KOREAN JOURNAL OF CROP SCIENCE (한국작물학회지)

The Korean Society of Crop Science (한국작물학회)
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Isolation of Isoflavones and Soyasaponins from the Germ of Soybean

콩 배아로 부터 Isoflavone과 Soyasaponin의 동시 분리

  • 김선림 (농촌진흥청 국립식량과학원) ;
  • 이재은 (농촌진흥청 국립식량과학원) ;
  • 김율호 (농촌진흥청 국립식량과학원) ;
  • 정건호 (농촌진흥청 국립식량과학원) ;
  • 김대욱 (농촌진흥청 국립식량과학원) ;
  • 이춘기 (농촌진흥청 국립식량과학원) ;
  • 김미정 (농촌진흥청 국립식량과학원) ;
  • 김정태 (농촌진흥청 국립식량과학원) ;
  • 이유영 (농촌진흥청 국립식량과학원) ;
  • 황태영 (농촌진흥청 국립식량과학원) ;
  • 이광식 (농촌진흥청 국립식량과학원) ;
  • 김욱한 (농촌진흥청 국립식량과학원) ;
  • 권영업 (농촌진흥청 국립식량과학원) ;
  • 김홍식 (충북대학교 농업생명환경대학 식물자원학과) ;
  • 정일민 (건국대학교 생명환경과학대학 응용생물과학과)
  • Received : 2013.01.17
  • Accepted : 2013.04.04
  • Published : 2013.06.30
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Abstract

The objective of present study was to simultaneously isolate of isoflavone and soyasaponin compounds from the germ of soybean seeds. Soy germ flours were defatted with hexane for 48h at room temperature, and methanolic extracts were prepared using reflux apparatus at $90^{\circ}C$ for 6h, two times. After extraction, extracts were separated with preparative RP-$C_{18}$ packing column ($125{\AA}$, $55-105{\mu}m$, $40{\times}150mm$), and collected 52 fractions were identified with TLC plate (Kieselgel 60 F-254) and HPLC, respectively. Among the identified isoflavone and soyasaponin fractions, isoflavone fractions were re-separated using a recycling HPLC with gel permeation column (Jaigel-W252, $20{\times}500mm$). Final fractions were air-dried, and the purified compounds of two isoflavones (ISF-1-1, ISF-1-2) and four soyasaponins (SAP-1, SAP-2, SAP-3, SAP-4) were obtained. Two isoflavone compounds (ISF-1-1, ISF-1-2) were acid-hydrolyzed for the identification of their aglycones, and confirmed by comparing with 12 types of isoflavone isomers. While the four kinds of soyasaponins were identified by using a micro Q-TOF mass spectrometer in the ESI positive mode with capillary voltage of 4.5kV, and dry temperature of $200^{\circ}C$. Base on the obtained results, it was conclude that ISF-1-1 is the mixture isomers of daidzin (43.4%), glycitin (47.0%), and genistin (9.6%), but ISF-1-2 is the single compound of genistin (99.8% <). On the other hand, soyasaponin SAP-1 is the mixture compounds of soyasaponin A-group (Aa, Ab, Ac, Ae, Af); SAP-2 is soyasaponin B-group (Ba, Bb, Bc) and E-group (Bd, Be); SAP-3 is soyasaponin B-group (Ba, Bb, Bc), E-group (Bd, Be), and DDMP-group (${\beta}g$); SAP-4 is soyasaponin B-group (Ba, Bb, Bc), E-group (Bd, Be), and DDMP-group (${\beta}g$, ${\beta}a$), respectively.

콩 관련 제품을 생산하는 과정에서 발생되는 부산물은 가축의 사료 및 퇴비 등에 이용되고 있으나 일부는 폐기물로 처리되어 추가 비용 및 각종 환경오염을 유발하는 등 사회적인 문제점으로 지적되고 있다. 따라서 본 연구에서는 가공 부산물로 다량 발생되는 콩 배아를 활용하여 생리활성물질인 isoflavone과 soyasaponin을 동시에 분리하는 방법을 개발하고자 실시하였다. 1. 콩 배아 methanol 추출물을 preparative $C_{18}$ column에 주입하고, 210 nm의 파장에서 0.5% 초산 용액 30%로부터 100% acetonitrile까지 분당 15mL의 유속으로 53분간 흘려주어 isoflavone과 soyasaponin 분획을 동시에 분리하였다. 2. Preparative $C_{18}$ column으로 분리된 isoflavone 및 soyasaponin 분획은 동결건조시켜 isoflavone 분말 ISF-1과 soyasaponin SAP-1, SAP-2, SAP-3 및 SAP-4의 분말을 얻었다. 3. Isoflavone 분획 ISF-1의 재분리는 젤투과 컬럼에서 100% acetonitrile을 분당 5 mL가 되도록 흘려주면서 254nm 파장에서 관찰하여 2종의 분획 ISF-1-1 및 ISF-1-2을 분리하였다. 4. 분리된 2종의 isoflavone 중 ISF-1-1은 그 조성이 daidzin, glycitin 및 genistin 이고, ISF-1-2는 genistin 단일물질이 주성분인 것으로 나타났다. 5. 분리된 4종의 soyasaponin 중 SP-1은 soyasaponin A계열인 Aa(MW: 1364), Ab(MW: 1436), Ac(MW: 1420), Ae(MW: 1202), Af(MW: 1274), SAP-2는 B계열인 Ba(MW: 958), Bb(MW: 942), Bc(MW: 912)와 E계열인 Bd(MW: 956), Be(MW: 940), SAP-3는 B계열인 Ba, Bb, Bc, E계열인 Bd, Be와 DDMP계열인 ${\beta}g$(MW: 1068), SAP-4는 B계열인 Ba, Bb, Bc, E계열인 Bd, Be와 DDMP계열인 ${\beta}g$, ${\beta}a$(MW: 1038)가 주성분임을 알 수 있었다.

Keywords

References

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