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中国精品科技期刊2020
陈艳萍,贺菊萍,刘意,等. 超声波-微波辅助提取杜仲叶多糖工艺优化及其体外抗凝血活性分析[J]. 食品工业科技,2023,44(17):202−211. doi: 10.13386/j.issn1002-0306.2022100189.
引用本文: 陈艳萍,贺菊萍,刘意,等. 超声波-微波辅助提取杜仲叶多糖工艺优化及其体外抗凝血活性分析[J]. 食品工业科技,2023,44(17):202−211. doi: 10.13386/j.issn1002-0306.2022100189.
CHEN Yanping, HE Juping, LIU Yi, et al. Optimization of Ultrasonic-Microwave Assisted Extraction of Polysaccharides from Eucommia ulmoides Leaves and Its Anticoagulant Activity in Vitro[J]. Science and Technology of Food Industry, 2023, 44(17): 202−211. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2022100189.
Citation: CHEN Yanping, HE Juping, LIU Yi, et al. Optimization of Ultrasonic-Microwave Assisted Extraction of Polysaccharides from Eucommia ulmoides Leaves and Its Anticoagulant Activity in Vitro[J]. Science and Technology of Food Industry, 2023, 44(17): 202−211. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2022100189.

超声波-微波辅助提取杜仲叶多糖工艺优化及其体外抗凝血活性分析

Optimization of Ultrasonic-Microwave Assisted Extraction of Polysaccharides from Eucommia ulmoides Leaves and Its Anticoagulant Activity in Vitro

  • 摘要: 为开发我国丰富的杜仲叶资源,研究超声波-微波辅助提取杜仲叶多糖的最优工艺条件及其理化性质和体外抗凝血活性。首先通过单因素实验确定超声波功率、提取温度、微波功率、料液比、提取时间等影响多糖得率因素的范围,然后通过Plackett-Burman试验筛选关键影响因素,再采用Box-Behnken试验对工艺条件进行优化,分析了所得杜仲叶精制多糖的分子量、单糖组成等理化性质及活化部分凝血酶原时间(APTT)、凝血酶原时间(PT)、凝血酶时间(TT)等抗凝血指标。结果表明,料液比、提取时间、提取温度为关键影响因素,最优提取工艺条件为超声波功率130 W、提取温度49 ℃、微波功率200 W、料液比1:30(g:mL)、提取时间20 min。在此条件下,杜仲叶多糖的实际得率为4.02%±0.03%,与理论得率4.08%接近。所得杜仲叶精制多糖的重均分子质量(Mw)为1653 kDa,单糖组成及摩尔比为果糖37.3%、葡萄糖35%、N-乙酰-D氨基葡萄糖14.6%、半乳糖8.6%、阿拉伯糖4.4%。分析结果显示,杜仲叶多糖为β型酸性多糖。与阴性对照相比,杜仲叶多糖能极显著延长APTT(P<0.01),而当杜仲叶多糖浓度为8 mg/mL时,PT、TT亦能极显著延长(P<0.01),说明杜仲叶多糖主要通过内源性途径影响凝血系统,同时也能够通过外源性、共同途径来影响凝血过程。此研究可为我国丰富的杜仲叶资源的高值化开发利用提供理论依据和技术参考。

     

    Abstract: To develop the rich resources of Eucommia ulmoides leaves in China, the optimal conditions of extracting polysaccharides from Eucommia ulmoides leaves by ultrasonic and microwave, the physicochemical properties and the anticoagulant activity in vitro of the extracted polysaccharides were studied. Firstly, the ranges of factors affecting the yield of polysaccharide, such as ultrasonic power, extraction temperature, microwave power, ratio of solid to liquid, extraction time, were determined by single factor experiment. The key factors were further screened by Plackett­Burman test, and the Box-Behnken test was used to optimize the process conditions. Finally, the physicochemical properties, such as molecular weight and monosaccharide composition, and the anticoagulant indices, such as activated partial prothrombin time (APTT), prothrombin time (PT), thrombin time (TT) of the refined Eucommia ulmoides leaf polysaccharides, were analyzed. The results showed that the ratio of solid to liquid, the extraction time and the extraction temperature were the key factors. The optimal extraction conditions were the ultrasonic power of 130 W, the extraction temperature of 49 ℃, the microwave power of 200 W, the solid-liquid ratio of 1 to 30 (g:mL) and the extraction time of 20 min. Under these conditions, the actual yield of Eucommia ulmoides leaf polysaccharides was 4.02%±0.03%, close to the theoretical yield of 4.08%. The weight-average molecular mass (Mw) of the refined polysaccharide was 1653 kDa, and the composition and molar ratio of monosaccharide were 37.3% fructose, 35% glucose, 14.6% n-acetyl-d-glucosamine, 8.6% galactose and 4.4% arabinose. The results showed that the polysaccharides of Eucommia ulmoides leaves was β-type acidic polysaccharide. Compared with the negative control, the Eucommia ulmoides polysaccharides significantly increased APTT (P<0.01), and significantly increased PT and TT (P<0.01) at the concentration of 8 mg/mL, indicating that it mainly affected coagulation system through endogenous pathway, could also affect the coagulation process through exogenous and common pathways. This study could provide theoretical basis and technical reference for the high-value utilization of Eucommia ulmoides leaves in China.

     

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