온라인 항산화 분석 시스템과 LC-NMR/MS를 활용한 천연물의 항산화 성분 분석
The Rapid Identification of the Radical Scavengers by using On-line Antioxidant Screening System and LC-NMR/MS
- 발행기관 강릉원주대학교 대학원
- 지도교수 전상일, 엄병헌
- 발행년도 2010
- 학위수여년월 2010. 2
- 학위명 석사
- 학과 및 전공 일반대학원 KIST강릉분원학.연협동과정
- 실제URI http://www.dcollection.net/handler/kangnung/000000006300
- 본문언어 한국어
초록/요약 도움말
The HPLC hyphenated techniques are increasingly used for phytochemical analysis. These techniques combine the separation power of HPLC with the structure and bioactivity information provided by NMR/MS and on-line antioxidant screening system. The major advantages over the traditional off-line techniques are rapidity and efficiency. Sophora flavescens Aiton (Leguminosae) is a Chinese herbal medicine. From Sophora Radix (dried root of SF) 9 prenyl-flavonoids were isolated by RP C18 prep-HPLC and identified by 1D, 2D NMR spectroscopy as Isoxanthohumol (1), Norkurarinol (2), Kurarinone (3), Leachianone A (4), Kushenol A (5), Kushenol T (6), 2′-Methoxy kurarinone (7), Sophoraflavanone G (8), Kuraridin (9). For the direct identification of the compounds present in Sophora Radix, hyphenated LC-NMR technique has been applied. 6 prenyl-flavonoids were identified as kushenol U (A), kurarinone (B), sophoraflavanone G (C), leachianone A (D), kuraridin (E), and kushenol A (F) by on-flow LC-NMR. The use of hyphenated HPLC techniques for the purpose of directly identifying the major antioxidants of Safflower (홍화, Carthamus tinctorius L.) seeds was investigated. Reversed-phase isocratic chromatography was performed using an acetonitrile-water solvent system on a C18 column. By on-line antioxidant screening system (ABTS radical scavenger system), the antioxidant activity of each peak was evaluated by comparing with the well known antioxidant Trolox. The major active peaks on HPLC chromatogram were identified as 8′-hydroxyarcgenin-4′-O-β-D-glucoside (1), N-(p-coumaroyl) serotonin (2), N-feruloylserotonin (3) through LC-NMR and LC-MS experiment. This study demonstrates on-line coupled HPLC-ABTS+ antioxidant screening system and HPLC-NMR/MS analysis make it possible for rapid identification of antioxidants in natural products.
more목차 도움말
I . 서론 1
1. 항산화 1
2. 초고속 구조/활성 분석 시스템- HTSEB(High throughput structure elucidation/bioassay) 4
2.1. 온라인 항산화 분석 시스템 (on-line antioxidant screening system) 4
2.3. NMR을 이용한 정량분석 (Quantitative NMR: Q-NMR) 10
3. 고삼 11
4. 홍화 13
5. 연구 목적 14
II . 고삼의 Prenyl-Flavonoid 성분 연구 18
1) 재료 및 방법 18
1. 재료 18
2. 시약 및 기기 18
3. Prep-HPLC를 활용한 고삼 성분 분리 20
3.1. 추출 및 분획 20
3.2. Prenyl flavonoids 성분 분리 정제 21
4. LC-NMR을 활용한 고삼 성분 분석 28
2) 결과 및 고찰 31
1. Prep-HPLC를 활용한 고삼 prenyl-flavonoid 성분의 구조 동정 31
Isoxanthohumol (1) 31
Norkurarinol (2) 36
Kurarinone (3) 41
Leachianone A (4) 46
Kushenol A (5) 50
Kushenol T (6) 54
2′-Methoxykurarinone (7) 59
Sophoraflavanone G (8) 64
Kuraridin (9) 68
2. LC-NMR을 활용한 고삼 prenyl-flavonoid 성분의 구조 동정 72
Kushenol U (A) 73
Kurarinone (B) 73
Sophoraflavanone G (C) 74
Leachianone A (D) 75
Kuraridin (E) 76
Kushenol A (F) 77
III . 홍화의 Phenol 화합물 성분 연구 80
1) 재료 및 방법 80
1. 재료 80
2. 시약 및 기기 80
3. 추출 82
4. On-line antioxidant screening system을 활용한 홍화종자의 항산화 성분 분석 82
5. Prep-HPLC를 활용한 홍화종자의 phenol 성분 분리 83
6. LC-NMR, LC-MS를 활용한 홍화종자의 항산화 성분 분석 85
2) 결과 및 고찰 86
1. On-line antioxidant screening system을 활용한 홍화종자의 항산화 성분 분석 87
2. Prep-HPLC를 활용한 홍화종자의 phenol 성분 구조 동정 88
N-(p-Coumaroyl) serotonin mono-β-D-glucopyranoside (1) 89
8′-Hydroxyarctigenin-4′-O-β-D-glucoside (2) 92
3. LC-NMR, LC-MS를 활용한 홍화종자의 항산화 성분 구조 동정 96
8′-hydroxyarcgenin-4′-O-β-D-glucoside (A) 96
N-(p-Coumaroyl) serotonin (B) 98
N-feruloylserotonin (C) 99
IV . 결론 104
V . 참고문헌 106
