Simultaneous determination and confirmation of melamine and cyanuric acid in animal feed by zwitterionic hydrophilic interaction chromatography and tandem mass spectrometry

We developed a new method to analyze animal feed and feed ingredients for melamine and cyanuric acid. The method is capable of extracting and detecting both melamine and cyanuric acid in a single procedure, whether present as free compounds or bound together as the melamine:cyanurate complex. A novel chromatographic system based on zwitterionic hydrophilic interaction chromatography (ZIC-HILIC) columns enables separation and detection of both compounds in one run. Samples are extracted with a strong aqueous acid which is then diluted to bring the concentration within the working range of the method. The method is applicable over the range of 0.5 to 50 micrograms/gram (microg/g). Samples at higher concentrations may be diluted into this range, which is equivalent to 3.6-360 ng/mL in the injection solvent. Analytes are detected using liquid chromatography/tandem mass spectrometry. The data confirm the presence of both compounds according to criteria recommended by the US FDA Center for Veterinary Medicine. The LC/MS/MS method provides an alternative to derivatization and gas chromatography-mass spectrometry for regulatory analysis of feed samples. Published in 2008 by John Wiley & Sons, Ltd.