Abstract:

A method for the determination of phenylbutazone and aminopyrine in beef has been developed by ultra high performance
liquid chromatography-high resolution mass spectrometry (UHPLC-HRMS). After a modified QuEChRES sample preparation
process, the samples were loaded onto an Acquity UPLC HSS T3 column (150 mm × 2.1 mm, 1.8 μm) and separated with
a gradient elution. The electrospray process was operated in the positive ion mode, and data were acquired in the full scan
mode. The calibration curves showed a good linearity in the range of 5.0–100.0 μg/kg, with correlation coefficients (r) equal
to or greater than 0.995. When the spiked levels were 5, 10 μg/kg and 20 μg/kg, the recovery rates of phenylbutazone and
aminopyrine in bovine ranged from 76.7% to 85.9%, with relative standard deviations (RSDs) of 5.6%–12.9%. The limits of
quantification (LOQs, RSN=10) for phenylbutazone and aminopyrine were 5.0 μg/kg and 1.0 μg/kg in beef, respectively. The
results indicate that the developed method is simple, rapid, sensitive and suitable for the qualitative and quantitative analysis
of phenylbutazone and aminopyrine in animal-derived foodstuffs including beef.

Key words: ultra performance liquid chromatography-high resolution mass spectrometry (UPLC-HRMS), phenylbutazone, aminopyrine, bovine