Journal
JOURNAL OF CHROMATOGRAPHY A
Volume 1398, Issue -, Pages 57-65Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.chroma.2015.04.018
Keywords
Knockout; HSCCC; DPPH-HPLC; Antioxidant; Mains doumeri
Funding
- National Natural Science Foundation of China [21275163]
- Collaboration and Innovation Center for Digital Chinese Medicine in Hunan
- Program of Survey and Monitoring of Chinese Medicines for National Drugs
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Due to the complexity of natural products, efficient identification of bioactive compounds, especially for minor compounds, would require a huge effort. Here, we developed an effective strategy based on combining major constituents' knockout with high-performance liquid chromatography-diode array detector-quadrupole time-of-flight tandem mass spectrometry (HPLC-DAD-QTOF-MS/MS) to comprehensively identify minor antioxidants in Mains doumeri, one of the longest known and most used tonic plant in Taiwan. First, five major compounds (I-V) in M. doumeri were knocked out by two-step step-wise high-speed countercurrent chromatography (HSCCC). Second, minor antioxidants were screened by 1,1-dipheny1-2-picrylhydrazyl radical-HPLC (DPPH-HPLC) assay. Third, structures of thirty minor antioxidants,including 11 dihydrochalcones, 4 flavanones, 3 flavonols, 2 flavones, 3 aurones and 7 phenolic acids, were unambiguously or tentatively identified by matching their characteristic UV spectra, accurate mass signals and key diagnostic fragment ions with standards or previously reported compounds. Twenty-six of them, as far as was known, were discovered from M. doumeri for the first time. The results indicated that the proposed method was a useful approach to explore minor bioactive compounds from complex natural products. (C) 2015 Elsevier B.V. All rights reserved.
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