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目的:评价新疆鼠尾草根中6,7-去氢罗列酮的体外抗血小板聚集作用并优选其提取工艺。方法:应用比浊法测定6,7-去氢罗列酮体外抗血小板聚集作用。在单因素试验基础上,以6,7-去氢罗列酮提取率为指标,通过正交设计法考察提取次数、料液比和提取时间对6,7-去氢罗列酮提取工艺的影响。采用HPLC测定6,7-去氢罗列酮含量,色谱条件为依利特C18色谱柱(4.6 mm×250 mm,5μm),流动相甲醇-0.02%甲酸(83∶17),流速1.0 mL·min-1,检测波长330 nm,柱温40℃。结果:6,7-去氢罗列酮高、中剂量组抑制凝血酶诱导的血小板聚集作用与生理盐水组比较均具有极显著性差异,高剂量组在15,30,45,60 min内由凝血酶诱导的血小板抑制聚集率分别为90%,75%,66%,64%。最佳提取工艺为加10倍量95%于85℃提取3次,每次3 h,新疆鼠尾草根中6,7-去氢罗列酮质量分数0.01%。结论:新疆鼠尾草根中6,7-去氢罗列酮表现出较强的抗血小板聚集作用,在此活性基础上优选的提取条件稳定可行。
OBJECTIVE: To evaluate in vitro anti-platelet aggregation of 6,7-drospirenone in sage root of Xinjiang and to optimize its extraction process. Methods: The turbidimetric method was used to determine the antiplatelet aggregation effect of 6,7-drospirenone in vitro. On the basis of single factor experiment, taking the extraction rate of 6,7-drospirenone as an index, the effects of extraction times, solid-liquid ratio and extraction time on the extraction of 6,7-drospirenone were investigated by orthogonal design. The content of 6,7-drospirenone was determined by HPLC. The chromatographic conditions were Elite C18 column (4.6 mm × 250 mm, 5 μm), mobile phase methanol-0.02% formic acid (83:17), flow rate 1.0 mL · min- 1, detection wavelength 330 nm, column temperature 40 ℃. Results: The platelet aggregation induced by thrombin in high and medium dose groups of 6, 7-drosplitazone group was significantly different from that of saline group. In the high dose group, Enzyme-induced platelet aggregation inhibition rates were 90%, 75%, 66%, 64%, respectively. The optimum extraction process was adding 10 times the amount of 95% at 85 ℃ extraction 3 times, each 3 h, Xinjiang Salvia root 6,7-drospirenone mass fraction of 0.01%. Conclusion: 6,7-Drospirenone in root of Salvia miltiorrhiza showed a strong anti-platelet aggregation effect. The optimal extraction conditions were stable and feasible.