Determination of clenbuterol in porcine tissues using solid-phase extraction combined with ultrasound-assisted dispersive liquid-liquid microextraction and HPLC-UV detection

A new pretreatment method, solid-phase extraction combined with dispersive liquid-liquid microextration (SPE-DLLME), was proposed in first time for the determination of clenbuterol (CLB) in porcine tissue samples. The tissue samples were firstly extracted by SPE, then its eluents were used as dispersant of the followed DLLME for further purification and enrichment of CLB. Various parameters (such as the type of SPE sorbent, the type and volume of elution solvent, the type and volume of extractant and dispersant, etc.) that affected the efficiency of the two steps were optimized. Good linearity of CLB was ranged from 0.19 μg/kg to 192 μg/kg with correlation coefficient (r2) of 0.9995. The limit of detection (LOD) was 0.07 μg/kg (S/N = 3) and the recoveries at three spiked levels were ranged from 87.9% to 103.6% with the relative standard deviation (RSD) less than 3.9% (n = 3). Under the optimized conditions, the enrichment factor (EF) for CLB could up to 62 folds. The presented method that combined the advantages of SPE and DLLME, had higher selectivity than SPE method and was successfully applied to the determination of CLB in tissue samples.