Optimization of Extraction Conditions for Bakkenolide B from the Leaves of Petasites japonicus by Using Response Surface Methodology

Optimal conditions for extraction of bakkenolide B from Petasites japonicus leaves were determined by using response surface methodology. A second-order Box-Behnken design representing three extraction temperatures (80, 100, $120^{\circ}C$), three extraction times (30, 45, 60 min), and three solvent pH's (5, 7, 9) was executed. The efficiency of the extraction conditions was defined using the ${\beta}$-hexosamidase assay by comparing both the bakkenolide B content and its anti-allergic activity expressed as extract inhibition on degranulation. The response surface plot described for the bakkenolide B content showed that the maximum content was predicted as 121.6 ${\mu}g/g$ with extraction conditions of $127.1^{\circ}C$, 46.6 min, and pH 7.7. Extraction temperature and time were important factors in determining bakkenolide B content. Using regression analysis, correlation between the inhibition effect of mast cell degranulation and bakkenolide B content was found to be low.

반응표면분석법에 의한 머위(Petasites japonicus)의 bakkenolide B추출공정 최적화

머위(Petasites japonicus) 잎에서 알레르기성 천식 억제 효능이 있는 bakkenolide B의 최적 추출공정을 반응표면 분석법을 이용하여 결정하였다. Box-Behnken design에 따라 설정한 추출온도(80, 100, $120^{\circ}C$), 추출시간(30, 45, 60 min) 및 추출용매의 pH (5, 7, 9)를 독립변수로 하고, 추출물의 특성 즉, bakkenolide B의 함량과 항알러지의 측정지표인 비만세포의 탈과립반응 억제효과를 종속변수로 하여 분석을 실시하였다. Bakkenolide B의 추출함량은 추출온도, 시간, pH의 유의적인 영향을 받았으며, 추출온도 $127.1^{\circ}C$, 시간 46.6분, 추출용매의 pH 7.7에서 최대치 121.6 ${\mu}g/g$이 예측되었다. 비만세포의 탈과립반응 억제효과의 회귀분석 결과는 유의성이 인정되지 않았고 낮은 상관계수를 보였다.