Simultaneous analysis of different classes of phytohormones in coconut ( Cocos nucifera L.) water using high-performance liquid chromatography and liquid chromatography–tandem mass spectrometry after solid-phase extraction

• Published in: Analytica chimica acta 2008-03, Vol.610 (2), p.274-281
• Main Authors: Ma, Zhen, Ge, Liya, Lee, Anna S.Y., Yong, Jean Wan Hong, Tan, Swee Ngin, Ong, Eng Shi
• Format: Article
• Language: English
• Subjects: Analytical chemistry; Chemistry; Chromatographic methods and physical methods associated with chromatography; Chromatography, High Pressure Liquid - methods; Coconut water; Cocos - chemistry; Exact sciences and technology; High-performance liquid chromatography; Liquid chromatography–tandem mass spectrometry; Other chromatographic methods; Phytohormones; Plant Growth Regulators - analysis; Plant Growth Regulators - classification; Reference Standards; Reproducibility of Results; Spectrometric and optical methods; Spectrometry, Mass, Electrospray Ionization - methods; Tandem Mass Spectrometry - methods
• ISSN: 0003-2670; 1873-4324
• DOI: 10.1016/j.aca.2008.01.045

Coconut ( Cocos nucifera L.) water, which contains many uncharacterized phytohormones is extensively used as a growth promoting supplement in plant tissue culture. In this paper, a high-performance liquid chromatography (HPLC) method was developed for the simultaneous determination of various classes phytohormones, including indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), abscisic acid (ABA), gibberellic acid (GA), zeatin (Z), N 6-benzyladenine (BA), α-naphthaleneacetic acid (NAA) and 2,4-dichlorophenoxyacetic acid (2,4-D) in young coconut water (CW). The analysis was carried out using a reverse-phase HPLC gradient elution, with an aqueous mobile phase (containing 0.1% formic acid, pH adjusted to 3.2 with triethylamine (TEA)) modified by methanol, and solute detection made at 265 nm wavelength. The method was validated for specificity, quantification, accuracy and precision. After preconcentration of putative endogenous phytohormones in CW using C 18 solid-phase extraction (SPE) cartridges, the HPLC method was able to screen for putative endogenous phytohormones present in CW. Finally, the identities of the putative phytohormones present in CW were further confirmed using independent liquid chromatography–tandem mass spectrometry (LC–MS/MS) equipped with an electrospray ionization (ESI) interface.