HPLC法测定并比较茵陈蒿煎剂与其配方颗粒中有效成分的含量
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篇名: HPLC法测定并比较茵陈蒿煎剂与其配方颗粒中有效成分的含量
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摘要: 目的:建立测定茵陈蒿煎剂与其配方颗粒中大黄素、大黄酸、芦荟大黄素、大黄酚、大黄素甲醚、栀子苷、绿原酸含量的方法,比较茵陈蒿煎剂与其配方颗粒中上述成分含量的差异。方法:采用高效液相色谱法(HPLC)测定煎剂中大黄素、大黄酸、芦荟大黄素、大黄酚、大黄素甲醚的含量:色谱柱为Ecosil C18,流动相为乙腈-0.5%磷酸(梯度洗脱),流速为1 ml/min,检测波长为254 nm,柱温为35 ℃,进样量为20 μl;采用HPLC法测定煎剂中栀子苷的含量:色谱柱为Ecosil C18,流动相为乙腈-水(13 ∶ 87,V/V),流速为1 ml/min,检测波长为238 nm,柱温为35 ℃,进样量为20 μl;采用HPLC法测定煎剂中绿原酸的含量:色谱柱为Ecosil C18,流动相为乙腈-0.05%磷酸(10 ∶ 90,V/V),流速为1 ml/min,检测波长为327 nm,柱温为35 ℃,进样量为20 μl。结果:大黄素、大黄酸、芦荟大黄素、大黄酚、大黄素甲醚、栀子苷、绿原酸检测质量浓度线性范围分别为2~20 μg/ml(r=0.996 5)、5.2~52.0 μg/ml(r=0.998 5)、2.6~26.0 μg/ml(r=0.999 9)、1.0~10.4 μg/ml(r=0.999 9)、1.0~10.0 μg/ml(r=0.999 8)、20~200 μg/ml(r=0.999 9)、20~200 μg/ml(r=0.999 9);精密度、稳定性、重复性试验的RSD<3%;回收率分别为 91.1%~96.9%(RSD=2.0%,n=6)、93.9%~96.1%(RSD=0.8%,n=6)、90.9%~93.4%(RSD=1.2%,n=6)、88.5%~92.7%(RSD=1.8%,n=6)、82.1%~87.9%(RSD=2.5%,n=6)、100.4%~102.0%(RSD=0.7%,n=6)、101.1%~102.2%(RSD=0.4%,n=6)。结论:该方法操作简便、稳定、重复性好,可用于茵陈蒿煎剂与其配方颗粒中有效成分含量的同时测定。茵陈蒿配方颗粒有效成分含量明显高于其煎剂。
ABSTRACT: OBJECTIVE: To establish a method for the contents determination of emodin, rhein, aloe-emodin, chrysophanol, physcion, geniposide, chlorogenic acid in Yinchenhao decoction and its dispensing granule, and to compare the difference among the above-mentioned ingredients. METHODS: HPLC was adopted to determine the contents of emodin, rhein, aloe-emodin, chrysophanol and physcion: the column was Ecosil C18 with mobile phase of acetonitrile-0.5% phosphoric acid (gradient elution) at a flow rate of 1 ml/min, the detection wavelength was 254 nm, column temperature was 35 ℃, and injection volume was 20 μl. When determining the content of geniposide by HPLC, the column was Ecosil C18 with mobile phase of acetonitrile - water (13 ∶ 87,V/V) at a flow rate of 1 ml/min, the detection wavelength was 238 nm, column temperature was 35 ℃, and injection volume was 20 μl. When determining the content of chlorogenic acid by HPLC, the column was Ecosil C18 with mobile phase of acetonitrile - 0.5% phosphoric acid (10 ∶ 90,V/V) at a flow rate of 1 ml/min, the detection wavelength was 327 nm, column temperature was 35 ℃, and injection volume was 20 μl. RESULTS: The linear range was 2-20 μg/ml for emodin (r=0.996 5), 5.2-52.0 μg/ml for rhein (r=0.998 5), 2.6-26.0 μg/ml for aloe-emodin (r=0.999 9), 1.0-10.4 μg/ml for chrysophanol (r=0.999 9), 1.0-10.0 μg/ml for physcion (r=0.999 8), 20-200 μg/ml for geniposide (r=0.999 9), 20-200 μg/ml for chlorogenic acid (r=0.999 9); RSDs of precision, stability and reproducibility tests were lower than 3%; recoveries were  91.1%-96.9%(RSD=2.0%,n=6)for emodin、93.9%-96.1%(RSD=0.8%,n=6)for rhein、90.9%-93.4%(RSD=1.2%,n=6)for aloe-emodin、88.5%-92.7%(RSD=1.8%,n=6)for chrysophanol、82.1%-87.9%(RSD=2.5%,n=6)for physcion,100.4%-102.0%(RSD=0.7%,n=6)for geniposide、101.1%-102.2%(RSD=0.4%,n=6) for chlorogenic acid. CONCLUSIONS: The method is simple, stable and reproducible, and can be used for the simultaneous contents determination of active ingredients in Yinchenhao decoction and its dispensing granule. The contents of active ingredients in Yinchenhao dispensing granule are obviously higher than those in its decoction.
期刊: 2016年第27卷第15期
作者: 林伟豪,陈连剑,高崇凯,吴婵娟
AUTHORS: LIN Weihao,CHEN Lianjian,GAO Chongkai,WU Chanjuan
关键字: 茵陈蒿煎剂;配方颗粒;高效液相色谱法
KEYWORDS: Yinchenhao decoction;Dispensing granule;HPLC
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