(-)-Epicatechin gallate (ECG) is a polyphenol compound of green tea exhibiting biological activities, such as antioxidant and anticancer effects. To examine the effect of ECG on porcine oocytes during in vitro maturation (IVM), oocytes were treated with 0-, 5-, 15-, and 25 μM ECG. After maturation, we investigated nuclear maturation, intracellular glutathione (GSH) and reactive oxygen species (ROS) levels and subsequent embryonic development after parthenogenetic activation (PA) and in vitro fertilization (IVF). After 42 hours of IVM, the 5 μM group exhibited significantly increased (p< 0.05) nuclear maturation (89.8%) compared with the control group (86.1%). However, the 25 μM group observed significantly decreased (p< 0.05) nuclear maturation (83.5%). In intracellular maturation assessment the 5-, 15-, and 25 μM groups had significantly increased (p< 0.05) GSH levels and decreased ROS levels compared with the controls. The 5- and 15 μM group showed significantly increased (p< 0.05) embryo formation rates and total cell number of blastocysts after PA (18% and 68.9, 15% and 85.1 vs. 12% and 59.5, respectively) compared with controls. Although the 25 μM group observed significantly lower blastocyst formation rates after PA (27.6% vs. 23.2%) than control group, the 5 μM group showed significantly increased blastocyst formation rates after PA (37.2% vs. 23.2%) compared to the control group. Furthermore, the 5 μM group measured significantly increased blastocyst formation rates (20.7% vs. 8.6%) and total cell number after IVF (88.3±1.5 vs. 58.0±3.6) compared to the control group. The treatment of 5 μM ECG during IVM affectively improved the porcine embryonic developmental competence by regulating intracellular oxidative stress during IVM.

본 연구는 낙지다리(Penthorum chinense Pursh) 추출물을 기능성 화장품소재로 개발하기 위해 (−)‑epicatechin gallate를 지표성분으로 선정하고, 품질관리를 위해 High Performance Liquid Chromatography (HPLC)를 이용하여 분석법을 개발하였다. 분석에 사용된 칼럼은 Unison US-C18 (4.6 × 250 mm, 5 μm, Imtakt, USA)을 사용하여 0.05% (v/v) trifluoroacetic acid (TFA)와 메탄올을 이동상 조건으로 컬럼 온도는 30 ℃ 에서 유속은 1.0 mL/min 로 검출파장은 280 nm에서 검출하였다. International Conference on Harmonization (ICH) 가이드라인(version 4, 2005)을 근거로 하여 특이성, 직선성, 정밀성, 정확성, 검출한계 및 정량한계를 분석하여 분석방법을 검증하였다. 검출한계 및 정량한계는 각각 0.11 mg/mL 및 0.33 mg/mL로 나타났으며, 검량곡선은 상관계수값이 0.9999로 양호한 직선성을 보였고, 정밀성 분석결과 도 0.6% 이하로 확인하였다. 또한, 회수율은 99.51 ~ 101.92% 범위로 정확성이 있음을 알 수 있다. 따라서, 본 분석법은 낙지다리 추출물의 지표성분의 분석법은 적합한 시험법임이 검증되었다.