刺梨果实发育过程中主要活性物质含量及其抗氧化性分析
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摘要
为了解刺梨果实中活性物质含量变化及其对抗氧化性能的贡献,本研究测定‘贵农5号’刺梨果实发育过程中总酚、总黄酮、总三萜、VC及超氧化物歧化酶(superoxide dismutase,SOD)5类主要活性物质含量,同时采用1,1-二苯基-2-三硝基苯肼、2,2’-联氮基-双-(3-乙基苯并噻唑啉-6-磺酸)二铵盐和铁离子还原体系分析体外抗氧化能力,并比较活性物质与抗氧化能力之间的相关性。结果显示,不同发育时期的刺梨果实中各活性物质的变化趋势不尽相同:VC含量随果实发育持续增加,花后100 d达到最大值;总酚、总黄酮和总三萜含量变化基本一致,均呈现先降低后升高的趋势,即幼果(花后20～40 d)和成熟果实中具有更高含量;而花后40～60 d的果实中SOD活性更强。总体上,3种抗氧化能力表现与总酚含量变化一致,相关性分析显示它们之间呈极显著正相关关系(P<0.01)。主成分分析显示5种活性物质对抗氧化能力贡献的顺序依次为总酚、总黄酮>VC>总三萜>SOD,其中前3种物质的累计贡献率达到80%以上。以上结果表明,刺梨果实的抗氧化能力是几类物质协同作用的结果,但其中总酚、总黄酮和VC起着决定性作用。
This study aimed to understand the changes in the contents of bioactive substances and their contributions to the antioxidant properties in the fruits of Rosa roxburghii(Guinong 5 cultivar). The contents of total phenolics, total flavonoids, total triterpenes and vitamin C and superoxide dismutase(SOD) at different developmental stages were determined. The antioxidant activity in vitro was evaluated by measuring scavenging capacities against 1,1-diphenyl-2-picrylhydrazyl(DPPH) and 2,2'-amino-di(2-ethyl-benzothiazoline sulphonic acid-6)ammonium salt(ABTS) radicals and ferric reducing antioxidant power(FRAP). We also investigated the correlation between the bioactive substance contents and antioxidant activity. The results showed that different bioactive substances in the fruits of Rosa roxburghii changed differently with developmental stage. The VC content increased to reach the maximum at 100 d after anthesis. The contents of total phenolics, total flavonoids and total triterpenes consistently decreased firstly and then increased; higher levels were observed in the young(20–40 d after anthesis) and ripe fruits. However, higher SOD activity was recorded at 40–60 d after anthesis. Overall, three antioxidant properties changed consistently with total phenolic content, showing a significantly positive correlation(P < 0.01). Principal component analysis(PCA) showed the contribution of five bioactive substances to antioxidant capacity was in the decreasing order of total phenolics, total flavonoids > VC > total triterpenes > SOD, and the cumulative contribution rate of the first three substances to the total variance was over 80%. All of the above results showed that the antioxidant capacity of the fruit was ascribed to the synergistic effects of several bioactive substances, and the total phenolics, total flavonoids and ascorbate played a decisive role.
This study aimed to understand the changes in the contents of bioactive substances and their contributions to the antioxidant properties in the fruits of Rosa roxburghii(Guinong 5 cultivar). The contents of total phenolics, total flavonoids, total triterpenes and vitamin C and superoxide dismutase(SOD) at different developmental stages were determined. The antioxidant activity in vitro was evaluated by measuring scavenging capacities against 1,1-diphenyl-2-picrylhydrazyl(DPPH) and 2,2'-amino-di(2-ethyl-benzothiazoline sulphonic acid-6)ammonium salt(ABTS) radicals and ferric reducing antioxidant power(FRAP). We also investigated the correlation between the bioactive substance contents and antioxidant activity. The results showed that different bioactive substances in the fruits of Rosa roxburghii changed differently with developmental stage. The VC content increased to reach the maximum at 100 d after anthesis. The contents of total phenolics, total flavonoids and total triterpenes consistently decreased firstly and then increased; higher levels were observed in the young(20–40 d after anthesis) and ripe fruits. However, higher SOD activity was recorded at 40–60 d after anthesis. Overall, three antioxidant properties changed consistently with total phenolic content, showing a significantly positive correlation(P < 0.01). Principal component analysis(PCA) showed the contribution of five bioactive substances to antioxidant capacity was in the decreasing order of total phenolics, total flavonoids > VC > total triterpenes > SOD, and the cumulative contribution rate of the first three substances to the total variance was over 80%. All of the above results showed that the antioxidant capacity of the fruit was ascribed to the synergistic effects of several bioactive substances, and the total phenolics, total flavonoids and ascorbate played a decisive role.
引文
[1]PHAM-HUY L A, HE H, PHAM-HUY C. Free radicals, antioxidants in disease and health[J]. International Journal of Biomedical Science,2008, 4(2):89-96.
[2]COOKE M S, EVANSM D, DIZDAROGLU M, et al. Oxidative DNA damage:mechanisms, mutation, and disease[J]. Faseb Journal Official Publication of the Federation of American Societies for Experimental Biology, 2003, 17(10):1195-214. DOI:10.1096/fj.02-0752rev.
[3]OBULESUM,VENUR,SOMASHEKHARR.Lipidperoxidation in Alzheimer’s disease:emphasis on metal-mediated neurotoxicity[J].Acta Neurologica Scandinavica, 2011, 124(5):295-301. DOI:10.1111/j.1600-0404.2010.01483.x.
[4]MAYNEST.Antioxidantnutrientsandchronicdisease:useof biomarkers of exposure and oxidative stress status in epidemiologic research[J].JournalofNutrition,2003,133(Suppl3):933S.DOI:10.1093/jn/133.3.933s.
[5]ALBARRACINSL,STABB,CASASZ,etal.Effectsofnatural antioxidants in neurodegenerative disease[J]. Nutritional Neuroscience,2012, 15(1):1-9. DOI:10.1179/1476830511Y.0000000028.
[6]KAWAGUCHIK,MATSUMOTOT,KUMZAZWAY.Effects of antioxidant polyphenols on TNF-alpha-related diseases[J]. Current Topics in Medicinal Chemistry, 2011, 11(14):1767-1779.DOI:10.2174/156802611796235152.
[7]SINGH B N, SHANKAR S, SRIVASTAVA R K. Green tea catechin,epigallocatechin-3-gallate(EGCG):mechanisms, perspectives and clinicalapplications[J].BiochemicalPharmacology,2011,82(12):1807-1821. DOI:10.1016/j.bcp.2011.07.093.
[8]安华明,刘明,杨曼,等.刺梨有机酸组分及抗坏血酸含量分析[J].中国农业科学, 2011, 44(10):2094-2100. DOI:10.3846/j.issn.0578-175.2011.10.014.
[9]鲁敏,安华明,赵小红.无籽刺梨与刺梨果实中氨基酸分析[J].食品科学, 2015, 36(14):118-121. DOI:10.7506/spkx1002-6630-2015140023.
[10]方修贵,李嗣彪,郑益清.刺梨的营养价值及其开发利用[J].食品工业科技, 2004, 25(1):137-138. DOI:10.3969/j.issn.1002-0306.2004.01.053.
[11]AGATI G, CEROVIC Z G, MARTA A D, et al. Optically-assessed preformed flavonoids and susceptibility of grapevine to Plasmopara viticolaunderdifferentlightregimes[J].FunctionalPlantBiology,2008, 35(1):77-84. DOI:10.1071/fp07178.
[12]安华明,樊卫国,刘庆林,等.刺梨果实和叶片发育过程中抗坏血酸和抗氧化酶的协同变化[J].园艺学报, 2007, 34(5):1293-1296.DOI:10.3321/j.issn:0513-353x.2007.05.038.
[13]戴支凯,余丽梅.刺梨的药理作用[J].中国药房,2007,18(21):1668-1669. DOI:10.3969/j.issn.1001-0408.2007.21.029.
[14]赵转地,张爱华,洪峰.刺梨及其产品的营养及保健药用价值研究进展[J].环境与职业医学, 2007, 24(1):82-84. DOI:10.3969/j.issn.1006-3617.2007.01.025.
[15]陈坤明,宫海军,王锁民.植物抗坏血酸的生物合成、转运及其生物学功能[J].西北植物学报, 2004, 24(2):329-336. DOI:10.3321/j.issn:1000-4025.2004.02.026.
[16]FAN Z L, WANG Z Y, LIU J R. Cold-field fruit extracts exert different antioxidant and antiproliferative activities in vitro[J]. Food Chemistry,2011, 129(2):402-407. DOI:10.1016/j.foodchem.2011.04.091.
[17]SHANMUGAMMK,ANHN,KUMARAP,etal.Targeted inhibitionoftumorproliferation,survival,andmetastasisby pentacyclictriterpenoids:potentialroleinpreventionandtherapy of cancer[J]. Cancer Letters, 2012, 320(2):158-170. DOI:10.1016/j.canlet.2012.02.037.
[18]蔡文国,吴卫,邵金凤,等. Folin-Ciocalteu法测定鱼腥草多酚的含量[J].食品科学, 2010, 31(14):201-204.
[19]ZHANGW,ZHAOX,SUNC,etal.Phenoliccompositionfrom different loquat(Eriobotrya japonica Lindl.)cultivars grown in China and their antioxidant properties[J]. Molecules, 2015, 20(1):542-555.DOI:10.3390/molecules20010542.
[20]杜薇,刘国文.刺梨总黄酮的含量测定及资源利用[J].食品科学,2003, 24(1):112-114. DOI:10.3321/j.issn:1002-6630.2003.01.030.
[21]罗锋,汪河滨,杨玲,等.超声-微波协同萃取法提取甘草黄酮的研究[J].食品研究与开发, 2006, 27(8):127-128. DOI:10.3969/j.issn.1005-6521.2006.08.040.
[22]吴媛琳,王世军,张社奇,等.修剪对苹果枝(梢)皮层总黄酮含量的影响[J].西北林学院学报, 2013, 28(6):103-107. DOI:10.3969/j.issn.1001-7461.2013.06.20.
[23]周巧霞,张经硕,顾明,等.测定山楂及提取物总三萜酸的含量[J].中国野生植物资源,2004,23(5):43-44.DOI:10.3969/j.issn.1006-9690.2004.05.015.
[24]张雁冰,王克让,刘宏民.马桑叶中总三萜酸的含量测定[J].时珍国医国药,2 0 0 6,1 7(4):5 2 9-5 3 0.D O I:1 0. 3 9 6 9/j.issn.1008-0805.2006.04.012.
[25]王乐乐,安华明. HPLC测定刺梨果实中维生素C含量方法的优化[J].现代食品科技, 2013, 29(2):397-400.
[26]王学奎.植物生理生化实验原理和技术[M].北京:高等教育出版社, 2006.
[27]LIXC,LINJ,GAOY,etal.Antioxidantactivityandmechanism ofRhizomaCimicifugae[J].ChemistryCentralJournal,2012,6(1):140-150. DOI:10.1186/1752-153x-6-140.
[28]RER,PELLEGRININ,PROTEGGENTEA,etal.Antioxidant activityapplyinganimprovedABTSradicalcationdecolorization assay[J]. Free Radical Biology and Medicine, 1999, 26(9/10):1231-1237. DOI:10.1016/s0891-5849(98)00315-3.
[29]BENZIE I, STRAIN J J. The ferric reducing ability of plasma(FRAP)as a measure of “antioxidant power”:the FRAP assay[J]. Analytical Biochemistry, 1996, 239(1):70-76. DOI:10.1006/abio.1996.0292.
[30]何照范,牛爱珍,向显衡,等.刺梨果实营养及其维生素C含量变化的研究[J].园艺学报, 1984(4):271-273.
[31]ROUTABOUL J M, KERHOAS L, DEBEAUJON I, et al. Flavonoid diversity and biosynthesis in seed of Arabidopsis thaliana[J]. Planta,2006, 224(1):96-107. DOI:10.1007/s00425-005-0197-5.
[32]XIANGQS,XIAOHF,GAOSB,etal.Invitroantioxidant properties of Rosa roxburghiia queous extracts[J]. Australian Journal of Crop Science, 2012, 6(5):854-860.
[33]HE J Y, ZHANG Y H, MA N, et al. Comparative analysis of multiple ingredientsinRosaroxburghii,andR.sterilis,fruitsandtheir antioxidant activities[J]. Journal of Functional Foods, 2016, 27:29-41.DOI:10.1016/j.jff.2016.08.058.
[34]杜国荣.猕猴桃、柿和苹果果实的抗氧化能力及其抗氧化活性成分的分析[D].杨凌:西北农林科技大学, 2009.
[35]ZHANGW,ZHAOX,SUNC,etal.Phenoliccompositionfrom different loquat(Eriobotrya japonica Lindl.)cultivars grown in China and their antioxidant properties[J]. Molecules, 2015, 20(1):542-555.DOI:10.3390/molecules20010542.
[36]XU H X, CHEN J W. Commercial quality, major bioactive compound content and antioxidant capacity of 12 cultivars of loquat(EriobotryajaponicaLindl.)fruits[J].JournaloftheScienceofFood&Agriculture, 2011, 91(6):1057-1063. DOI:10.1002/jsfa.4282.
[37]吴媛琳,赵听,张凯煜,等.枇杷不同部位主要有效成分含量及抗氧化活性比较[J].西北林学院学报, 2015, 30(1):196-201.DOI:10.3969/j.issn.1001-7461.2015.01.32.
[2]COOKE M S, EVANSM D, DIZDAROGLU M, et al. Oxidative DNA damage:mechanisms, mutation, and disease[J]. Faseb Journal Official Publication of the Federation of American Societies for Experimental Biology, 2003, 17(10):1195-214. DOI:10.1096/fj.02-0752rev.
[3]OBULESUM,VENUR,SOMASHEKHARR.Lipidperoxidation in Alzheimer’s disease:emphasis on metal-mediated neurotoxicity[J].Acta Neurologica Scandinavica, 2011, 124(5):295-301. DOI:10.1111/j.1600-0404.2010.01483.x.
[4]MAYNEST.Antioxidantnutrientsandchronicdisease:useof biomarkers of exposure and oxidative stress status in epidemiologic research[J].JournalofNutrition,2003,133(Suppl3):933S.DOI:10.1093/jn/133.3.933s.
[5]ALBARRACINSL,STABB,CASASZ,etal.Effectsofnatural antioxidants in neurodegenerative disease[J]. Nutritional Neuroscience,2012, 15(1):1-9. DOI:10.1179/1476830511Y.0000000028.
[6]KAWAGUCHIK,MATSUMOTOT,KUMZAZWAY.Effects of antioxidant polyphenols on TNF-alpha-related diseases[J]. Current Topics in Medicinal Chemistry, 2011, 11(14):1767-1779.DOI:10.2174/156802611796235152.
[7]SINGH B N, SHANKAR S, SRIVASTAVA R K. Green tea catechin,epigallocatechin-3-gallate(EGCG):mechanisms, perspectives and clinicalapplications[J].BiochemicalPharmacology,2011,82(12):1807-1821. DOI:10.1016/j.bcp.2011.07.093.
[8]安华明,刘明,杨曼,等.刺梨有机酸组分及抗坏血酸含量分析[J].中国农业科学, 2011, 44(10):2094-2100. DOI:10.3846/j.issn.0578-175.2011.10.014.
[9]鲁敏,安华明,赵小红.无籽刺梨与刺梨果实中氨基酸分析[J].食品科学, 2015, 36(14):118-121. DOI:10.7506/spkx1002-6630-2015140023.
[10]方修贵,李嗣彪,郑益清.刺梨的营养价值及其开发利用[J].食品工业科技, 2004, 25(1):137-138. DOI:10.3969/j.issn.1002-0306.2004.01.053.
[11]AGATI G, CEROVIC Z G, MARTA A D, et al. Optically-assessed preformed flavonoids and susceptibility of grapevine to Plasmopara viticolaunderdifferentlightregimes[J].FunctionalPlantBiology,2008, 35(1):77-84. DOI:10.1071/fp07178.
[12]安华明,樊卫国,刘庆林,等.刺梨果实和叶片发育过程中抗坏血酸和抗氧化酶的协同变化[J].园艺学报, 2007, 34(5):1293-1296.DOI:10.3321/j.issn:0513-353x.2007.05.038.
[13]戴支凯,余丽梅.刺梨的药理作用[J].中国药房,2007,18(21):1668-1669. DOI:10.3969/j.issn.1001-0408.2007.21.029.
[14]赵转地,张爱华,洪峰.刺梨及其产品的营养及保健药用价值研究进展[J].环境与职业医学, 2007, 24(1):82-84. DOI:10.3969/j.issn.1006-3617.2007.01.025.
[15]陈坤明,宫海军,王锁民.植物抗坏血酸的生物合成、转运及其生物学功能[J].西北植物学报, 2004, 24(2):329-336. DOI:10.3321/j.issn:1000-4025.2004.02.026.
[16]FAN Z L, WANG Z Y, LIU J R. Cold-field fruit extracts exert different antioxidant and antiproliferative activities in vitro[J]. Food Chemistry,2011, 129(2):402-407. DOI:10.1016/j.foodchem.2011.04.091.
[17]SHANMUGAMMK,ANHN,KUMARAP,etal.Targeted inhibitionoftumorproliferation,survival,andmetastasisby pentacyclictriterpenoids:potentialroleinpreventionandtherapy of cancer[J]. Cancer Letters, 2012, 320(2):158-170. DOI:10.1016/j.canlet.2012.02.037.
[18]蔡文国,吴卫,邵金凤,等. Folin-Ciocalteu法测定鱼腥草多酚的含量[J].食品科学, 2010, 31(14):201-204.
[19]ZHANGW,ZHAOX,SUNC,etal.Phenoliccompositionfrom different loquat(Eriobotrya japonica Lindl.)cultivars grown in China and their antioxidant properties[J]. Molecules, 2015, 20(1):542-555.DOI:10.3390/molecules20010542.
[20]杜薇,刘国文.刺梨总黄酮的含量测定及资源利用[J].食品科学,2003, 24(1):112-114. DOI:10.3321/j.issn:1002-6630.2003.01.030.
[21]罗锋,汪河滨,杨玲,等.超声-微波协同萃取法提取甘草黄酮的研究[J].食品研究与开发, 2006, 27(8):127-128. DOI:10.3969/j.issn.1005-6521.2006.08.040.
[22]吴媛琳,王世军,张社奇,等.修剪对苹果枝(梢)皮层总黄酮含量的影响[J].西北林学院学报, 2013, 28(6):103-107. DOI:10.3969/j.issn.1001-7461.2013.06.20.
[23]周巧霞,张经硕,顾明,等.测定山楂及提取物总三萜酸的含量[J].中国野生植物资源,2004,23(5):43-44.DOI:10.3969/j.issn.1006-9690.2004.05.015.
[24]张雁冰,王克让,刘宏民.马桑叶中总三萜酸的含量测定[J].时珍国医国药,2 0 0 6,1 7(4):5 2 9-5 3 0.D O I:1 0. 3 9 6 9/j.issn.1008-0805.2006.04.012.
[25]王乐乐,安华明. HPLC测定刺梨果实中维生素C含量方法的优化[J].现代食品科技, 2013, 29(2):397-400.
[26]王学奎.植物生理生化实验原理和技术[M].北京:高等教育出版社, 2006.
[27]LIXC,LINJ,GAOY,etal.Antioxidantactivityandmechanism ofRhizomaCimicifugae[J].ChemistryCentralJournal,2012,6(1):140-150. DOI:10.1186/1752-153x-6-140.
[28]RER,PELLEGRININ,PROTEGGENTEA,etal.Antioxidant activityapplyinganimprovedABTSradicalcationdecolorization assay[J]. Free Radical Biology and Medicine, 1999, 26(9/10):1231-1237. DOI:10.1016/s0891-5849(98)00315-3.
[29]BENZIE I, STRAIN J J. The ferric reducing ability of plasma(FRAP)as a measure of “antioxidant power”:the FRAP assay[J]. Analytical Biochemistry, 1996, 239(1):70-76. DOI:10.1006/abio.1996.0292.
[30]何照范,牛爱珍,向显衡,等.刺梨果实营养及其维生素C含量变化的研究[J].园艺学报, 1984(4):271-273.
[31]ROUTABOUL J M, KERHOAS L, DEBEAUJON I, et al. Flavonoid diversity and biosynthesis in seed of Arabidopsis thaliana[J]. Planta,2006, 224(1):96-107. DOI:10.1007/s00425-005-0197-5.
[32]XIANGQS,XIAOHF,GAOSB,etal.Invitroantioxidant properties of Rosa roxburghiia queous extracts[J]. Australian Journal of Crop Science, 2012, 6(5):854-860.
[33]HE J Y, ZHANG Y H, MA N, et al. Comparative analysis of multiple ingredientsinRosaroxburghii,andR.sterilis,fruitsandtheir antioxidant activities[J]. Journal of Functional Foods, 2016, 27:29-41.DOI:10.1016/j.jff.2016.08.058.
[34]杜国荣.猕猴桃、柿和苹果果实的抗氧化能力及其抗氧化活性成分的分析[D].杨凌:西北农林科技大学, 2009.
[35]ZHANGW,ZHAOX,SUNC,etal.Phenoliccompositionfrom different loquat(Eriobotrya japonica Lindl.)cultivars grown in China and their antioxidant properties[J]. Molecules, 2015, 20(1):542-555.DOI:10.3390/molecules20010542.
[36]XU H X, CHEN J W. Commercial quality, major bioactive compound content and antioxidant capacity of 12 cultivars of loquat(EriobotryajaponicaLindl.)fruits[J].JournaloftheScienceofFood&Agriculture, 2011, 91(6):1057-1063. DOI:10.1002/jsfa.4282.
[37]吴媛琳,赵听,张凯煜,等.枇杷不同部位主要有效成分含量及抗氧化活性比较[J].西北林学院学报, 2015, 30(1):196-201.DOI:10.3969/j.issn.1001-7461.2015.01.32.