黑玉米中黑色素和氨基酸的研究
|
摘要
黑玉米是玉米的一个新品种,其籽粒角质层不同程度地沉淀黑色素,外观乌黑发亮。黑玉米不仅色泽独特,而且营养丰富,富含水溶性黑色素、多种氨基酸和各种人体必需的微量元素。本课题以黑玉米为研究对象,对黑玉米中黑色素的提取工艺、组成、抗氧化性及营养成分氨基酸进行了研究。研究内容主要包括以下三个部分:1.黑玉米中黑色素的提取工艺及组成研究;2.黑玉米和黄玉米提取物的抗氧化性研究;3.毛细管电泳-电化学法检测黑玉米皮中三种氨基酸的含量。
黑玉米色素属天然花青苷色素。通过正交实验确定了黑色素的最佳提取条件:95%的乙醇溶液、料液比1:30、pH=1、提取时间45 min;并采用液质联用(LC-MS)技术对山东和山西两种黑玉米中花青苷的种类进行了确定,其中山东黑玉米中主要含有两种花青苷,分别是矢车菊和天竺葵花青素;山西黑玉米中主要含有三种花青苷,分别是矢车菊、天竺葵和芍药花青素。
为了研究其抗氧化活性和比较抗氧化效果,本实验以常见的黄玉米与黑玉米为原材料,进行了一系列抗氧化性能实验。用不同极性的溶剂对玉米中的抗氧化性物质进行超声提取,并对所得提取物进行清除DPPH·自由基实验,得出黄玉米和黑玉米各自的不同溶剂提取物中抗氧化性最强的部分;以两种玉米提取物中抗氧化性最强的部分为材料,进行邻苯三酚自氧化抑制实验和抑制猪油自氧化实验,并对两种玉米提取物中抗氧化性最强的部分进行总黄酮含量的测定。结果表明:黄玉米和黑玉米各自的不同溶剂提取物中抗氧化性最强的部分均为80%乙醇提取物;黄玉米乙醇提取物中总黄酮含量为16.2mg/8g,黑玉米乙醇提取物中总黄酮含量为51.5mg/8g,三种抗氧化实验均说明黑玉米抗氧化性比黄玉米好,具有较好的清除自由基和抗氧化的能力。
采用OPA为柱前衍生化试剂,用毛细管电泳-电化学检测(CE-ED)的方法测定了黑玉米中亮氨酸、缬氨酸和丙氨酸的含量。以直径为300μm的碳圆盘电极为工作电极,研究了运行缓冲液pH值、检测电压、分离电压和进样时间对三种氨基酸分离和检测的影响,得到了最佳电泳条件:100mmol/L硼砂(pH7.8)为运行缓冲液、检测电压0.95V(vs.SCE)、分离电压18kV、进样时间6s。在优化条件下,三组分可在18 min内完全分离,检出限分别为2.5×10-4 g/L、2.5×10-4 g/L、5.0×10-4 g/L。该方法准确、简单易行,样品处理简单,为氨基酸分离测定提供了一种快速灵敏的检测方法。
Black corn is a new corn species, which color is different from other corn, mainly because there is a plenty of black pigment in its epidermis. It is rich in water-soluble black pigment , a variety of essential amino acids and various trace elements. Taking black corn as experimental material, studied the extraction and composition of black corn pigment, evaluated antioxidative activities of corn by a series of antioxidative tests and determined the contents of amino acids in black corn epidermis. The thesis mainly includes the following three parts :1. study on extraction technology and composition of black corn pigment. 2. study on the antioxidative activities of extracts from black corn and yellow corn. 3. determination of three amino acids in black corn epidermis by capillary electrophoresis with electrochemical detection (CE-ED).
Firstly, black corn pigment is a natural anthocyanin pigment. The effects of different factors on extraction of pigment with orthogonal test were investigated. The optimal parameters of extraction of black corn pigment were as fellows: ethanol–water (95:5, v/v) as extracting solvent, solid-liquid ratio 1:30, extracting time 45 min. Liquid chromatography-mass spectrometry (LC-MS) were applied to determine the components of black corn pigments. The results showed that there were two major anthocyanins in ShanDong sample: cyanidin and pelargonin, and three major anthocyanins in ShanXi sample: cyanidin, pelargonin, and paeonidin.
Secondly, to compare the antioxidative activity with other common corn, we chose yellow corn and black corn as the raw materials to carry on a series of antioxidative tests. The different fractions of corn antioxidative extracts were obtained using solvents with different polarities. The antioxidative activity of these extracts were assessed for 1,1-Diphenyl-2-picrylhydrazyl(DPPH)free radical scavenging activity to find the strongest antioxidative fractions in both corns. Taking the strongest antioxidative fractions as experimental objects, their antioxidiative activities were investigated by two different methods in scavenging superoxide anion (O2-) generated by pyrogallol autoxidant system and restrainning lard oxidation. The total flavonoids contents of ethanolic extract from the both corns were determined. The results showed that ethanol-water antioxidative extract fraction had the strongest antioxidative capacities among the five fractions in both corns .The total flavonoids contents of ethanolic extract were 16.2 mg/8g and 51.5 mg/8g for yellow corn and black corn, respectively. Three antioxidative experiments shown that the antioxidative activity of black corn was better than that of yellow corn.
Finally, using o-phthaldehyde (OPA) as a pre-column derivative agent, the contents of leucine, valine and alanine in black corn epidermis were determined by CD-ED in this paper. A 300μm diameter carbon disk electrode was used as the working electrode, the effects of several factors such as pH of running buffer, separation voltage, injection time, and detection potential were investigated to acquire the optimum conditions. The optimum CE-ED conditions were as fellows: applied potential to the working electrode at 0.95V (vs.SCE), 100 mmol/L borate solution (pH 7.8) as running buffer, injection time 6s, the analytes could be well separated within 18 min at a separation voltage of 18 kV in a 100 mmol/L borax running buffer . The detection limits (S/N=3) were 2.5×10-4 g/L、2.5×10-4 g/L、5.0×10-4 g/L, for leucine, valine and alanine, respectively. This method is accurate and feasible, the sample preparation is simple, which provided a fast and sensitive determination method for amino acids.
黑玉米色素属天然花青苷色素。通过正交实验确定了黑色素的最佳提取条件:95%的乙醇溶液、料液比1:30、pH=1、提取时间45 min;并采用液质联用(LC-MS)技术对山东和山西两种黑玉米中花青苷的种类进行了确定,其中山东黑玉米中主要含有两种花青苷,分别是矢车菊和天竺葵花青素;山西黑玉米中主要含有三种花青苷,分别是矢车菊、天竺葵和芍药花青素。
为了研究其抗氧化活性和比较抗氧化效果,本实验以常见的黄玉米与黑玉米为原材料,进行了一系列抗氧化性能实验。用不同极性的溶剂对玉米中的抗氧化性物质进行超声提取,并对所得提取物进行清除DPPH·自由基实验,得出黄玉米和黑玉米各自的不同溶剂提取物中抗氧化性最强的部分;以两种玉米提取物中抗氧化性最强的部分为材料,进行邻苯三酚自氧化抑制实验和抑制猪油自氧化实验,并对两种玉米提取物中抗氧化性最强的部分进行总黄酮含量的测定。结果表明:黄玉米和黑玉米各自的不同溶剂提取物中抗氧化性最强的部分均为80%乙醇提取物;黄玉米乙醇提取物中总黄酮含量为16.2mg/8g,黑玉米乙醇提取物中总黄酮含量为51.5mg/8g,三种抗氧化实验均说明黑玉米抗氧化性比黄玉米好,具有较好的清除自由基和抗氧化的能力。
采用OPA为柱前衍生化试剂,用毛细管电泳-电化学检测(CE-ED)的方法测定了黑玉米中亮氨酸、缬氨酸和丙氨酸的含量。以直径为300μm的碳圆盘电极为工作电极,研究了运行缓冲液pH值、检测电压、分离电压和进样时间对三种氨基酸分离和检测的影响,得到了最佳电泳条件:100mmol/L硼砂(pH7.8)为运行缓冲液、检测电压0.95V(vs.SCE)、分离电压18kV、进样时间6s。在优化条件下,三组分可在18 min内完全分离,检出限分别为2.5×10-4 g/L、2.5×10-4 g/L、5.0×10-4 g/L。该方法准确、简单易行,样品处理简单,为氨基酸分离测定提供了一种快速灵敏的检测方法。
Black corn is a new corn species, which color is different from other corn, mainly because there is a plenty of black pigment in its epidermis. It is rich in water-soluble black pigment , a variety of essential amino acids and various trace elements. Taking black corn as experimental material, studied the extraction and composition of black corn pigment, evaluated antioxidative activities of corn by a series of antioxidative tests and determined the contents of amino acids in black corn epidermis. The thesis mainly includes the following three parts :1. study on extraction technology and composition of black corn pigment. 2. study on the antioxidative activities of extracts from black corn and yellow corn. 3. determination of three amino acids in black corn epidermis by capillary electrophoresis with electrochemical detection (CE-ED).
Firstly, black corn pigment is a natural anthocyanin pigment. The effects of different factors on extraction of pigment with orthogonal test were investigated. The optimal parameters of extraction of black corn pigment were as fellows: ethanol–water (95:5, v/v) as extracting solvent, solid-liquid ratio 1:30, extracting time 45 min. Liquid chromatography-mass spectrometry (LC-MS) were applied to determine the components of black corn pigments. The results showed that there were two major anthocyanins in ShanDong sample: cyanidin and pelargonin, and three major anthocyanins in ShanXi sample: cyanidin, pelargonin, and paeonidin.
Secondly, to compare the antioxidative activity with other common corn, we chose yellow corn and black corn as the raw materials to carry on a series of antioxidative tests. The different fractions of corn antioxidative extracts were obtained using solvents with different polarities. The antioxidative activity of these extracts were assessed for 1,1-Diphenyl-2-picrylhydrazyl(DPPH)free radical scavenging activity to find the strongest antioxidative fractions in both corns. Taking the strongest antioxidative fractions as experimental objects, their antioxidiative activities were investigated by two different methods in scavenging superoxide anion (O2-) generated by pyrogallol autoxidant system and restrainning lard oxidation. The total flavonoids contents of ethanolic extract from the both corns were determined. The results showed that ethanol-water antioxidative extract fraction had the strongest antioxidative capacities among the five fractions in both corns .The total flavonoids contents of ethanolic extract were 16.2 mg/8g and 51.5 mg/8g for yellow corn and black corn, respectively. Three antioxidative experiments shown that the antioxidative activity of black corn was better than that of yellow corn.
Finally, using o-phthaldehyde (OPA) as a pre-column derivative agent, the contents of leucine, valine and alanine in black corn epidermis were determined by CD-ED in this paper. A 300μm diameter carbon disk electrode was used as the working electrode, the effects of several factors such as pH of running buffer, separation voltage, injection time, and detection potential were investigated to acquire the optimum conditions. The optimum CE-ED conditions were as fellows: applied potential to the working electrode at 0.95V (vs.SCE), 100 mmol/L borate solution (pH 7.8) as running buffer, injection time 6s, the analytes could be well separated within 18 min at a separation voltage of 18 kV in a 100 mmol/L borax running buffer . The detection limits (S/N=3) were 2.5×10-4 g/L、2.5×10-4 g/L、5.0×10-4 g/L, for leucine, valine and alanine, respectively. This method is accurate and feasible, the sample preparation is simple, which provided a fast and sensitive determination method for amino acids.
引文
[1]田春莲,陈建华,李立君等.黑玉米色素的提取工艺及特性研究[J].中国林副特产, 2003, 3: 30-31.
[2]王霞,高云.黑甜玉米中黑色素提取及纯化工艺研究[J].食品科学, 2004, 25 : 198-200.
[3]周波,王晓红,郭连营.玉米紫色植株花色苷色素延缓衰老的功能[J].中国临床康复,2006,10(19):138-140
[4]周波,王晓红,陈丽丽等.玉米紫色植株色素体外抗氧化活性实验研究[J].现代食品科技,2006,23(4):23-25
[5]张钟,郭元新,李凤霞.黑糯玉米芯色素清除O2-·和·OH的体外实验研究[J].食品与发酵工业,2006,32(11):36-38
[6]卢艳杰,姚惠源.黄体素、玉米黄素及其生理功能研究现状[J].食品与发酵工业, 2003,29(2): 81-84.
[7]庞志申.花色苷研究概况[J].北京农业科学,2000,18(5): 37-39
[8]马自超,庞业珍.天然色素化学及生产工艺学[M].北京:中国林业出版社. 1994
[9] J.B, H.B等编,戴伦凯,谢玉如等译.黄酮类化合物[M].北京:科学出版社. 1983,4.
[10] Victor Hong, Ronald E. Wrolstad. Characterization of Anthocyanins-containing Colorants and Fruits Juices by HPLC/Photodiode Array Detection[J]. J. Agric. Food Chem. 1990, 38(3): 698-708
[11] Victor Hong, Ronald E. Wrolstad. Use of HPLC Separation/Photodiode Array Detection for Characterization of Anthocyanins[J]. J. Agric. Food Chem. 1990, 38(3): 708-715
[12]谭仁祥.植物成分功能[M].北京:科学出版社, 2003.
[13]曹平.天然抗氧化剂抑制油脂氧化的研究进展[J].中国油脂, 2005, 30(7):49-53.
[14]彭辉.油脂、含油食品的氧化、抗氧化以及抗氧化剂[J].食品工业, 2004, 3:34-36.
[15]游见明,刘达玉.茶多酚对棕榈油抗氧化性的研究[J].广州食品工业科技, 2004, 20(4):38-39.
[16]李海波.茶多酚的应用与研究[J].食品研究与开发, 2004, 25(2):12-15.
[17]张红雨,郑纪庆,姜洪君.茶多酚抗氧化作用的理论探讨[J].中国油脂, 1998, 6:45-46.
[18]沈建福,张英,徐维娅.竹叶黄酮糖苷的水解及其苷元的抗氧化性能研究[J].中国粮油学报, 2001, 16(4):14-17.
[19]王卫良,李普选,周宇雄. L-抗坏血酸硬脂酸酯-新型营养抗氧化剂的合成研究[J].中国油脂, 2005, 30(2): 56-58.
[20]李庆,姚凯,谭敏.新型天然抗氧化剂-鞣花酸[J].四川食品与发酵, 2001, 34(4): 10-13.
[21]张德权,台建祥,付勤.生物类黄酮德研究及应用状况[J].食品与发酵工业, 1999 (6): 52-57.
[22] Lin C C, Wu S J, Chang C H, et al. Antioxidant activity of Cinnamomun cassia[J]. Phytotherapy Research, 2003, 17: 726-730.
[23]熊皓平,杨伟丽,张友胜等.天然植物抗氧化剂的研究进展[J].天然产物研究与开发, 2001, 13(5): 75-79.
[24]朱宏莉,韦海洪等.竹叶总黄酮的提取和纯化工艺的研究[J].食品科学, 2005,26(8): 158-161.
[25]周才琼等.金银花花叶中总黄酮类化合物的最佳提取工艺研究[J].西南农业大学学报, 2003, 25(3): 63-67.
[26]郭青枝,赵二劳,陈婕.正交试验优选苦瓜黄酮超声波提取工艺[J].酿酒科技, 2007, 1: 36-37.
[27]张健,蔡宝国,杜福彦.山绿茶中黄酮德超声波提取研究[J].食品工业, 2006, 1: 49-51.
[28]李巧玲.微波萃取技术在天然食用色素提取上的应用[J].食品科技, 2003(10): 60-64.
[29]王琪,李洪玲.木兰属植物花蕾总黄酮含量测定及提取工艺研究[J].现代中药研究与实践, 2003, 17(2): 32-33.
[30]汪茂山,谢培山等.天然有机化合物提取分离与结果鉴定[M].北京:化学工业出版社, 2004, 31-35.
[31]朱自强编著.超临界流体技术-原理和应用[M].北京:化学工业出版社, 2000.
[32]孙婷.超临界CO2法萃取银杏叶黄酮及其含量测定的研究[J].中国食品学报, 2005, 5(3): 126-129.
[33]凌关庭.抗氧化食品与健康[M].北京:化学工业出版社, 2004, 330-345.
[34]曾佑伟.玫瑰鲜花抗氧化能力及其活性成分的研究[硕士学位论文].华南示范大学, 2004
[35] Yoshida T, Mori K, Hatano T, et al. Studies on inhibition mechanism of autoxidation by tannins and flavonoids[J]. ChemPharm Bull, 1989, 37(7): 1919.
[36] Mellors A, Tappel A L. The inhibition of mitochondrial peroxidation by ubiquinone and ubiquinol[J]. The Journal of Biological Chemistry, 1966, 241(19): 4353
[37] Cotelle N, Bernierj L, Catteauj P, et al. Antioxidant properties of hydroxy-flavones[J]. Free Radical Biology & Medicine, 1998, 20(1): 35.
[38]许申鸿,杭瑚.一种筛选自由基清除剂的简便方法[J].中草药, 2000, 31(2): 96-97.
[39]汪洁,王立强,石岩.线性聚丙烯酰胺凝胶毛细管电泳地迁移特性[J].分析化学, 2008, 36(3): 330-334.
[40]丁祥欢,高乐怡,韩茹等.毛细管电泳-电化学检测食醋和酱油中氨基酸含量[J].华东师范大学学报, 2002, 1: 56-60.
[41]黄颖,段建平,杨明灿等.毛细管电泳法测定黄瓜和西红柿中的谷胱甘肽[J].色谱, 2003, 21(5): 510-512.
[42]张玉奎,陈农,王磊等.几种标准蛋白质的毛细管区带电泳的分离及其迁移行为[J].分析化学,1994, 22(8): 798-800.
[43]黄宝美,张爱华,姚程炜.高效毛细管电泳电导法拆分苯丙氨酸对映体[J].分析测试学报,2007, 26(2): 258-260.
[44]孙汉文,赵伟,何攀.毛细管电泳有效分离和同时检测8种氟喹诺酮[J].河北大学学报,2008, 28(2): 152-157.
[45] Brown R S, Luongj H T, Szolaro H J, et al. Cyclodextrin-modified capillary electrophoresis: Determination of polycyclic aromatic hydrocarbons in contaminated soils[J]. Anal Chem,1996, 68: 287-292.
[46] Belin G K, Erim F B, Clacar F. Capillary electrokinetic separation of polycyclic aromatic hydrocarbons using cetylpyridium bromide [J]. Poly Arom Comp, 2004, 24: 343-352.
[47]鲁京兰,申凤善,太俊哲.黑玉米中黑色素的提取[J].延边大学农学学报, 2002, 24(2): 96-98.
[48]王新广,罗先群,云芸.海南黑糯玉米色素的提取及其稳定性的研究[J].中国食品添加剂, 2003, 5: 19-21.
[49]陈连文,刘敬兰,师建华.黑糯玉米色素的提取及其稳定性研究[J].食品科技, 2001, 4 : 39-40
[50]张兰杰,辛广,张维华.双波长分光光度法测定黑玉米花粉中总黄酮的含量[J].食品科学, 2006, 27: 230-232.
[51]张翼伸.怎样研究植物多糖[J].生命科学, 1999, 19(6): 296-297.
[52]郑晶泉.抗氧化剂抗氧化实验研究进展[J].国外医学卫生学分册, 2000, 27(1): 37-40.
[53]张名位,郭宝江,池建伟等.黑米皮提取物的体外抗氧化作用与成分分析[J].中国粮油学报, 2005, 20(6): 49.
[54]金杰,李志西,张锋等.桑椹醋提取物对二苯代苦味酰基自由基(DPPH·)的清除作用[J].西北农林科技大学学报, 2006, 34(3): 135.
[55]侯同刚,李天择,黄淑霞.毛细管电泳在氨基酸分析中的最新进展[J].食品科技, 2004, 2: 85-87.
[56]张晖,徐永,姚惠源.纸层析法定量测定米胚芽中的γ-氨基丁酸[J].无锡轻工大学学报, 2004, 23 (2) :101
[57] Saikusa T. Accumulation of gamma aminobutyric acid (GABA) in the rice germ during water soaking[J]. Biosci Biotesh Biochem, 1994, 58(12): 2291-2292.
[58] Lindroth P, Mopper K. High-performance liquid chromatography determination of subpicomole of amino acids by fluorescence derivatization with o-phthaldialdedyde[J]. Annl Chem, 1975, 51(10): 1667-1669.
[59]Liu H J. Determination of amino acids by precolumn derivatization with 6-aminoquinolyl-N-hydroxysucciniomidyl carbamate and high- performance liquid chromatography with ultraviolet detection[J]. J Chromarogr, 1994, 670(1): 59-62.
[60]张红漫,于文涛,欧阳平凯等.气相色谱-质谱法测定中药沙苑子中的氨基酸[J].氨基酸和生物资源, 2004, 26 (2): 62-64.
[61]王安群. LC-MS法测定发芽糙米等产品中的γ-氨基丁酸[J].中国科技信息, 2006, 4: 81-83.
[62]常碧影,刘洪基,闫惠文等.柱前衍生高效液相色谱法测定氨基酸[J].分析化学, 1995, 23 (1): 100-102.
[63] Olivia P, Isable M F, Eulalia M, et al. Effect of temperature on evolution of free amino acid and biogenic amine contents during storage of Azeitao cheese[J]. Food chemistry, 2001, 75: 287-291.
[64]曹玉华,汪云.毛细管电泳电化学检测法测定胡黄连中香草酸和阿魏酸的含量[J].分析测试学报, 2003, 22(6): 95-97.
[2]王霞,高云.黑甜玉米中黑色素提取及纯化工艺研究[J].食品科学, 2004, 25 : 198-200.
[3]周波,王晓红,郭连营.玉米紫色植株花色苷色素延缓衰老的功能[J].中国临床康复,2006,10(19):138-140
[4]周波,王晓红,陈丽丽等.玉米紫色植株色素体外抗氧化活性实验研究[J].现代食品科技,2006,23(4):23-25
[5]张钟,郭元新,李凤霞.黑糯玉米芯色素清除O2-·和·OH的体外实验研究[J].食品与发酵工业,2006,32(11):36-38
[6]卢艳杰,姚惠源.黄体素、玉米黄素及其生理功能研究现状[J].食品与发酵工业, 2003,29(2): 81-84.
[7]庞志申.花色苷研究概况[J].北京农业科学,2000,18(5): 37-39
[8]马自超,庞业珍.天然色素化学及生产工艺学[M].北京:中国林业出版社. 1994
[9] J.B, H.B等编,戴伦凯,谢玉如等译.黄酮类化合物[M].北京:科学出版社. 1983,4.
[10] Victor Hong, Ronald E. Wrolstad. Characterization of Anthocyanins-containing Colorants and Fruits Juices by HPLC/Photodiode Array Detection[J]. J. Agric. Food Chem. 1990, 38(3): 698-708
[11] Victor Hong, Ronald E. Wrolstad. Use of HPLC Separation/Photodiode Array Detection for Characterization of Anthocyanins[J]. J. Agric. Food Chem. 1990, 38(3): 708-715
[12]谭仁祥.植物成分功能[M].北京:科学出版社, 2003.
[13]曹平.天然抗氧化剂抑制油脂氧化的研究进展[J].中国油脂, 2005, 30(7):49-53.
[14]彭辉.油脂、含油食品的氧化、抗氧化以及抗氧化剂[J].食品工业, 2004, 3:34-36.
[15]游见明,刘达玉.茶多酚对棕榈油抗氧化性的研究[J].广州食品工业科技, 2004, 20(4):38-39.
[16]李海波.茶多酚的应用与研究[J].食品研究与开发, 2004, 25(2):12-15.
[17]张红雨,郑纪庆,姜洪君.茶多酚抗氧化作用的理论探讨[J].中国油脂, 1998, 6:45-46.
[18]沈建福,张英,徐维娅.竹叶黄酮糖苷的水解及其苷元的抗氧化性能研究[J].中国粮油学报, 2001, 16(4):14-17.
[19]王卫良,李普选,周宇雄. L-抗坏血酸硬脂酸酯-新型营养抗氧化剂的合成研究[J].中国油脂, 2005, 30(2): 56-58.
[20]李庆,姚凯,谭敏.新型天然抗氧化剂-鞣花酸[J].四川食品与发酵, 2001, 34(4): 10-13.
[21]张德权,台建祥,付勤.生物类黄酮德研究及应用状况[J].食品与发酵工业, 1999 (6): 52-57.
[22] Lin C C, Wu S J, Chang C H, et al. Antioxidant activity of Cinnamomun cassia[J]. Phytotherapy Research, 2003, 17: 726-730.
[23]熊皓平,杨伟丽,张友胜等.天然植物抗氧化剂的研究进展[J].天然产物研究与开发, 2001, 13(5): 75-79.
[24]朱宏莉,韦海洪等.竹叶总黄酮的提取和纯化工艺的研究[J].食品科学, 2005,26(8): 158-161.
[25]周才琼等.金银花花叶中总黄酮类化合物的最佳提取工艺研究[J].西南农业大学学报, 2003, 25(3): 63-67.
[26]郭青枝,赵二劳,陈婕.正交试验优选苦瓜黄酮超声波提取工艺[J].酿酒科技, 2007, 1: 36-37.
[27]张健,蔡宝国,杜福彦.山绿茶中黄酮德超声波提取研究[J].食品工业, 2006, 1: 49-51.
[28]李巧玲.微波萃取技术在天然食用色素提取上的应用[J].食品科技, 2003(10): 60-64.
[29]王琪,李洪玲.木兰属植物花蕾总黄酮含量测定及提取工艺研究[J].现代中药研究与实践, 2003, 17(2): 32-33.
[30]汪茂山,谢培山等.天然有机化合物提取分离与结果鉴定[M].北京:化学工业出版社, 2004, 31-35.
[31]朱自强编著.超临界流体技术-原理和应用[M].北京:化学工业出版社, 2000.
[32]孙婷.超临界CO2法萃取银杏叶黄酮及其含量测定的研究[J].中国食品学报, 2005, 5(3): 126-129.
[33]凌关庭.抗氧化食品与健康[M].北京:化学工业出版社, 2004, 330-345.
[34]曾佑伟.玫瑰鲜花抗氧化能力及其活性成分的研究[硕士学位论文].华南示范大学, 2004
[35] Yoshida T, Mori K, Hatano T, et al. Studies on inhibition mechanism of autoxidation by tannins and flavonoids[J]. ChemPharm Bull, 1989, 37(7): 1919.
[36] Mellors A, Tappel A L. The inhibition of mitochondrial peroxidation by ubiquinone and ubiquinol[J]. The Journal of Biological Chemistry, 1966, 241(19): 4353
[37] Cotelle N, Bernierj L, Catteauj P, et al. Antioxidant properties of hydroxy-flavones[J]. Free Radical Biology & Medicine, 1998, 20(1): 35.
[38]许申鸿,杭瑚.一种筛选自由基清除剂的简便方法[J].中草药, 2000, 31(2): 96-97.
[39]汪洁,王立强,石岩.线性聚丙烯酰胺凝胶毛细管电泳地迁移特性[J].分析化学, 2008, 36(3): 330-334.
[40]丁祥欢,高乐怡,韩茹等.毛细管电泳-电化学检测食醋和酱油中氨基酸含量[J].华东师范大学学报, 2002, 1: 56-60.
[41]黄颖,段建平,杨明灿等.毛细管电泳法测定黄瓜和西红柿中的谷胱甘肽[J].色谱, 2003, 21(5): 510-512.
[42]张玉奎,陈农,王磊等.几种标准蛋白质的毛细管区带电泳的分离及其迁移行为[J].分析化学,1994, 22(8): 798-800.
[43]黄宝美,张爱华,姚程炜.高效毛细管电泳电导法拆分苯丙氨酸对映体[J].分析测试学报,2007, 26(2): 258-260.
[44]孙汉文,赵伟,何攀.毛细管电泳有效分离和同时检测8种氟喹诺酮[J].河北大学学报,2008, 28(2): 152-157.
[45] Brown R S, Luongj H T, Szolaro H J, et al. Cyclodextrin-modified capillary electrophoresis: Determination of polycyclic aromatic hydrocarbons in contaminated soils[J]. Anal Chem,1996, 68: 287-292.
[46] Belin G K, Erim F B, Clacar F. Capillary electrokinetic separation of polycyclic aromatic hydrocarbons using cetylpyridium bromide [J]. Poly Arom Comp, 2004, 24: 343-352.
[47]鲁京兰,申凤善,太俊哲.黑玉米中黑色素的提取[J].延边大学农学学报, 2002, 24(2): 96-98.
[48]王新广,罗先群,云芸.海南黑糯玉米色素的提取及其稳定性的研究[J].中国食品添加剂, 2003, 5: 19-21.
[49]陈连文,刘敬兰,师建华.黑糯玉米色素的提取及其稳定性研究[J].食品科技, 2001, 4 : 39-40
[50]张兰杰,辛广,张维华.双波长分光光度法测定黑玉米花粉中总黄酮的含量[J].食品科学, 2006, 27: 230-232.
[51]张翼伸.怎样研究植物多糖[J].生命科学, 1999, 19(6): 296-297.
[52]郑晶泉.抗氧化剂抗氧化实验研究进展[J].国外医学卫生学分册, 2000, 27(1): 37-40.
[53]张名位,郭宝江,池建伟等.黑米皮提取物的体外抗氧化作用与成分分析[J].中国粮油学报, 2005, 20(6): 49.
[54]金杰,李志西,张锋等.桑椹醋提取物对二苯代苦味酰基自由基(DPPH·)的清除作用[J].西北农林科技大学学报, 2006, 34(3): 135.
[55]侯同刚,李天择,黄淑霞.毛细管电泳在氨基酸分析中的最新进展[J].食品科技, 2004, 2: 85-87.
[56]张晖,徐永,姚惠源.纸层析法定量测定米胚芽中的γ-氨基丁酸[J].无锡轻工大学学报, 2004, 23 (2) :101
[57] Saikusa T. Accumulation of gamma aminobutyric acid (GABA) in the rice germ during water soaking[J]. Biosci Biotesh Biochem, 1994, 58(12): 2291-2292.
[58] Lindroth P, Mopper K. High-performance liquid chromatography determination of subpicomole of amino acids by fluorescence derivatization with o-phthaldialdedyde[J]. Annl Chem, 1975, 51(10): 1667-1669.
[59]Liu H J. Determination of amino acids by precolumn derivatization with 6-aminoquinolyl-N-hydroxysucciniomidyl carbamate and high- performance liquid chromatography with ultraviolet detection[J]. J Chromarogr, 1994, 670(1): 59-62.
[60]张红漫,于文涛,欧阳平凯等.气相色谱-质谱法测定中药沙苑子中的氨基酸[J].氨基酸和生物资源, 2004, 26 (2): 62-64.
[61]王安群. LC-MS法测定发芽糙米等产品中的γ-氨基丁酸[J].中国科技信息, 2006, 4: 81-83.
[62]常碧影,刘洪基,闫惠文等.柱前衍生高效液相色谱法测定氨基酸[J].分析化学, 1995, 23 (1): 100-102.
[63] Olivia P, Isable M F, Eulalia M, et al. Effect of temperature on evolution of free amino acid and biogenic amine contents during storage of Azeitao cheese[J]. Food chemistry, 2001, 75: 287-291.
[64]曹玉华,汪云.毛细管电泳电化学检测法测定胡黄连中香草酸和阿魏酸的含量[J].分析测试学报, 2003, 22(6): 95-97.