本文選題：樹莓 + 黃酮��； 參考：《中北大學(xué)》2017年碩士論文

【摘要】：樹莓(Rubus idaeus)為多年生小灌木,兼木草兩性。其果實營養(yǎng)成分和生物活性成分含量豐富,被美國人稱為“生命之果”。本研究利用大孔吸附樹脂法對樹莓干果粗提物進行純化,研究了抗生素與提取物的配伍抑菌作用,測定了水溶、醇溶、煮沸和高溫高壓四種不同條件下樣品的總多酚和總黃酮含量,并利用HPLC鑒定了其中的主要黃酮成分和含量變化,最后對高溫高壓和水溶條件下樣品的抗氧化活性和抑菌活性進行分析。結(jié)果表明:(1)通過靜態(tài)吸附和解吸試驗,確定XDA-6為純化樹莓粗提物的最佳樹脂,吸附率為83.7%,解吸率為83.0%;結(jié)合靜態(tài)與動態(tài)吸附解吸試驗,確定最佳純化條件為:原液以6 BV/h的流速上樣(大孔樹脂的吸附濃度為14.50 mg/mL),60%乙醇以4 BV/h的流速進行洗脫,洗脫液用量約為5 BV。純化后黃酮含量35.8%較粗提物黃酮含量16.2%提高了1.21倍(以下將純化前的提取物記為“粗黃酮”,純化后的記為“純黃酮”)。對大腸桿菌和金黃色葡萄球菌的最小抑菌濃度(Minimal Inhibitory Concentration,MIC)由粗黃酮的8 mg/m L下降到純黃酮的2 mg/mL;對小麥赤霉菌和棉花枯萎菌的菌絲抑制率表現(xiàn)為低濃度下提高多,高濃度下提高少的趨勢,但是真菌菌絲抑制效率的提高遠(yuǎn)遠(yuǎn)低于對細(xì)菌的抑制效率的提高幅度。(2)通過研究四種抗生素與黃酮的配伍抑菌效果,得出四種抗生素與黃酮配伍均有協(xié)同抑菌效果。氨芐青霉素與黃酮配伍抑制金黃色葡萄球菌和大腸桿菌兩種實驗菌的協(xié)同效果不明顯,對沙門氏菌有較好的協(xié)同抑菌效果,且青霉素的抑菌濃度降至原濃度的1/10。而紅霉素結(jié)果與氨芐青霉素正好相反,與黃酮配伍抑制金黃色葡萄球菌和大腸桿菌兩種實驗菌的協(xié)同效果明顯,而對沙門氏菌協(xié)同抑菌效果不明顯。鹽酸四環(huán)素和硫酸鏈霉素與黃酮配伍抑制三種菌的協(xié)同效果均很好,FICI值均小于0.25,最低可達到0.1。(3)四種條件下樣品中總多酚和總黃酮含量的變化趨勢為:高溫高壓沸水浴乙醇水,其中以純黃酮樣品的變化最為明顯。高溫高壓后,粗黃酮和純黃酮的總多酚含量分別達到331.02 mg/g,395.82 mg/g,相對于水溶條件下的含量分別提高了50.21%,67.07%;總黃酮含量分別增加73.23 mg,166.73 mg,較水溶條件提高了38.72%,86.92%。具體到化學(xué)成分,黃酮樣品中以維生素C和兒茶素為主,含有少量槲皮素和蘆丁,山奈酚含量低于檢出線。粗黃酮和純黃酮高溫高壓后,維生素C分別減少18.45 mg/g(27.26%),34.24 mg/g(35.20%);兒茶素分別增加9.29 mg/g(12.54%),56.53 mg/g(59.02%);蘆丁分別減少0.2 mg/g(57.14%),0.25 mg/g(22.32%),槲皮素和其他未標(biāo)明的峰變化量極小。(4)高溫高壓后,樣品的抗氧化能力均減弱。清除DPPH自由基實驗中,粗黃酮的IC50值由28.36μg/mL升高到38.91μg/m L;純黃酮的IC50值由22.93μg/mL升高到35.84μg/m L。清除ABTS自由基實驗中,粗黃酮的IC50值由14.07μg/m L升高到21.46μg/mL;純黃酮的IC50值由10.29μg/mL升高到21.3μg/m L。兩組實驗均說明樣品抗氧化能力降低,最高降幅超過50%。高溫高壓后,樣品的抑菌活性顯著提高。粗黃酮高溫高壓后對大腸桿菌的抑菌率提高10%左右;在2 mg/mL時,對沙門氏菌的抑菌率由83.6%上升到100%,完全抑制;對金黃色葡萄球菌的抑菌率由62.3%上升到100%,提高了60.5%。純黃酮高溫高壓后對大腸桿菌抑菌率提高了約20%;在1 mg/mL時,對沙門氏菌和金黃色葡萄球菌已完全抑制,達到了殺菌劑量級,分別提高了138%和100%。綜上所述,純化與高溫高壓條件均可有效提高提取物的總多酚和總黃酮含量,間接提高了其抑菌活性,表明熱處理過程不僅有利于樹莓提取物的保存還有利于其酚類成分的富集。黃酮與抗生素配伍抑菌時可降低抗生素的用量,為解決抗生素的濫用問題提供新的思路。
[Abstract]:The raspberry (Rubus idaeus) is a perennial shrub and the hermaphrodite of wood grass. The content of its nutritive and bioactive components is rich. It is called "the fruit of life" by the American. This study used macroporous adsorption resin to purify the crude extract of raspberry, and studied the antibacterial effect of antibiotics and extracts. The content of total polyphenols and total flavonoids in four different conditions under boiling and high temperature and high pressure, and the changes of the main flavonoids and content were identified by HPLC. Finally, the antioxidant activity and antibacterial activity of the samples under high temperature and high pressure and water soluble conditions were analyzed. The results showed that: (1) by static adsorption and desorption test, XDA-6 was determined. The best resin to purify raspberry is 83.7% and the desorption rate is 83%. Combining static and dynamic adsorption desorption experiments, the optimum purification conditions are as follows: the original solution is 6 BV/h of the flow rate (the adsorption concentration of macroporous resin is 14.50 mg/mL), 60% ethanol is eluted with 4 BV/h flow speed, and the eluant dosage is about 5 BV. after the purification of Huang Tonghan. The content of the flavonoid content of the crude extract 35.8% was 1.21 times higher than that of the crude extract (below the purified extract as "crude flavonoids", and the purified one was recorded as "pure flavonoids"). The minimum inhibitory concentration (Minimal Inhibitory Concentration, MIC) of Escherichia coli and Staphylococcus aureus (MIC) decreased from 8 mg/m L of crude flavonoids to 2 mg/mL of pure flavonoids; The mycelial inhibition rate of wheat and cotton Fusarium wilt showed a tendency to increase in low concentration and less in high concentration, but the enhancement of fungal mycelium inhibition efficiency was much lower than that of bacteria inhibition efficiency. (2) the compatibility of four antibiotics with flavonoids was studied, and the compatibility of four antibiotics with flavonoids was obtained. The synergistic effect of ampicillin with flavonoids was not obvious, and the synergistic effect of two experimental strains of Staphylococcus aureus and Escherichia coli was not obvious, and Salmonella had better synergistic bacteriostasis effect, and the inhibitory concentration of penicillin decreased to 1/10. of the original concentration, while erythromycin results were opposite to ampicillin and were compatible with flavonoids. The synergistic effect of inhibiting two kinds of experimental bacteria of Staphylococcus aureus and Escherichia coli was obvious, but the synergistic effect of Salmonella was not obvious. The synergistic effect of tetracycline hydrochloride and streptomycin sulfate with flavonoids was good, the FICI value was less than 0.25 and the lowest could reach the total polyphenols and total yellow in the samples of 0.1. (3) and four conditions. The change trend of the content of ketone was as follows: high temperature and high pressure boiling water bath ethanol water, the most obvious change of the YISHION flavone samples. After high temperature and high pressure, the total polyphenols content of the crude flavonoids and pure flavonoids reached 331.02 mg/g, 395.82 mg/g respectively. The content of the total flavonoids increased by 50.21%, 67.07%, respectively, compared with the water soluble conditions, and the total flavonoids content increased by 73.23 mg, 166.7 respectively. 3 mg, higher than water solubility, 38.72%, 86.92%. specific to chemical composition, vitamin C and catechin in flavonoids, containing a small amount of quercetin and rutin, and the content of kaempferol is lower than the detection line. After high temperature and high temperature and high pressure of crude flavonoids and pure flavonoids, vitamin C is reduced by 18.45 mg/g (27.26%), 34.24 mg/g (35.20%), and catechin increases 9.29 mg, respectively. /g (12.54%), 56.53 mg/g (59.02%); rutin decreased by 0.2 mg/g (57.14%), 0.25 mg/g (22.32%), quercetin and other unlabeled peaks. (4) the antioxidant capacity of the samples decreased after high temperature and high pressure. In the DPPH free radical scavenging experiment, the IC50 value of the crude flavonoids increased from 28.36 Mu to 38.91 u g/m L, and the IC50 value of pure flavonoids was from 22.93 mu g. In the 35.84 micron g/m L. scavenging experiment of ABTS free radical, the IC50 value of the crude flavonoids increased from 14.07 mu L to 21.46 mu g/mL, and the IC50 value of the pure flavonoids from 10.29 mu g/mL to 21.3 mu g/m L. two groups showed that the antioxidant capacity of the samples decreased and the highest decrease exceeded the high temperature and high pressure. The antibacterial activity of the samples was significantly increased. The high temperature of the crude flavonoids was high. The bacteriostasis rate of Escherichia coli increased by about 10% after high pressure. At 2 mg/mL, the bacteriostasis rate of Salmonella increased from 83.6% to 100%, completely inhibited, the bacteriostasis rate of Staphylococcus aureus increased from 62.3% to 100%, and increased by high temperature and pressure of 60.5%. pure flavonoids by 20%, and Salmonella and Salmonella at 1 mg/mL. Staphylococcus aureus was completely inhibited and reached the magnitude of bactericide, which increased by 138% and 100%., respectively. Purification and high temperature and high pressure can effectively improve the content of total polyphenols and total flavonoids in the extract, and indirectly improve its antibacterial activity. It shows that the heat treatment process is not only beneficial to the preservation of the extract of raspberry but also to its phenols. The combination of flavonoids and antibiotics can reduce the dosage of antibiotics and provide a new way to solve the problem of abuse of antibiotics.

【學(xué)位授予單位】：中北大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：TQ914.1

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