香茹抗蝕骨細胞分化活性之萃取條件最適化研究

本研究以回應曲面法 (Response surface methodology; 簡稱RSM)來優化香茹草最佳萃取條件，其操作參數包括：萃取時間、攪拌速率、酒精濃度、溫度及固液比，目標函數為：萃取回收率、抗蝕骨細胞活性及結合此兩個參數之期望函數。抗蝕骨細胞以抗酒石酸磷酸酶抑制濃度(IC)來檢測其蝕骨細胞被香茹萃取物抑制的程度。本研究首先針對抗蝕骨細胞活性利用部分因子實驗設計及陡升實驗來搜尋最佳操作範圍。隨後，以陡升實驗所找到的最佳條件作為中心點進行中心混成實驗；並將所得實驗數據分別以統計迴歸分析得到三個二階方程式，據此分別預測出最佳萃取操作條件。經因子影響效應分析發現，針對此三個目標函數之最大影響因子皆為酒精濃度。當以萃取率作為目標函數時，可得最佳萃取率(20.3%)，但抗骨質疏鬆之效果較不佳(IC = 77.0 g/ml)；若以抗骨質疏鬆活性作為目標函數時，其抑制活性最佳(IC = 46.0 g/ml)，但萃取率則降低至12.1%。在以結合此兩個參數之期望函數作為目標函數時，可得萃取率為19.9%，抗骨質疏鬆抑制活性IC = 63.1 g/ml之折衷萃取條件，可兼顧生產成本及保健活性。將此萃取物與未經最佳化的萃取物相比，在濃度為25 μg/ml時，抗酒石酸磷酸酶活性之百分比值由80.0%大幅下降至47.1%，且萃取率由14.7% 提升至19.9%。

關鍵字

香茹；抗蝕骨細胞生成；骨質疏鬆；萃取條件最適化；回應曲面

並列摘要

Response surface methodology (RSM) was applied to optimize the extraction conditions for anti-osteogenesis activity of Glossogyne tenuifolia. Five operation factors were investigated, including extraction time, agitation rate, alcohol concentration, temperature and solid-solution ratio (SSR). The extraction yield, the anti-osteogenesis activity and the desirability that combines these two indexes were the three objective functions. The anti-osteogenesis activity of G. tenuifolia extract was assayed by tartrate resistant acid phosphatase (TRAP) activity and expressed by the inhibition concentration (IC). At first, the experiments of fractional factorial design (FFD) and the path of steepest ascent were performed to search for the optimum operation area for TRAP activity. Then, the central composite designed (CCD) experiments were carried out to search the optimal extraction conditions. Three second-order equations were obtained by regression via each of the three objective functions, and the equations were used to predict the optimum points. The factor analysis from these equations showed that the most important factor on each of these three objective indexes was all the alcohol concentration. When the extraction yield was the objective function, the highest yield (20.3%) was obtained, but the anti-osteogenesis activity was low (IC = 77.0 g/ml). When the anti-osteogenesis activity was the objective function, the highest anti-osteogenesis activity (IC = 46.0 g/ml) was obtained, but the extraction yield was low (12.1%). If the desirability function was used as the optimization index, a compromise result (yield = 19.9%, IC = 63.1 g/ml), that may take into account the production cost and health activity, was obtained. To compare this extract with a non-optimized extract, the TRAP activity decreased from 80% to 47.1% at a dose of 25 μg/ml, and the extraction yield increased from 14.7% to 19.9%.

並列關鍵字

Glossogyne tenuifolia ； anti-osteogenesis ； osteoporosis ； optimization of extraction conditions ； response surface methodology

參考文獻

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香茹抗蝕骨細胞分化活性之萃取條件最適化研究

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