| Peony is famous in China for it not only has highly ornamental value, but also has alot of health functions. The peony seeds are rich in oil, in which unsaturated fatty acid,such as linolenic acid, accounted for most. Now the peony seed oil has become a valuablenew food resource and has got large-scale development and utilization. The seed cake ofpeony is the production remains of peony seed oil, but up to now there is no systemresearch about it. In the present paper, taking this biological resource as raw materials, thechemical composition of it was analyzed, and six main compounds were isolated andidentified successfully. Then basing on the six references, a suit of method to quantitatethose compositions in seed cake of peony by high performance liquid chromatographic wasestablished and a method to extract and purify the compositions by macroporousadsorption resin was also developed, which provided a scientific basis for thecomprehensive development and utilization of the natural products resources of peonyseeds.The effective components were extracted from1kg seed cakes of peony with30%ethanol, preliminary purified by macroporous adsorption resin, then fine prepared bypreparative HPLC with methanol as the mobile phase. Six main ingredients were got,respectively142,310,61,549,127, and549mg. Their purity detected by peak areapercentage method was all over95%. Finally, identified the structures of these chemicalcomponents with LC-MS and1H-NMR spectroscopy, and they were6’-O-β-D-glucopyran-osylalbiflorin, albiflorin, β-gentiobiosylpaeoniflorin, paeoniflorin, benzoic acid andpaeoniflorigenone, respectively.Taking the six paeoniflorins as references, a method to analysis the paeoniflorins inseed cakes of peony by high performance liquid chromatographic method was established,and these six paeoniflorins were separated and determined well in17min. Column:Kromasil C18(200×4.6mm,5μm); mobile phase: A:500μL/L formic acid; B: methanolcontaining500μL/L formic acid; the gradient elution condition (min/B%) was:0/40,4/40,8/60,15/60,16/40,20/40; flow:0.8mL/min; temperature:30℃; diode array detector(DAD) at230nm; injection quantity:10μL. And the external standard calibration curveswere used in quantification. The paeoniflorins in seed cakes of peony were separated wellunder these conditions. The relative standard deviations (RSD) of repeated measurement were between1.91%2.72%. The detection limits were between2.8ng16.1ng. What ismore, between the linearity range of0.8ng4000ng, the correlation coeffeicent (r) was0.99920.9999. Standard addition recovery rate was between93.0%100.0%.Taking the four major components in seed cakes of peony (paeoniflorin, albiflorin,6’-O-β-D-glucopyranosylalbiflorin and β-gentiobiosylpaeoniflorin) as indexes, thepurification technology of macroporous resin adsorption was optimized. HPD-200A resinwas chose as the more suitable resin. The optimal process for the purification was asfollows: the concentration of sample solution was8.0mg (paeoniflorin)/mL, the flow ratewas1/16BV (bed volume)/min and the loading capacity was4.5BV; then eluted with50%ethanol for4BV with elution flow rate of1/16BV/min. In these optimum conditions, thecontent of paeoniflorin, albiflorin,6’-O-β-D-glucopyranosylalbiflorin and β-gentiobiosylp-aeoniflorin was reached to32.3%,16.5%,8.02%and6.63%, respectively. |
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