Determination of 16 sulfonamide antibiotics residues in human urine with liquid chromatography-tandem mass spectrometry

Abstract

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 C ₁₈ 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.

Abstract

【摘 要】 目的 建立同时测定尿液中 16 种磺胺类抗菌药物多残留的超高效液相色谱 – 串联质谱（UPLC-MS/MS）分析方法。 方法 尿液经酶解处理后，Oasis PRiME HLB（200 mg、6 mL）固相萃取柱净化，采用 Waters BEH C ₁₈ 色谱柱（2.1 × 100 mm，1.7 μm），水（含 0.01 mol/L 乙酸铵）和甲醇（含 0.01 mol/L 乙酸铵）作为流动相进行梯度洗脱，电喷雾离子源电离，正离子扫描，多反应监测模式（MRM）进行定性和定量分析。 结果 16 种磺胺类抗菌药物的线性范围为 5 ～ 1 000 ng/mL，相关系数均大于 0. 99。方法定量下限为 5 ng/mL，检出限为 0.3 ～ 1.7 ng/mL，3 个加标水平回收率为 71.6 % ～ 97.3 %，相对标准偏差（RSD）为 1.4 % ～ 11.7 %。 结论 超高效液相色谱 – 串联质谱法操作简便、具有较高的灵敏度、特异度和精确性，适用于尿中多种痕量磺胺类抗菌药物残留量的同时测定。