Development and validation of a high performance liquid chromatography method for the identification and quantification of clenbuterol in bovine liver

摘要

A methodology for the determination of clenbuterol in bovine liver by high performance liquid chromatography was developed. Extraction was performed with acetonitrile and isopropanol. The extract was analyzed using a reverse phase column C18, a UV detection at 214 nm, and NaH2PO4 0.05M (pH 3.0)/acetonitrile (85:15, v/v) as the mobile phase. The retention time for clenbuterol was 24.82 min. The best-fit line of the standard calibration curve was y = 1340.8563x - 1539.7319, and r2=0.9997 (p0.05). The detection and quantification limits showed values of 0.9 and 1.8 ng/mL, respectively. The mean recovery of clenbuterol were 111.7, 82.0 and 84.8%, for the concentration levels prepared (26.2, 104.9 and 209.8 ng/mL, respectively), which are considered suitable for the proposed method. The method accuracy was measured by the relative standard deviation, which was less than 4.74%. Seventeen liver samples purchased in public markets of México City were analyzed, of which 11 of them had concentrations above the MRL for clenbuterol with a median of 17.68ppb. This method is accurate and reliable due to its limit of detection, chromatographic separation efficiency, precision, accuracy, sensitivity and stability.