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花叶万年青功能成分提取条件优化与活性探究

花匠小妙招
2024-12-09 00:19

摘要：

为优化花叶万年青中多糖与生物碱类成分的提取工艺,并探究其多糖提取物的抗氧化活性,以及生物碱的杀虫和抑菌活性,以花叶万年青叶为材料,通过正交设计试验优化多糖与生物碱类成分的提取工艺。杀虫试验以虫体浸渍法,探究花叶万年青生物碱提取物对棉铃虫、小菜夜蛾、大麦虫的触杀效果;用滤纸片法探究花叶万年青生物碱提取物对大肠埃希菌、金黄色葡萄球菌、铜绿假单胞菌、粪肠球菌、枯草芽孢杆菌、痢疾志贺氏菌、伤寒沙门氏菌的抑菌作用;通过测定对1,1-二苯基-2-三硝基苯肼(DPPH)自由基、2,2'-联氮-双-3-乙基苯并噻唑啉-6-磺酸(ABTS)自由基、羟基自由基(·OH)清除能力和对Fe3+的还原能力,研究花叶万年青叶多糖的抗氧化活性。结果表明,花叶万年青叶多糖的最佳提取工艺为料液比1∶ 30(g·mL-1),提取次数3次,提取时间1 h,提取温度90 ℃。花叶万年青多糖的抗氧化活性研究结果显示,当多糖质量浓度为5.0 mg·mL-1时,对DPPH自由基、ABTS自由基、羟基自由基的清除率分别为51.78%、48.60%、40.86%,Fe3+还原力达到0.338 6,且随多糖质量浓度的不断增加,抗氧化活性也逐渐增强,但均低于维生素C。花叶万年青叶片生物碱的提取中各因素的影响大小为超声温度>超声时间>液料比>乙醇体积分数,最佳工艺为超声温度66 ℃,超声时间80 min,料液比1∶30(g·mL-1),乙醇体积分数90%。杀虫试验结果表明,花叶万年青生物碱提取物对3种虫的触杀效果有显著差异,处理36 h后棉铃虫、小菜夜蛾、大麦虫的死亡率分别为92.22%、96.67%、96.67%。抑菌试验结果表明:12.5 mg·mL-1生物碱提取物对不同菌种的抑制效果有所不同,对大肠埃希菌、铜绿假单胞菌抑菌活性最强,抑菌环直径可达10.95、10.65 mm;其次是粪肠球菌、金黄色葡萄球菌、枯草芽孢杆菌、痢疾志贺氏菌,抑菌环直径分别为8.00、8.20、8.85、8.25 mm;而伤寒沙门氏菌表现出不敏感或敏感度不明显。

关键词: 多糖, 生物碱, 抑菌活性, 杀虫活性, 抗氧化活性, 花叶万年青

Abstract:

In order to optimize the extraction process of polysaccharides and alkaloids from Dieffenbachia picta Lodd, and explore the antioxidant activity of its polysaccharide extract, and the insecticidal and bacteriostatic activity of alkaloids, Dieffenbachia picta was used as material, the extraction processes of polysaccharides and alkaloids were optimized by orthogonal design. Insecticidal test was carried out by the insecticidal impregnation method, to explore the contact killing effect of alkaloid extract of D. picta on Helicoverpa armigera, Plutella xylostella and Zophobas atratus; Filter paper method was used to explore the bacteriostatic effect of alkaloid extract of D. picta on Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Enterococcus faecalis, Bacillus subtilis, Shigella dysentery and Salmonella typhi. The antioxidant activity of polysaccharide from Dieffenbachia picta Lodd was studied by measuring the ability of scavenging DPPH, ABTS, hydroxyl radical (·OH) and reducing ability to Fe3+. The results showed that the optimal extraction process of polysaccharide from D. picta were the solid-liquid ratio 1∶30 (g·mL-1), extraction for 3 times, extraction time 1 h, extraction temperature 90 ℃. The antioxidant activity of polysaccharide from D. picta show that when the polysaccharide concentration was 5.0 mg·mL-1, the scavenging rates of DPPH radical, ABTS radical and hydroxyl (·OH) radical were 51.78%, 48.60% and 40.86%, respectively, and the reducing power of Fe3+ reached 0.338 6. With the continuous increase of polysaccharide concentration, the antioxidant activity also gradually increased, but they were lower than those in the control group. The influence of each factor on the extraction of alkaloids from the leaves of D. picta was ultrasonic temperature>ultrasonic time>solid-liquid ratio>ethanol volume fraction. The optimal extraction process of alkaloids from the leaves of D. picta were ultrasonic temperature 66 ℃, ultrasonic time 80 min, solid-liquid ratio 1∶ 30 (g·mL-1), ethanol volume fraction 90%. The results of the insecticidal test showed that the contact toxicity of the alkaloids from D. picta leaves to the three insects was significantly different. After 36 h of treatment, the mortality rates of H. armigera, P. xylostella and Z. atratus worm were 92.22%, 96.67% and 96.67%, respectively. The results of bacteriostatic test showed that 12.5 mg·mL-1 alkaloid extract had different inhibitory effects on different strains. It had the strongest antibacterial activity against E. coli and P. aeruginosa, and the diameter of bacteriostatic ring could reach 10.95 mm and 10.65 mm, followed by E. faecalis, S. aureus, B. subtilis and S. dysentery, the diameters of bacteriostatic rings were 8.00, 8.20, 8.85 and 8.25 mm, respectively. Salmonella typhi showed insensitive or insensitive.

Key words: polysaccharide, alkaloid, bacteriostatic acitivity, insecticidal activity, antioxidant activity, Dieffenbachia picta Lodd

中图分类号:

S567.239