【摘要】：本文以龍眼加工廢棄物龍眼核為研究對象,對龍眼核主要化學(xué)成分進行了測定;通過不同極性溶劑萃取、大孔樹脂結(jié)合Sephadex LH-20凝膠對龍眼核多酚進行分離純化,采用高效液相色譜(HPLC)和高效液相-電噴霧質(zhì)譜(HPLC-ESI-MS)初步分析了龍眼核多酚的主要組成成分;在此基礎(chǔ)上對龍眼核多酚各級組分進行了抗氧化、抑菌活性的初步研究,同時探究了龍眼核多酚對小鼠燙傷愈合的影響,主要結(jié)果如下:(1)主要化學(xué)成分分析結(jié)果表明,龍眼核中的淀粉含量最高,其次是蛋白質(zhì)和還原糖,粗纖維、脂肪含量相對較低,且含有較多的多酚類物質(zhì),故可作為一種良好的營養(yǎng)源。(2)液-液萃取法分離龍眼核多酚的研究結(jié)果表明,乙酸乙酯萃取物的多酚含量最高,達到693.51±9.52 mg/g;在不同自由基體系中,4種萃取物均對?O2-的清除能力最弱,但對ABTS?+、DPPH?、?OH均表現(xiàn)出較強的清除效果,其中乙酸乙酯萃取物的抗氧化活性最強;除水層剩余物對沙門氏菌和大腸桿菌均無明顯抑制作用外,4種萃取物對沙門氏菌、大腸桿菌、金黃色葡萄球菌均具有不同強度的抑制作用,其中乙酸乙酯萃取物的抑菌活性最為突出。(3)通過靜態(tài)試驗比較了7種大孔吸附樹脂對龍眼核多酚的吸附及解吸效果,篩選出AB-8為最佳樹脂,其吸附率和解析率分別為69.65%和94.47%。龍眼核多酚的最佳純化工藝條件為:p H值3.5,上樣濃度2 mg/m L,上樣流速2 m L/min,洗脫劑70%乙醇,洗脫流速2 m L/min。在此條件下,龍眼核多酚的回收率為75.65%,產(chǎn)品得率為4.93%,純度為65.84%。(4)采用不同濃度的乙醇溶液對AB-8樹脂上吸附的龍眼核多酚進行梯度洗脫,發(fā)現(xiàn)各洗脫部分的多酚組成和含量存在一定差異,表明AB-8樹脂可用于龍眼核粗多酚的初步分離;體外抗氧化、抑菌實驗結(jié)果表明,相同多酚濃度下,30%乙醇洗脫物的抗氧化和抑菌活性總體上均優(yōu)于其他洗脫部分。Sephadex LH-20凝膠對30%乙醇洗脫物的分離效果較好,能獲得5個組分,經(jīng)HPLC和HPLC-ESI-MS初步分析表明,龍眼核多酚的主要活性成分為柯里拉京、鞣花酸等化合物。(5)通過創(chuàng)面愈合率、生化指標(biāo)測定及組織病理學(xué)分析來評價創(chuàng)面的愈合質(zhì)量,結(jié)果表明:中、高劑量組在燙傷愈合前期對創(chuàng)面收縮有一定的促進作用,之后不同時間點的愈合率與生理鹽水組相比無明顯差異;一定劑量的龍眼核多酚溶液對創(chuàng)面組織和血清中蛋白質(zhì)和羥脯氨酸的含量變化有一定影響;在炎性細(xì)胞浸潤、成纖維細(xì)胞和上皮細(xì)胞增生、毛細(xì)血管生成、肉芽組織形成等方面對組織病理學(xué)進行評價,龍眼核多酚各劑量組與生理鹽水組比較均無明顯差異。因此,龍眼核多酚對小鼠深Ⅱ度燙傷愈合有一定的影響,但無明顯促進作用。
[Abstract]:In this paper, the main chemical constituents of longan nucleus were determined, and the polyphenols in longan nucleus were separated and purified by macroporous resin and Sephadex LH-20 gel with different polarity solvent extraction. High performance liquid chromatography (HPLC) and high performance liquid phase electrospray ionization mass spectrometry (HPLC-ESI-MS) were used to analyze the main components of polyphenols in longan nucleus. The main results were as follows: (1) the main chemical component analysis showed that the starch content in longan nucleus was the highest, followed by protein and reducing sugar, crude fiber and fat content was relatively low. It can be used as a good nutrient source. (2) the results of separation of polyphenols from longan nucleus by liquid-liquid extraction showed that the content of polyphenols in ethyl acetate extract was the highest. In different free radical systems, the four extracts had the weakest scavenging ability to? O2-, but they showed strong scavenging effects on ABTS? and DPPH?,?OH, among which ethyl acetate extract had the strongest antioxidant activity. Except that the water layer residue had no obvious inhibition on Salmonella and Escherichia coli, the four extracts had different inhibitory effects on Salmonella, Escherichia coli and Staphylococcus aureus. The antibacterial activity of ethyl acetate extract was the most prominent. (3) the adsorption and desorption of polyphenols in longan nucleus by seven macroporous adsorption resins were compared by static test. The best resin was AB-8, the adsorption rate and resolution rate were 69.65% and 94.47% respectively. The optimum purification conditions of polyphenols in longan nucleus were as follows: p H value 3.5, sample concentration 2 mg/m / L, sample flow rate 2 mL / min, eluant 70% ethanol, elution rate 2 mL / min. Under these conditions, the recovery of polyphenols in longan nucleus was 75.65, the yield of the product was 4.93 and the purity was 65.84. (4) gradient elution of longan polyphenols adsorbed on AB-8 resin was carried out with different concentration of ethanol solution. It was found that there were some differences in the composition and content of polyphenols among the eluting parts, which indicated that AB-8 resin could be used for the primary separation of crude polyphenols in longan nucleus, and the results of antioxidation and bacteriostatic experiments in vitro showed that, At the same concentration of polyphenols, the antioxidant and bacteriostatic activities of 30% ethanol elution were better than those of other elution parts. Sephadex LH-20 gel had a better separation effect on 30% ethanol elution, and five components could be obtained. The results of HPLC and HPLC-ESI-MS analysis showed that: 1. The main active components of polyphenols in longan nucleus are Corilagin, tannic acid, etc. (5) the wound healing quality is evaluated through wound healing rate, biochemical index measurement and histopathological analysis. In the high dose group, the wound contraction was promoted at the early stage of scald healing, but the healing rate at different time points was not significantly different from that in the saline group. The changes of protein and hydroxyproline contents in wound tissue and serum were affected by a certain dose of longan nucleus polyphenol solution, and in inflammatory cell infiltration, fibroblast and epithelial cell proliferation, capillary angiogenesis, The histopathology was evaluated in granulation tissue formation and so on. There was no significant difference between each dosage group and normal saline group in the evaluation of histopathology of longan nucleus polyphenols. Therefore, polyphenols in longan nucleus have a certain effect on the healing of deep second degree scald in mice, but have no obvious promoting effect.
【學(xué)位授予單位】：華南農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：R151;TS209

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