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  • Title: A simple and high-throughput method for multiresidue and multiclass quantitation of antimicrobials in pangasius (Pangasionodon hypophthalmus) fillet by liquid chromatography coupled with tandem mass spectrometry.
    Author: Bortolotte AR, Daniel D, de Campos Braga PA, Reyes FGR.
    Journal: J Chromatogr B Analyt Technol Biomed Life Sci; 2019 Aug 15; 1124():17-25. PubMed ID: 31176266.
    Abstract:
    The development and validation of a throughput method for the determination of 25 antibacterial drugs (two β-lactams, eight quinolones, two macrolides, five sulfonamides, trimethoprim, four tetracyclines and three amphenicols) in pangasius fish muscle by LC-MS/MS were performed. A simple, efficient and fast extraction procedure was developed using acetonitrile and a 0.1 M EDTA solution as solvents for extraction. All compounds were determined in a single run, and chromatographic separation was achieved using a Zorbax SB C18 column with a mobile phase comprised of purified water +0.1% formic acid (A) and acetonitrile +0.1% formic acid (B) in a linear gradient program. The method was validated aαording to the requirements of European Decision 2002/657/EC. To quantify the analytes, matrix-matched analytical curves were constructed with spiked blank tissues and showed linearity (r2) higher than 0.99. For all analytes, the precision and accuracy were determined at the levels of 3 ng/g (low), 10 ng/g (low-middle), 50 ng/g (high-middle) and 100 ng/g (high). The precision (CV%) was lower than 18.6% and the accuracy (determined as recovery) was between 65% and 119%. The limit of quantitation was 3.0 ng/g, with the exception of chloramphenicol, which was 0.3 ng/g, and amoxicillin and doxycycline, which were 10 ng/g. The method was successfully applied to analyze pangasius muscle samples from Vietnam available at the Brazilian retail market, and 5 out of 40 samples showed the presence of low-residue levels of enrofloxacin and, consequently, must be considered out of conformity. It is recommended that competent authorities should avoid the commercialization of pangasius fillet contaminated with residues of this veterinary drug.
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