HPLC-DAD法同时测定参芪十一味颗粒的11种成分

Simultaneous determination of 11 constituents in Shenqi Shiyiwei Granule by HPLC-DAD

目的建立HPLC-DAD法同时测定参芪十一味颗粒(SSG)中23-乙酰泽泻醇B、阿魏酸、毛蕊花糖苷、人参皂苷Rg1、人参皂苷Re、人参皂苷Rb1、天麻素、大黄酚、橙黄决明素、毛蕊异黄酮葡萄糖苷和金丝桃苷的方法。方法采用RP-HPLC法,色谱柱为Waters XBridge-C18(250 mm×4.6 mm,5.0μm);流动相为甲醇-乙腈-水(15∶80∶5)和乙腈-0.1%磷酸水溶液(10∶90),梯度洗脱,体积流量1.0 m L/min;柱温35℃,进样量10μL。结果 23-乙酰泽泻醇B、阿魏酸、毛蕊花糖苷、人参皂苷Rg1、人参皂苷Re、人参皂苷Rb1、天麻素、大黄酚、橙黄决明素、毛蕊异黄酮葡萄糖苷和金丝桃苷11种成分能够达到很好分离;其线性范围分别为0.4~8.0μg/m L(r=0.999 2)、0.2~4.0μg/m L(r=0.999 5)、0.2~4.0μg/m L(r=0.999 5)、0.1~2.0μg/m L(r=0.999 6)、0.1~2.0μg/m L(r=0.999 7)、0.1~2.0μg/m L(r=0.999 4)、0.5~10μg/m L(r=0.999 2)、0.6~12μg/m L(r=0.999 2)、0.4~8.0μg/m L(r=0.999 4)、1.0~20μg/m L(r=0.999 6)、0.8~16μg/m L(r=0.999 3),平均加样回收率分别为98.1%、98.1%、99.1%、98.3%、99.5%、99.9%、98.5%、100.4%、101.6%、99.7%、101.2%,RSD分别为0.9%、1.6%、1.6%、1.8%、1.5%、0.6%、0.7%、0.8%、0.4%、0.9%、1.1%(n=6)。9批次供试品中23-乙酰泽泻醇B、阿魏酸、毛蕊花糖苷、人参皂苷Rg1、人参皂苷Re、人参皂苷Rb1、天麻素、大黄酚、橙黄决明素、毛蕊异黄酮葡萄糖苷和金丝桃苷质量浓度分别为0.081~0.089、0.261~0.269、0.060~0.069、0.038~0.047、0.030~0.037、0.042~0.049、0.420~0.428、0.141~0.151、0.178~0.189、0.107~0.117、0.069~0.078 mg/g,结果表明本品各批次之间差异较小。结论本方法操作简便,测定结果准确可靠,可用于SSG的质量控制。

Objective To establish HPLC-DAD method for the simultaneous determination of 23-acetate alisol B, ferulic acid, verbascoside, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, gastrodin, chrysophanol, aurantio-obtusin, calycosin 7-O-β-Dglucopyranoside, and hyperoside in Shenqi Shiyiwei Granule（SSG）. Methods The chromatographic separation was achieved on an Waters XBridge-C18（250 mm × 4.6 mm, 5.0 μm） column with methanol-acetonitrile-water（15∶80∶5） and methanol-0.1% phosphoric acid（10∶90） as mobile phases for gradient elution, at the flow rate of 1.0 m L/min; The column temperature was 35 ℃.Results The linear ranges of 23-acetate alisol B, ferulic acid, verbascoside, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, gastrodin, chrysophanol, aurantio-obtusin, calycosin 7-O-β-D-glucopyranoside, and hyperoside were 0.4—8.0 μg/m L（r = 0.999 2）, 0.2—4.0 μg/m L（r = 0.999 5）, 0.2—4.0 μg/m L（r = 0.999 5）, 0.1—2.0 μg/m L（r = 0.999 6）, 0.1—2.0 μg/m L（r = 0.999 7）, 0.1—2.0 μg/m L（r = 0.999 4）, 0.5—10 μg/m L（r = 0.999 2）, 0.6—12 μg/m L（r = 0.999 2）, 0.4—8.0 μg/m L（r = 0.999 4）, 1.0—20 μg/m L（r = 0.999 6）, and 0.8—16 μg/m L（r = 0.9993）. The average recoveries（n = 6） were 98.1%（RSD = 0.9%）, 98.1%（RSD = 1.6%）, 99.1%（RSD = 1.6%）, 98.3%（RSD = 1.8%）, 99.5%（RSD = 1.5%）, 99.9%（RSD = 0.6%）, 98.5%（RSD = 0.7%）, 100.4（RSD = 0.8%）, 101.6%（RSD = 0.4%）, 99.7%（RSD = 0.9%）, and 101.2%（RSD = 1.1%）, respectively. The contents of nine batches of 23-acetate alisol B, ferulic acid, verbascoside, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, gastrodin, chrysophanol, aurantio-obtusin, calycosin 7-O-β-D-glucopyranoside, and hyperoside were 0.081—0.089, 0.261—0.269, 0.060—0.069, 0.038—0.047, 0.030—0.037, 0.042—0.049, 0.420—0.428, 0.141—0.151, 0.178—0.189, 0.107—0.117, and 0.069—0.078 mg/g. The results showed that there was little difference among the batches. Conclusion The method is accurate, sensitive, credible, and repeatable. It can be applied to the quality control of SSG.