摘要
目的:建立同时测定参芪延肾颗粒中淫羊藿药材中朝藿定A1、朝藿定A、朝藿定B、朝藿定C、淫羊藿苷及宝藿苷Ⅰ6个黄酮类成分含量的方法。方法:采用高效液相色谱(HPLC)法,色谱柱为Waters Symmetry C18,流动相为乙腈-水(梯度洗脱),流速为1.0 mL/min,柱温为25℃,检测波长为270 nm,进样量为10μL。在外标法的基础上分别采用多点校正法及斜率校正法建立各成分与淫羊藿苷(参照物)的相对校正因子(fk/s以计算各成分含量;比较基于HPLC法的外标法与多点校正法及斜率校正法所得4批参芪延肾颗粒样品中6个黄酮类成分含量的差异,验证一测多评法的可行性及准确性。结果:朝藿定A,、朝藿定A、朝藿定B、朝藿定C、淫羊藿苷及宝藿苷Ⅰ检测质量浓度线性范围分别为2.03~50.80、4.34~108.60、2.26~56.40、4.14~103.60、4.24~106.00、1.78~44.60μg/mL(r均为0.999 5);检测限分别为65.80、71.49、74.26、68.79、70.56、86.09 ng/mL;定量限分别为196.62、213.63、223.72、208.46、215.96,255.88 ng/mL;精密度、稳定性(24 h)、重复性试验的RSD<2%(n=6);平均回收率为96.03%~99.04%(RSD为0.65%~1.04%,n=6);采用多点校正法,朝藿定A1、朝藿定A、朝藿定B、朝藿定C、宝藿苷Ⅰ的fk/s分别为0.837、0.818、0.845、0.831、1.387;采用斜率校正法,朝藿定A1、朝藿定A、朝藿定B、朝藿定C、宝藿苷Ⅰ的fk/s分别为0.835、0.815、0.851、0.829、1.419。采用一测多评两种校正法与外标法所得含量结果比较,无显著性差异(P>0.05)。结论:建立的HPLC-一测多评法结果准确,可应用于参芪延肾颗粒中淫羊藿药材中朝藿定A,、朝藿定A、朝藿定B、朝藿定C、淫羊藿苷及宝藿苷Ⅰ6个黄酮类成分的质量控制。
OBJECTIVE:To establish a HPLC method for simultaneous determination of 6 flavonoids in Epimedium brevicornu from Shenqi yanshen granules, such as epimedin A1, epimedin A, epimedin B, epimedin C, icariin and baohuoside Ⅰ. METHODS:HPLC method was adopted. The determination was performed on Waters Symmetry C18 column with mobile phase consisted of acetonitrile-water(gradient elution)at the flow rate of 1.0 mL/min;the column temperature was 25 ℃,and detection wavelength was 270 nm. The sample size was 10 μ L. Relative correction factors (fk/s) of each component to icariin (reference substance)were established by multi-point correction method and slope correction method on the basis of external standard method to calculate the contents of each component. The contents of 6 flavonoids in 4 batches of Shenqi yanshen granules determined by HPLC external standard method were compared with by multi-point correction method and slope correction method. Feasibility and accuracy of quantitative analysis of multi-components by single-marker(QAMS)were validated. RESULTS:The linear range of epimedin A1,epimedin A,epimedin B,epimedin C,icariin and baohuoside Ⅰ were 2.03-50.80 μg/mL(r=0.999 5),4.34-108.60 μ g/mL(r=0.999 5),2.26-56.40 μ g/mL(r=0.999 5),4.14-103.60 μ g/mL(r=0.999 5),4.24-106.00 μ g/mL(r=0.999 5), 1.78-44.60 μ g/mL(r=0.999 5),respectively,the limits of detection were 65.80,71.49,74.26,68.79,70.56,86.09 ng/mL, respectively;the limits of quantification were 196.62,213.63,223.72,208.46,215.96,255.88 ng/mL,respectively;RSDs of precision, stability (24 h), reproducibility tests were less than 2%(n=6), respectively. The average recoveries were 96.03%-99.04%(RSDs were 0.65%-1.04%, n=6). By multi-point correction method,fk/s of epimedin A1,epimedin A,epimedin B,epimedin C and baohuoside Ⅰ were 0.837, 0.818,0.845,0.831,1.387,respectively;by slope correction method,fk/s of them were 0.835,0.815,0.851,0.829,1.419, respectively. There was no significant difference in content determination results between two correction methods of QAMS and external standard (P>0.05). CONCLUSIONS:Established HPLC- QAMS method is accurate and suitable for the quality control of epimedin A1,epimedin A,epimedin B, epimedin C,icariin and baohuoside Ⅰ in E. brevicornu from Shenqi yanshen granules.
作者
沈洁
王琴
熊维建
徐冲
SHEN Jie;WANG Qin;XIONG Weijian;XU Chong(Dept. of Pharmacy,Chongqing Hospital of TCM,Chongqing 400021,China)
出处
《中国药房》
CAS
北大核心
2019年第17期2327-2331,共5页
China Pharmacy
基金
国家自然科学基金资助项目（No.81603611）
重庆市社会民生科技创新专项项目（No.cstc2016shms-zd10001）
重庆市科研院所绩效激励引导专项项目（No.cstc2017jxjl130025）
重庆市卫生和计划生育委员会中医药科技项目（No.ZY201702003）
