Abstract
Capillary electrophoresis (CE) coupled with mass spectrometry (MS) or tandem mass spectrometry (MS/MS) is reported as an alternative and potentially useful method for the simultaneous analysis of various classes of phytohormones with diversified structures, including indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), abscisic acid (ABA), gibberellic acid (GA), zeatin (Z), N6-benzyladenine (BA), α-naphthaleneacetic acid (NAA) and 2,4-dichlorophenoxyacetic acid (2,4-D). The key to the CE-MS/MS analysis was based on electroosmotic flow reversal using a cationic polymer-coated capillary. Under optimum conditions, a baseline separation of eight phytohormones was accomplished within 30 min using 60 mM ammonium formate/formic acid buffer of pH 3.8 with −20 kV as the separation voltage. The accessibility of MS/MS together with the characterization by migration properties obtained by CE allows for the development of CE-MS/MS as an emerging potential method for the analysis of different classes of phytohormones in a single run. The utility of the CE-MS/MS method was demonstrated by the comprehensive screening of phytohormones in coconut (Cocos nucifera L.) water after pre-concentration and purification through solid-phase extraction (SPE) cartridge. IAA, ABA, GA and Z were detected and quantified in the purified coconut water extract sample.
Highlights
Phytohormones are a group of naturally occurring organic compounds that play crucial roles in mediating plant growth through a whole range of developmental processes
Since the cationic polymer-coated capillary was used in the current study, the reverse electroosmotic flow (EOF) was highly suppressed in the alkaline medium
This work demonstrated that capillary electrophoresis (CE)-mass spectrometry (MS) or CE-MS/MS can be used to separate different classes of phytohormones with a special cationic polymer-coated capillary
Summary
Phytohormones are a group of naturally occurring organic compounds that play crucial roles in mediating plant growth through a whole range of developmental processes These structurally diverse compounds mainly include auxins, cytokinins, abscisic acid, gibberellins and ethylene [1,2,3]. This problem was overcome by employing cationic polymer-coated capillary to reverse the EOF [19] By using these coated capillaries with positively charged surface, it was possible to create an EOF constantly towards the anode (MS side) without adding any buffer modifiers. Mixed-phase SPE cartridges that combine reversed-phase and anion-exchange chromatography separation were obviously the ideal pre-concentration and sample clean-up method for phytohormones [24]. In our current work, Mixed-Mode Anion-eXchange (MAX) SPE was employed as an efficient approach to pre-concentrate and purify phytohormones within a biological matrix. Four phytohormones (IAA, ABA, GA and Z) in young coconut (Cocos nucifera L.) water were successfully identified and quantified
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