壓縮膨發處理對五葉松多酚萃取及其抗氧化活性之評估研究

Evaluation of compressional-puffing pretreatment for antioxidant phenolic compounds extraction from Pinus morrisonicola

江拜石(Pai-Shih Chiang)

指導教授 ： 李篤中

國立臺灣大學/工學院/化學工程學研究所/碩士(2016年)

https://doi.org/10.6342/NTU201601000

摘要

臺灣五葉松在許多研究中已被指出具有生物益性且富含高含量的抗氧化成分；壓縮膨發法是一種常見的食物加工處理方法，操作簡單亦環保，可有效的破壞食物的外殼結構，因此本研究旨在藉由對五葉松進行壓縮膨發預處理來提升其溶劑萃取效率並對製程最佳化。除了多酚的基本定量分析，本研究利用掃描式電子顯微鏡(SEM)和水銀測孔儀(MIP)分析比較五葉松在預處理前後的形態變化；並利用高效液相層析儀(HPLC)和傅立葉紅外光譜儀(FTIR)對五葉松萃取物進行定性分析。
實驗結果指出壓縮膨發法可以有效地破壞五葉松的細胞壁，並在表面形成奈米孔洞，再將數據以二次反應方程式回歸，得出預處理對表面的破壞可有效加速初始萃取速率。另外，藉由HPLC和FTIR的分析，得到五葉松的主要成份為兒茶素，進一步的分析更得到為了有效的萃取出兒茶素，溶劑萃取應控制在攝氏70度以下，以避免多酚化合物的熱分解及離子化。
除了預處理，本研究亦利用反映曲面法對溶劑萃取五葉松中的多酚成份及其抗氧化能力進行最佳化實驗。反映曲面法得到二次回歸式其p value (<0.01)跟R2 (>0.98)都顯示出實驗結果得出的模型具有良好的回歸，而模型得出的最佳萃取環境為：36.1%酒精水溶液、攝氏70度的萃取溫度和50 mL/g的溶劑物料比。最後，以反映曲面法得出的最佳萃取環境對預處理過後的五葉松進行萃取處理，所得到的萃取物出比未進行預處理的五葉松萃取物含有5倍的多酚含量及3倍的抗氧化能力，而且，跟市售的酵素產品必較亦有相當的競爭力。

關鍵字

臺灣五葉松 ； 多酚萃取 ； 抗氧化能力 ； 壓縮膨發預處理 ； 反映曲面法

並列摘要

Pinus morrisonicola has been reported bio-benefit and contains high amount of antioxidant bioactive compounds. This study is aimed to increase the extraction yield and efficiency of the antioxidant phenolic compounds from Pinus morrisonicola. A simple, environmentally friendly and reactant-saving compressional-puffing process is applied to the pretreatment of the extraction process. Scanning electron microscopy and mercury intrusion porosimetry are used to characterize the morphological changes. Also, high-performance liquid chromatography and Fourier transform infrared spectroscopy are used to analyze and characterize phenolic compounds of the pine needles extracts. The results show that compressional-puffing leads to a considerable destruction of the cell walls of the pine needles’ with the appearance of numerous nanopores. By fitting the data to a second-order kinetic extraction model has led to a 9 fold increase of the initial extraction rate. By HPLC and FTIR, the major components of the pine needles extracts are identified as catechin. The suitable extraction conditions for catechin are aqueous ethanol as solvent in a range of 25 to 50 % (v/v) while the extraction temperature is maintained less than 70 ºC in order to prevent the decomposition and ionization of phenolic compounds.
Besides the utilization of the pretreatment, the optimum extraction conditions for the phenolic compounds from pine needles are determined by using response surface methodology with a central-composite design. The effects of ethanol concentration, extraction temperature and liquid-solid ratio on the extraction yield (total flavonoids content and total phenolic content) and antioxidant activity (DPPH scavenging activity, FRAP and ABTS scavenging activity) are investigated. A quadratic model afforded a superior ‘fit’ of the experimental data with R2 > 0.98 and p value < 0.01. The optimum conditions to maximise the extraction yields and antioxidant activity of the pine needles extracts are: 36.1 % (v/v) of ethanol, 70 ºC and 50 mL/g of liquid-solid ratio. Under these conditions the extraction of pine needles, pretreated by compressional-puffing, is conducted and the result show a 5 fold increase in extraction yield of phenolic compounds and a 3 fold increase in antioxidant activity. In addition, the pine needles extracts are also compared with commercial enzyme products and show competitiveness in both total phenolic content and antioxidant activity.

並列關鍵字

Pinus morrisonicola ； Phenolic compounds extraction ； Antioxidant activity ； Compressional-puffing pretreatment ； catechin ； Response surface methodology

參考文獻

[2] Z. M. Kong, Z. T. Liu, and B. Y. Ding, “Study on the Antimutagenic Effect of Pine Needle Extract,” Mutation Research Letters, vol. 347, no. 3, pp. 101–104, Aug. 1995.

[3] G.-C. Yen, P.-D. Duh, D.-W. Huang, C.-L. Hsu, and T. Y.-C. Fu, “Protective effect of pine (Pinus morrisonicola Hay.) needle on LDL oxidation and its anti-inflammatory action by modulation of iNOS and COX-2 expression in LPS-stimulated RAW 264.7 macrophages,” Food and Chemical Toxicology, vol. 46, no. 1, pp. 175–185, Jan. 2008.

[4] A. Schäfer, Z. Chovanová, J. Muchová, K. Sumegová, A. Liptáková, Z. Ďuračková, and P. Högger, “Inhibition of COX-1 and COX-2 activity by plasma of human volunteers after ingestion of French maritime pine bark extract (Pycnogenol),” Biomedicine & Pharmacotherapy, vol. 60, no. 1, pp. 5–9, Jan. 2006.

[5] H. Song, R. Yang, W. Zhao, W. Katiyo, X. Hua, and W. Zhang, “Innovative Assistant Extraction of Flavonoids from Pine ( Larix olgensisHenry) Needles by High-Density Steam Flash-Explosion,” J. Agric. Food Chem., vol. 62, no. 17, pp. 3806–3812, Apr. 2014.

[6] M.-C. Cheng, W.-H. Chang, C.-W. Chen, W.-W. Li, C.-Y. Tseng, and T.-Y. Song, “Antioxidant Properties of Essential Oil Extracted from Pinus morrisonicola Hay Needles by Supercritical Fluid and Identification of Possible Active Compounds by GC/MS,” Molecules, vol. 20, no. 10, pp. 19051–19065, Oct. 2015.