title: An in-advance stable isotope labeling strategy for relative analysis of multiple acidic plant hormones in sub-milligram Arabidopsis thaliana seedling and a single seed.
abstract: A sensitive and reliable in-advance stable isotope labeling strategy was developed for simultaneous relative quantification of 8 acidic plant hormones in sub-milligram amount of plant materials. Bromocholine bromide (BETA) and its deuterated counterpart D9-BETA were used to in-advance derivatize control and sample extracts individually, which were then combined and subjected to solid-phase extraction (SPE) purification followed by UPLC-MS/MS analysis. Relative quantification of target compounds was obtained by calculation of the peak area ratios of BETA/D9-BETA labeled plant hormones. The in-advance stable isotope labeling strategy realized internal standard-based relative quantification of multiple kinds of plant hormones independent of availability of internal standard of every analyte with enhanced sensitivity of 1-3 orders of magnitude. Meanwhile, the in-advance labeling contributes to higher sample throughput and more reliability. The method was successfully applied to determine 8 plant hormones in 0.8mg DW (dry weight) of seedlings and 4 plant hormones from single seed of Arabidopsis thaliana. The results show the potential of the method in relative quantification of multiple plant hormones in tiny plant tissues or organs, which will advance the knowledge of the crosstalk mechanism of plant hormones.
authors.affiliation: National Center for Plant Gene Research (Beijing), Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, 1-2 Beichen West Road, Beijing 100101, China.
authors.forename: Xiaohong
authors.initials: X
authors.lastname: Sun
authors.affiliation: National Center for Plant Gene Research (Beijing), Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, 1-2 Beichen West Road, Beijing 100101, China; Department of Pharmaceutical Analysis, Heilongjiang University of Chinese Medicine, Harbin 150040, China.
authors.forename: Yue
authors.initials: Y
authors.lastname: Ouyang
authors.affiliation: National Center for Plant Gene Research (Beijing), Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, 1-2 Beichen West Road, Beijing 100101, China.
authors.forename: Jinfang
authors.initials: J
authors.lastname: Chu
authors.affiliation: Department of Pharmaceutical Analysis, Heilongjiang University of Chinese Medicine, Harbin 150040, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Yan
authors.affiliation: Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai 200032, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Yu
authors.affiliation: Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai 200032, China.
authors.forename: Xiaoqiang
authors.initials: X
authors.lastname: Li
authors.affiliation: Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai 200032, China. Electronic address: yangj@sioc.ac.cn.
authors.forename: Jun
authors.initials: J
authors.lastname: Yang
authors.affiliation: National Center for Plant Gene Research (Beijing), Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, 1-2 Beichen West Road, Beijing 100101, China. Electronic address: cunyuyan@genetics.ac.cn.
authors.forename: Cunyu
authors.initials: C
authors.lastname: Yan
mesh_terms: D017360:Arabidopsis / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D007553:Isotope Labeling / Q000379:methods; D010937:Plant Growth Regulators / Q000032:analysis* / Q000737:chemistry; D015203:Reproducibility of Results; D036226:Seedlings / Q000737:chemistry*; D012639:Seeds / Q000737:chemistry; D052616:Solid Phase Extraction; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D010937:Plant Growth Regulators
keywords: Arabidopsis thaliana; LC–MS/MS; Plant hormones; Relative quantification; Stable isotope labeling