茶树紫色芽叶花青苷组分分析及结构推测

doi:10.13305/j.cnki.jts.2014.03.010

摘要/Abstract

摘要： 通过筛选流动相、优化洗脱梯度,建立了茶树紫色芽叶花青素HPLC的分析方法,并从提取的花青素粗制品中分离出9个主要色谱峰;运用HPLC-DAD-MS/MS联用技术,获得了其中7个组分的紫外-可见光谱信息、质谱数据,结合文献资料,初步推测了茶树紫色芽叶花青素中5个主要组分,分别为：飞燕草-3-半乳糖苷、矢车菊-3-半乳糖苷、飞燕草-3-芸香糖苷、矢车菊-3-芸香糖苷和天竺葵-3-芸香糖苷,此外,采用外标法对各组分进行了定量分析,并获得了粗提物,纯度为32%。该方法简便快捷,重现性好,可用于茶树紫色芽叶中花青素组分的分析与鉴定。

关键词: 茶树紫色芽叶, 花青素, 组分, 高效液相色谱

Abstract: A rapid method for analysis of anthocyanins in purple buds and leaves of tea plant was established by high-performance liquid chromatography (HPLC) based on the screening of mobile phase and the optimization of elution gradient. Using the established HPLC method, nine major chromatographic peaks were separated from crude extracts of anthocyanins from purple buds and leaves of tea plant. Further, the ultraviolet-visible spectrum and mass spectrum data of seven out the nine components were obtained by HPLC-DAD-MS/MS analysis. Combined with the literature data, five major components of anthocyanins were identified as delphindin-3-galactoside, cyanidin-3-gal-actoside, delphindin-3-rutinoside, cyanidin-3-rutinoside, and pelargonium-3-rutinoside. Additionally, each component was quantitatively analyzed using an external standard method, and crude extracts of anthocyanins were obtained at the purity of 32%. The proposed method is convenient and rapid with high accuracy and reproducibility, which can be applied for determining anthocyanins in purple buds and leaves of tea plant.

Key words: purple leaves of tea plant, anthocyanins, component, HPLC

中图分类号:

李智, 王日为, 张丽霞, 韩晓阳. 茶树紫色芽叶花青苷组分分析及结构推测[J]. 茶叶科学, 2014, 34(3): 279-287. doi: 10.13305/j.cnki.jts.2014.03.010.

LI Zhi, WANG Riwei, ZHANG Lixia, HAN Xiaoyang. Analysis and Structural Identification of Anthocyanin Components in Purple Buds and Leaves of Tea Plant[J]. Journal of Tea Science, 2014, 34(3): 279-287. doi: 10.13305/j.cnki.jts.2014.03.010.

参考文献 27

[1]	陈兴琰. 茶树育种学[M]. 2版. 北京: 中国农业出版社, 1986: 89.
[2]	赵先明, 王孝仕, 杜晓. 茶树紫色芽叶的呈味特征及降低苦涩味的研究[J]. 茶叶科学, 2009, 29(5): 372-378.
[3]	吴华玲, 何玉媚, 李家贤, 等. 11个红紫芽茶树新品系的芽叶特性和生化成分研究[J]. 植物遗传资源学报, 2012, 13(1): 42-47.
[4]	安徽农学院. 茶叶生物化学[M]. 北京: 中国农业出版社, 1995: 93-94.
[5]	杨兴荣, 包云秀, 黄玫. 云南稀有茶树品种“紫娟”的植物学特征和品质特征[J]. 茶叶, 2009(1): 17-18.
[6]	王日为, 张丽霞. 茶叶中花青素类物质研究进展[J]. 茶叶科学技术, 2002, 4(32): 14-17.
[7]	吴朝霞. 葡萄籽原花青素分离提纯、组分鉴定及抗氧化性研究[D]. 沈阳: 沈阳农业大学, 2005: 30-32.
[8]	Wang H C, Prior R L.Oxygen radical absorbing capacity of[J]. Agric Food Chem, 1997, 45(3): 304-309.
[9]	Youdim K A, Mcdonal D J, Kalt W, et a1. Potential role dietary flavonoids in reducing microvascular endothelium vulnerabilit to oxidative and inflammatory insults[J]. Nutr Biochem, 2002, 13(2): 282-288.
[10]	Prior R L, Wu X.Anthocyanins: structural characteristics that result in unique metabolic patterns and biologica activities[J]. Free Radic Res, 2006, 40(10): 1014-1028.
[11]	萧力争, 苏小倩, 李勤, 等. 紫芽品种茶树春梢芽叶生化成分分析[J]. 福建农林大学学报: 自然科学版, 2009, 38(1): 30-33.
[12]	李双伶, 郭俊凌, 杜晓, 等. 茶树紫色芽叶中花青素的提取-层析分离及初步鉴定[J]. 安徽农业科学, 2009, 37(28): 13799-13802.
[13]	吕海鹏, 费旭元, 梁名志, 等. 茶树特异品种“紫娟”中的花青素组分分析[J]. 食品科学, 2012, 33(22): 203-206.
[14]	龚家顺, 隋华嵩, 彭春秀, 等. “紫娟”晒青绿茶色素的 HPLC-ESI-MS分离鉴定及其稳定性研究[J]. 茶叶科学, 2012, 32(2): 179-188.
[15]	L C Kerio, F N Wachira, J K Wanyoko, et al. Characterization of anthocyanins in Kenyan teas: Extraction and identiﬁcation[J]. Food Chemistry, 2012, 131(1): 31-38.
[16]	Snyder L R.Practical HPLC method development [M]. 张玉奎, 译. 2nd ed. 北京: 华文出版社, 2001: 498-506.
[17]	Panagiotis Arapitsas, Per J R, Sjo berg, et al. Characterisation of anthocyanins in red cabbage using high resolution liquid chromatography coupled with photodiode array detection and electrospray ionization-linear ion trap mass spectrometry[J]. Food Chemistry,2008(1092): 198-226.
[18]	F Pomar, M Novo, A Masa.Varietal differences among the anthocyanin profiles of 50 red table grape cultivars studied by high performance liquid chromatography[J]. Journal of Chromatography A, 2005(1094): 34-41.
[19]	杨正宪, 蔡正宗, 桑葚之. Anthocyanins结构鉴定[J]. 台湾食品科学, 1994, 21(5): 319-330.
[20]	Farah S Hosseinian, Wende Li, Trust Beta.Measurement of anthocyanins and other phytochemicals in purple wheat[J]. Food Chemistry, 2008, 109: 916-924.
[21]	Romina Daniela Di Paola-Naranjo, José Sánchez-Sánchez, Ana Mar´ıa González-Paramás. Liquid chromatographic-mass spectrometric analysis of anthocyanin composition of dark blue bee pollen from Echium plantagineum[J]. Journal of Chromatography A, 2004, 1054: 205-210.
[22]	Ana C Manhita, Dora M Teixeira, Cristina T da Costa. Application of sample disruption methods i n the extraction of anthocyanins from solid or semi-solid vegetable samples[J]. Journal of Chromatography A, 2006, 1129: 14-20.
[23]	Norihiko Terahara, Yoshiyuki Takeda, Atsushi Nesumi, et al. Anthocyanins from red flower tea (Benibana-cha), Camellia sinensis[J]. Phytochemistry, 2001(56): 359-361.
[24]	Glenda A Macz-Pop, Julia′n C Rivas-Gonzalo, Jose′ J Pe′rez-Alonso. Natural occurrence of free anthocyanin aglycones in beans (Phaseolus vulgaris L.)[J]. Food Chemistry, 2006, 94: 448-456.
[25]	F Pomar, M Novo, A Masa.Varietal differences among the anthocyanin profiles of 50 red table grape cultivars studied by high performance liquid chromatography[J]. Journal of Chromatography A, 2005, 1094: 34-41.
[26]	Qingguo Tian, M Monica Giusti, Gary D Stoner. Screening for anthocyanins using high-performance liquid chromatography coupled to electrospray ionization tandem mass spectrometry with precursor-ion analysis, product-ion analysis, common-neutral-loss analysis, and selected reaction monitoring[J]. Journal of Chromatography A, 2005, 1091: 72-82.
[27]	Lin L Z, Chen P, James M, et al. NEW phenolic components and chromatographic profiles of green and fermented teas[J]. J Aric Food Chem, 2008, 56(17): 8130-8140.