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Boar taint is an off-odour that can occur when meat or fat from entire male pigs is heated. Most of the currently available analytical methods are not capable of detecting the three known boar taint compounds (indole. skatole and androstenone) simultaneously, which renders their analysis often labour-intensive and time-consuming as separate analyses are required. In this study a validated U-HPLC-HR-Orbitrap-MS analysis method is described for the quantitative determination of the three boar taint compounds in fat. The sample pre-treatment involves a melting step followed by extraction with methanol and clean-up consisting of a freezing step and solid phase extraction (HLB cartridges). The analytes are then chromatographically separated and detected with an Exactive (TM) high-resolution mass spectrometer. Due to the absence of guidelines for the analysis of boar taint in fat, the Commission Decision 2002/657/EC  and ISO 17025  guidelines were used as guideline for validation of the developed detection method. This resulted in limits of detection and limits of quantification between 2.5 and 7 mu g kg(-1) and between 5 and 10 mu g kg(-1) for the three compounds, respectively, which is far below the threshold values set at 100 mu g L-1 for indole, 200 mu g L-1 for skatole and 1000 mu g L-1 for androstenone in pig fat samples. The method obtained for the three compounds a repeatability (RSD) lower then 12.7% and a within-laboratory reproducibility (RSD) lower than 16.9 The recovery of the three compounds ranged between 99 and 112 and an excellent linearity (R-2 >= 0.99) was found. In the future, this method may be extended with other compounds that turn out to be correlated with boar taint. (C) 2012 Elsevier B.V. All rights reserved.