Citation: ZHANG Yanmei, KANG Jingwu. Screening of the active ingredients in natural products by capillary electrophoresis and high performance liquid chromatography-mass spectrometry[J]. Chinese Journal of Chromatography, ;2013, 31(7): 640-645. doi: 10.3724/SP.J.1123.2013.03013 shu

Screening of the active ingredients in natural products by capillary electrophoresis and high performance liquid chromatography-mass spectrometry

ZHANG Yanmei ,
KANG Jingwu ^,

1.
Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai 200032, China

Corresponding author: KANG Jingwu,
Received Date: 5 March 2013

Fund Project: 国家自然科学基金项目(21175146,20975109). (21175146,20975109)

A new strategy for screening the crude natural extracts and quickly identifying the bioactive compounds was developed. In combination with high performance liquid chromatography-mass spectrometry (HPLC-MS), the biologically active compounds, such as the enzyme inhibitors, in the crude natural extract can be quickly identified by capillary electrophoresis (CE)-based activity assay. The crude natural extracts were assayed by a CE-based enzyme inhibitor screening method, and the active extract was isolated by HPLC-MS/MS with a semipreparative column. Then, each eluted component was assayed again with the CE-based assay method. Finally, the structures of the identified active compounds were elucidated by MS/MS analysis. Acetylcholinesterase (ACHE), its substrate acetylthiocholine chloride (AThCh), as well as the crude extract of Rhizoma coptidis were utilized for the proof of the methodology. Seven isoquinoline alkaloids, namely jatrorrhizine, epiberberine, columbamine, coptisine, corysamine, palmatine and berberine were identified to be active as the inhibitors of ACHE. Their IC₅₀ values were 40, 442, 38, 182, 419, 54 and 16 μ mol/L, respectively. Compared with the traditional screening methods, the method is characterized with several advantages, such as extremely low sample and reagent consumption, high speed of analysis, high sensitivity of detection, high throughput in terms of preparation of the natural products by HPLC. Overall, the results demonstrate that the method is valuable for the screening of the bioactive compounds in the crude natural extracts.
- capillary electrophoresis (CE),
- high performance liquid chromatography-mass spectrometry (HPLC-MS),
- active ingredients,
- natural products,
- drug screening
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