Study On The Extraction, Separation And Active Filter From Hedyotis Diffusa And Fructus Schisandrae

Today, natural medicines have been received more attention because of their good therapeutic effects and safety with little side effect. But the development is constrained by their disadvantages, such as containing complex chemical compositions, underlying mechanism is unclear, and so on. Therefore, it is important to identify the active compounds and evaluate their biological activity.In this paper, the inhibit mechanization of compounds in Hedyotis diffusa and Fructus schisandrae against alpha-glycosidase has been studied by high performance liquid chromatography(HPLC), liquid chromatography tandem electrospray mass spectrometry(LC-ESI MS) and ultrafiltration mass spectrometry(UF-MS). Their inhibitory activity against alpha-glycosidase has been discussed, the main active compounds have been isolated and purified by high-speed counter-current chromatography(HSCCC), and the potential anti-diabetic â…¡ activity by Hedyotis diffusa and Fructus schisandrae has been demonstrated. The results are shown below:Identifing the compounds in crude extract from Hedyotis diffusa by LC-ESI MS. Nineteen compounds have been identified, including Asperuloside, 1,3-Dihydroxy-2-methyl anthraquinone, Ferulic acid diglucoside, 1,7-Dihydroxy-6-methoxyl-2-methyl anthraquinone, 2-Hydroxyl-3-methyl anthraquinone, Ferulic acid derivative, 2-Methyl-3-methoxy anthraquinone, 2-Hydroxyl-1-methoxy anthraquinone, Methoxy-cinnamoyl hexoside, Caffeic acid derivetive, 2-Methyl-3-hydroxyl anthraquinone, 2-Hydroxyl-3-methyl-1-methoxyl anthraquinone, 2-Hydroxyl-1-methoxy anthraquinone, Lupenyl acetate.Ultrafiltration liquid chromatography with photo-diode array detection coupled to electraspray ionization tandem mass spectrometry(ultrafiltration LC-DAD-ESI-MS) was used to screen alpha-glycosidase inhibitors from Hedyotis diffusa extract. As a result, ten compounds were identified as 2-hydroxy-3-methyl anthraquinone, ferulic acid derivatives, 2-methyl-3-methoxy anthraquinone, 2-hydroxy-1-methoxy anthraquinone, methoxy cinnamon acyl indican, coffee derivative, 2-methyl-3-hydroxy anthraquinone, 2-hydroxy-3-methyl-1-methoxy anthraquinone and 2-hydroxy-1-methoxy anthraquinone. A separation by HSCCC of activity oriented was carried out based on the solvent system containing n-hexane-ethyl acetate-ethanol-water(5:5:5:5, v/v/v/v). 2-Hydroxyl-3-methyl-1-methoxyl anthraquinone, 2-Methyl-3-hydroxyl anthraquinone, 2-Hydroxyl-3-methyl anthraquinone and compound x have been isolated with purities of 97.71%, 95.96%, 87.60% and 93.66%, determined by HPLC.In addition, oleanolic acid(OA) and ursolic acid(UA) are two important compounds in Hedyotis diffusa with good activities, such as liver-protection, anti-inflammation, hypoglycemic, anti-HIV and anti-tumor. So it is meaningful to simultaneously determinate OA and UA in herbs. But it is difficult because OA and UA are isomeride. In this work, an UPLC-MS/MS method for simultaneous determination of OA and UA in herbs has been developed. Ten herbs have been analyzed using this method. The result indicated that the concentration of OA and UA had good linear relationship with peak area in the range of 0.5-50 ng/mL and the correlation coefficients(r2) were 0.9998 and 0.9997, respectively. The recoveries of OA and UA at three spiked levels were 101.1% and 100.8%, with RSDs(n=9) of 1.8% and 0.04%, respectively. The method showed the advantages of simpleness, high accuracy and good reproducibility, and could be used for detection content of OA and UA in the Chinese herb.Identifing the compounds in crude extract from Fructus schisandrae by LC-ESI MS. Eight compounds have been identified, including schizandrin, schisandrol, angeloyl gomisin H, gomisin D, crotonoyl Gomisin P, schisantherin A and schizandrol B. Ultrafiltration LC-DAD-ESI-MS was used to screen alpha-glycosidase inhibitors and lactate dehydrogenase inhibitors from Fructus schisandrae extract. As a result, schizandrin, schisandrol, angeloyl gomisin H, gomisin D and crotonoyl Gomisin P shows good alpha-glycosidase inhibition activity. Schizandrin, schisandrol and angeloyl gomisin H shows good lactate dehydrogenase inhibition activity.