Highly sensitive and quantitative profiling of acidic phytohormones using derivatization approach coupled with nano-LC–ESI-Q-TOF-MS analysis

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2012-09, Vol.905, p.67-74
• Main Authors: Chen, Ming-Luan, Fu, Xiao-Meng, Liu, Jia-Qi, Ye, Tian-Tian, Hou, Sheng-Yu, Huang, Yun-Qing, Yuan, Bi-Feng, Wu, Yan, Feng, Yu-Qi
• Format: Article
• Language: English
• Subjects: Acidic phytohormones; chromatography; Chromatography, Liquid - instrumentation; Chromatography, Liquid - methods; Derivatization; detection limit; electrospray ionization mass spectrometry; Ethylene Glycols - chemistry; gibberellins; Hydrogen-Ion Concentration; Ionization; Leaves; Liquid-liquid extraction; Methacrylates - chemistry; Nano-LC–ESI-Q-TOF-MS; Nanostructure; Nanotechnology; Oryza - chemistry; Plant Extracts - analysis; Plant Extracts - chemistry; Plant Growth Regulators - analysis; Plant Growth Regulators - chemistry; plant hormones; Plant Leaves - chemistry; Poly(methacrylic acid-co-ethylene glycol dimethacrylate) monolith; Profiling; quantitative analysis; Regression Analysis; Reproducibility of Results; Rice; Sensitivity and Specificity; solid phase extraction; Spectrometry, Mass, Electrospray Ionization - methods; Standard deviation; Tandem Mass Spectrometry - methods; Trapping
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2012.08.005

► Using polymer monolithic column, 15 targeted hormones are well separated. ► High sensitivity of pg/mL is obtained, combined with online trapping process. ► Tandem SPE followed by LLE is developed for purification of hormones from matrix. ► Developed method is applied in the analysis of acidic phytohormones in 5mg rice. In current study, we developed a highly sensitive method for the quantitative profiling of acidic phytohormones. Tandem solid-phase extraction (SPE) and liquid–liquid extraction (LLE) was employed to efficiently purify acidic phytohormones, which were further derived by 3-bromoactonyltrimethylammonium bromide (BTA) to increase the ionization efficiency in electrospray ionization-mass spectrometry detection. Additionally, fifteen BTA-derived acidic phytohormones, including ten gibberellins (GAs), were well separated with a salt gradient on poly(methacrylic acid-co-ethylene glycol dimethacrylate) (MAA-co-EDMA) monolithic column. By employing online trapping system, the signal intensities of the analytes were significantly improved. The limits of detection (LODs, Signal/Noise=3) of targeted phytohormones ranged from 1.05 to 122.4pg/mL, which allowed the highly sensitive determination of low abundant acidic phytohormones with tiny amount plant sample. Good reproducibility was obtained by evaluating the intra- and inter-day precisions with relative standard deviations (RSDs) less than 10.9 and 11.9%, respectively. Recoveries of the target analytes from spiked rice leave samples ranged from 88.3 to 104.3%. By employing the method developed here, we were able to simultaneously determine 11 endogenous acidic phytohormones from only 5mg of rice leave sample, which dramatically decreased the required sample amount (three orders of magnitude lower) for the profiling of low abundant acidic phytohormones compared to previous reports. Taken together, the method provided a good solution for the highly sensitive and quantitative profiling of endogenous acidic phytohormones.