A simple, selective and sensitive procedure for determining nine widely used aminoglycoside antibiotics (AGs) in bovine whole milk is presented. It is based on matrix solid-phase dispersion with heated water, at 70 degrees C, as extractant followed by liquid chromatography (LC)-tandem mass spectrometry (MS) using an electrospray ion source. After acidification and filtration, 0.2 ml of the aqueous extract was injected into the LC column. MS data acquisition was performed in the multi reaction monitoring mode, selecting two (three, when possible) precursor ion > product ion transitions for each target compound. Analyte recoveries ranged between 70 and 92%. Using ammosidine (an AG not used in veterinary medicine) as surrogate internal standard, the accuracy of the method at three spike levels varied between 80 and 107% with R.S.D. not larger than 11%. The limits of quantification were between 2 ng/ml (apramycin) and 13 ng/ml (streptomycin). They are well below the tolerance levels set by both the European Union and the U.S. Food and Drug Administration.