董方　张群峰　阮建云

摘要： 基于超高效液相色譜-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。

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

中图分类号： O658;TS272 文献标识码： A 文章编号： 1000-4440（2021）01-0204-09

Determination of flavonol glycosides in teas by ultra-high performance liquid chromatography combined with triple four-bar tandem mass spectrometry

DONG Fang1，2， ZHANG Qun-feng2， RUAN Jian-yun 2

（1.Institute of Horticulture， Jiangxi Academy of Agricultural Sciences， Nanchang 330200， China;2.Tea Research Institute， Chinese Academy of Agricultural Sciences， Hangzhou 310008， China）

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.

Key words： tea;flavonol glycosides;ultra-high performance liquid chromatography combined with triple four-bar tandem mass spectrometry;mass spectrometry multiple reaction monitoring;optimization

黄酮醇糖苷（Flavonols glycosides，FGs）是茶叶中除儿茶素外较为重要的多酚类物质之一，大都由黄酮醇苷元（杨梅素、槲皮素、山柰酚等）或黄酮苷元（芹菜素等）与糖分子（葡萄糖、半乳糖、芸香糖等）结合形成O-糖苷[1-2]。已有研究发现，FGs不仅对茶叶中的苦涩味及茶汤的光泽度、亮度具有重要作用[3-6]，而且具有类黄酮物质的生理学共性，在生物体中发挥着抗氧化、清除活性氧自由基及抵御紫外线、抗菌抑菌等作用[7-8]。

国内外有很多FGs检测方法的报道，目前常用的检测方法主要包括紫外分光光度法（UV spectrophotometer）、高效液相色谱法（HPLC）、液相色谱-质谱检测法（LC-MS）、超高效液相色谱法（UPLC）、毛细管电泳法（CE）及电化学检测法（ED）等[9]。……