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A dose-response study was carried out to examine the carryover of zearalenone (ZEN), deoxynivalenol (DON) and their metabolites into bovine milk. Therefore, a feeding trial with 30 dairy cows fed with three different levels of Fusarium (FUS) toxin-contaminated maize was performed. A control group (0.02mg ZENkg(-1) dry matter (DM) and 0.07 DONkg(-1) DM) was compared with two groups fed contaminated diets. The first diet contained 0.33mg ZENkg(-1) DM and 2.62mg DONkg(-1) DM (group FUS-50) and the second diet contained 0.66mg ZENkg(-1) DM and 5.24mg DONkg(-1) DM (group FUS-100). For milk sample analysis, a new cost-efficient sample preparation method was developed for the simultaneous determination of ZEN, DON and their metabolites. The method comprised the separation of the milk fat followed by an SPE clean-up on Oasis HLB and a LC-MS/MS measurement. The less toxic metabolite de-epoxy-DON had the highest detected concentration (5.6ngml(-1) milk) in the milk samples obtained from the feeding trial. Additionally, ZEN (up to 0.29ngml(-1)), alpha-zearalenol (up to 0.17ngml(-1)), beta-zearalenol (up to 0.95ngml(-1)) and DON (up to 2.5ngml(-1)) were detected in these samples. The milk toxin concentrations of cows fed the control diet were significantly lower compared with cows fed the contaminated diet. The calculated carryover rates ranged between 0 and 0.0075 for ZEN and metabolites and between 0 and 0.0017 for DON independent of exposure. It can be concluded that dietary toxin concentrations in the feed below or close to the current guidance values do not pose a risk for consumers due to negligible carryover rates.