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Simple and rapid determination of N 6 ‐(Δ 2 ‐isopentenyl)adenine, zeatin, and dihydrozeatin in plants using on‐line cleanup liquid chromatography coupled with hybrid quadrupole‐Orbitrap high‐resolution mass spectrometry
Author(s) -
Ai LianFeng,
Yue ZhiLiang,
Ma YuSong,
Wang JianChang,
Zhang ShengWei,
Sun Ying
Publication year - 2015
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201401321
Subject(s) - chromatography , chemistry , analyte , orbitrap , elution , detection limit , mass spectrometry , analytical chemistry (journal) , resolution (logic) , extraction (chemistry) , high performance liquid chromatography , artificial intelligence , computer science
A simple and rapid method was developed for the determination of three free cytokinins, namely, N 6 ‐(Δ 2 ‐isopentenyl)adenine, zeatin, and dihydrozeatin, in plants using TurboFlow on‐line cleanup liquid chromatography combined with hybrid quadrupole‐Orbitrap high‐resolution mass spectrometry. The samples were extracted using acetonitrile, and then the extract was purified on a C 18 ‐p column, in which the sample matrix was removed and the analytes were retained. Subsequently, the analytes were eluted from the extraction column onto the analytical column (Hypersil Gold C 18 column) prior to chromatographic separation and hybrid Q‐Orbitrap detection using the targeted‐MS 2 scan mode. The linearity was satisfactory with a correlation coefficient of >0.999 at concentrations ranging from 5–5000 pg/mL. The limits of quantification for the analytes ranged from 4.2–5.2 pg/mL. The intra‐ and inter‐day average recoveries of analytes fortified at three levels ranged from 85.4–108.2%, and the intra‐ and inter‐day relative standard deviations ranged from 4.04–8.57%. The method was successfully applied for the determination of free cytokinins in different tissue samples of Oryza sativa and Arabidopsis thaliana .