Determination of Plant Hormones in Fertilizers by High-Performance Liquid Chromatography with Photodiode Array Detection: Method Development and Single-Laboratory Validation

Abstract

A simple and reliable high-performance liquid chromatographic method that uses photodiode array detection was developed for the simultaneous determination of 12 native and synthetic plant hormones, i.e., plant growth regulators (PGRs), in fertilizers, such as 1-naphthol, 2,4-dichlorophenoxyacetic acid, 4-(2,4-dichlorophenoxy)butyric acid, 4-chlorophenoxyacetic acid, indole-3-acetic acid, 4-(3-indolyl)butyric acid, dichlorprop, (4-chloro-2-methylphenoxy)acetic acid, -naphthaleneacetic acid, 1-naphthaleneacetamide, -naphthoxyacetic acid, and thidiazuron. The method was experimentally validated for routine regulatory application, and the following analytical parameters were assessed for all PGRs studied: linearity; specificity; precision (relative standard deviation) and accuracy, both measured at 3 concentration levels (0.1, 0.05, and 0.01, w/w); ruggedness; limit of detection; and limit of quantification. Results were satisfactory for all method validation parameters tested and for all PGRs studied, demonstrating the suitability of the method for the determination of PGRs in fertilizers. The uncertainty of measurement was also estimated at 3 concentration levels for all PGRs by using the approach of the International Organization for Standardization, described in its Guide to the Expression of Uncertainty in Measurement. The method was applied to 20 samples of liquid fertilizer with declared biostimulant properties.

Liquid Chromatographic Determination of Endogenous Phytohormones in Vegetable Samples Based on Chemical Derivatization with 6-Oxy(acetylpiperazine) Fluorescein

“…The calibration ranges were 0.05−10 μM for 2,4-D and 0.1−10 μM for IBA and NAA. The detection limits for IBA, NAA, and 2,4-D were 14.8, 7.24, and 4.43 nM, respectively, which were lower than or equivalent to those of phytohormones obtained by other methods − .…”

“…To date, various analytical techniques have been developed for the determination of exogenous phytohormones, such as gas chromatography−mass spectrometry (GC-MS) − , liquid chromatography−mass spectrometry (LC-MS) − , capillary electrophoresis with UV detection (CE-UV) , , capillary electrophoresis with fluorescence detection (CE-FD) , high-performance liquid chromatography with UV detection (HPLC-UV) − , high-performance liquid chromatography with chemiluminescence detection (HPLC-CL) , and enzyme-linked immunosorbent assay (ELISA) . Although these methods make great contributions to the analysis of phytohormones, some limitations are still found in the application of these methods.…”

Supercritical Algal Extracts: A Source of Biologically Active Compounds from Nature

“…Liquid chromatography used for plant hormone analysis does not require sample derivatization. Popular methods of detection connected with LC or HPLC are UV-V is detectors, diode array and fluorescence detection, and especially MS that allow determining the chemical structure of the analytes [67]. The potential of this method is being multiplied when tandem mass detector is used [68].…”

Development of a Rapid LC-DAD/FLD Method for the Simultaneous Determination of Auxins and Abscisic Acid in Plant Extracts

“…Various analytical techniques have been developed for endogenous and exogenous phytohormone determination: gas chromatography–mass spectrometry (GC-MS), liquid chromatography–mass spectrometry (LC-MS), , capillary electrophoresis with UV (CE-UV) or fluorescence (CE-FLD) detection, high-performance liquid chromatography with UV (HPLC-UV), − fluorescence (HPLC-FLD) or chemiluminescence (HPLC-CL) detection, and enzyme-linked immunosorbent assay (ELISA) . Although these methods have contributed greatly to phytohormone analysis, some limitations persist in their application.…”