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Journal of Natural Medicines

, Volume 72, Issue 1, pp 73–79 | Cite as

Two flavone C-glycosides as quality control markers for the manufacturing process of ephedrine alkaloids-free Ephedra Herb extract (EFE) as a crude drug preparation

  • Naohiro Oshima
  • Takuro Maruyama
  • Tadatoshi Yamashita
  • Nahoko Uchiyama
  • Yoshiaki Amakura
  • Sumiko Hyuga
  • Masashi Hyuga
  • Shunsuke Nakamori
  • Hiroaki Takemoto
  • Yoshinori Kobayashi
  • Takashi Hakamatsuka
  • Hiroshi Odaguchi
  • Toshihiko Hanawa
  • Yukihiro GodaEmail author
Original Paper

Abstract

As part of our continuing study of ephedrine alkaloids-free Ephedra Herb extract (EFE) in pursuit of its approval as a crude drug preparation, we identified two quantitative markers for the quality control of the manufacturing process of EFE and sought to establish cost-effective and simple methods for quantitative analyses. We analysed Ephedra Herb extracts grown in different habitats and collection years by liquid chromatography/high-resolution mass spectrometry (LC/HRMS) and detected two notable peaks common to each extract. These peaks were identified as vicenin-2 (1) and isovitexin 2″-O-rhamnoside (2). Quantitative analyses using the isocratic condition of LC/MS showed that the content percentages of 1 and 2 in EFE were 0.140–0.146% and 0.350–0.411%, respectively. We concluded that 1 and 2 were adequate quality control markers for quantitative analysis of EFE. Furthermore, we quantitatively analysed apigenin (3), an aglycon common to 1 and 2, and found that the conversion factors of 1 to 3 and 2 to 3 were 1.3 and 1.5, respectively. Therefore, we concluded that 3 was a secondary standard for quantifying the contents of 1 and 2 in EFE. A series of results obtained from this study will be valuable for the quality control of EFE.

Keywords

Ephedra Herb Ephedrine alkaloids-free Ephedra Herb extract Quality control markers Flavone C-glycosides Apigenin 

Abbreviations

EFE

Ephedrine alkaloids-free ephedra herb extract

LC/HRMS

Liquid chromatography/high-resolution mass spectrometry

Notes

Acknowledgements

This research is partially supported by the Research and Development of New Drugs project of the Japan Agency for Medical Research and Development (AMED).

Compliance with ethical standards

Conflict of interest

We have applied for a patent under the regulation of the Patent Cooperation Treaty (PCT).

Supplementary material

11418_2017_1111_MOESM1_ESM.tif (78 kb)
Fig. 1S UV/PDA chromatogram (λ = 337 nm) of EH extract and mass chromatograms at m/z 593 of EH extract and an isolated compounds. (A) EH extract (λ = 337 nm), (B) EH extract (m/z 593), (C) vicenin-2 (1) (m/z 593). (TIFF 78 kb)
11418_2017_1111_MOESM2_ESM.tif (76 kb)
Fig. 2S UV/PDA chromatogram (λ = 337 nm) of EH extract and mass chromatograms at m/z 577 of EH extract and the isolated compounds. (A) EH extract (λ = 337 nm), (B) EH extract (m/z 577), (C) compound 2 (m/z 577). (TIFF 76 kb)
11418_2017_1111_MOESM3_ESM.tif (154 kb)
Fig. 3S Mass spectra of vicenin-2 (1) (left, EFE; right, authentic compound). ESI (+), positive ion mode; ESI (–), negative ion mode. (TIFF 154 kb)
11418_2017_1111_MOESM4_ESM.tif (188 kb)
Fig. 4S Mass spectra of isovitexin 2”-O-rhamnoside (2) (left, EFE; right, isolated compound). ESI (+), positive ion mode; ESI (–), negative ion mode. (TIFF 187 kb)
11418_2017_1111_MOESM5_ESM.tif (114 kb)
Fig. 5S UV spectra of vicenin-2 (1), isovitexin 2”-O-rhamnoside (2) and apigenin (3). (TIFF 114 kb)

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Copyright information

© The Japanese Society of Pharmacognosy and Springer Japan KK 2017

Authors and Affiliations

  • Naohiro Oshima
    • 1
    • 7
  • Takuro Maruyama
    • 2
  • Tadatoshi Yamashita
    • 3
  • Nahoko Uchiyama
    • 2
  • Yoshiaki Amakura
    • 4
  • Sumiko Hyuga
    • 5
  • Masashi Hyuga
    • 2
  • Shunsuke Nakamori
    • 5
    • 6
  • Hiroaki Takemoto
    • 5
    • 6
  • Yoshinori Kobayashi
    • 5
    • 6
  • Takashi Hakamatsuka
    • 2
  • Hiroshi Odaguchi
    • 5
  • Toshihiko Hanawa
    • 5
  • Yukihiro Goda
    • 2
    Email author
  1. 1.Department of Pharmaceutical SciencesInternational University of Health and WelfareOhtawaraJapan
  2. 2.Division of DrugsNational Institute of Health SciencesTokyoJapan
  3. 3.TOKIWA Phytochemical Co., Ltd.SakuraJapan
  4. 4.Department of Pharmacognosy, College of Pharmaceutical SciencesMatsuyama UniversityMatsuyamaJapan
  5. 5.Oriental Medicine Research CenterKitasato UniversityTokyoJapan
  6. 6.Department of Pharmacognosy, School of PharmacyKitasato UniversityTokyoJapan
  7. 7.Faculty of Pharmaceutical SciencesTokyo University of ScienceNodaJapan

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