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A quantitative method is described for the characterization of alkylamine ethoxylate (ANEO) surfactants in commercial herbicide formulations using reversed-phase high-performance liquid chromatography with electrospray ionization mass spectrometric detection. The proposed protocol allows the simultaneous determination of a full range of oligomers which were monitored as [M-H](+) ions in positive ion mode. The instrumental detection limits (LODs) were 0.5 µg. The method developed was successfully applied to the determination of ANEOs in a number of glyphosate formulations. Both the total concentration of these surfactants and the oligomer distribution varied between the different commercial herbicides analysed. The precision of the determination of the oligomers in samples ranged from 0.5 to 15%.