Objective To develop a confirmative method detecting 16 sulfonamide antibiotics residues in human urine simultaneously with ultra performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS).

Methods Urine samples were analyzed following incubation with β-glucuronidase, and then purified with an Oasis PRiME HLB (200 mg, 6 mL) solid-phase extraction (SPE) cartridge. The UPLC separation was performed on an Waters BEH C₁₈ column (2.1 × 100 mm, 1.7 μm) utilizing a gradient elution program of methanol (containing 0.01 mol/L ammonium acetate) and water (containing 0.01mol/L ammonium acetate) as the mobile phase. Identification and quantification were achieved using electrospray ionization (ESI) in positive mode with multiple reaction monitoring (MRM).

Results Good linearities of the analyses were achieved over the concentration of 5 – 1 000 ng/mL, with the correlation coefficients above 0.99. The method quantitation limits (MQL) for target compounds were 5 ng/mL. The detection limits of the method were 0.3 – 1.7 ng/mL. Average recoveries of 16 analytes at three spiked levels ranged from 71.6% to 97.3% with the relative standard deviations (RSDs) of 1.4% – 11.7%.

Conclusion The established UPLC-MS/MS method is of simplify operation and high sensitivity, selectivity and precision, and is applicable to the measurement of trace amounts of sulfonamide antibiotics residues in human urine.