Validation of a Method for the Determination of Theobromine in Feed Materials and Compound Feeds by Liquid Chromatography with UV Detection or with Tandem Mass Spectrometry

Kirstin M Gray a and D Thorburn Burnsb
LGC, Teddington, Middlesex, TW11 0LY, UK
b    Institute for Global Food Security, The Queen’s University of Belfast, Belfast BT9 5HN, Northern Ireland, UK


An inter-laboratory validated HPLC separation coupled with UV detection method for the determination of theobromine in animal feed materials and in compounded feeds is described using 7-(β-hydroxyethyl) theophylline as the internal standard. If more forensically robust identification of the presence of theobromine is required, the same HPLC separation can be coupled to a tandem mass spectrometer and theobromine-d3 used as the internal standard. At the time of publishing, the method described in this paper is in the process of a formal vote before being published as a CEN standard.

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