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Li C, Zhao Y, Guo Z, et al. Effective 2D-RPLC/RPLC enrichment and separation of micro-components from Hedyotis diffusa Willd. and characterization by using ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry. J Pharm Biomed Anal. 2014;99:35-44doi: 10.1016/j.jpba.2014.06.020.
Li, C., Zhao, Y., Guo, Z., Zhang, X., Xue, X., & Liang, X. (2014). Effective 2D-RPLC/RPLC enrichment and separation of micro-components from Hedyotis diffusa Willd. and characterization by using ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry. Journal of pharmaceutical and biomedical analysis, 9935-44. https://doi.org/10.1016/j.jpba.2014.06.020
Li, Cunman, et al. "Effective 2D-RPLC/RPLC enrichment and separation of micro-components from Hedyotis diffusa Willd. and characterization by using ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry." Journal of pharmaceutical and biomedical analysis vol. 99 (2014): 35-44. doi: https://doi.org/10.1016/j.jpba.2014.06.020
Li C, Zhao Y, Guo Z, Zhang X, Xue X, Liang X. Effective 2D-RPLC/RPLC enrichment and separation of micro-components from Hedyotis diffusa Willd. and characterization by using ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry. J Pharm Biomed Anal. 2014 Oct;99:35-44. doi: 10.1016/j.jpba.2014.06.020. Epub 2014 Jul 09. PMID: 25061712.
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J Pharm Biomed Anal. 2014 Oct;99:35-44. doi: 10.1016/j.jpba.2014.06.020. Epub 2014 Jul 09.

Effective 2D-RPLC/RPLC enrichment and separation of micro-components from Hedyotis diffusa Willd. and characterization by using ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry.

Journal of pharmaceutical and biomedical analysis

Cunman Li, Yanyan Zhao, Zhimou Guo, Xiuli Zhang, Xingya Xue, Xinmiao Liang

Affiliations

  1. Hebei Research Centre of Analysis and Testing, Hebei University of Science and Technology, Shijiazhuang 050018, China.
  2. Pharmacy College, Dalian Medical University, Dalian 116044, China.
  3. Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China.
  4. Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China. Electronic address: [email protected].

PMID: 25061712 DOI: 10.1016/j.jpba.2014.06.020

Abstract

An effective method aiming at enrichment and analysis of micro-components in traditional Chinese medicine (TCM) was developed. One fraction (fraction E) from the extract of Hedyotis diffusa Willd. was selected as test sample, which was isolated by using the XAD-4 macroporous resin. To study the micro-components, a two-dimensional reverse-phase liquid chromatography (2D-RPLC/RPLC) method was developed, comprising Click OEG and C18 stationary phases as the first and second dimensions, respectively. Of the eight sub-fractions isolated from the first dimension, three sub-fractions (fractions II-IV) containing micro-components were further separated with the second dimension. The 2D-RPLC/RPLC system was proved to possess high orthogonality. Furthermore, the micro-components were characterized by using ultra-performance liquid chromatography-diode array detector/quadrupole time-of-flight mass spectrometry (UPLC-DAD/Q-TOF MS) with electrospray ionization (ESI) source. With the optimized separation and characterization method, a large number (>400) of micro-components were enriched and detected from the extracts of H. diffusa Willd., the majority of which has not been isolated from the herb before. Among these isolated micro-components, 38 compounds involving 24 phenylpropanoids, 7 flavonoids and 7 iridoid glucosides (IGs), were identified or tentatively identified from the H. diffusa extracts on the basis of spectral data of the authentic standards and the fragmentation characteristics information available in literatures. The proposed method made it possible to effectively screen and analyze the micro-components in TCMs or other complex natural medicines.

Copyright © 2014 Elsevier B.V. All rights reserved.

Keywords: 2D-RPLC/RPLC; Click OEG stationary phase; Hedyotis diffusa Willd.; Micro-component; UPLC–DAD/Q-TOF MS

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