本文關(guān)鍵詞： 藥桑葉 HPLC-MS-MS 黃酮類化合物 抗氧化活性　出處：《塔里木大學(xué)》2016年碩士論文　論文類型：學(xué)位論文

【摘要】：本研究以新疆特色植物藥桑葉為原材料,對(duì)其化學(xué)成分、藥桑葉中總黃酮提取工藝、醇提物乙酸乙酯和正丁醇相黃酮類化合物進(jìn)行分離鑒定及黃酮單體間體外抗氧化活性等進(jìn)行研究。1、采用系統(tǒng)化學(xué)預(yù)試法對(duì)藥桑葉所含化學(xué)成分進(jìn)行初步研究。結(jié)果發(fā)現(xiàn)藥桑葉中含有:糖類、酚類、有機(jī)酸類、黃酮類、揮發(fā)油類、生物堿類、甾體類、萜類等多種化學(xué)成分。2、在單因素的基礎(chǔ)上采用正交實(shí)驗(yàn)方法,研究藥桑葉中黃酮類化合物的提取條件,建立了適宜的提取工藝:料液比1:25、超聲時(shí)間45 min、乙醇濃度80%、回流時(shí)間60 min。該工藝條件下總黃酮的得率為2.24%。3、采用色譜分離技術(shù)從藥桑葉分離出11個(gè)黃酮類化合物,通過核磁共振(NMR)和高效液相-串聯(lián)質(zhì)譜(HPLC-MS-MS)鑒定其結(jié)構(gòu)為蘆丁(1)、異槲皮苷(2)、金絲桃苷(3)、山奈酚-3-O-β-D-吡喃葡萄糖(4)、草質(zhì)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�；�)-吡喃半乳糖苷-7-O-α-吡喃阿拉伯糖苷(6)、紅景天素(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)首次從新疆藥桑葉中分離得到。4、以自由基清除率為評(píng)價(jià)指標(biāo),采用1,1-二苯基-2-三硝基苯肼(DPPH)和2,2-聯(lián)氮基-雙-(3-乙基苯并噻唑啉-6-磺酸)二氨鹽(ABTS)自由基清除法對(duì)乙酸乙酯相分離得到的單體化合物進(jìn)行抗氧化活性評(píng)價(jià)。結(jié)果表明,從乙酸乙酯相分離得到4個(gè)單體化合物,分別為蘆丁、異槲皮苷、金絲桃苷、山奈酚-3-O-β-D-吡喃葡萄糖,對(duì)DPPH自由基清除率的半數(shù)抑制濃度(IC50)分別為0.0083 mg/mL、0.0237 mg/mL、0.0060 mg/mL、0.0510 mg/mL,對(duì)ABTS自由基清除率IC50分別為0.0172 mg/mL、0.0210 mg/mL、0.0120mg/mL、0.0870 mg/mL。4個(gè)單體化合物均具有很強(qiáng)的抗氧化活性,且抗氧化活性順序?yàn)?金絲桃苷蘆丁異槲皮苷山奈酚-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,評(píng)價(jià)各黃酮單體的抗氧化活性,并篩選出抗氧化活性較好的四種單體化合物—蘆丁、金絲桃苷、草質(zhì)素-7-O-(3″-O-β-D-葡萄糖基)-α-L-鼠李糖苷、紅景天素,進(jìn)行抗氧化協(xié)同作用研究,發(fā)現(xiàn)紅景天素在清除自由基方面和其他化合物具有拮抗作用,其他三種黃酮單體化合物相互結(jié)合具有協(xié)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-偽 -L-rhamnoside, quercetin -3-O- 尾 -pyrrolidene 2 "-Oacetyl. Galactoside -7-O- 偽 -pyran arabinopyranoside 6o, salidrosin 7, kaempferol -3-O- 尾 -buten2 "-O- 偽 -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 偽 -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.
【學(xué)位授予單位】：塔里木大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R29
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本文編號(hào)：1516986