摘要
目的:建立淫羊藿总黄酮提取物的高效液相色谱(HPLC)指纹图谱,并建立同时测定其中8种成分含量的方法。方法:采用HPLC法。色谱柱为ZORBAX Eclispse SB-C_(18),流动相为乙腈-水(梯度洗脱),流速为1.0 mL/min,柱温为30℃,检测波长为270nm,进样量为5μL。以淫羊藿苷峰为参照峰,绘制5批样品的HPLC指纹图谱,采用《中药色谱指纹图谱相似度评价系统(2004 A版)》进行相似度评价,确定共有峰。结果:5批样品的HPLC图谱中有22个共有峰,相似度均大于0.99,并指认淫羊藿属苷A、朝藿定A1、朝藿定A、朝藿定B、朝藿定C、淫羊藿苷、鼠李糖基淫羊藿次苷Ⅱ、宝藿苷Ⅰ等8个共有峰。上述8种成分进样量线性范围分别为0.019 5～0.292 5μg(r=0.999 9)、0.028 2～0.423 0μg(r=0.999 6)、0.050 5～0.757 5μg(r=0.999 9)、0.066 9～1.003 5μg(r=0.999 9)、0.089 6～1.344 0μg(r=0.999 9)、0.16～2.40μg(r=0.999 9)、0.026 8～0.402 0μg(r=0.999 8)、0.027 24～0.408 60μg(r=0.999 8);定量限分别为390.00、564.00、506.00、535.20、448.00、426.68、643.20、544.80 ng/mL,检测限分别为97.50、141.00、126.25、133.80、112.00、106.67、160.80、136.20 ng/mL;精密度、稳定性、重复性试验的RSD均小于3%;加样回收率分别为97.64%～103.79%(RSD=2.00%,n=6)、96.41%～99.10%(RSD=1.12%,n=6)、96.56%～103.56%(RSD=2.67%,n=6)、96.10%～99.57%(RSD=1.20%,n=6)、99.34%～104.18%(RSD=1.70%,n=6)、100.35%～105.37%(RSD=1.93%,n=6)、98.76%～102.83%(RSD=1.60%,n=6)、96.30%～101.10%(RSD=1.80%,n=6)。结论:所建HPLC指纹图谱可为淫羊藿总黄酮提取物的质量控制提供参考;所建含量测定方法简便、准确、重复性好,可用于同时测定淫羊藿总黄酮提取物中8种成分的含量。
OBJECTIVE:To establish HPLC fingerprint of total flavonoid extract from Epimedium brevicornu,and to determine the contents of 8 components.METHODS:HPLC method was adopted.The separation was carried out on ZORBAX Eclispse SB-C18 column with mobile phase consisted of acetonitrile-water(gradient elution)at the flow rate of 1.0 mL/min.The column temperature was 30℃,and detection wavelength was set at 270 nm.The sample size was 5μL.Using icariin peak as reference,HPLC fingerprints of 5 batches of sample were drawn.The similarity evaluation was performed by using Similarity Evaluation System of Chromatogram Fingerprint of TCM(2004 A edition)to determine common peaks.RESULTS:There were 22 common peaks in HPLC chromatogram of 5 batches of sample,with the similarity above 0.99.Eight peaks were identified,such as icariin A,epimedin A1,epimedin A,epimedin B,epimedin C,icariin,rhamnosy licarisidⅡand baicin I.The linear range of eight components were 0.019 5-0.292 5μg(r=0.999 9),0.028 2-0.423 0μg(r=0.999 6),0.050 5-0.757 5μg(r=0.999 9),0.066 9-1.003 5μg(r=0.999 9),0.089 6-1.344 0μg(r=0.999 9),0.16-2.40μg(r=0.999 9),0.026 8-0.402 0μg(r=0.999 8),0.027 24-0.408 60μg(r=0.999 8),respectively.The limits of quantitation were 390.00,564.00,506.00,535.20,448.00,426.68,643.20,544.80 ng/mL;the limits of detection were 97.50,141.00,126.25,133.80,112.00,106.67,160.80,136.20 ng/mL,respectively.RSD of precision,stability and reproducibility tests were all lower than 3%.The recoveries were 97.64%-103.79%(RSD=2.00%,n=6),96.41%-99.10%(RSD=1.12%,n=6),96.56%-103.56%(RSD=2.67%,n=6),96.10%-99.57%(RSD=1.20%,n=6),99.34%-104.18%(RSD=1.70%,n=6),100.35%-105.37%(RSD=1.93%,n=6),98.76%-102.83%(RSD=1.60%,n=6),96.30%-101.10%(RSD=1.80%,n=6),respectively.CONCLUSIONS:Established of HPLC fingerprint can provide reference for quality control of total flavonoid extract from E.brevicornu.Established method is simple,accurate and reproducible,and can be used for simultaneous determination of 8 components in total flavonoid extract from E.brevicornu.
作者
牛晓静
鲁静
孙广科
段晓颖
徐立然
NIU Xiaojing;LU Jing;SUN Guangke;DUAN Xiaoying;XU Liran(The First Affiliated Hospital of Henan University of TCM,Zhengzhou 450008,China)
出处
《中国药房》
CAS
北大核心
2018年第24期3376-3380,共5页
China Pharmacy
基金
河南省基础与前沿技术研究项目(No.152300410105)
河南中医学院科研苗圃工程项目(No.MP2015-03)
