一测多评法同时测定扶正固本颗粒中6种成分的含量

仝立国 牛艳艳 王若瑜 吉海杰 宋美卿 冯玛莉 夏召弟 汪欣文

摘 要 目的：建立扶正固本顆粒中2，3，5，4′-四羟基二苯乙烯基葡萄糖苷、黄芩苷、淫羊藿苷、汉黄芩苷、黄芩素、汉黄芩素等6种成分含量的测定方法。方法：采用高效液相色谱法。色谱柱为Dikma Diamonsil C18，流动相为乙腈-0.1%磷酸水溶液（梯度洗脱），检测波长为275 nm（0～8 min）、320 nm（8～9 min）和275 nm（9～33 min），流速为1.0 mL/min，柱温为25 ℃，进样量为10 μL。以黄芩苷为基准物，采用多点校正法和斜率校正法分别计算另外5种成分的相对校正因子（fk/s），并采用保留时间差值法对待测成分进行色谱峰定位，比较上述两种一测多评法所得计算值与外标法实测值的差异。结果：2，3，5，4′-四羟基二苯乙烯基葡萄糖苷、黄芩苷、淫羊藿苷、汉黄芩苷、黄芩素、汉黄芩素检测进样量的线性范围分别为0.053～2.12、0.163～6.52、0.059～2.36、0.021 6～0.864、0.03～1.2、0.021～0.84 μg（r>0.999），精密度、稳定性（12 h）、重复性试验的RSD均小于3%;平均加样回收率为98.72%～99.82%（RSD为0.89%～1.24%，n=9）。以黄芩苷为基准物，2，3，5，4′-四羟基二苯乙烯基葡萄糖苷、淫羊藿苷、汉黄芩苷、黄芩素、汉黄芩素多点校正法的fk/s分别为1.172、0.528、1.479、1.820、2.534，斜率校正法的fk/s分别为1.234、0.550、1.559、1.939、2.664;3种方法测得10批扶正固本颗粒中6种成分含量的RSD为0.29%～2.77%（n=10）;两种一测多评法测得结果与外标法的Pearson相关系数均不低于0.999 9（P<0.001）。结论：成功建立了可用于同时测定扶正固本颗粒中6种成分含量的一测多评法。

关键词 一测多评法;扶正固本颗粒;2，3，5，4′-四羟基二苯乙烯基葡萄糖苷;黄芩苷;淫羊藿苷;汉黄芩苷;黄芩素;汉黄芩素;含量

ABSTRACT   OBJECTIVE： To establish the method for content determination of 6 components in Fuzheng guben granules， such as 2，3，5，4′-tetrahydroxystilbene glucoside，baicalin，icariin，scutellarin，baicalein and wogonin. METHODS： HPLC method was adopted. The determination was performed on Dikma Diamonsil C18 column with mobile phase consisted of  acetonitrile-0.1% phosphoric acid aqueous solution （gradient elution） at the flow rate of 1.0 mL/min. The detection wavelengths were set at 275 nm （0-8 min）， 320 nm（8-9 min） and 275 nm（9-33 min）. The column temperature was set at 25 ℃， and sample size was 10 μL. With baicalin as reference material， the relative correction factors （fk/s） of other five components were calculated by multi-point correction method and slope correction method; the retention time difference method was used to locate the chromatographic peaks; the calculation values obtained by above 2 QAMS were compared with measured values of external standard method. RESULTS： The linear range of 2，3，5，4′-tetrahydroxystilbene glucoside， baicalin， icariin， scutellarin， baicalein and wogonin were 0.053-2.12， 0.163-6.52， 0.059-2.36， 0.021 6-0.864， 0.03-1.2， 0.021-0.84 μg（r>0.999）， respectively. RSDs of precision， stability （12 h） and reproducibility tests were all lower than 3%. Average recoveries were 98.72%-99.82%（RSDs were 0.89%-1.24%，n=9）. Using baicalin as reference material， fk/s of multi-point correction method for 2，3，5，4′-tetrahydroxystilbene glucoside， icariin， scutellarin， baicalein and wogonin were 1.172， 0.528， 1.479， 1.820 and 2.534， respectively; fk/s of slope correction method were 1.234， 0.550， 1.559， 1.939， 2.664. RSDs of 6 components in 10 batches of Fuzheng guben granules by 3 methods were 0.29%-2.77%（n=10）， respectively. Pearson correlation coefficient was not lower than 0.999 9 （P<0.001） in measured values between QAMS and external standard method. CONCLUSIONS： QAMS method is established successfully for simultaneous determination of 6 components in Fuzheng guben granules.

KEYWORDS   QAMS; Fuzheng guben granules; 2，3，5，4′-tetrahydroxystilbene glucoside; Baicalin; Icariin; Scutellarin; Bai- calein; Wogonin; Content

中图分类号 R944.2+7;R927.2 文献标志码 A 文章编号 1001-0408（2021）02-0225-06

DOI 10.6039/j.issn.1001-0408.2021.02.17

扶正固本顆粒由黄芩、淫羊藿、何首乌等8味中药组成，具有益气养阴、凉血解毒的功效，可作为食管癌以及胃寒气阴两虚兼热毒症患者放、化疗的合并用药，其制剂标准被收载于国家药品监督管理局国家药品标准[WS-5250（B-0250）-2002]中。目前……