[1]董方,张群峰,阮建云.超高效液相色谱-三重四级杆串联质谱法测定茶叶中黄酮醇糖苷种类及含量[J].江苏农业学报,2021,(01):204-212.[doi:doi:10.3969/j.issn.1000-4440.2021.01.027]
　DONG Fang,ZHANG Qun-feng,RUAN Jian-yun.Determination of flavonol glycosides in teas by ultra-high performance liquid chromatography combined with triple four-bar tandem mass spectrometry[J].,2021,(01):204-212.[doi:doi:10.3969/j.issn.1000-4440.2021.01.027]
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超高效液相色谱-三重四级杆串联质谱法测定茶叶中黄酮醇糖苷种类及含量()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2021年01期
页码:
204-212
栏目:
加工贮藏·质量安全
出版日期:
2021-02-28

文章信息/Info

Title:
Determination of flavonol glycosides in teas by ultra-high performance liquid chromatography combined with triple four-bar tandem mass spectrometry
作者:
董方1; 2; 张群峰2; 阮建云2
（1.江西省农业科学院园艺研究所,江西南昌330200；2.中国农业科学院茶叶研究所,浙江杭州310008）
Author(s):
DONG Fang1; 2; ZHANG Qun-feng2; RUAN Jian-yun2
(1.Institute of Horticulture, Jiangxi Academy of Agricultural Sciences, Nanchang 330200, China;2.Tea Research Institute, Chinese Academy of Agricultural Sciences, Hangzhou 310008, China）
关键词:
茶叶; 黄酮醇糖苷; 超高效液相色谱-三重四级杆串联质谱法; 质谱多反应监测; 优化
Keywords:
tea; flavonol glycosides; ultra-high performance liquid chromatography combined with triple four-bar tandem mass spectrometry; mass spectrometry multiple reaction monitoring; optimization
分类号:
O658；TS272
DOI:
doi:10.3969/j.issn.1000-4440.2021.01.027
文献标志码:
A
摘要:
基于超高效液相色谱-三重四级杆串联质谱（UPLC-QqQ-MS/MS）联用技术对茶叶中黄酮醇糖苷（FGs）种类及含量进行测定并对提取方式和检测条件进行优化。用75%（体积分数）甲醇水溶液提取目标化合物，再用Waters Acquity HSS T3色谱柱（粒径为1.8 μm，长度×内径为100.0 mm×2.1 mm）分离目标化合物，以含有0.1%（体积分数）甲酸的乙腈溶液为流动相进行梯度洗脱，采用电喷雾正离子源（ESI+）与质谱多反应监测（MRM）方法对成品茶中15种FGs进行定量测定。结果表明，在0.1～20.0 μg/ml的质量浓度范围内FGs的基质标准曲线的线性关系良好，其检测限（LOD）为0.08～0.23 μg/L，定量限（LOQ）为0.25～0.76 μg/L，相对标准偏差（RSD）为1.30%～2.80%，出峰时间为3.60～7.40 min。与现有最优检测技术相比，超高效液相色谱-三重四级杆串联质谱法具有分析速度快、灵敏度高、稳定性好等特点，可为提高茶叶品质成分的检测效率提供参考。
Abstract:
The extraction methods and detection conditions of flavonol glycosides (FGs) in teas were optimized based on ultra-high performance liquid chromatography combined with triple four-bar tandem mass spectrometry (UPLC-QqQ-MS/MS) technology, and the varieties and contents were determined. The target compounds were extracted with 75% (volume fraction) methanol aqueous solution, separated on Waters Acquity HSS T3 column (particle size was 1.8 μm, length × inner diameter was 100.0 mm × 2.1 mm), and gradient elution was carried out using acetonitrile solution containing 0.1% (volume fraction) formic acid as moving phase. Positive electrospray ionization (ESI+) and mass spectrometry multiple reaction monitoring (MRM) methods were used to quantitatively determine 15 flavonol glycosides in made teas. The results indicated that the matrix standard curve of FGs showed a good linearity in the concentration range of 0.1-20.0 μg/ml, the limit of detection (LOD) was 0.08-0.23 μg/L, the limit of quantity (LOQ) was 0.25-0.76 μg/L, the relative standard deviation (RSD) was 1.30%-2.80%, and the peak out time was 3.60-7.40 min. Compared with the existing optimal detection technology, UPLC-QqQ-MS/MS method has fast analysis speed, high sensitivity and good stability, which can provide reference for improving the detection efficiency of tea quality components.

参考文献/References:

[1]MATSUBARA Y, KUMAMOTO H, IIZUKA Y, et al. Studies on physiologically active substances in citrus peel. Part Ⅱ. Structure and hypotensive effect of flavonoid glycosides in Citrus unshiu peelings[J]. Agricultural and Biological Chemistry, 1985, 49: 909-914.
[2]刘阳,陈根生,许勇泉,等. 冲泡过程中西湖龙井茶黄酮苷类浸出特性及滋味贡献分析[J].茶叶科学,2015, 35(3): 217-224.
[3]WU Y H, JIANG X L, ZHANG S X， et al. Quantification of flavonol glycosides in Camellia sinensis by MRM mode of UPLC-QqQ-MS/MS[J]. Journal of Chromatography B, 2016, 1017: 10-17.
[4]林杰,段玲靓,吴春燕,等. 茶叶中的黄酮醇类物质及对感官品质的影响[J].茶叶,2010, 36(1): 14-18.
[5]戴前颖,夏涛,高丽萍,等. 绿茶汤呈色物质研究进展[J].安徽农业大学学报,2011, 38(6): 887-891.
[6]朱博,夏涛,高丽萍,等. 绿茶茶汤中黄酮醇及其苷类的测定方法以及对茶汤色度的影响[J].食品与发酵工业, 2009,35(2):146-150.
[7]WANG S Y, BOWMAN L, DING M. Methyl jasmonate enhances antioxidant activity and flavonoid content in blackberries (Rubus sp.) and promotes antiproliferation of human cancer cells[J]. Food Chemistry, 2008, 107(3): 1261-1269.
[8]FERREYRA M L F, RIUS S P, CASATI P. Flavonoids:biosynthesis, biological functions, and biotechnological applications[J].Frontiers in Plant Science,2012, 3: 1-10.
[9]方芳,王凤忠. 植物黄酮醇的检测方法研究进展[J].食品工业科技, 2018, 39(11): 327-332.
[10]王智聪,沙跃兵,余笑波,等. 超高效液相色谱-二极管阵列检测-串联质谱法测定茶叶中15种黄酮醇糖苷类化合物[J].色谱, 2015, 33(9): 974-980.
[11]张维冰,王智聪,张凌怡. 超高效液相色谱-光电二极管阵列检测-串联四级杆质谱法测定红洋葱中黄酮醇及其糖苷类化合物[J].分析化学,2014, 42(3): 415-422.
[12]ZHAO Y Y, GUO H Z, CHEN Y G, et al. Simultaneous quantification of flavonol glycosides, terpene lactones, polyphenols and carboxylic acids in Ginkgo biloba leaf extract by UPLC-QTOF-MSE based metabolomic approach[J]. Journal of Pharmaceutical Sciences, 2017, 26(11): 789-804.
[13]KIM J C, SHIM Y S. Method validation of analytical method for 12 flavonol glycosides in foods using ultra high-performance liquid chromatography coupled with photodiode array detection[J]. Food Acience and Biotechnology, 2016, 25(3): 659-664.
[14]JIANG H Y, ENGELHARDT U H, THRNE C, et al. Determination of flavonol glycosides in green tea, oolong tea and black tea by UHPLC compared to HPLC[J]. Food Chemistry, 2015, 183: 30-35.
[15]ZHANG Q F, SHI Y Z, MA L F, et al. Metabolomic analysis using ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry (UPLC-Q-TOF MS) uncovers the effects of light intensity and temperature under shading treatments on the metabolites in tea[J]. PLoS One, 2014, 9(11): e112572.
[16]DE VOS R C, MOCO S, LOMMEN A, et al. Untargeted large-scale plant metabolomics using liquid chromatography coupled to mass spectrometry[J]. Nature Protocols, 2007, 2(4): 778-791.
[17]VRHOVSEK U, MASUERO D, GASPEROTTI M, et al. A versatile targeted metabolomics method for the rapid quantification of multiple classes of phenolics in fruits and beverages[J]. Journal of Agricultural and Food Chemistry, 2012, 60(36): 8831-8840.
[18]戴伟东,解东超,吕美玲,等. 黄酮醇糖苷与茶树品种适制性关系[J].食品科学, 2017, 38(16): 104-109.
[19]吴春燕,须海荣, HRITIER J,等.不同茶树品种中黄酮苷含量的测定[J].茶叶科学, 2012, 32(2): 122-128.
[20]RIO D D, STEWART A J, MULLEN W, et al. HPLC-MSn analysis of phenolic compounds and purine alkaloids in green and black tea[J]. Journal of Agricultural and Food Chemistry, 2004, 52(10): 2807-2815.
[21]BENVENUTI M E, O’CONNOR A. Melamine, ammeline and cyanuric acid analysis by UPLC/MS/MS and UPLC/PDA[EB/OL].
[2020-01-02]. http://www.waters.com/wa-ters/library.htm.
[22]王旭堂,刁志祥,张培杨,等. 超高液相色谱荧光检测法测定鹌鹑蛋和鸽蛋中甲砜霉素残留量[J].江苏农业学报, 2020, 36(1): 206-211.
[23]陈丛瑾,黄克瀛,李德良,等. 植物中黄酮类化合物的提取方法研究概况[J].生物质化学工程, 2007,41(3):42-46.
[24]梁滨, 周青. UV-B辐射对植物类黄酮影响的研究进展[J].中国生态农业学报, 2007, 15(3): 191-194.
[25]骆耀平. 茶树栽培学[M]. 5版.北京:中国农业出版社, 2015: 88-100.
[26]宋楚君,范方媛,龚淑英,等. 不同产地红茶的滋味特征及主要贡献物质[J].中国农业科学, 2020, 53(2): 383-394.
[27]郭世昌,常有礼,胡非,等. 纬度和海拔高度对云南地面紫外线强度影响的数值试验[J].云南地理环境研究,2004,16(1):9-13.
[28]江昌俊. 茶树育种学[M].北京:中国农业出版社, 2005: 93-95.
[29]陈宗懋,甄永苏. 茶叶的保健功能[M].北京:科学出版社, 2014: 35-56.
[30]PETERSON J, DWYER J, BHAGWAT S, et al. Major flavonoids in dry tea[J]. Journal of Food Composition and Analysis, 2005, 18(6): 487-501.

备注/Memo

备注/Memo:
收稿日期：2020-05-14基金项目：国家重点研发计划项目（2016YFD0200900）；国家茶产业技术体系建设专项（CARS-19）作者简介：董方(1992-)，男，陕西西乡人，硕士研究生，研究方向为茶树栽培生理与生态。（E-mail）18305811752@163.com通讯作者：张群峰，（E-mail）hill@tricaas.com
更新日期/Last Update: 2021-03-15