Extraction,Purification,Identification And Inhibitory Effect On HepG2 Cells Of Monoterpene Glycosides From Paeonia Suffruticosa Seed Meal

Paeonia suffruticosa(P.suffruticosa)seed meal is a by-product of oil processing of P.suffruticosa,which is rich in monoterpene glycosides.At present,P.suffruticosa seed meal is mainly disposed in the form of discard and has not been effectively utilized.It is of great significance to study the extraction,purification and inhibitory effect on HepG2 cells of monoterpene glycosides from P.suffruticosa seed meal for the efficient utilization of P.suffruticosa seed meal.In this paper,P.suffruticosa seed meal was taken as the research object.The monoterpene glycosides in P.suffruticosa seed meal were extracted by ultrasonic assisted ethanol extraction process,and the crude extract was purified by macroporous resin.The main compounds of P.suffruticosa seed meal extract were identified by UPLC-Q-TOF-MS/MS.On this basis,the inhibitory effect of monoterpene glycoside extract from P.suffruticosa seed meal on HepG2 cells was discussed.The results are as follows:(1)The optimal extraction process of monoterpene glycosides from P.suffruticosa seed meal was determined by single factor tests and response surface tests:ethanol concentration,33%;ultrasonic temperature,55℃;ultrasonic power,400 W;liquid-material ratio,33:1;ultrasonic time,44 min.The extraction yield of monoterpene glycosides from P.suffruticosa seed meal under this condition was 121.03 mg/g.(2)Macroporous resins were used to purify monoterpene glycosides from P.suffruticosa seed meal.Among the eight macroporous resins,the purification performance of LSA-900C macroporous resin was best,and the optimal purification process was sample concentration of 4 mg/mL,sample solution pH of 5,sample flow rate of 1.5 mL/min,sample volume of 70 mL,ethanol concentration of 50%,elution flow rate of 2.5 mL/min,and elution volume of 110 mL.Under this condition,the purity of monoterpene glycosides from P.suffruticosa seed meal increased from 20.50%to 71.20%.(3)The monoterpene glycoside extract from P.suffruticosa seed meal was identified by UPLC-Q-TOF-MS/MS,including:oxy paeoniflorin,isomaltose paeoniflorin,albiflorin,6’-O-β-D-glucopyranoside,paeoniflorin and Mudanpioside i.The main monoterpene glycosides were albiflorin and paeoniflorin,and the contents in P.suffruticosa seed meal were determined by high performance liquid chromatography to be 15.24 mg/g and 14.12 mg/g,respectively.(4)The inhibitory effect of monoterpene glycoside extract from P.suffruticosa seed meal on HepG2 cells was discussed.The results showed that monoterpene glycosides of P.suffruticosa seed meal could significantly inhibit the cell viability of HepG2 cells,destroy the normal morphology of HepG2 cells,inhibit the invasion and cloning ability of HepG2 cells.The mechanism may be that monoterpene glycoside extract could induce HepG2 cells mitochondrial-dependent apoptosis by regulating Bcl-2/Caspase-3 signaling pathway and block HepG2 cells in G1 phase.