HPLC法同时测定杜仲叶中8种成分的含量
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篇名: HPLC法同时测定杜仲叶中8种成分的含量
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摘要: 目的:建立同时测定杜仲叶中桃叶珊瑚苷、京尼平苷酸、儿茶素、绿原酸、车叶草苷、芦丁、异槲皮苷和紫云英苷等8种成分的含量测定方法。方法:采用高效液相色谱法。色谱柱为Agilent ZORBAX SB-C18,流动相为乙腈-0.1%磷酸溶液(梯度洗脱),流速为1.0 mL/min,检测波长为203 nm(桃叶珊瑚苷、儿茶素)、239 nm(京尼平苷酸、车叶草苷)、220 nm(绿原酸)、354 nm(芦丁、异槲皮苷)、266 nm(紫云英苷),进样量为5 μL。结果:桃叶珊瑚苷、京尼平苷酸、儿茶素、绿原酸、车叶草苷、芦丁、异槲皮苷和紫云英苷的线性范围分别为0.812~6.090 μg(r=0.999 3)、0.438~3.285 μg(r=0.999 2)、0.045~0.336 μg(r=0.999 2)、0.882~6.615 μg(r=0.999 3)、0.097~0.726 μg(r=0.999 1)、0.064~0.483 μg(r=0.999 3)、0.048~0.360 μg(r=0.999 1)、0.014~0.108 μg(r=0.999 7);精密度、重复性和稳定性试验的RSD均小于3.5%(n=6);平均加样回收率分别为101.60%、103.06%、99.77%、96.93%、98.17%、96.75%、98.97%、99.60%,RSD分别为1.42%、2.65%、2.78%、2.05%、2.26%、0.93%、2.79%、3.08%(n=6)。不同采集时间、不同种植品种的12批杜仲叶样品中桃叶珊瑚苷、京尼平苷酸、儿茶素、绿原酸、车叶草苷、芦丁、异槲皮苷和紫云英苷的含量范围分别为10.903~17.245、5.578~7.892、0.198~0.440、13.890~19.782、1.008~1.547、1.102~2.396、0.267~0.701、0.150~0.412 mg/g,含量波动较大;杜仲皮样品中桃叶珊瑚苷、京尼平苷酸、儿茶素、绿原酸、芦丁、松脂醇二葡萄糖苷的含量分别为0.299、0.123、0.580、0.112、0.026、1.961 mg/g。结论:本方法操作简便、重复性好、准确度高,可用于评价杜仲叶的质量;不同采集时间、不同种植品种的杜仲的不同药用部位(皮、叶)中的成分组成及含量存在明显的差异。
ABSTRACT: OBJECTIVE: To establish content determination method for simultaneously determining aucubin,geniposidic acid,catechin,chlorogenic acid,asperuloside,rutin,isoquercitrin and astragalin in Eucommia ulmoides leaves. METHODS:HPLC method was adopted. The determination was performed on Agilent ZORBAX SB-C18 column with the mobile phase consisted of acetonitrile-0.1% phosphoric acid(gradient elution) at the flow rate of 1.0 mL/min. The detection wavelength was set at 203 nm for aucubin and catechin,239 nm for geniposidic acid and asperuloside,220 nm for chlorogenic acid,354 nm for rutin and isoquercitrin,and 266 nm for astragalin. The sample size was 5 μL. RESULTS: The linear range of aucubin, geniposidic acid, catechin, chlorogenic acid, asperuloside, rutin, isoquercitrin and astragalin were 0.812-6.090 μg(r=0.999 3),0.438-3.285 μg(r=0.999 2),0.045-0.336 μg(r=0.999 2),0.882-6.615 μg(r=0.999 3),0.097-0.726 μg(r=0.999 1),0.064-0.483 μg(r=0.999 3),0.048-0.360 μg(r=0.999 1) and 0.014-0.108 μg(r=0.999 7),respectively. RSDs of precision, stability and reproducibility tests were all lower than 3.5%(n=6). The average recovery rates were 101.60%,103.06%,99.77%,96.93%,98.17%,96.75%,98.97% and 99.60%,with RSDs of 1.42%,2.65%,2.78%,2.05%,2.26%,0.93%,2.79% and 3.08%,respectively(n=6). The contents of aucubin, geniposide, catechin, chlorogenic acid, plantain, rutin, isoquercetin and astragaloside in 12 batches of E. ulmoides leaves from different collection time and planting varieties were 10.903-17.245, 5.578-7.892, 0.198-0.440, 13.890-19.782, 1.008-1.547, 1.102-2.396, 0.267-0.701, 0.150-0.412 mg/g, respectively. The content fluctuated greatly. The contents of aucubin, geniposide, catechin, chlorogenic acid, rutin and pinoresinol diglucoside in cortex of E. ulmoides were 0.299, 0.123, 0.580, 0.112, 0.026,1.961 mg/g, respectively. CONCLUSIONS: The method is simple, reproducible and accurate. It can be used to evaluate the quality of E. ulmoides leaves. There are obvious differences in composition and content of components in different medicinal parts (cortex, leaves) of E. ulmoides.
期刊: 2019年第30卷第24期
作者: 张留记,李宁,屠万倩,李向阳,王昌华,邵战波
AUTHORS: ZHANG Liuji,LI Ning,TU Wanqian,LI Xiangyang,WANG Changhua,SHAO Zhanbo
关键字: 杜仲叶;高效液相色谱法;桃叶珊瑚苷;京尼平苷酸;儿茶素;绿原酸;车叶草苷;芦丁;异槲皮苷;紫云英苷;含量测定
KEYWORDS: Eucommia ulmoides leaves;HPLC;Aucubin;Geniposidic acid;Catechin;Chlorogenic acid;Asperuloside;Rutin; Isoquercitrin; Astragalin; Content determination
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