Sensitivity enhancement of aminoglycosides in hydrophilic interaction liquid chromatography with tandem mass spectrometry by post-column addition of trace sodium acetate in methanol

Abstract

ABSTRACTThe development of a sensitive and accurate analytical method for monitoring aminoglycosides in food, environmental, and clinical samples is needed for many purposes. This study found that the responses of sodiated and protonated aminoglycosides in hydrophilic interaction chromatography with tandem mass spectrometry were enhanced upon addition of sodium acetate in methanol (5 mg L−1 at a flow rate of 0.2 mL min−1) as a post-column reagent. The sensitivities of sodiated spectinomycin, kanamycin, gentamicins, neomycin, and amikacin were significantly higher than those of the protonated molecules. Streptomycin and dihydrostreptomycin only formed protonated molecules, suggesting the preferential ionisation of the guanidine moieties in these aminoglycosides. The limits of quantification of these aminoglycosides were 0.19–2.5 ng mL−1. Notably, this is the first quantification of aminoglycosides that uses the sodiated molecules. The enhancement technique enables us to eliminate a concentration step from the clean-up process from food samples. We also proposed a rapid analytical method for residual aminoglycosides in milk and meat samples; validation showed good accuracy and precision of this method at the Japanese maximum residual limits of aminoglycosides (40–500 µg kg−1). The application of this method to contaminated bovine tissues revealed remarkably high residual levels of kanamycin. This technique will be useful for the sensitive detection of aminoglycosides not only in food, but also in environmental samples and human plasma.