Abstract:

An immunoaffinity column cleanup-high performance liquid chromatography (HPLC) method was established for
the simultaneous determination of residues of chloramphenicol, zearalanol and its analogs (α-zeranol, β-zeranol, α-zearalenol,
β-zearalenol, zearalanone, and zearalenone) in milk. Samples were cleaned up and enriched on an immunoaffinity column.
The chromatographic separation was performed on a Cloversil-C18 column using acetonitrile-methanol-water as the mobile
phase and ultraviolet (UV) detection was performed at 265 nm. The recoveries of seven target compounds at different
spiked levels ranged from 74% to 101% and the relative standard deviations were less than 7%. The limits of detection
were 0.02 μg/L for chloramphenicol, 0.03 μg/L for α-zearalanol, β-zearalanol, α-zearalenol, and β-zearalenol, 0.04 μg/L for
zearalanone, and 0.05 μg/L for zearalenone. The proposed method is efficient, stable, reliable and accurate, and can be used
for the determination of trace residues of chloramphenicol, zearalanol and its analogs in milk.

Key words: milk, chloramphenicol, zearalanol, immunoaffinity column, high performance liquid chromatography (HPLC)