Isolation and Purification of Tyrosinase Inhibitors from the Seeds of Thuja orientalis L.
식물추출물로부터 보다 안전하고 효과있는 새로운 tyrosinase 저해제를 개발하기 위한 연구의 일환으로 먼저 기존의 spectrophotometric assay를 개선한 새로운 TLC 검색방법을 사용하여 여러 식물추출물을 검색한 결과 백자인의 메탄올추출물이 강한 tyrosinase 저해활성을 갖고 있음을 알 수 있었다. 따라서 백자인의 메탄올추출물로부터 tyrosinase 저해제를 분리하기 위하여 먼저 탈지 메탄올추출물을 에테르, 에틸아세테이트
Previously, the methanolic extracts of thirty Korean medicinal plant seeds were screened for tyrosinase inhibitors using a rapid and simple TLC method, which was superior to a conventional spectrophotometrical in vitro assay. As a result, the methanolic extracts of Thuja orientalis seeds was found to have strong tyrosinase inhibitory activity. To isolate active tyrosinase inhibitors, the seeds were defatted with n-hexane under reflux, and then extracted twice with methanol under reflux at 90. The methanolic extract was evaporated to a small volumn in vacuo, and then successively fractionated with ether, ethyl acetate and n-butanol. The ether extract showing significant tyrosinase inhibitory activity was solubilized with 5% NaHCOand then acidified with 6N HCI. The ether souble acidic fraction was successively ohromatographed on silica gel, Sephadex LH-20 and preparative TLC. Among four compounds isolated, two of them showed stronger tyrosinase inhibitory activity, comparable to that of L-ascorbic acid (IC50/=28/). These results suggest that Thuja orientalis seeds may be useful as potential sources of antibrowning agents in fruits and vegetables, and anti-melanoma agents in cosmetics and phamaceuticals.