Rapid Determination of Ribavirin Residue in Poultry Meat through Purification by Solid-phase Extraction and Analysis by Ultra-performance Liquid Chromatography-tandem Mass Spectrometry
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Abstract:
A rapid analysis method was established for determining the ribavirin residue in poultry meat through purification by solid-phase extraction and analysis by ultra-performance liquid chromatography-tandem mass spectrometry. After enzymatic hydrolysis with acid phosphatase at 37 ℃, the samples were extracted with acetonitrile and purified with an EMR-Lipid solid phase extraction column to remove liposoluble substances. The purified sample solution was subjected to separation using a BEH Z-HILIC column (100 mm×2.1 mm, 1.7 μm) and a mobile phase consisting of acetonitrile and 0.1% aqueous formic acid solution (containing 5 mmol/L ammonium formate) for analysis by tandem mass spectrometer in positive electrospray ionization mode with multiple reaction monitoring (MRM). An isotope-labeled internal standard method was used for quantitative analysis. The pretreat conditions, HPLC conditions and mass spectrometry parameters were systematically investigated. This method showed good linearity in the range of 0.05~5 μg/L with the limit of quantification (LOQ) being 2.0 μg/kg. When the spiking levels were 2.0, 4.0 and 20.0 μg/kg, the recoveries of ribavirin were in the range of 94.5%~112.0% with the relative standard deviations (n=6) being 1.4%~3.2%. Compared with the standard method, this method is simple and rapid, thus suitable for batch analysis of ribavirin residue in poultry meat.
