Simultaneous determination of ten phytohormones in five parts of Sargasum fusiforme (Hary.) Seichell by high performance liquid chromatography-triple quadrupole mass spectrometry

Abstract

A method for the simultaneous determination of indole-3-acetic acid, N6-(2-isopentenyl) adenosine, N6-(2-isopentenyl) adenine, trans-zeatin riboside, zeatin, strigolactone, abscisic acid, salicylic acid, gibberellin A3 and jasmonic acid in five different parts of main branch, lateral branch, primary leaf, secondary leaf and stem of Sargasum fusiforme (Hary.) Seichell was established by high performance liquid chromatography-triple quadrupole mass spectrometry (HPLC-TQMS). The samples were extracted with methanol/water/formic acid (15 : 4 :1, v/v/v) (containing 0.5% 2, 6-di-tert-butyl-4-methylphenol, BHT) after vacuum freeze-drying. The separation was performed on a Hypersil Gold C18 column by using methanol and water as mobile phases with gradient elution. The analytes were detected by tandem mass spectrometry in selected reaction monitoring (SRM) mode. The electrospray ionization (ESI) source was used for the quantitative analysis in the positive mode or negative mode. Under the optimized conditions, the correlation coefficients (r) of the ten phytohormones were from 0. 9989 to 1. 0000 in the linear ranges. The detection limits of the ten phytohormones were 0. 001 2-4. 651 2 μ/L. The average recoveries were 72. 24% -91. 31% with the relative standard deviations not more than 6. 59%. In the five parts of fresh Sargasum fusiforme (Hary.) Seichell samples, the contents of the ten phytohormones were from not detected to 4 041. 431 ng/g. This method has good sensitivity, precision, recovery, and can be used to simultaneously determine the phytohormones.