新疆药桑叶黄酮类化合物提取分离及协同抗氧化活性研究

发布时间：2018-02-17 02:17

  本文关键词： 药桑叶 HPLC-MS-MS 黄酮类化合物 抗氧化活性　出处：《塔里木大学》2016年硕士论文　论文类型：学位论文

【摘要】：本研究以新疆特色植物药桑叶为原材料,对其化学成分、药桑叶中总黄酮提取工艺、醇提物乙酸乙酯和正丁醇相黄酮类化合物进行分离鉴定及黄酮单体间体外抗氧化活性等进行研究。1、采用系统化学预试法对药桑叶所含化学成分进行初步研究。结果发现药桑叶中含有:糖类、酚类、有机酸类、黄酮类、挥发油类、生物碱类、甾体类、萜类等多种化学成分。2、在单因素的基础上采用正交实验方法,研究药桑叶中黄酮类化合物的提取条件,建立了适宜的提取工艺:料液比1:25、超声时间45 min、乙醇浓度80%、回流时间60 min。该工艺条件下总黄酮的得率为2.24%。3、采用色谱分离技术从药桑叶分离出11个黄酮类化合物,通过核磁共振(NMR)和高效液相-串联质谱(HPLC-MS-MS)鉴定其结构为芦丁(1)、异槲皮苷(2)、金丝桃苷(3)、山奈酚-3-O-β-D-吡喃葡萄糖(4)、草质素-7-O-(3″-O-β-D-葡萄糖基)-α-L-鼠李糖苷(5)、槲皮素-3-O-β-(2″-O-乙酰基)-吡喃半乳糖苷-7-O-α-吡喃阿拉伯糖苷(6)、红景天素(7)、山奈酚-3-O-β-(2″-O-α-L-吡喃鼠李糖基)-葡萄糖醛酸苷(8)、鼠李素-3-O-半乳糖苷(9)、槲皮素-3′-O-β-D-葡萄糖苷(10)、槲皮素-3-O-β-D-吡喃葡萄糖苷(11),其中化合物(3)~(10)首次从新疆药桑叶中分离得到。4、以自由基清除率为评价指标,采用1,1-二苯基-2-三硝基苯肼(DPPH)和2,2-联氮基-双-(3-乙基苯并噻唑啉-6-磺酸)二氨盐(ABTS)自由基清除法对乙酸乙酯相分离得到的单体化合物进行抗氧化活性评价。结果表明,从乙酸乙酯相分离得到4个单体化合物,分别为芦丁、异槲皮苷、金丝桃苷、山奈酚-3-O-β-D-吡喃葡萄糖,对DPPH自由基清除率的半数抑制浓度(IC50)分别为0.0083 mg/mL、0.0237 mg/mL、0.0060 mg/mL、0.0510 mg/mL,对ABTS自由基清除率IC50分别为0.0172 mg/mL、0.0210 mg/mL、0.0120mg/mL、0.0870 mg/mL。4个单体化合物均具有很强的抗氧化活性,且抗氧化活性顺序为:金丝桃苷芦丁异槲皮苷山奈酚-3-O-β-D-吡喃葡萄糖。5、采用1,1-二苯基-2-三硝基苯肼(DPPH)自由基清除法,评价各黄酮单体的抗氧化活性,并筛选出抗氧化活性较好的四种单体化合物—芦丁、金丝桃苷、草质素-7-O-(3″-O-β-D-葡萄糖基)-α-L-鼠李糖苷、红景天素,进行抗氧化协同作用研究,发现红景天素在清除自由基方面和其他化合物具有拮抗作用,其他三种黄酮单体化合物相互结合具有协同增效作用。
[Abstract]:In this study, the chemical constituents and extraction process of total flavonoids from mulberry leaves of Xinjiang characteristic plant were studied. Ethanol extract ethyl acetate and flavonoids in n-butanol phase were isolated and identified, and the antioxidant activity between flavonoids was studied in vitro. 1. The chemical constituents of mulberry leaves were studied by systematic chemical pretest. Fruit found that mulberry leaves contain: sugar, Phenols, organic acids, flavonoids, volatile oils, alkaloids, steroids, terpenes, and other chemical constituents. The extraction conditions of flavonoids in mulberry leaves were studied by orthogonal experiment on the basis of single factor. The optimum extraction process was established: the ratio of material to liquid was 1: 25, ultrasonic time was 45 min, ethanol concentration was 80 and reflux time was 60 min. The yield of total flavonoids was 2.24.3. 11 flavonoids were isolated from mulberry leaves by chromatographic separation. The structures were identified by NMR and HPLC-MS-MS as rutin 1, isoquercetin 2, hyperoside 3, kaempferol-3-O- 尾-D- glucopyranoside 4, oxalin-7-O-3 "-O7-O-3" -O- 尾 -glucosyl-伪 -L-rhamnoside, quercetin -3-O- 尾 -pyrrolidene 2 "-Oacetyl. Galactoside -7-O- 伪 -pyran arabinopyranoside 6o, salidrosin 7, kaempferol -3-O- 尾 -buten2 "-O- 伪 -L-pyrrolidene-glucuronoside 8, rhamnetin -3-O- galactosine 9, quercetin -3O- O- 尾 -D- glucoside 10, quercetin -3-O- 尾 -Dpyranoside 11, in which rhamnose -3-O- galactosine 9, quercetin -3O- O- 尾 -D- glucoside, quercetin -3-O- 尾 -Dpyranoside 11. 4 was isolated from mulberry leaves in Xinjiang for the first time, and the free radical scavenging rate was used as the evaluation index. The antioxidant activity of monomers separated from ethyl acetate was evaluated by the radical scavenging method of 1-diphenyl-2-trinitrophenylhydrazine (DPPHH) and 2-diazo-2-diazo-bis-3-ethylbenzothiazoline-6-sulfonic acid) (ABTSs). Four monomers were isolated from ethyl acetate phase. They were rutin, isoquercetin, hyperoside, kaempferol -3-O- 尾 -Dpyran glucose. The median inhibitory concentration (IC50) on the scavenging rate of DPPH free radical was 0.0083 mg / mL 0.0237 mg / mL 0.060 mg / mL 0.0510 mg / mL, and 0.0172 mg / mL IC50 0.0210 mg / mL 0.0210 mg 路mL ~ (-1) ~ 0.0120 mg / mL ~ 0.0870 mg/mL.4, respectively. The order of antioxidant activity was as follows: hyperoside rutin isoquercetin kaempferol-3-O- 尾-D- pyran glucose. 5. The antioxidant activity of each flavonoid monomer was evaluated by the scavenging method of 1-diphenyl-2-trinitrophenylhydrazine (DPPHs) free radical. Four kinds of monomer compounds with good antioxidant activity, rutin, hyperoside, herbicide -7-O-f3 "-O- 尾 -Dglucosyl 伪 -L-rhamnoside and salidrogenin, were selected to study the synergistic effect of antioxidation. It was found that salidrosine had antagonistic effect on scavenging free radical with other compounds, and the other three flavonoids had synergistic effect.
【学位授予单位】：塔里木大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R29
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本文编号：1516986