검색 상세

비파인(枇杷仁)용매분획물의 생리활성 및 유효성분 분석에 관한 연구

A Study on Physiological Activities and Analysis of Bioactive Compounds in the Extract and its Fractions from Seed of Eriobotrya japonica Lindley

초록/요약

본 연구에서는 천연물질인 비파인(枇杷仁)이 가지고 있는 항산화 활성과 항균 및 항염증 활성을 중심으로 생리활성을 검증하고, 활성에 관련된 유효성분의 활용에 관한 기초자료를 수집하기 위해 수행하였다. 비파인을 80% MeOH로 추출하여 농축한 후 이를 여러 용매로 분획하고 분리한 다음 이 분획물들의 항산화, 항균 및 항염증활성을 측정하였다. 비파인 추출물과 분획물의 추출 수율은 aqueous fraction이 59.79%로 가장 높았으며 butanol fraction 16.62%, chloroform fraction 11.03%, MeOH extract 9.38%, ethyl acetate fraction 4.56% 의 순서로 나타났다. 비파인 메탄올 추출물과 분획물의 유효성분 함량을 비교한 결과 total phenolic compound는 ethyl acetate fraction에서 381.55mg/g, flavonoid 함량은 chloroform fraction에서 6.32mg/g으로 가장 높게 측정되었다. 비파인 추출물과 분획물을 HPLC로 분석한 결과, methanol 추출물에서 amygdalin이 2.33%, chlorogenic acid가 1.05%, benzaldehyde가 1.10% 검출되었다. 비파인 추출물과 분획물의 생리활성 가운데 DPPH radical 소거능은 ethyl acetate fraction과 chloroform fraction이 0.15mg/ml의 농도에서 각각 49.00%, 28.07%의 높은 활성을 보였다. NO 소거능과 생성 저해능은 추출물과 분획물 모두에서 농도 의존적인 활성을 보였다. Raw 264.7 세포주에 대한 세포생존율을 측정한 결과 전반적으로 양호한 생존율을 보임으로써 세포독성은 우려할 만한 수준이 아님을 알 수 있었다. 항균활성은 ethyl acetate fraction이 Staphylococcus aureus, Bacillus cereus, Staphylococcus epidermidis 균주에 대해 강한 활성을 나타냈으며, 이로써 비파인 ethyl acetate fraction을 항산화활성에 기초한 천연 항염·항균제로 개발할 수 있는 가능성을 확인하였다.

more

초록/요약

In recent years, cultivation of Eriobotrya japonica is on the increase around the southern area in Korea. The purpose of this study was to investigate a physiological activity of the methanol extract and its fractions from the seed of Eriobotrya japonica. Moreover, we suggested the condition to analyze five compound simultaneously in Eriobotrya japonica seed such as amygdalin, benzoic acid, caffeic acid, chlorogenic acid and benzaldehyde using HPLC with C18 column and UV-detector. In DPPH radical scavenging ability at 0.15 mg/ml, radical scavenging ability of the methanol extract, ethyl acetate fraction and chloroform fraction were 11.07%, 49.00% and 28.07% respectively. Radical scavenging ability of ascorbic acid used as positive control in same concentration was 70.03%. The ethyl acetate fraction exhibited most effective scavenging ability than others. The total polyphenol contents of the methanol extract, ethyl acetate and butanol fractions were 57.31 mg/g, 381.55 mg/g and 68.96 mg/g. And the flavonoid contents of the methanol extract, EtOAc and BuOH fractions were 1.7 mg/g, 4.88 mg/g and 5.02 mg/g, respectively. Amygdalin content in the methanol extract and butanol fraction was 2.33% and 6.05%. Benzoic acid content in the methanol extract and chloroform fraction was 0.53% and 2.00%. Caffeic acid content in the methanol extract and ethyl acetate fraction was 0.10% and 0.29%. Chlorogenic acid content in the methanol extract and butanol fraction was 1.05% and 2.96%. Benzaldehyde content in the methanol extract and chloroform fraction was 1.10% and 2.08%. In antibacterial activity by disc diffusion assay, the methanol extract did not showed antibacterial activites in tested concentrations(0.5 mg/disc ～ 5.0 mg/disc) against Staphylococcus aureus and Pseudomonas aeruginosa. Moreover, aqueous fraction did not showed antibacterial activites in tested concentrations against all tested bacteria. However, the ethyl acetate fraction showed the highest antibacterial activity against all tested bacteria. In antibacterial activities of compounds in seed of E. japonica, benzoic acid, caffeic acid and chlorogenic acid showed more strong antibacterial activity than other compounds. In addition, the ethyl acetate fraction was found to be the best inhibitor of nitric oxide(NO) production than other fractions and the methanol extract. It was the results caused by the highest caffeic acid content in ethyl acetate fraction. On the other hand, in the Raw264.7 cell viability measurement by MTT assay, the methanol extract and every fractions were showed a moderate viabilities. As a result, benzoic acid showed the strongest antibacterial activity and caffeic acid was found to be the best inhibitor of NO production. Moreover, the ethyl acetate fraction of the methanol extract with high content of benzoic acid and caffeic acid was found to be a highest effective fraction in the methanol extract from seed of Eriobotrya japonica.

more

목차

LIST OF TABLES ………………………………………………………………… i
LIST OF FIGURES ……………………………………………………………… ii
ABBREVIATIONS …………………………………………………………………ⅲ
국문초록 ……………………………………………………………………………… iv
I. 서론 ………………………………………………………………………………… 1
II. 재료 및 방법 …………………………………………………………………… 3
1. 재료 ……………………………………………………………………………… 3
(1) 식물 재료 ……………………………………………………………………… 3
(2) 세포주 …………………………………………………………………………… 3
(3) 시약 및 배지 ………………………………………………………………… 3
2. 실험방법 ………………………………………………………………………… 4
(1) 추출 및 분획…………………………………………………………………… 4
(2) 성분 분석 ……………………………………………………………………… 4
1) Total phenolic compound 및 flavonoid 함량 분석 …………………… 5
2) HPLC에 의한 활성성분 분석 ……………………………………………… 5
(3) 항산화활성 측정……………………………………………………………… 5
1) DPPH radical 소거능 측정 ……………………………………………… 5
2) Nitric oxide 소거능 측정 ………………………………………………… 10
(4) 항균활성 측정………………………………………………………………… 10
(5) 세포독성 및 항염증 활성 측정 ………………………………………………11
1) 세포주 배양 ……………………………………………………………………11
2) MTT assay에 의한 세포생존율 측정 ……………………………………11
3) Nitric oxide 생성 억제 활성 측정……………………………………………11
(6) 통계분석 ……………………………………………………………………… 12
III. 결과 및 고찰 …………………………………………………………………… 14
1. 비파인 추출물과 분획물의 추출수율 및 성분함량 ………………………… 14
1) 추출 및 분획수율 …………………………………………………………… 14
2) Total phenolic compound 및 flavonoid 함량 ………………………… 16
3) 유효성분 함량 ……………………………………………………………… 18
2. 비파인 추출물과 분획물의 생리활성………………………………………… 20
(1) 항산화 활성 ………………………………………………………………… 20
1) DPPH radical 소거능 ……………………………………………………… 20
2) Nitric oxide 소거능 ………………………………………………………… 22
(2) Disc diffusion assay 에 의한 항균활성 ……………………………………24
1) 비파인 추출물 및 분획물의 항균활성 …………………………………… 24
2) 비파인 유효성분의 항균활성 ……………………………………………… 26
(3) 세포독성 및 항염증 활성 …………………………………………………… 28
1) MTT assay에 의한 Raw264.7 세포생존율 ……………………………… 28
2) Nitric oxide 생성 억제 활성 ……………………………………………… 31
영문초록 ……………………………………………………………………………… 34
참고문헌 ……………………………………………………………………………… 37

more
반출 Meta View 목록