Citation: WU Can, XU Pingcui, YAO Weixuan, SHOU Dan, ZHU Yan, WANG Nani. Rapid screening of anti-osteoporosis active ingredients from Liuwei Dihuang Decoction by osteoblast membrane chromatography/ultra-high performance liquid chromatography-time of flight mass spectrometry[J]. Chinese Journal of Chromatography, ;2019, 37(3): 305-312. doi: 10.3724/SP.J.1123.2018.11022 shu

Rapid screening of anti-osteoporosis active ingredients from Liuwei Dihuang Decoction by osteoblast membrane chromatography/ultra-high performance liquid chromatography-time of flight mass spectrometry

WU Can ^1, ,
XU Pingcui ² ,
YAO Weixuan ³ ,
SHOU Dan ² ,
ZHU Yan ¹ ,
WANG Nani ^1,2

1.
Department of Chemistry, Zhejiang University, Hangzhou 310007, China;
2.
Department of Medicine, Zhejiang Academy of Traditional Chinese Medicine, Hangzhou 310007, China;
3.
Department of Criminal Science and Technology, Key Laboratory of Drug Prevention and Control Technology of Zhejiang Province, Zhejiang Police College, Hangzhou 310053, China

Received Date: 15 November 2018

Fund Project: National Natural Science Foundation of China (No. 81603252)

A method was established for determination of potential anti-osteoporosis ingredients from Liuwei Dihuang Decoction by osteoblast cell membrane chromatography/ultra-performance liquid chromatography/time-of-flight mass spectrometry (CMC/UPLC-TOF/MS). The osteoblasts were used as the source of cell membrane for the preparation of CMC stationary phase. An osteoblast CMC column (10 mm×2 mm) was prepared by coating silica gel (0.05 g) with cell membrane. The active ingredients in the aqueous extract of Liuwei Dihuang Decoction (90 g/L) were first screened by CMC. Water was used as the mobile phase, and the flow rate was 0.20 mL/min. Then, the eluates of the CMC were qualitatively analyzed by UPLC-TOF/MS using a WATERS ACQUITY UPLC BEH C18 column (10 mm×2 mm). Acetonitrile-water was used as the mobile phase at a flow rate of 0.40 mL/min. This method could quickly and selectively identify 16 potential anti-osteoporotic active ingredients from Liuwei Dihuang Decoction. Due to high catalpol content in Liuwei Dihuang Decoction and its good affinity with CMC column, catapol was selected for in vivo and in vitro pharmacological examinations. It was found that catalpol (1-10 μmol/L) could significantly promote the proliferation of osteoblasts in a dose-dependent manner. It also significantly increased the area of bone staining in osteoporosis zebrafish model. The osteoblast CMC/UPLC-TOF/MS method could quickly screen the anti-osteoporosis active ingredients in complex traditional Chinese medicine prescriptions, and had the advantages of simple operation, high efficiency and high sensitivity.
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