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A sensitive method based on liquid chromatography-tandem mass spectrometry has been developed for the determination of triptorelin levels in beagle dog plasma. Plasma samples were applied to Oasis (R) HLB solid-phase extraction (SPE) cartridges. Extracted samples were evaporated under a stream of nitrogen and then reconstituted with 100 mu l methanol:water:formic acid (60:40:0.08, v/v/v). The separation was achieved on a Venusil MP-C18 column (2.1 mm x 50 mm, 3 p,m, Agela) with a gradient elution. Detection utilized a Qtrap5500 system operated in the positive ion mode with multiple reaction monitoring of the analyte at m/z 656.5 249.1 and of the I.S. at m/z 510.8 -> 120.1. The proposed method was validated by assessing the specificity, linearity, precision and accuracy, recovery, matrix effects, and stability. Linear calibration curves were obtained in the concentration range of 0.01-10 ng/ml (the correlation coefficients were above 0.995). The lower limit of quantification (LLOQ) of the method was 0.01 ng/ml. The method was successfully applied to a pharmacokinetic study of a slow release triptorelin formulation in beagle dogs following a single intramuscular injection. Crown Copyright (C) 2012 Published by Elsevier B.V. All rights reserved.