用HPLC法测定不同地区三种苍耳子中绿原酸的含量

Determination of chlorogenic acid in three species of cocklebur fruits from different regions of China by HPLC method

目的：建立测定中药苍耳子中活性成分绿原酸的HPLC法，考察国内不同产地3种苍耳子中绿原酸的含量。方法：采用Zorbax SB-C18色谱柱（250mm×4．6mm，5μm），流动相为甲醇和0．5％磷酸水溶液，梯度洗脱，检测波长为327nm，流速为0．8mL／min，柱温为25℃。结果：绿原酸回归方程为Y=2．1357X-120，r=0．9998（n=5），线性范围为200～1000ng，平均回收率为（98．58±1．16）％（n=9）。苍耳（Xanthium sibiricum）11个样品中的绿原酸含量差异较大，但普遍高于蒙古苍耳（X．mongolicum）和近无刺苍耳（X．sibiricum var．subinerrne）样品。海南定安苍耳绿原酸含量最高，达（0．969±0．010）％。结论：本研究建立的HPLC法简便、准确、专属性强，为苍耳子质量的评价提供了可靠依据。国产苍耳子绿原酸含量的种问差异和种内变异均较明显，海南定安苍耳可以作为苍耳子药材的优良种质资源。

Objective： To establish a HPLC method for the determination of chlorogenic acid in cocklebur fruits, and to investigate the content of chlorogenic acid in three species of cocklebur fruits collected from different regions of China. Methods： The determination was performed on Zorbax SB-C18 column （250 mm× 4.6 mm,5 μm） with the temperature set at 25 ℃. The mobile phase consisted of methanol-0.5 % phosphoric acid solution at a flow rate of 0.8 mL/min. The detection wavelength was 327 nm. Results： The regression equation of chlorogenic acid was Y=2. 135 7 X--120（r=0. 999 8,n=5）. The linear range of chlorogenic acid was 200-1 000 ng and the average recovery was （98.58±1.16）%（n=9）. The contents of chlorogenic acid in Xanthium sibiricum samples from 11 locations were conspicuously different, and they were higher than those in X. mongolicum and X. sibiricum var. subinerme samples. The content of chlorogenic acid was highest in cocklebur fruits from Ding＇an in Hainan Province, which was （0. 969±0. 010）%. Conclusion： The method is proved to be simple, accurate and specific, and offers dependable evidence for the quality evaluation of cocklebur fruits. The interspecies and intraspecies differences of the content of chlorogenic acid are considerable. The cocklebur fruits from Ding＇an in Hainan Province can be considered as the resource of high-grade medical material of cocklebur fruits.