The Separation And Identification Of Monoterpene Glycoside And Oligostilbene Compound Of Paeonia Lactiflora Pall Seed And Their Effect On The Lipid Accumulation In HepG2 Cells

Paeonia lactiflora Pall is a kind of traditional Chinese ornamental plant widely cultivated in China.Studies demonstrated that seed have abundant levels of oils,monoterpene glycosides and oligostilbene compounds,and the research value of the seed of Paeonia lactiflora Pall will be significant.The chemical constituents of seed,and the identification,extraction and separation of monoterpene glycoside and oligostilbene compound in the seed kernels and hulls of Paeonia lactiflora Pall in this paper.Improving lipid accumulation in HepG2 cells of monoterpene glycoside and oligostilbene compound had been deeply explored,which provide a theoretical basis for the comprehensive development and utilization of Paeonia lactiflora Pall seed.The main research contents were as follows:First,shell and kernel percentage,seed oil content and fatty acid composition,and main chemical components and content of seed kernels and hulls of Paeonia lactiflora Pall were evaluated.The results showed that the oil content of Paeonia lactiflora seed kernel was33.7%,while the oil content of Paeonia lactiflora seed shell was 3.07%.The seed oil had abundant levels of unsaturated fatty acids,including oleic,linoleic and α-linolenic acids.Moreover,multiple beneficial dietary phytochemicals(tocopherol,phytosterols,and squalene)are contained in Paeonia lactiflora Pall seed oil.The seed shell accounted for 32.02%,and seed kernel represented 67.98% of the mass of the seeds.The results also showed that the seed kernels contain high concentrations of proteins and monoterpene glycosides,whilst the seed hulls contain high concentrations of cellulose,lignin,and oligostilbene compounds.Then,the ultra-high performance liquid chromatography of quadrupole time of flight-tandem mass spectrometry was used to qualitatively and quantitatively analyze the bioactive compounds extracts in seed kernels and hulls of Paeonia lactiflora Pall.And a total of 96 compounds were tentatively identified,including 24 monoterpene glycosides,16 oligostilbene compounds,and several phenolic acid,flavone and iridoid glycosides.Paeoniflorin(1779.61±10.33 mg/100 g)was the predominant monoterpene glycoside in seed kernels,whilst hulls had the highest concentrations of sufruticosol A(791.9 ± 25.09 mg/100 g)and trans-ε-viniferin(607.4 ± 16.22 mg/100 g).Thirdly,we extracted the monoterpene glycosides and oligostilbene compounds from Paeonia lactiflora Pall seed kernels and hulls by ultrasound,and extraction conditions were optimized by single factor experiment.The results showed that under the optimal conditions as extraction time 30 min,ethanol volume fraction of 70% and solid-to-liquid ratio of 1:30(g/m L),the manximum extraction yield of monoterpene glycoside reached 4.24%;the AB-8macroporous resin was selected for purification of monoterpene glycoside crude extract,and the optimum desorption conditions were as follows: the p H of 5,the loading concentration of1.5 mg/m L,the eluting with 30% ethanol.The optimal extraction conditions of oligostilbene compounds were as follows: extraction time 30 min,ethanol volume fraction of 50% and solid-to-liquid ratio of 1:30(g/m L).Under these conditions,the maximum extraction yield of oligostilbene compound reached 5.56%;the D101 macroporous resin was selected for purification of oligostilbene compound crude extract,and the optimum desorption conditions were as follows: the p H of 4,the loading concentration of 1.5 mg/m L,the eluting with 30%ethanol.Finally,the effects of monoterpene glycoside and oligostilbene compound on the lipid accumulation in HepG2 cells were investigated,and the results of experimented showed that monoterpene glycoside and oligostilbene compound can reduce the levels of total triglycerides,cholesterol,and LDL-c in HepG2 cells.But there is no significant leves of improvement of HDL-c in HepG2 cells.The TG contents of high dose group in HepG2 cells treated by the model group decreased by 83.19% and 84.61%,respectively,the TC contents decreased by 78.18% and 80.37%,respectively,and the LDL-c decreased by 78.18% and80.37%,respectively.