HSCCC-based strategy for preparative separation of in vivo metabolites after administration of an herbal medicine: Saussurea laniceps, a case study.
This paper reports a novel strategy based on high-speed counter-current chromatography (HSCCC) technique to separate in vivo metabolites from refined extract of urine after administration of an herbal medicine. Saussurea laniceps (SL) was chosen as a model herbal medicine to be used to test the feasibility of our proposed strategy. This strategy succeeded in the case of separating four in vivo metabolites of SL from the urine of rats. Briefly, after oral administration of SL extract to three rats for ten days (2.0 g/kg/d), 269.1 mg of umbelliferone glucuronide (M1, purity, 92.5%), 432.5 mg of scopoletin glucuronide (M2, purity, 93.2%), 221.4 mg of scopoletin glucuronide (M3, purity, 92.9%) and 319.0 mg of scopoletin glucuronide (M4, purity, 90.4%) were separated from 420 mL of the rat urine by HSCCC using a two-phase solvent system composed of methyl tert-butyl ether-n-butanol-acetonitrile-water (MTBE-n-BuOH-ACN-H2O) at a volume ratio of 10:30:11:49. The chemical structures of the four metabolites, M1 to M4, were confirmed by MS and (1)H, (13)C NMR. As far as we know, this is the first report of the successful separation of in vivo metabolites by HSCCC after administration of an herbal medicine.
School of Chinese Medicine, Hong Kong Baptist University, Kowloon Tong, Hong Kong Special Administrative Region, China.|Institute of Research and Continuing Education (Shenzhen), Hong Kong Baptist University, Shenzhen, China.
Tao
T
Yi
School of Chinese Medicine, Hong Kong Baptist University, Kowloon Tong, Hong Kong Special Administrative Region, China.
Lin
L
Zhu
State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Technology, Taipa, Macau Special Administrative Region, China.
Guo-Yuan
GY
Zhu
School of Chinese Medicine, Hong Kong Baptist University, Kowloon Tong, Hong Kong Special Administrative Region, China.|Institute of Research and Continuing Education (Shenzhen), Hong Kong Baptist University, Shenzhen, China.
Yi-Na
YN
Tang
School of Chinese Medicine, Hong Kong Baptist University, Kowloon Tong, Hong Kong Special Administrative Region, China.|Institute of Research and Continuing Education (Shenzhen), Hong Kong Baptist University, Shenzhen, China.
Jun
J
Xu
School of Chinese Medicine, Hong Kong Baptist University, Kowloon Tong, Hong Kong Special Administrative Region, China.
Jia-Yi
JY
Fan
School of Chinese Medicine, Hong Kong Baptist University, Kowloon Tong, Hong Kong Special Administrative Region, China.
Zhong-Zhen
ZZ
Zhao
School of Chinese Medicine, Hong Kong Baptist University, Kowloon Tong, Hong Kong Special Administrative Region, China.|Institute of Research and Continuing Education (Shenzhen), Hong Kong Baptist University, Shenzhen, China.
Hu-Biao
HB
Chen
D000818:Animals; D002845:Chromatography; D008297:Male; D008517:Phytotherapy; D010936:Plant Extracts / Q000737:chemistry* / Q000493:pharmacokinetics* / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D030441:Saussurea / Q000737:chemistry*
D010936:Plant Extracts