Simultaneous Determination of Twelve Endogenous Hormones in Woody Plants by Ultra Performance Liquid Chromatography and Its Application
An ultra performance liquid chromatography method(UPLC)was developed for the simultaneous determination of twelve kinds of endogenous hormones(indole acetonitrile,indole pyruvate,indole acetamide,tryptamine,3-indoleacetic acid,zeatin,trans-zeatin riboside,abscisic acid,gibberellin A1,gibberellin A3,gibberellin A4,and gibberellin A7)in Woody plant.The woody plant tissues were extracted with 80%methanol.Purification of samples was performed with a MCX solid phase extraction column,the chromatography was performed on a ZORBAX EP-C18 column with gradient elution of methanol/0.1%glacial acetic acid at the column temperature of 30 ℃ and detection wavelength of 254 nm and 210 nm.The limits of detection(LODs)and quantification(LOQs)were in the range of 0.002-0.029 ng·mL-1 and 0.006-0.088 ng·mL-1.The recoveries of each substance under different spiking levels were 78.1%-116.8%and 82.9%-114.2%,and the relative standard deviations(RSD)were 0.3%-4.7%and 0.2%-4.2%.The results showed that the content of abscisic acid was the highest(768.88 ng g-1 and 366.27 ng·g-1),and the contents of GA3,GA4 and GA7 were lower.The established method could be used to purify and detect various hormones in 84K Poplar and Betula platyphylla Suk,and it provided a good reference for the determination of hormones in other woody plants.
PretreatmentUltra performance liquid chromatographyExternal standard methodEndogenous hormoneWoody plant