摘要
牛膝现行的质量标准难以有效控制其质量。本研究运用双外标结合校正因子,同时测定牛膝中牛膝皂苷C、牛膝皂苷D、β-蜕皮甾酮、25R-牛膝甾酮和25S-牛膝甾酮的含量。采用Agilent Poroshell 120 EC-C18(150 mm×4.6 mm,2.7μm)色谱柱,流动相为0.1%磷酸乙腈-0.1%磷酸水溶液梯度洗脱,流速0.8 mL·min^(-1),柱温35℃,进样量5μL,分析时间30 min。分别以β-蜕皮甾酮和牛膝皂苷D为对照,计算25R-牛膝甾酮、25S-牛膝甾酮和牛膝皂苷C的相对校正因子(relative correction factors,RCF)和相对保留时间(relative retention time,RRT),25R-牛膝甾酮、25S-牛膝甾酮和牛膝皂苷C的相对校正因子分别为1.116、1.056和0.8881,不同条件下RCF稳定性良好。分别使用外标法和双外标校准法对20批不同来源牛膝样品的含量进行计算,两种方法所得结果相对误差(relative error,RE)小于5%,表明建立的双外标校准法同时测定牛膝中5种成分准确可行。含量测定结果显示牛膝2种皂苷和3种甾酮的含量分别为0.597%~1.916%和0.044%~0.150%,皂苷含量约为甾酮的10倍,提示牛膝皂苷是牛膝药材质量的关键成分。该研究结果为牛膝药材提供了科学可行的整体质量评价方法。
Saponins and sterones are two main characteristic components in Achyranthis Bidentatae Radix.In order to control the quality of Achyranthis Bidentatae Radix more effectively,a high-performance liquid chromatography(HPLC)method was established by using double external standards calibration method(DESCM)for simultaneous determination of the contents of achyranthoside C,achyranthoside D,β-ecdysterone,25R-inokosterone and 25S-inokosterone in Achyranthis Bidentatae Radix.Chromatographic separation was achieved on an Agilent Poroshell 120 EC-C18(150 mm×4.6 mm,2.7μm)using 0.1%phosphoric acid in water and 0.1%phosphoric acid in acetonitrile as mobile phase.The flow rate was 0.8 mL·min^(-1)and the column temperature was set as 35℃,the injection volume was 5μL and the total analytical time was 30 min.β-Ecdysterone was used as the reference to calculate the relative correction factors(RCF)and relative retention time(RRT)of 25Rinokosterone and 25S-inokosterone,achyranthoside D was used for achyranthoside C.The RCFs of 25Rinokosterone,25S-inokosterone,and achyranthoside C were 1.116,1.056,and 0.8881,respectively.The double external standards calibration method(DESCM)and external standard method(ESM)were used to calculate the contents of five ingredients in Achyranthis Bidentatae Radix samples from different sources and the variation between the results was within acceptable limits(RE≤5%).The results showed that the contents of two saponins and three sterones of Achyranthis Bidentatae Radix were 0.597%-1.916%and 0.044%-0.150%respectively.The total content of saponins was about 10 times that of sterones.In conclusion,the established DESCM allowed simultaneous determination of five ingredients(achyranthoside C,achyranthoside D,β-ecdysterone,25R-inokosterone,and 25S-inokosterone)in Achyranthis Bidentatae Radix,providing a scientific and feasible overall quality evaluation method for Achyranthis Bidentatae Radix.
作者
吴冰筱
金武燮
张留记
谷丽华
李林楠
杨莉
王峥涛
WU Bing-xiao;KIM Moo-seob;ZHANG Liu-ji;GU Li-hua;LI Lin-nan;YANG Li;WANG Zheng-tao(The MOE Key Laboratory for Standardization of Chinese Medicines and the SATCM Key Laboratory for New Resources and Quality Evaluation of Chinese Medicines,Institute of Chinese Materia Medica,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Shanghai R&D Center for Standardization of Chinese Medicines,Shanghai 201203,China;Henan Engineering Center for Development of Genuine Medicinal Materials,Henan Academy of Traditional Chinese Medicine,Zhengzhou 450004,China)
出处
《药学学报》
CAS
CSCD
北大核心
2022年第6期1868-1873,共6页
Acta Pharmaceutica Sinica
基金
国家药典委员会标准基金资助项目(2018Z003)
上海市技术标准基金资助项目(21DZ2202100)
上海市中医药三年行动计划(ZY(2021-2023)-0215)。
关键词
牛膝
牛膝皂苷D
β-蜕皮甾酮
双外标校准
高效液相色谱
质量评价
Achyranthis Bidentatae Radix
achyranthoside D
β-ecdysterone
double external standards calibration
HPLC
quality evaluation
