桂花乙醇萃取物的抗氧化活性及主成分结构的鉴定
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摘要
桂花富含天然抗氧化成分,在天然抗氧化剂领域具有良好的开发应用潜力。本研究以制取香料浸膏后的桂花残渣为原料,以乙醇为溶剂提取桂花中的抗氧化物质,得到桂花乙醇粗提物,并对该粗提物开展溶解性能、pH稳定性和热稳定性等理化性质分析。随后我们运用硅胶柱层析技术,对该粗提物进行极性分级,并对不同分级产物开展体外抗氧化活性的比较分析。在此基础上,进一步对具有良好抗氧化活性的分级产物开展了初步的结构鉴定,结果表明:桂花乙醇萃取物在溶解度参数大于10的溶剂中有较大的溶解度;其在中性、酸性和温度低于60℃的条件下较为稳定;此乙醇粗提物经硅胶柱层析分离可以得到3个分级产物。体外抗氧化活性评价结果表明,分级产物中组分1(Frac1)具有良好的1,1-二苯基-2-三硝基苯肼(DPPH)自由基清除和还原能力,而组分3(Frac3)抑制亚油酸氧化的能力最强。高效液相色谱-质谱联用分析结果表明,Frac1中活性成分主要为黄酮甙类物质,而Frac3中的主要活性组分为双黄酮类物质。
Osmanthus fragrans is rich in natural antioxidants,which has great potential in the fields of natural antioxidants. In this study,the osmanthus residue after extracting the flavor extract was used as the raw material,and antioxidant substances in the residue were extracted with ethanol. The ethanol extract of osmanthus residue( EEOR)was obtained and characterized by solubility test,p H stability test and heat stability test,respectively. Subsequently,silica gel column chromatography technology was used to separate different fractions of EEOR by polarity difference,and their antioxidant activities in vitro were compared afterwards. Furthermore,the preliminary structural identification of fractions with good antioxidant activity was carried out. Results showed that EEOR had greater solu-bility in solvents with solubility parameters greater than 10,and it was stable and neutral,acidic pH with the temperature lower than 60 ℃. EEOR was separated by silica gel column chromatography to give three fractions. The results of antioxidant activity evaluation in vitro indicated that component 1( Frac1) had good 1,1-diphenyl-2-picrylhydrazyl( DPPH) radical scavenging and reducing ability,while component 3( Frac3) had the strongest ability to inhibit the oxidation process of linoleic acid. High performance liquid chromatography-mass spectrometry analysis showed that the active components in Frac1 were mainly flavonoids,while the main active components in Frac3 were biflavonoids.
Osmanthus fragrans is rich in natural antioxidants,which has great potential in the fields of natural antioxidants. In this study,the osmanthus residue after extracting the flavor extract was used as the raw material,and antioxidant substances in the residue were extracted with ethanol. The ethanol extract of osmanthus residue( EEOR)was obtained and characterized by solubility test,p H stability test and heat stability test,respectively. Subsequently,silica gel column chromatography technology was used to separate different fractions of EEOR by polarity difference,and their antioxidant activities in vitro were compared afterwards. Furthermore,the preliminary structural identification of fractions with good antioxidant activity was carried out. Results showed that EEOR had greater solu-bility in solvents with solubility parameters greater than 10,and it was stable and neutral,acidic pH with the temperature lower than 60 ℃. EEOR was separated by silica gel column chromatography to give three fractions. The results of antioxidant activity evaluation in vitro indicated that component 1( Frac1) had good 1,1-diphenyl-2-picrylhydrazyl( DPPH) radical scavenging and reducing ability,while component 3( Frac3) had the strongest ability to inhibit the oxidation process of linoleic acid. High performance liquid chromatography-mass spectrometry analysis showed that the active components in Frac1 were mainly flavonoids,while the main active components in Frac3 were biflavonoids.
引文
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[26]许海棠,赵彦芝,张金彦,等.响应面法提取瓶尔小草总黄酮及其抗氧化活性[J].食品科技,2017,42(2):215-220.
[27]王卫记,罗建光,孔令义. HPLC-ESI-MS~n分析酸枣仁有效部位的化学成分[J].中国中药杂志,2009,34(21):2768.
[2]丁立新,张宇,宗希明,等.桂花不同溶剂提取物对氧自由基清除作用的ESR研究[J].中国现代应用药学,2010,27(1):10-13.
[3]张璟琳,李娟,黄明泉,等. 3种桂花浸膏的挥发性物质分析[J].食品安全质量检测学报,2016,7(4):1554-1560.
[4]张雪松,裴建军,赵林果,等.不同酶处理对桂花浸膏及精油成分的影响[J].现代食品科技,2017,33(4):254-263.
[5]王丽梅,余龙江,崔永明,等.桂花黄酮的提取纯化及抑菌活性研究[J].天然产物研究与开发,2008,20(4):717-720.
[6]汪海波,靳熙茜.β-葡萄糖苷酶水解制备桂花浸膏的新工艺研究[J].食品科学,2008,29(11):324-327.
[7]靳熙茜,汪海波,王枫,等.超声波辅助提取桂花总黄酮的工艺研究[J].武汉轻工大学学报,2009,28(4):21-24.
[8]靳熙茜,汪海波.桂花总黄酮提取及其体外抗氧化性能研究[J].粮食与油脂,2009,163(11):42-45.
[9]陈培珍,马春华,刘俊劭,等.桂花精油提取工艺优化及其成分分析[J].粮食与油脂,2016,29(10):54-57.
[10]夏必帮,鲁韦韦,李蕤.桂花精油提取工艺及其抗氧化性的研究[J].氨基酸和生物资源,2016,38(1):49-52.
[11]胡春弟,张锦红,徐亮,等.桂花总黄酮的提取及抗氧化能力研究[J].食品与机械,2009,25(2):71-73.
[12] Tu,Y. G.,Sun,Y. Z.,Tian,Y. G.,et al. Physicochemical characterisation and antioxidant activity of melanin from the muscles of Taihe Black-bone silky fowl(Gallus gallus domesticus Brisson)[J]. Food Chemistry,2009,114(4):1345-1350.
[13] Pan,Y.,Zhu,Z.,Huang,Z.,et al. Characterisation and free radical scavenging activities of novel red pigment from Osmanthus fragrans'seeds[J]. Food Chemistry,2009,112(4):909-913.
[14]杨小兰,郑海鹰,胡仕屏,等.啤酒花超临界CO2萃余物中总黄酮的提取与抗氧化活性研究[J].食品科学,2008,29(11):137-141.
[15] Isabelcfr,F.,Edmur,A.,Jo2ocm,B.,et al. Antioxidant activity of Portuguese honey samples:Different contributions of the entire honey and phenolic extract[J]. Food Chemistry,2009,114(4):1438-1443.
[16] Lu,Y.,Foo,L. Y. Antioxidant and radical scavenging activities of polyphenols from apple pomace[J]. Food Chemistry,2000,68(1):81-85.
[17]邓胜国,邓泽元,黄丽.荷叶黄酮体外抗氧化活性的研究[J].食品科技,2006,31(7):285-287.
[18]郭庆彬.芦苇叶黄酮的提取、纯化及抗氧化性研究[D].陕西:西北农林科技大学,2008.
[19] Pages,G.,Delaurent,C.,Phan-TanLuu,R.,et al. Different chemometric approaches to optimize the assay of St. John's Wort active ingredients[J]. Chemometrics&Intelligent Laboratory Systems,2007,86(2):159-167.
[20]徐曌.荷叶中黄酮类化合物的提取、分离纯化工艺及其抗氧化性研究[D].武汉:武汉理工大学,2008.
[21] Orhan,D. D.,Aslan,M.,Aktay,G.,et al. Evaluation of hepatoprotective effect of Gentiana olivieri herbs on subacute administration and isolation of active principle[J]. Life Sciences,2003,72(20):2273-2283.
[22] Luo,W.,Zhao,M.,Yang,B.,et al. Identification of bioactive compounds in Phyllenthus emblica L. fruit and their free radical scavenging activities[J]. Food Chemistry,2009,114(2):499-504.
[23]谭莉,陈瑞战,彭雨沙,等.蓝莓花青素提取工艺优化及抗氧化活性评价[J].食品工业,2017,38(8):136-141.
[24]吴培熙.聚合物共混改性[M].北京:中国轻工业出版社,1996.
[25]乔建卫,裴广庆,闻莉,等.黄芩黄酮抗氧化稳定性的研究[J].食品工业科技,2014,35(7):66-68.
[26]许海棠,赵彦芝,张金彦,等.响应面法提取瓶尔小草总黄酮及其抗氧化活性[J].食品科技,2017,42(2):215-220.
[27]王卫记,罗建光,孔令义. HPLC-ESI-MS~n分析酸枣仁有效部位的化学成分[J].中国中药杂志,2009,34(21):2768.