UPLC法同时测定复方龙胆碳酸氢钠片中10种大黄蒽醌类成分及土大黄苷的含量

Simultaneous determination of 10 rhubarb anthraquinones and rhaponticin in Compound gentian and sodium bicarbonate tablets by UPLC

目的 建立同时测定复方龙胆碳酸氢钠片中10种大黄蒽醌类成分和伪品特征成分土大黄苷含量的方法,用于含大黄复方制剂的质量评价。方法 采用超高效液相色谱(UPLC)法对8个生产企业40批次复方龙胆碳酸氢钠片中10种大黄蒽醌类成分(芦荟大黄素-8-O-葡萄糖苷、大黄酸-8-O-β-D-葡萄糖苷、大黄酚-8-O-β-D-葡萄糖苷、大黄素-8-O-葡萄糖苷、大黄素甲醚-8-O-β-D-葡萄糖苷、芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚)和土大黄苷的含量进行测定。色谱柱为Agilent Eclipse Plus C18,流动相为乙腈-0.1%磷酸溶液(梯度洗脱),流速为0.3 mL/min,柱温为30℃,进样量为5μL。结合主成分分析与聚类分析对含量测定结果进行综合分析,并对不同企业样品进行质量评价。结果 上述11种成分在各自检测质量浓度范围内线性关系均良好(r≥0.999 3),精密度、重复性、稳定性试验的RSD均小于3%(n均为6),平均加样回收率为96.82%~98.92%(RSD≤1.74%,n=6);含量分别为0.011 7~0.252 0、0~0.323 3、0.131 3~1.236 6、0.081 1~1.056 2、0.015 2~0.189 8、0.001 8~0.152 3、0~0.255 2、0.001 9~0.223 4、0.054 3~0.303 0、0.022 7~0.172 2、0~2.835 9 mg/g。3个主成分的累计方差贡献率为95.533%;40批次样品可分为4类:a和d企业的样品分别自成一类,f、b、g、e企业的样品为一类,c、h企业的样品为一类。8家生产企业样品中的大黄蒽醌类成分含量存在较大差异,且有1家企业的样品检出了土大黄苷。结论 所建UPLC法稳定、可靠,可用于复方龙胆碳酸氢钠片中10种大黄蒽醌类成分及土大黄苷的含量测定。

OBJECTIVE To establish a method for the simultaneous determination of 10 rhubarb anthraquinones in Compound gentian sodium bicarbonate tablets and the content of rhaponticin,which are the characteristic components of artifacts,and to use the method to evaluate the quality of compound preparation containing Rheum officinale.METHODS The ultra-performance liquid chromatography(UPLC) method was adopted to determine the contents of 10 rhubarb anthraquinones(aloe-emodin-8-O-glucoside,rheinic acid-8-O-β-D-glucoside,emodin-8-O-glucoside,chrysophanol-8-O-β-D-glucoside,emodin monomethyl ether-8-O-β-D-glucoside,aloe-emodin,rheinic acid,emodin,chrysophanol,emodin monomethyl ether) and rhaponticin in 40 batches of Compound gentian sodium bicarbonate tablets from 8 manufacturers.The determination was performed on the Agilent Eclipse Plus C18 column with a mobile phase consisted of acetonitrile-0.1% phosphoric acid solution(gradient elution) at a flow rate of 0.3 mL/min;the column temperature was set at 30 ℃,and the injection volume was 5 μL.Combining principal component analysis and cluster analysis to synthesize the results of content determination,the quality of samples from different manufacturers was evaluated.RESULTS All of above 11 components showed favorable linear relationships with peak areas in their respective mass concentration ranges(r≥0.999 3),with RSDs of precision,repeatability and stability less than 3%(n=6);average sample recoveries ranged 96.82%-98.92%(RSD≤1.74%,n=6);their contents were 0.011 7-0.252 0,0-0.323 3,0.131 3-1.236 6,0.081 1-1.056 2,0.015 2-0.189 8,0.001 8-0.152 3,0-0.255 2,0.001 9-0.223 4,0.054 3-0.303 0,0.022 7-0.172 2,0-2.835 9 mg/g,respectively.The cumulative variance contribution of the first three principal components was 95.533%;the 40 batches of samples can be clustered into 4 categories:samples from enterprises a and d were in a category of their own,samples from enterprises f,b,g and e were in a category,and samples from enterprises c and h were in a category.There were large differences in the content of rhubarb anthraquinone in the samples from 8 manufacturers,and rhaponticin was only detected in the sample from one enterprise.CONCLUSIONS Established UPLC method is stable and reliable;it can be used for the content determination of 10 rhubarb anthraquinones and rhaponticin in Compound gentian sodium bicarbonate tablets.