longan Lour.on activity ofα-glucosidase was a result of coefficient of multiple elements,correlation peaks mainly gathered at the parts of ethyl acetate and n-butyl alcohol.（2）The applicability and feasibility of the QAMS method established in this study were evaluated to be favorable for quality control of Dimocarpus longan.As a new model of quality control,it can provide one more choice of multi-components quality control methods in the absence of standard substances or instruments.（3）There is a certain antioxidant activity in the drug serum,EFD drug serum contains substances with antioxidant activity,and it is preliminarily determined that ethyl gallate,quercetin,luteolin and other components may be part of the substance basis of anti-type 2 diabetes of leaves of Dimocarpus longan Lour.