A method of hepatocyte extraction conjugated with HPLC is established for screening potential active components in Chinese medicines--probing Herba Artemisiae Scopariae as an exemplifying approach.
In order to establish an effective and quick method for screening potential bioactive compounds in Traditional Chinese Medicines (TCMs), hepatocytes were employed for extracting either bifendate, a clinical medicine for liver diseases, or chemicals in Herba Artemisiae Scopariae (A. Scopariae), a commonly used traditional Chinese medicine for remedying liver diseases such as hepatitis induced by viruses, chemicals or alcohol. After hepatocyte extraction the compounds were analyzed by HPLC, therefore this method was referrred to as hepatocyte extraction conjugated with HPLC (HE-HPLC). In the first part of this study, HE-HPLC showed that bifendate was extracted by hepatocytes and detected by HPLC-DAD which indicated the feasibility of this method. Then in the second part of the study, the potential active components in the A. scopariae extract were studied using HE-HPLC. Six chemicals in the A. scopariae extract, which could bind to hepatocytes in vitro, were detected by HPLC-DAD and three were identified as 7-hydroxy-coumarin (7-OH-C), capillartemisin A and 7-methoxy-coumarin, respectively. In vitro assays showed that 7-OH-C protected HL-7702 hepatocytes from H₂O₂ injury. The results indicated that these compounds could be extracted by hepatocytes, could be detected by HPLC and more importantly were bioactive. It is suggested that HE-HPLC is a useful method for screening potent active components in Chinese medicines used to treat liver diseases.
National Standard Laboratory of Pharmacology for Chinese Medicine, Nanjing University of Chinese Medicine, Nanjing 210029, China. hongmin72@hotmail.com
Min
M
Hong
Hong-Yu
HY
Ma
Xiang-Rui
XR
Wu
Yong-Qing
YQ
Hua
Quan
Q
Zhu
Hong-Wei
HW
Fan
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D022781:Hepatocytes / Q000737:chemistry*; D008516:Medicine, Chinese Traditional