Membrane-supported liquid–liquid–liquid microextraction combined with anion-selective exhaustive injection capillary electrophoresis-ultraviolet detection for sensitive analysis of phytohormones

• LPME and on-column sample stacking in CE were combined for the high enrichment of phytohormones.
• A method of MS-LLLME-ASEI-CE–UV was developed for trace phytohormones quantification.
• The developed method provides high sensitivity, wide linear range, and good sample cleanup ability.
• It is suitable for the determination of trace analytes in complicated real samples by CE–UV.

A novel method based on off-line membrane-supported liquid–liquid–liquid microextraction (MS-LLLME) combined with on-column anion-selective exhaustive injection (ASEI) capillary electrophoresis–ultraviolet (CE–UV) detection was established for the analysis of seven phytohormones (abscisic acid (ABA), jasmonic acid (JA), 2,4-dichlorophenoxyacetic acid (2,4-D), 1-naphthalene acetic acid (NAA), indole-3-acetic acid (IAA), salicylic acid (SA) and gibberellic acid (GA)). In MS-LLLME, the target phytohormones were extracted from the acid donor phase to the alkaline acceptor phase, and the acceptor solutions were directly analyzed by ASEI-CE–UV. Under the optimal experimental conditions, the analytical performance of the method was evaluated. The limits of detection (LODs) of ABA, JA, 2,4-D, NAA, IAA, SA and GA were determined to be 1.00, 2.21, 0.33, 0.17, 0.67, 0.05 and 16.5 ng/mL, respectively. The relative standard deviations (RSDs, n = 7) ranged from 4.7% to 12.9%, and the enrichment factors were in the range of 307 to 20,160. The proposed method was successfully applied for the determination of multiple phytohormones in banana, cabbage and cucumber extracts, and ABA, IAA and SA were detected in these samples. The recoveries for the spiked samples were in the range of 79.0 to 116.4%. The proposed method was demonstrated to be suitable for the simultaneous quantification of multiple phytohormones with high sensitivity and good sample cleanup ability.