Simultaneous determination of gibberellic acid, indole-3-acetic acid and abscisic acid in wheat extracts by solid-phase extraction and liquid chromatography-electrospray tandem mass spectrometry.
A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for simultaneous determination of three representative phytohormones in plant samples: gibberellic acid (GA(3)), indole-3-acetic acid (IAA) and abscisic acid (ABA). A solid-phase extraction (SPE) pretreatment method was used to concentrate and purify the three phytohormones of different groups from plant samples. The separation was carried out on a C(18) reversed-phase column, using methanol/water containing 0.2% formic acid (50:50, v/v) as the isocratic mobile phase at the flow-rate of 1.0 mL min(-1), and the three phytohormones were eluted within 7 min. A linear ion trap mass spectrometer equipped with electrospray ionization source was operated in negative ion mode. Selective reaction monitoring (SRM) was employed for quantitative measurement. The SRM transitions monitored were as 345-->239, 301 for GA(3), 174-->130 for IAA and 263-->153, 219 for ABA. Good linearities were found within the ranges of 5-200 microg mL(-1) for IAA and 0.005-10 microg mL(-1) for ABA and GA(3). Their detection limits based on a signal-to-noise ratio of three were 0.005 microg mL(-1), 2.2 microg mL(-1) and 0.003 microg mL(-1) for GA(3), IAA and ABA, respectively. Good recoveries from 95.5% to 102.4% for the three phytohormones were obtained. The results demonstrated that the SPE-LC-MS/MS method developed is highly effective for analyzing trace amounts of the three phytohormones in plant samples.
Hou
Shengjie
S
Key Lab of Bioorganic Phosphorus Chemistry & Chemical Biology, Ministry of Education, Department of Chemistry, Tsinghua University, Beijing 100084, China.
Zhu
Jiang
J
Ding
Mingyu
M
Lv
Guohua
G
D000040:Abscisic Acid / Q000032:analysis*; D002138:Calibration; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D002845:Chromatography / Q000379:methods; D002853:Chromatography, Liquid / Q000379:methods*; D005875:Gibberellins / Q000032:analysis*; D007210:Indoleacetic Acids / Q000032:analysis*; D007477:Ions; D008956:Models, Chemical; D010937:Plant Growth Regulators / Q000032:analysis*; D015203:Reproducibility of Results; D052616:Solid Phase Extraction / Q000379:methods; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D014908:Triticum / Q000378:metabolism*
D005875:Gibberellins; D007210:Indoleacetic Acids; D007477:Ions; D010937:Plant Growth Regulators; C030737:indoleacetic acid; D000040:Abscisic Acid; C007842:gibberellic acid