Method development and validation: solid Phase extraction-ultra performance liquid chromatography-tandem mass...

Pirlimycin, a lincosamide antibiotic, is one of the most commonly used antibiotics for the treatment of mastitis in dairy cows. Assessment of pirlimycin loadingto the environment via fecal and urinary excretion is critical to develop efficient management strategies to reduce environmental pollution by the livestock industry. Therefore, the aim of this study was to develop and validate an analytical method to identify and quantify pirlimycin in bovine feces and urine. Samples were extracted with methanol- phosphate buffer and cleaned up by SPE before analysis for pirlimycin using UPLC-MS/MS. This method was sensitive (LOQ 1.47 ng/g wet feces, 0.90 ng/mL urine), accurate (recovery, 80-108%), and precise (repeatability, 2.3-13%; reproducibility, 2.3-14%) for both bovine feces and urine. With the application of this method to samples collected in the first 10 h and then every 24 h for 120 h following intramammary dosing (50 mg/cow; n = 3 cows), pirlimycin was detected at 40.5-287 ng/g and 46.1-254 ng/mL in feces and urine, respectively. This robust, sensitive, and accurate method can be used to assess the fate and environmental impact of antibiotics used on farms.