a tlc-hplc method for determination of thiamphenicol in pig, chicken, and fish feedstuffs

An effective thin-layer chromatography (tlc) purification procedure coupled to high-performance liquid chromatography (hplc) method was developed for the determination of thiamphenicol (tap) in pig, chicken, and fish feedstuffs. the feedstuff samples were extracted with ethyl acetate, defatted with n-hexane saturated with acetonitrile, and further purified by tlc. the chromatographic separation was performed on a waters symmetry c18 column using an isocratic procedure with acetonitrile-water (21.7:78.3, v/v) at 0.6 ml/min. the ultraviolet (uv) detector was set at a wavelength of 225 nm. the tap concentrations in feedstuff samples were quantified using a standard curve. good linear correlations (y = 162,630x − 2381.7, r > 0.9998) were achieved within the concentration range of 0.05–10.00 μg/ml. the recoveries of tap spiked at levels of 1, 10, and 100 μg/g ranged from 82.0 to 114.9% with the intra-day and inter-day relative standard deviation (rsd) less than 9.0%. the limit of detection (lod) and limit of quantitation (loq) were 0.02 and 0.03 mg/kg for pig feedstuffs, 0.02 and 0.04 mg/kg for chicken feedstuffs, and 0.02 and 0.03 mg/kg for fish feedstuffs, respectively. this reliable, simple, and cost-effective method could be applied to the routine monitoring of tap in animal feedstuffs.