摘要
【目的】建立可同时测定6种植物激素(GA3、iPAde、6-BA、IAA、SA、ABA)的液相色谱-串联质谱联用法(Liquid chromatography-mass spectrometry/mass spectrometry,LC-MS/MS).【方法】采用C18色谱柱(150 mm×2.1 mm,1.8μm)分离和梯度洗脱,质谱采用电喷雾离子源负模式(ESI-),在多反应监测(Multi-reaction monitor,MRM)模式下进行定性分析,标准品标准曲线定量.【结果】在流动相:B(甲醇)∶A(0.1%甲酸),梯度洗脱:0~1 min 10%~40%(B)、1~2.5 min 40%~45%(B)、2.5~5 min 45%~80%(B)、5~5.1 min 80%~10%(B)、5.1~7.1 min 10%~10%(B),检测波长290 nm,柱温35℃,流速0.3 mL/min,离子源温度350℃,干燥气流速11 L/min,雾化器压力35 psi和毛细管电压4 kV等条件下,标准品检出限为1.0μg/L,在1.0~100μg/L质量浓度范围内线性关系良好,相关系数R2≥0.99.采用上述检测方法,马铃薯、油菜和苜蓿3种植物叶片中6种内源激素均有较好的出峰和分离度.【结论】该LC-MS检测方法快速、灵敏和准确,可进行多种植物激素的同时测定.
【Objective】To establish a method of simultaneous determination of six plant hormones(GA3,iPAde,6-BA,IAA,SA and ABA)by LC-MS/MS.【Method】The C18 column(150 mm×2.1 mm,1.8μm)and elution gradient,with electro spray ion source negative mode(ESI-)used in MS,combined with multi-reaction monitor(MRM)and standard curve carried out to the qualitative and quantitative analysis.【Result】With the binary solvent system consisting of 100%methanol(eluent B)and 0.1%formic acid in H2O(eluent A)as follow:0~1 min 10%~40%(B),1~2.5 min 40%~45%(B),2.5~5 min 45%~80%(B),5~5.1 min 80%~10%(B)and 5.1~7.1 min 10%~10%(B),as well as detection wavelength(290 nm),column temperature(35℃),flow speed(0.3 mL/min),ion source temperature(350℃),drying gas flow rate(11 L/min),atomizer pressure(35 psi)and capillary voltage(4kV),the limit of detection was 1.0μg/L,the linear relationship was good in the concentration range 1.0~100μg/L,and the correlation coefficient was R2≥0.99.Under the above condition,the six plant hormones in the leaves of Solanum tuberosum,Brassica napus and Medicago sativa presented better peak and separation.【Conclusion】The established method with the advantages of rapid,sensitive and specific can be simultaneously applied in determining multiple hormones in plants.
作者
张丹
孙萍
陈思瑾
幸华
芮仕立
牛早霞
栗孟飞
ZHANG Dan;SUN Ping;CHEN Si-jin;XING Hua;RUI Shi-li;NIU Zao-xia;LI Meng-fei(College of Life Science and Technology,Gansu Provincial Key Lab of Arid Land Crop Science,Gansu Agricultural University,Lanzhou 730070,China;Gansu Dingxi Station of Agricultural Ecology and Resource Protection Technology,Dingxi,743000,China)
出处
《甘肃农业大学学报》
CAS
CSCD
北大核心
2020年第4期98-103,共6页
Journal of Gansu Agricultural University
基金
国家自然科学基金项目(81560617)
甘肃农业大学“伏羲杰出人才培育计划”(Gaufx-02J04)
干旱生境作物学重点实验室开放基金(GSCS-2018-1,GSCS-2018-2).
关键词
液相色谱-串联质谱联用法
植物内源激素
同时测定
liquid chromatography-tandem mass spectrometry
endogenous phytohormones
simultaneous determination
