直读显色快速检测绿茶中的茶氨酸
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摘要
本研究首先基于溶胶-凝胶方法成功构筑了茚三酮/纳米二氧化钛纳米材料,将其作为纳米显色剂打印在高效薄层板上茶氨酸所对应的比移值(Rf)处。以乙醇:冰乙酸展开体系对适当脱色处理后的茶叶浸出物进行快速薄层直读显色分析。结果表明,在优化后的展开条件下该直读条带能够有效地分离茶叶提取物中多种氨基酸,所构筑的新型纳米显色剂对目标氨基酸茶氨酸实现了灵敏的直读显色,自显色后条带色斑均匀清晰,其颜色深浅度与其浓度之间有明确的半定量关系。本研究构建的直读显色条带具有安全、简便快捷、成本低等优点,在农产品品质鉴定与现场快速检测氨基酸等领域有着广泛的应用前景。
In this study, ninhydrin/titanium dioxide nanomaterials were constructed by using Sol-Gel method. These nanomaterial were then used as direct-reading colorimetric materials and printed on the surface loci of thin layer plate where theanine would migrate according to its retardation factor(Rf). In addition, ethanol: acetic acid developing system was chosen as developing reagent, the extraction solution of tea after being decolorized by ethyl acetate was analyzed by thin layer chromatography(TLC) via direct-reading colorimetric reaction. The results showed that the current direct-reading strip could effectively separate several amino acids in tea extraction under optimized conditions. The new constructed colorimetric nanomaterials had achieved a sensitive direct-reading coloration results for the target amino acid theanine, and the color spots on the strip were uniform and clear after self-coloration. Moreover, its color depth had a clear semi-quantitative relationship with its concentration. The current colorimetric strip has a serial of advantages including safety, easy-operation, convenient and low cost, and has a broad application prospect in fields such as quality identification of agricultural products and on-site rapid detection of amino acids.
In this study, ninhydrin/titanium dioxide nanomaterials were constructed by using Sol-Gel method. These nanomaterial were then used as direct-reading colorimetric materials and printed on the surface loci of thin layer plate where theanine would migrate according to its retardation factor(Rf). In addition, ethanol: acetic acid developing system was chosen as developing reagent, the extraction solution of tea after being decolorized by ethyl acetate was analyzed by thin layer chromatography(TLC) via direct-reading colorimetric reaction. The results showed that the current direct-reading strip could effectively separate several amino acids in tea extraction under optimized conditions. The new constructed colorimetric nanomaterials had achieved a sensitive direct-reading coloration results for the target amino acid theanine, and the color spots on the strip were uniform and clear after self-coloration. Moreover, its color depth had a clear semi-quantitative relationship with its concentration. The current colorimetric strip has a serial of advantages including safety, easy-operation, convenient and low cost, and has a broad application prospect in fields such as quality identification of agricultural products and on-site rapid detection of amino acids.
引文
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[13]Wood PL,Khan MA,Moskal JR.Neurochemical analysis of amino acids,polyamines and carboxylic acids:GC-MS quantitation of t BDMS derivatives using ammonia positive chemical ionization[J].Journal of Chromatography B,2006,831:313-319.
[14]Thippeswamy R,Gouda KGM,Rao DH,et al.Determination of theanine in commercial tea by liquid chromatography with fluorescence and diode array ultraviolet detection[J].Journal of Agricultural and Food Chemistry,2006,54(19):7014-7019.
[15]杨四润,张冬莲,方乔慧,等.γ-氨基丁酸茶薄层检测方法的建立[J].食品安全质量检测学报,2012,3(1):33-39.
[16]王督,张文,李培武,等.胶体金免疫层析法快速定量分析粮油农产品中黄曲霉毒素B1[J].中国油料作物学报,2014,36(4):529-532.
[17]张锂.薄层层析酶化学法测定马拉硫磷残留[J].兰州工业学院学报,2017,24(6):82-85.
[18]刘顺字,郭淑贞,林双娣,等.酶抑制法在农药残留快速检测过程中的应用分析[J].湖南农业科学,2017(7):85-87,91.
[19]杨丽,尤丽,叶金秀,等.纸层析分离鉴定氨基酸实验的改进[J].云南民族大学学报(自然科学版),2011,20(3):229-231
[20]王海龙,崔智慧,岳秀兰.氨基酸薄层层析茚三酮显色改良[J].包头医学院学报,2003,19(4):312-360.
[21]马雪泷,邹鹏飞,王荡强,等.纸层析-分光光度法检测茶氨酸[J].植物生理学报,2012,48(4):413-417.
[22]张宜明,叶海鹏,李顺,等.茚三酮/纳米二氧化钛复合物及其制备方法和用途:CN201510817477.3[P].2016-01-27[2017-12-05].http://www.wanfangdata.com.cn/details/detail.do?_type=pat ent&id=CN201510817477.3.
[23]阮宇成,王月根.茶氨酸的简化定量法[J].茶叶通讯.1981(3):41-45.
[24]张丹丹,叶小辉,赵峰,等.基于游离氨基酸组分的白茶滋味品质研究[J].福建农业学报,2016,31(5):515-520.
[2]Wang HF,Tsai YS,Lin ML,et al,Comparison of bioactive components in GABA tea and green tea produced in Taiwan[J].Food Chemistry 2006,96(4):648-653.
[3]Scharbert S,Hofmann T.Molecular definition of black tea taste by means of quantitative studies,taste reconstitution,and omission experiments[J].Journal of Agricultural and Food Chemistry,2005,53:5377-5384.
[4]毛清黎.茶叶氨基酸的研究进展[J].氨基酸和生物资源,1989(4):16-21.
[5]程启坤,阮宇成,王月根,等.绿茶滋味化学鉴定法[J].茶叶科学,1985,5(1):7-17.
[6]刘爽,杨停,谭俊峰,等.绿茶滋味定量描述分析及其化学成分的相关性研究[J].中国农学通报,2014,30(24):40-46.
[7]Ruan JY,Gerendas J,Hardter R,et al.Effect of nitrogen form and root-zone p H on growth and nitrogen uptake of tea(Camellia sinensis)plants[J].Annals of Botany,2007,99(2):301-310.
[8]Deng WW,Ogita S,Ashihara H,Biosynthesis of theanine(gamma-ethylamino-L-glutamic acid)in seedlings of Camellia sinensis[J].Phytochemistry Letters,2008,1(2):115-119.
[9]Oh K,Kato T,Xu HL.Transport of nitrogen assimilation in xylem vessels of green tea plants fed with NH4-N and NO3-N[J].Pedosphere,2008,18(2):222-226.
[10]Ruan J,Haerdter R,Gerendas J.Impact of nitrogen supply on carbon/nitrogen allocation:a case study on amino acids and catechins in green tea[Camellia sinensis(L.)O.Kuntze]plants[J].Plant Biology,2010(12):724-734.
[11]Sasaoka K,Kito M,Onishi Y.Some properties of theanine synthesizing enzyme in tea seedlings[J].Agricultural and Biological Chemistry,1965,29(11):984-988.
[12]黄建琴.氨基酸在茶叶制造中的转化机理及对茶叶品质的影响[J].氨基酸和生物资源,1992(1):26-29.
[13]Wood PL,Khan MA,Moskal JR.Neurochemical analysis of amino acids,polyamines and carboxylic acids:GC-MS quantitation of t BDMS derivatives using ammonia positive chemical ionization[J].Journal of Chromatography B,2006,831:313-319.
[14]Thippeswamy R,Gouda KGM,Rao DH,et al.Determination of theanine in commercial tea by liquid chromatography with fluorescence and diode array ultraviolet detection[J].Journal of Agricultural and Food Chemistry,2006,54(19):7014-7019.
[15]杨四润,张冬莲,方乔慧,等.γ-氨基丁酸茶薄层检测方法的建立[J].食品安全质量检测学报,2012,3(1):33-39.
[16]王督,张文,李培武,等.胶体金免疫层析法快速定量分析粮油农产品中黄曲霉毒素B1[J].中国油料作物学报,2014,36(4):529-532.
[17]张锂.薄层层析酶化学法测定马拉硫磷残留[J].兰州工业学院学报,2017,24(6):82-85.
[18]刘顺字,郭淑贞,林双娣,等.酶抑制法在农药残留快速检测过程中的应用分析[J].湖南农业科学,2017(7):85-87,91.
[19]杨丽,尤丽,叶金秀,等.纸层析分离鉴定氨基酸实验的改进[J].云南民族大学学报(自然科学版),2011,20(3):229-231
[20]王海龙,崔智慧,岳秀兰.氨基酸薄层层析茚三酮显色改良[J].包头医学院学报,2003,19(4):312-360.
[21]马雪泷,邹鹏飞,王荡强,等.纸层析-分光光度法检测茶氨酸[J].植物生理学报,2012,48(4):413-417.
[22]张宜明,叶海鹏,李顺,等.茚三酮/纳米二氧化钛复合物及其制备方法和用途:CN201510817477.3[P].2016-01-27[2017-12-05].http://www.wanfangdata.com.cn/details/detail.do?_type=pat ent&id=CN201510817477.3.
[23]阮宇成,王月根.茶氨酸的简化定量法[J].茶叶通讯.1981(3):41-45.
[24]张丹丹,叶小辉,赵峰,等.基于游离氨基酸组分的白茶滋味品质研究[J].福建农业学报,2016,31(5):515-520.