Validating a High-performance Liquid Chromatography method for the quantification of gibberellic acid in germinating maize seeds

Abstract

Gibberellic acid is a phytohormone that triggers the germination of seeds in a state of dormancy. Through the quantification of this hormone, the physiological condition of seeds of economic importance can be studded. In this work we validated a High-Performance Liquid Chromatography method to quantify gibberellic acid in germinated maize (Zea mays L.) seeds. Chromatographic conditions included the use of a C-18 reversed-phase column, acetonitrile-formic acid (1:9 %) as the mobile phase, flow of 0.5 mL min−1 , and detection at 195 nm. We evaluated our method for seven analytical parameters. The method was linear for gibberellic acid concentrations from 1.0 mg·kg−1 to 50.0 mg·kg−1 . The method’s limits were 0.3 mg·kg−1 and 1.0 mg·kg−1 for detection and quantification, respectively. The method was highly precise; we obtained variable but low relative standard deviations (2.62 % - 12.66 %) for the studied gibberellic acid concentrations. We assessed accuracy through recovery percentages, ranging from 52.85 % - 63.68 %, for three gibberellic acid concentrations. We conclude that our analytical method can be used to measure gibberellic acid during the early stages of maize germination. In addition, the method could be used for the analysis of other types of plant matrices.

Keywords

• high-performance liquid chromatography; dynamic sonication-assisted solvent extraction (dsase); phytohormones; gibberellic acid; validation; concentration; quantification.