HPLC-DAD法同时测定更年安片中6个活性成分的含量

目的: 建立同时分析测定更年安片中6个活性成分(2,3,5,4’-四羟基二苯乙烯-2-O-β-D-葡萄糖苷、毛蕊花糖苷、仙茅苷、丹皮酚、哈巴俄苷、五味子醇甲)的方法。 方法: 采用HPLC-DAD法,色谱柱为奥泰Alltima ODS(250 mm×4.6 mm,5 μm),以乙腈为流动相A,0.05%磷酸溶液为流动相B,梯度洗脱,流速1.0 mL·min^-1,检测波长为254 nm(测定五味子醇甲)、284 nm(测定仙茅苷、丹皮酚、哈巴俄苷)、330 nm(测定2,3,5,4’-四羟基二苯乙烯-2-O-β-D-葡萄糖苷、毛蕊花糖苷)。 结果: 2,3,5,4’-四羟基二苯乙烯-2-O-β-D-葡萄糖苷、毛蕊花糖苷、仙茅苷、丹皮酚、哈巴俄苷、五味子醇甲线性范围分别为0.0404～1.616 μg(r=0.9999),0.0194～0.774 μg(r=0.9998),0.0202～0.810 μg(r=0.9997),0.0201～0.804 μg(r=0.9999),0.0206～0.826 μg(r=0.9998),0.0398～1.592 μg(r=0.9999)。平均加样回收率(n=6)分别为97.8%,98.2%,97.9%,98.3%,101.2%,97.5%;RSD分别为1.2%,1.1%,0.95%,1.1%,0.97%,1.1%。 结论: 该方法操作简单,重复性好,为评价和监控更年安片的质量提供可靠的方法。

Objective: To establish a method for simultaneous determination of six active ingredients(2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside, acteoside, curculigoside, paeonol, harpagoside, schisandrin). Methods: The determination was carried out with Alltima ODS column(250 mm×4.6 mm, 5 μm); The mobile phase consisted of acetonitrile(A)-0.05% phosphoric acid solution(B) with gradient elution at a flow rate of 1.0 mL·min^-1; The detection wavelengths were 254 nm for schisandrin, 284 nm for curculigoside, paeonol and harpagoside, 330 nm for 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside and acteoside. Results: The calibration curves were linear in the ranges of 0.0404-1.616 μg for 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside(r=0.9999), 0.0194-0.774 μg for acteoside (r=0.9998), 0.0202-0.810 μg for curculigoside(r=0.9997), 0.0201-0.804 μg for paeonol (r=0.9999), 0.0206-0.826 μg for harpagoside(r=0.9998), 0.0398-1.592 μg for schisandrin(r=0.9999); The average recoveries(n=6) were 97.8%, 98.2%, 97.9%, 98.3%, 101.2%, and 97.5%; RSDs were 1.2%, 1.1%, 0.95%, 1.1%, 0.97%, and 1.1%, respectively. Conclusion: The method is simple and repeatable,which can be applied to the quality control of Gengnian’an tablets.

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