大黄提取物中五种蒽醌类活性成分的同步检测方法  

Simultaneous Detection Method of Five Anthraquinone Active Components in Rhubarb Extract

作　　者：胡晓钰 刘霁 吴琳 王祥艳 白鸿 郭怀兰[1] 樊柏林[2] 刘春霞[2] HU Xiao-yu;LIU Ji;WU Lin;WANG Xiang-yan;BAI Hong;GUO Huai-lan;FAN Bo-lin;LIU Chun-xia(College of Pub-lic Health,Hubei University of Medicine,Shiyan,Hubei 442000;Hubei Provincial Key Laboratory for Applied Toxicology,Hubei Provincial Centers for Disease Control and Prevention,Wuhan,Hubei 430000;State Administration for Market Regu-lation Food Review Center,Beijing 100000,China)

机构地区：[1]湖北医药学院公共卫生与健康学院,湖北十堰442000 [2]湖北省疾病预防控制中心应用毒理湖北省重点实验室,湖北武汉430000 [3]国家市场监督管理总局食品审评中心,北京100000

出　　处：《湖北医药学院学报》2023年第4期349-354,共6页Journal of Hubei University of Medicine

基　　金：国家中医药行业科研专项(201307012);湖北省公共卫生领军人才培养计划(鄂卫通2021第73号);湖北省重点研发计划(2021ACB003)。

摘　　要：目的:建立高效液相色谱-紫外分析法(HPLC-UV)同步检测大黄提取物中五种蒽醌类活性成分。方法:优化蒽醌类活性成分同步检测方法的溶剂、色谱分离条件,并从标准曲线线性、精密度、准确度、回收率、稳定性及重现性等方面对方法进行了验证,采用验证的方法检测了大黄提取物中五种蒽醌类成分的含量。结果:分别采用氯仿、二甲基亚砜、甲醇作为不同对照品储备液溶剂,采用甲醇∶水(50∶50,V∶V)作为对照品工作溶液溶剂,采用甲醇配制样品溶液。最佳色谱分离条件:采用Symmetry C_(18)(5μm,4.6 mm×250 mm)色谱柱,采用二元泵等度洗脱方式,流动相A为含0.2%甲酸的水溶液,流动相B为乙腈(A∶B体积比为15∶85),流速1.30mL/min,柱温30℃,检测波长254 nm,进样体积20.0μL。方法曲线拟合度好,R2均大于0.9999;6份平行样品芦荟大黄素、大黄素、大黄酸、大黄酚和大黄素甲醚的RSD分别为1.10%、1.05%、0.74%、0.84%和0.82%,回收率为94.54%~98.70%;室温放置22 h以及4℃条件下储存20 d基本稳定,自动进样器温度下存储24 h重新进样重现性良好。采用建立的方法测得大黄冻干粉样品中芦荟大黄素、大黄素、大黄酸、大黄酚和大黄素甲醚的含量分别为0.900、0.639、3.506、0.780、0.304 mg/g。结论:本研究建立了可同步检测芦荟大黄素、大黄素、大黄酸、大黄酚和大黄素甲醚等五种蒽醌类活性成分的高效液相色谱-紫外分析法(HPLC-UV),经验证其具有良好的线性关系、准确性、精密度、稳定性和重现性。Objective To establish a high performance liquid chromatography-ultraviolet(HPLC-UV)method for simultaneous determination of five anthraquinone active components in rhubarb extract.Methods In this study,the solvent and chromatographic separation conditions of the simultaneous detection method of anthraquinone active ingredients were optimized.The method was verified from the aspects of standard curve linearity,precision,accuracy,recovery rate,stability and reproducibility.The content of five anthraquinones in rhubarb extract was detected by the validated method.Results Chloroform,dimethyl sulfoxide and methanol were used as solvents for stock solutions of different reference substances,methanol∶water(50:50,V∶V)was used as reference working solution solvent,and methanol was used to prepare sample solution.The optimum chromatographic separation conditions were as follows:Symmetry C_(18)(5μm,4.6 mm×250 mm)chromatographic column was used,binary pump was used for isocratic elution,mobile phase A was an aqueous solution containing 0.2%formic acid,mobile phase B was acetonitrile(mobile phase A:mobile phase B volume ratio was 15∶85),the flow rate was 1.30 mL/min,the column temperature was 30℃,the detection wavelength was 254 nm,and the injection volume was 20.0μL.The curve fitting of the method was good,and R2 was greater than 0.9999.The relative standard deviations(RSDs)of aloe-emodin,emodin,rhein,chrysophanol and physcion in 6 parallel samples were 1.10%,1.05%,0.74%,0.84%and 082%,respectively.The recovery rate was 94.54%~98.70%.It was basically stable after storage at room temperature for 22 h and 4℃for 20 d.The reproducibility of reinjection was good after storage for 24 h at the temperature of automatic sampler.The established method was used to detect the contents of aloe-emodin,emodin,rhein,chrysophanol and physcion in rhubarb freeze-dried powder samples with the results of 0.900,0.639,3.506,0.780,0.304 mg/g respectively.Conclusion A high performance liquid chromatography-ultraviolet analysis(HPLC-UV)m

关 键 词：芦荟大黄素 大黄素 大黄酸 大黄酚 大黄素甲醚 HPLC-UV 

分 类 号：R284.2[医药卫生—中药学]