Development and validation of an LC-MS/MS method for the quantification of tiamulin, trimethoprim, tylosin, sulfadiazine and sulfamethazine in medicated feed

Abstract

A new multi-compound method for the analysis of veterinary drugs, namely tiamulin, trimethoprim, tylosin, sulfadiazine and sulfamethazine was developed and validated in medicated feeds. After extraction, the samples were centrifuged, diluted in Milli-Q water, filtered and analysed by high performance liquid chromatography coupled to tandem mass spectrometry. The separation of the analytes was performed on a biphenyl column with a gradient of 0.1% formic acid in acetonitrile and 0.1% formic acid in Milli-Q water. Quantitative validation was done in accordance with the guidelines laid down in European Commission Decision 2002/657/EC. Method performances were evaluated by the following parameters: linearity (R < 0.99), precision (repeatability <14% and within-laboratory reproducibility <24%), recovery (73.58-115.21%), sensitivity, limit of detection (LOD), limit of quantification (LOQ), selectivity and expanded measurement uncertainty (k. = 2). The validated method was successfully applied to the 2 medicated feeds obtained from the interlaboratory studies and feed manufactures from Spain in August 2017. In these samples, tiamulin, tylosin and sulfamethazine were detected at the concentration levels declared by the manufacturers. The developed method can therefore be successfully used to routinely control the content and homogeneity of these antibacterial substances in medicated feed. Abbreviations AAFCO - Association of American Feed Control Officials; TYL - tylosin; TIAM - tiamulin fumarate; TRIM - trimethoprim; SDZ - sulfadiazine; SMZ - sulfamethazine; UV - ultraviolet detector; FLD - fluorescence detector; HPLC - high performance liquid chromatography; MS/MS - tandem mass spectrometry; LOD - limit of detection; LOQ - limit of quantification; CV - coefficient of variation; SD - standard deviation; U - uncertainty.

Pharmacokinetics of a sulfadiazine and trimethoprim suspension in neonatal foals

“…Samples were shipped on dry ice and stored at −80°C until assayed. A validated liquid chromatography–mass spectrometry (LC‐MS) assay modified from a previously reported method (Patyra et al., 2018) was used for the determination of trimethoprim/sulfadiazine plasma concentrations at each time point. Samples were chromatographically separated using a Shimadzu Prominence HPLC System with a HTC‐PAL Autosampler fitted with a Waters SunFire C18 column (4.6 × 50 mm).…”

Analysis of Transfer of Tiamulin to Animal Tissue after Oral Administration: An Important Factor for Ensuring Food Safety and Environmental Protection

“…The method is advantageous due to the low quantification limit of 0.002 µg/g [ 4 ]. Tiamulin content was also successfully determined in medicated feed using a different gradient of 0.1% formic acid in acetonitrile and 0.1% formic acid in ultrapure water and a biphenyl column [ 5 ].…”

Quantum dots‐based fluorescence immunoassay for detection of tiamulin in pork

“…There are many methods to detect TML by instruments, including liquid chromatography tandem–mass spectrometry (LC–MS/MS) (Patyra, Nebot, Gavilan, Cepeda, & Kwiatek, 2018), high‐performance liquid chromatography (HPLC) (De Baere et al, 2015; Krasucka, Mitura, Cybulski, Kos, & Pietroń, 2010), ultra‐performance liquid chromatography (UPLC), and so forth (Sun et al, 2017; Yang et al, 2014). Although these assay methods have low detection limits and good accuracy, they require expensive equipment and well‐trained operators.…”