In house validation of a reference method for the determination of boar taint compounds by LC-MSMS

Abstract

Meat from male pigs may develop an off-flavour, commonly known as boar taint. For that reason male piglets are surgically castrated at young age to avoid the potential off-flavour formation. \nAnimal welfare concerns have triggered research into alternatives to surgical castration of male piglets with the long-term goal of abandoning it by 1 January 2018.\nThe ALCASDE study has shown that the agreement of testing results for androstenone and skatole in pig fat produced in several laboratories was not satisfactory and one of the conclusions of this project was that in a follow-on project a standardised reference method should be elaborated to rectify this shortcoming.\nIn the suggested reference method, the 3 marker compounds (skatole, andostrenone and indole), for boar taint are quantified in pork fat by isotope dilution liquid chromatography tandem mass spectrometry (LC MS/MS). The fat is separated from the ground pork fat tissue sample via melting and centrifugation thereafter. The fat is spiked with isotopically labelled standards and prepared for size exclusion chromatography (SEC). The SEC purified sample is evaporated nearly to dryness and after addition of an injection standard analysed by LC-MS/MS in selected reaction monitoring mode. \nThis method has been validated to obtain the method performance characteristics.\nWhen summarising the method performance characteristics and comparing them to requirements for official food control methods in the area of food contaminants it can be concluded that the method is fit for its intended purpose.\nThe recovery rates are between 106 % and 115% and are therefore within the range of 60 % to 120 %.\nThe repeatability standard deviation ranges from 7 % to 18 % and the intermediate precision between 9 % and 18 %, all below the set limit of 20 %.\nThe measurement uncertainty (U) is below 30 % for all analytes. The method has proven to be robust and free from matrix interferences.\nThe method is sensitive enough to determine the off-flavour compounds at the sensory threshold values.