본 연구에서는 Bovine bile에서 항산화 물질분획을 분리하고 이에 따른 항산화 효소들의 여러 활성효과를 측정하였으며 GC/FID에 의한 항산화성 물질의 패턴을 확인하였다. 항산화력은 Butylated hydroxy toluene(BHT)에 비해 Bovine bile의 EtOAc추출물(BBE)이 51.2%로 나타났고 xanthine oxidase를 측정 48.3%의 저해효과를 보였으며 glutatione-S-transferase의 활성은 85.7%의 활성이 나타났다. MTT방법에 의한 세포생존율의 조사에서는 6mM glutathione 투여에서 95%라는 생존율과 비교하여 20mg, 60mg의 BBE를 처리했을 때 56%와 65%, 140mg처리에서는 78%의 세포생존율이 나타났다. Bovine bile EtOAc추출물을 TLC로 분석한 결과 Rf값이 0.72에서 항산화 활성이 나타났고 이 분획을 methylation과 trimethysilyl(TMS) 유도체화를 시켜서 Gas Chromatography(FID)와 mass spectrum을 통한 성분 분석을 하고 bile acid의 구성분과 비교하여 계통분석을 실시한 결과 Coprostan-3ol로 추정되었다.
This study was carried out to extract the natural antioxidants from Bovine bile and to investigate their effects on various antioxidant activities. It also characterized the patterns of antioxidants by GC/FID and GC/MS. The antioxidative activities and chemical structure of the antioxidant were elucidated by examining the effects of biological activity and the analysis of GC/MS. The antioxidant materials extracted from bovine bile were isolated and purified by silica gel column chromatography and TLC. It was confirmed that there were effects of antioxidants such as Xanthine Oxidase(XO) and Glutathione-S-Transferase(GSH-T) on antioxidative activities. When they were compared with BHT, bile extracts showed the relative effects of 51.2% on the antioxidant activity, the inhibition effects of 48.3% on XO activity, and the synergism effects of 85.7% on the GSH-T activity. According to the results of investigation at neuron cell of mouse, the rate of cell activity in the treatment of 6mM glutathione was 96%, While it in the treatment of 140mg of bile extract was 78%. Based on the TLC analysis of EtOAc extracts from the Bovine bile, the antioxidant activity appeared at $R_f$ value, 0.72. These results suggested that the antioxidant may be coprostan 3-ol.
