| In order to disclose material basis of the anti-tumor activity of D.hainanensis, we isolated the chemistry constituents from the D.hananensis by various chromatographic methods, such as Si gel, Sephadex LH-20, macroporous resin, MPLC, recrystallization, etc.8compounds were isolated from the leaves and stems of the D.hananensis,6compounds were isolated from the D.hananensis fruits and obtained multiple liposoluble constituents in separation process. The obtained compounds were identified on the basis of UV, IR, NMR, MS, GC data, comparison with standard samples, measurement of physicochemical characters and related reference. The analysis and identified results as the following:8compounds from the ethyl acetate etract of leaves and stems of D.hananensis were identified as:4β-hydroxy-23-nor-friedel-3-one (1), friedelin (2), Diethylhexyl phthalate (3), a-Amyrin (4), β-Amyrin (5), Lupeol (6), Friedelane-3,7-dione (7) and β-sitosterol (8).2compounds from the petroleum ether extract of leaves and stems of D.hananensis were identified as:friedelin (A),23-nor-friedel-3-one (B). Compound1and B were new compounds, compound3was isolated for the first time from Drypetes, compounds2,4,5,6,7,8were isolated for the first time from species D.hananensis. The liposoluble constituents from D.hananensis stems and leaves, which contained high content of saturated fatty acids, unsaturated fatty acids and Bis (2-ethylhexyl) phthalate (DEHP), also contains alcohols, paraffins, olefins, silicones, carboxylic acids and ketones.6compounds, e.g. β-sitosterol(Ⅰ), stigmasterol(Ⅱ), friedelin(III), palmitic acid(IV),β-daucosterol(V), Vanillin(VI) were isolated from the fruits of D.hananensis. Compounds I-VI were isolated for the first time from the D.hananensis fruits. The chloroform liposoluble extract I from the D.hananensis fruits was contained DEHP and alkanes, mainly; lipophilic extract II contained olefin, alkanes, esters, as well as plant alcohol mainly.We discovered considerable DEHP in the liposoluble constituents from the leaves and stems of D.hananensis. In order to exclude the using of plastic containers in the separation process, which could cause the sample contain large amounts of DEHP, a methodology of diethylhexyl phthalate quantitative analysis in leaves and stems of Drypetes hainanensis by high performance liquid chromatography (HPLC) was established. The results were optimized by sample pre-treatment and the variation of chromatographic detection conditions. The retention time of peak was used to qualitatively evaluate the results, and the ex-situ measurement was applied for quantitative analysis. Using YMC-Sil column (250mm×4.6mm,5μm), the condition was:the inject volume was10μL, the mobil phase was CH2Cl2, the flow rate was1.0mL/min, the detection wavelength was275nm. When the content of DEHP is between0.05～0.80μg (R=0.9973), there was a good linear relationship and the accuracy and precision were good. The average recovery of DEHP was99.23%with RSD of1.22%. The content of DEHP in leaves and stems of D.hananensis was10.93(±0.21) μg/g.Compounds1,2and7have the similar structure, and they were evaluated for their growth inhibitory activity against human cell lines BEL-7402, A-549, and HL-60by MTT and SRB, in which adriamycin was used as the reference substance. However, the inhibition rates are all below50%. Compound7showed significant cytotoxicity, which was comparable to compound1and2(P<0.05) against all three cell lines. Compund7(Friedelane-3,7-dione) was keto derivative, it was observed that the ketone groups at C-3and C-7in7considerably increased the cytotoxicity against tumor cell lines. In addition, the literature data showed that: the compounds4,5and6have significant anti-tumor activity, which inferred the compounds4-6are the pharmacodynamic material basis of the anti-tumor activity of D.hainanensis. |
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