基于一测多评法对正心降脂片中7种成分的质量控制研究

Quality control for 7 components in Zhengxin Jiangzhi Tablets based on quantitative analysis of multi-components with a single-marker

目的建立一测多评法(QAMS)同时测定正心降脂片中木犀草素-7-O-β-D-葡萄糖醛酸苷、芹菜素-7-O-β-D-葡萄糖醛酸苷、橙黄决明素、黄决明素、决明素、毛蕊异黄酮葡萄糖苷和芒柄花苷的含量,验证此方法在制剂中应用的准确性和可行性。方法采用Shimadzu C18(250 mm×4.6 mm,5μm)色谱柱;柱温为25℃;流动相A:乙腈-甲醇(9∶1),流动相B:1.0 mL·L^-1甲酸溶液,梯度洗脱;流速为1.0 mL·min^-1;检测波长分别为345 (检测木犀草素-7-O-β-D-葡萄糖醛酸苷和芹菜素-7-O-β-D-葡萄糖醛酸苷),284(检测橙黄决明素、黄决明素和决明素)和254 nm(检测毛蕊异黄酮葡萄糖苷和芒柄花苷)。以正心降脂片为研究对象,选定橙黄决明素为内参物,建立其与木犀草素-7-O-β-D-葡萄糖醛酸苷、芹菜素-7-O-β-D-葡萄糖醛酸苷、黄决明素、决明素、毛蕊异黄酮葡萄糖苷和芒柄花苷的相对校正因子,计算待测成分含量,同时将外标法实测值与一测多评法的计算值进行比对,以验证一测多评法的准确性和可靠性。结果木犀草素-7-O-β-D-葡萄糖醛酸苷、芹菜素-7-O-β-D-葡萄糖醛酸苷、橙黄决明素、黄决明素、决明素、毛蕊异黄酮葡萄糖苷和芒柄花苷的线性范围分别为8.57~214.25,5.91~147.75,2.73~68.25,1.18~29.50,0.79~19.75,3.07~76.75和1.89~47.25μg·mL^-1,r值均在0.999 2~0.999 7范围内;平均回收率分别为99.59%,98.95%,97.82%,98.00%,97.65%,96.99%和100.02%;RSD值分别为0.74%,1.12%,1.28%,0.99%,1.44%,1.01%和0.88%(n=9)。木犀草素-7-O-β-D-葡萄糖醛酸苷、芹菜素-7-O-β-D-葡萄糖醛酸苷、橙黄决明素、黄决明素、决明素、毛蕊异黄酮葡萄糖苷和芒柄花苷含量的一测多评法计算值和外标法实测值比较差异无统计学意义。结论一测多评法可同时测定正心降脂片中7种成分的含量,该方法简便、重复性好、准确度高。

Objective To establish a quantitative analysis of multi-components by single-marker(QAMS) method for the simultaneous determination of luteolin-7-O-β-D-glucuronide,apigenin-7-O-β-D-glucuronide,aurantio-obtusin,chrysoobtusin,obtusin,calycosin 7-O-β-D-glucopyranoside and ononin in Zhengxin Jiangzhi Tablets,and validate its accuracy and feasibility for application in the preparation.Methods The analysis was performed on a 25 ℃ thermostatic Shimadzu C18(250 mm×4.6 mm,5 μm) chromatographic column with the mobile phase comprising of acetonitrile-methanol(9∶1)(A) and 1.0 mL·L^-1formic acid solution(B) at a flow rate of 1.0 mL·min^-1 in a gradient elution manner.The detection wavelength was set at 345 nm for luteolin-7-O-β-D-glucuronide and apigenin-7-O-β-D-glucuronide;284 nm for aurantio-obtusin,chrysoobtusin and obtusin;254 nm for calycosin 7-O-β-D-glucopyranoside and ononin.By using Zhengxin Jiangzhi Tablets as the research object,aurantio-obtusin as the internal reference substance,the relative correlation factors of luteolin-7-O-β-D-glucuronide,apigenin-7-O-β-D-glucuronide,chrysoobtusin,obtusin,calycosin7-O-β-D-glucopyranoside and ononin were established,and the content of each component was calculated.The measured values of the external standard method were compared with the calculated results of correction factor in order to test the accuracy of the established method.Results Luteolin-7-O-β-D-glucuronide,apigenin-7-O-β-D-glucuronide,aurantio-obtusin,chrysoobtusin,obtusin,calycosin 7-O-β-D-glucopyranoside and ononin had good relations within the ranges of 8.57-214.25,5.91-147.75,2.73-68.25,1.18-29.50,0.79-19.75,3.07-76.75 and 1.89-47.25 μg·mL^-1(r=0.999 2-0.999 7).The average recoveries were 99.59%,98.95%,97.82%,98.00%,97.65%,96.99% and 100.02%;RSDs were 0.74%,1.12%,1.28%,0.99%,1.44%,1.01% and 0.88%(n=9),respectively.The results of luteolin-7-O-β-D-glucuronide,apigenin-7-O-β-D-glucuronide,aurantio-obtusin,chrysoobtusin,obtusin,calycosin 7-O-β-D-glucopyranoside and ononin were no significant difference between external standard method and QAMS method.Conclusion The QAMS method which can simultaneously determine the content of 7 components in Zhengxin Jiangzhi Tablets is simple,and shows good repeatability and high accuracy.