紫草药材的HPLC指纹图谱研究及其β-乙酰氧基异戊酰阿卡宁的含量测定

目的:研究紫草药材的高效液相色谱(HPLC)指纹图谱,并测定其中β-乙酰氧基异戊酰阿卡宁的含量。方法:色谱柱为ACE C18(250 mm×4.6 mm,5μm),流动相为乙腈-0.1%甲酸溶液(70∶30,V/V),检测波长为516 nm,流速为1.0 ml/min。结果:所得紫草药材的HPLC指纹图谱分离度、精密度、重复性和稳定性均良好;10批紫草药材指纹图谱的相似度均>0.950。β-乙酰氧基异戊酰阿卡宁的进样量在0.3⁹.0μg范围内与其峰面积积分值呈良好的线性关系(r=0.999 9);平均加样回收率为101.40%,RSD=4.08%(n=6);不同产地紫草药材中β-乙酰氧基异戊酰阿卡宁的含量差异较大。结论:β-乙酰氧基异戊酰阿卡宁可作为紫草药材含量测定和指纹图谱研究的指标性成分。所建方法可以为2015年版《中国药典》紫草药材标准的修订提供一定的依据。

Analysis of HPLC Fingerprints and Content Determination of β-acetoxyisovaleryalkannin in Arnebiae Radix

OBJECTIVE： To study HPLC fingerprints of Arnebiae Radix, and to determine the contents of β-acetoxyisovaleryal- kannin. METHODS： The determination was performed on ACE C18 column （250 mm×4.6 mm, 5μm） with mobile phase consisted of acetonitrile-0.1% formic acid （70 ： 30, V/V） at the flow rate of 1.0 ml/min. The detection wavelength was set at 516 nm. RESULTS： HPLC fingerprints of Arnebiae Radix showed good separation, precision, reproducibility and stability; the similarity of the fingerprints in 10 batches of Amebiae Radix was more than 0.950. The linear range of fl-acetoxyisovaleryalkannin were 0.3-9.0 μg （r=0.999 9） with an average recovery of 101.40% （RSD=4.08%, n=6）. The contents of β-acetoxyisovaleryalkannin from different habitats were significantly different from each other. CONCLUSIONS：β-acetoxyisovaleryalkannin can be good chemical marker for the content determination and fingerprint study of Arnebiae Radix. It can provide basis for the verification standards of Amebiae Radix in Chinese Pharmacopeia （2015 edition）.