摘要
目的:建立同时测定六神丸中6个蟾蜍二烯内酯类成分含量的一测多评法(QAMS),并验证其准确性和可行性。方法:采用高效液相色谱法,使用Waters Xbridge C18(250 mm×4.6 mm,5μm)色谱柱,以乙腈-0.2%磷酸水溶液为流动相,梯度洗脱,流速1 mL·min-1,柱温30℃,检测波长296 nm。以华蟾酥毒基为内参物,建立其与远华蟾毒精、蟾毒它灵、华蟾毒它灵、蟾毒灵、脂蟾毒配基5个成分的相对校正因子(RCF),并进行含量计算,实现一测多评;同时采用外标法(ESM)测定六神丸中蟾酥的6个成分的含量,比较一测多评法计算值与外标法实测值的差异。结果:在一定线性范围内,华蟾酥毒基与远华蟾毒精、蟾毒它灵、华蟾毒它灵、蟾毒灵、脂蟾毒配基的RCF值分别为0.897、0.965、1.023、0.844、0.964,且在不同实验条件下重现性良好(RSD <5.0%),一测多评法测得15批样品中华蟾酥毒基、远华蟾毒精、蟾毒它灵、华蟾毒它灵、蟾毒灵、脂蟾毒配基的含量范围分别为2.26~9.27 mg·g-1、0.95~2.06 mg·g-1、1.12~2.50 mg·g-1、1.83~3.48 mg·g-1、1.10~2.81mg·g-1、0.90~3.65 mg·g-1,与外标法实测值无显著性差异。结论:所建立的同时测定六神丸中的6个蟾蜍二烯内酯类化合物的一测多评法准确可靠,可用于六神丸中多个成分的定量分析质量评价。
Objective:To determine six bufadienolides in Liushen pills simultaneously with a quantitative analysis of multi-components by single-marker(QAMS)and accuracy and validate feasibility of the method.Methods:A Waters Xbridge C18(250 mm×4.6 mm,5 μm)column was used in the high performance liquid chromatography(HPLC)assay.Acetonitrile-0.2% phosphoric acid aqueous solution with gradient elution was employed as the mobile phase.The flow rate was 1 mL·min-1,the column temperature was maintained at 30 ℃ and the detection wavelength was 296 nm.Cinobufagin was used as the internal reference substance to calculate the relative correction factors(RCFs)of elocinobufagin,bufotalin,cinobufotalin,bufalin and resibufogenin and evaluate their contents accordingly.Moreover,the external standard method(ESM)was used to quantify the six components in Liushen pills.The results of the two methods were compared.Results:In certain linear range,RCFs of telocinobufagin,bufotalin,cinobufotalin,bufalin and resibufogenin with reference to cinobufagin were 0.897,0.965,1.023,0.844 and 0.964 respectively.The results were repeatable under different conditions with RSDs less than 5%.The content ranges of cinobufagin,telocinobufagin,bufotalin,cinobufotalin,bufalin and resibufogenin by the QAMS method were 2.26-9.27 mg·g-1,0.95~2.06 mg·g-1,1.12~2.50 mg·g-1,1.83~3.48 mg·g-1,1.10~2.81 mg·g-1,0.90~3.65 mg·g-1.Significant difference was not found between the the multi-evaluation method and the external standard method.Conclusion:The established QAMS can quantify six bufadienolides in Liushen pills accurately and reliably.This developed method can be used to determine multiple components in Liushen pills.
作者
丁梦磊
张雯
张越
李俊松
李全
王恒斌
李文
王洪兰
狄留庆
DING Meng-lei;ZHANG Wen;ZHANG Yue;LI Jun-song;LI Quan;WANG Heng-bin;LI Wen;WANG Hong-lan;DI Liu-qing(Nanjing University of Chinese Medicine,Nanjing 210023,China;Jiangsu Engineering Research Center for Efficient Delivery System of TCM,Nanjing 210023,China;Leiyunshang Pharmaceutical Co.Limited,Suzhou 215003,China;The Affiliated Nanjing Hospital of Nanjing University of Chinese Medicine,Nanjing,210001,China)
出处
《药物分析杂志》
CAS
CSCD
北大核心
2019年第11期2003-2009,共7页
Chinese Journal of Pharmaceutical Analysis
基金
国家中医药管理局中药标准化项目(ZYBZH-C-JS-30)
江苏省科技成果转化专项资金项目(BA2016104)
江苏省产学研合作项目(FZ20140109)
