变波长RP-HPLC法同时测定调经丸中12种成分

Simultaneous determination of 12 chemical components in Tiaojing Pills by RP-HPLC under variable wavelengths

目的建立同时测定调经丸中芍药苷、毛蕊花糖苷、阿魏酸、盐酸益母草碱、橙皮苷、丹皮酚、黄芩苷、川续断皂苷VI、柠檬苦素、白术内酯I、白术内酯III、茯苓酸12种成分的RP-HPLC方法。方法采用RP-HPLC法,色谱柱为Venusil MP C18（250 mm×4.6 mm,5.0μm）;流动相为乙醇-乙腈（40∶60,A）和0.2%磷酸水溶液（B）,梯度洗脱,体积流量0.8m L/min;柱温45℃。结果 12种指标成分芍药苷、毛蕊花糖苷、阿魏酸、盐酸益母草碱、橙皮苷、丹皮酚、黄芩苷、川续断皂苷VI、柠檬苦素、白术内酯I、白术内酯III、茯苓酸分别在0.5~50.0 mg/L（r=0.999 5）、0.1~10.0 mg/L（r=0.999 1）、0.2~20.0 mg/L（r=0.999 2）、0.3~30.0 mg/L（r=0.999 3）、4.0~400.0 mg/L（r=0.999 8）、0.2~20.0 mg/L（r=0.999 1）、0.6~60.0 mg/L（r=0.999 4）、1.5~150.0 mg/L（r=0.999 8）、7.0~700.0 mg/L（r=0.999 9）、0.5~50.0 mg/L（r=0.999 3）、0.5~50.0 mg/L（r=0.999 4）、1.0~100.0 mg/L（r=0.999 6）与峰面积呈良好的线性关系;精密度良好,RSD≤0.97%;重复性良好,RSD≤1.25%;加样回收率在98.5%~103.5%,RSD≤1.24%。供试品溶液在室温条件下24 h内稳定,RSD≤1.09%。12批次供试品中芍药苷、毛蕊花糖苷、阿魏酸、盐酸益母草碱、橙皮苷、丹皮酚、黄芩苷、川续断皂苷VI、柠檬苦素、白术内酯I、白术内酯III、茯苓酸质量分数分别为4.328~4.688、0.033~0.054、0.073~0.091、0.177~0.199、0.243~0.283、0.043~0.069、1.144~1.173、0.037~0.061、0.094~0.126、0.127~0.157、0.155~0.179、0.285~0.327 mg/g。结论所建立的方法快速、灵敏度高、准确度高、专属性好,为调经丸的质量控制提供依据。

Objective To establish a RP-HPLC method to determine the contents of paeoniflorin, acteoside, ferulic acid, leonurine hydrochloride, hesperidin, paeonol, baicalin, asperosaponin VI, limonin, atractylenolide I, atractylenolide III, and pachymic acid in Tiaojing Pills. Methods The determination was performed on a Venusil MP-C18 column（250 mm × 4.6 mm, 5.0 μm） with ethanol-acetonitrile（40∶60, A） and 0.2% phosphoric acid（B） as mobile phases for gradient elution, at the flow rate of 0.8 m L/min; The column temperature was 45 ℃. Results The nine components were well separated and showed good linearity, such as paeoniflorin 0.5—50.0 mg/L（r = 0.999 5）, acteoside 0.1—10.0 mg/L（r = 0.999 1）, ferulic acid 0.2—20.0 mg/L（r = 0.999 2）, leonurine hydrochloride 0.3—30.0 mg/L（r = 0.999 3）, hesperidin 4.0—400.0 mg/L（r = 0.999 8）, paeonol 0.2—20.0 mg/L（r = 0.999 1）, baicalin 0.6—60.0 mg/L（r = 0.999 4）, asperosaponin VI 1.5—150.0 mg/L（r = 0.999 8）, limonin 7.0—700. 0 mg/L（r = 0.999 9）, atractylenolide I 0.5—50. 0 mg/L（r = 0.999 3）, atractylenolide III 0.5—50. 0 mg/L（r = 0.999 4）, and pachymic acid 1.0—100. 0 mg/L（r = 0.999 6）. The precision was good, RSD ≤ 0.97%, the repeatability was good in terms of RSD ≤ 1.25% and the recovery rate was 98.5%—103.5%（RSD ≤ 1.24%）. Test solution was stable at room temperature within 24 h. The contents of twelve batches of the paeoniflorin, acteoside, ferulic acid, leonurine hydrochloride, hesperidin, paeonol, baicalin, asperosaponin VI, limonin, atractylenolide I, atractylenolide III and pachymic acid were 4.328—4.688, 0.033—0.054, 0.073—0.091, 0.177—0.199, 0.243—0.283, 0.043—0.069, 1.144—1.173, 0.037—0.061, 0.094—0.126, 0.127—0.157, 0.155—0.179, and 0.285—0.327 mg/g, respectively. Conclusion The method is rapid and has high sensitivity, high accuracy, and good specificity, It can be applied to the quality control of Tiaojing Pills.