UPLC-MS/MS法快速分析换锦花中加兰他敏的动态积累变化  

Rapid Analysis of Dynamic Accumulation of Galanthamine in Lycoris sprengeri by UPLC-MS/MS Method

作　　者：李青竹[1] 蔡友铭[1] 张永春[1] 郑玉红[2] 杨柳燕[1] Li Qingzhu;Cai Youming;Zhang Yongchun;Zheng Yuhong;Yang Liuyan(Shanghai Key Laboratory of Protected Horticultural Technology,Forestry and Pomology Research Institute,Shanghai Academy of Agricultural Sciences,Shanghai,201106;Institute of Botany,Jiangsu Province and Chinese Academy of Sciences,Nanjing,210014)

机构地区：[1]上海市农业科学院林木果树研究所上海市设施园艺技术重点实验室,上海201106 [2]江苏省中国科学院植物研究所,南京210014

出　　处：《分子植物育种》2019年第3期1011-1018,共8页Molecular Plant Breeding

基　　金：上海市农委攻关项目(沪农科攻字(2016)第5-3号);上海市农委应用基础项目(沪农科攻字(2016)第6-1-6号);上海市农委产业体系建设专项(沪农科产字(2018)第8号);上海市农委攻关项目(沪农科攻字(2014)第1-2号)共同资助;上海市科委"科技创新行动计划"社会发展领域重点研发项目(17DZ1201802)

摘　　要：本研究以超高效液相色谱-串联四极杆/线性离子阱质谱(UPLC-QTRAP-MS/MS)为基础,建立了1种快速准确检测石蒜属植物中加兰他敏含量的方法。提取液超声波辅助提取离心后,经Waters ACQUITY UPLC BEH C18(150 mm×2.1 mm, 1.7μm)色谱柱分离,以含0.1%甲酸的水溶液和乙腈为流动相进行等度洗脱,利用正离子多反应监测模式(MRM)进行分析,使分析时间大幅缩短,提高分析准确度。在0.5～100μg/m L浓度范围内线性关系良好,相关系数达0.999 9,可在3分钟以内的程序时间内得到检测结果,回收率为95.20%～98.92%,相对标准偏差为1.03%～1.75%,经方法学确认本方法准确可靠。应用此方法测得石蒜属换锦花鳞茎周年不同月份加兰他敏含量介于163.86～2 065.79μg/g干重之间,7月积累最高,加兰他敏积累与生长过程密切相关,总体表现为在休眠期积累,不同的生长时期加兰他敏在换锦花各个器官的分配变化较大。本研究揭示了换锦花加兰他敏的动态积累规律,并为研究加兰他敏的合成代谢调控提供了准确的分析手段。This work established an efficient analytical method based on ultra-performance liquid chromatography- quadrupole linear ion trap mass spectrometry (UPLC-QTRAP-MS/MS)for rapid determination of galanthamine in Lycoris spp.After ultrasonic-assisted extraction and centrifugation,the sample was separated on a Waters ACQUITY UPLC BEH C18 (150 mm×2.1 mm, 1.7μm)with a mobile phase of acetonitrile-water (0.1% formic acid)at isocratic elution program.The target compounds galanthamine was analyzed in the positive ion multiple reaction monitoring (MRM)mode with shorter time and improved accuracy.As a result,good linearities was obtained in a certain concentration ranges (0.5~100μg/m L)with correlation coefficients of 0.999 9 within 3 min.The average recoveries were 95.20%~98.92%,and relative standard deviations were 1.03%~1.75%.The accuracy and reliability were confirmed by methodology assessment.Using this method,we found that the monthly dynamic variation of galanthamine contents in L.sprengeri ranged from 163.86~2 065.79 p,g/g dry weight,and the highest accumulation period was July,indicating that accumulation of galanthamine had close relationship with its growth stage.Generally,galanthamine was highly accumulated at the dormancy stage and its accumulation pattern among different tissues varied a lot during different growth stages.This work showed the dynamic accumulation pattern of galanthamine in Lycoris sprengeri,which could provide a reliable and effective technique for the galanthamine accumulation detection and metabolic regulation in Lycoris spp.

关 键 词：换锦花 超高效液相色谱-串联二级质谱 加兰他敏 动态积累 

分 类 号：S682.29[农业科学—观赏园艺]