QuEChERS-高效液相色谱-串联质谱法同时测定水产养殖环境沉积物中磺胺类、喹诺酮类、大环内酯类抗生素

建立了QuEChERS结合高效液相色谱-串联质谱同时测定水产养殖环境沉积物中磺胺类、喹诺酮类、大环内酯类等26种抗生素残留的分析方法。沉积物经20 mL乙腈-磷酸盐缓冲液(1∶1,V/V)超声提取,0.15 g乙二胺四乙酸二钠(Na2EDTA)络合除杂,氯化钠(NaCl)盐析,无水硫酸镁(MgSO4)、N-丙基乙二胺(PSA)、C18吸附剂和石墨化碳黑(GCB)填料净化,然后,通过Ultimate XB-C18色谱柱分离,采用4 mmol/L乙酸铵-0.1%甲酸水溶液和0.1%甲酸甲醇梯度洗脱,电喷雾正离子模式下,以多反应监测模式(MRM)监测,外标法定量。结果表明:在1.0~200μg/L范围内,26种抗生素的线性相关系数均在0.99以上,方法定量限为1~2μg/kg;在3个浓度添加水平下,平均回收率为60.4%~113%,相对标准偏差为1.5%~13.6%。该方法具有良好的精密度和准确度、灵敏度高、适用范围广,可在一定程度上实现这26种抗生素的多残留快速测定。采用该方法对某养殖区的20个养殖池塘沉积物进行检测,磺胺甲基嘧啶、磺胺二甲嘧啶、磺胺甲噁唑、磺胺嘧啶、红霉素、罗红霉素、克拉霉素均有检出,其中磺胺嘧啶、磺胺二甲嘧啶、磺胺甲噁唑的检出率较高。

Simultaneous Determination of Sulfonamides,Quinolones and Macrolides Antibiotics Residues in Sediment from Aquaculture Environment by QuEChERS-HPLC-MS/MS

The presence of sulfonamides, quinolones and macrolides antibiotic residues in environment is a major concern for both aquatic ecosystems and human health, particularly because they can induce drug-resistant bacteria and resistance genes. An efficient method was developed for the determination of sulfonamides, quinolones and macrolides antibiotics in sediment from aquaculture environment by QuEChERS (quick, easy, cheap, effective, rigged and safe)-high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). The target analytes in sediments were ultrasonically extracted with 20 mL phosphate buffer solution and acetonitrile mixture (1∶1,V/V) and salted out with sodium chloride (NaCl), followed by 0.15 g Na₂EDTA complexation. After centrifugation, the supernatant extract was cleaned up with magnesium sulfate (MgSO₄), primary secondary amine (PSA), C18 sorbent and graphitized carbon black. The analytes were separated on an Ultimate XB-C18 column (2.1 mm×150 mm×5 μm) by gradient elution with 4 mmol/L ammonium acetate solution containing 0.1% formic acid-0.1% formic acid methanol as mobile phase. The formic acid played a key role in mobile phase as modifier to improve the ionization efficiency of antibiotics. The column temperature was 40 ℃ and the flow rate was 0.3 mL/min. The analytes was performed using electrospray ionization (ESI) in positive ion mode and multiple reaction monitoring (MRM), quantified with external standard method. In order to achieve more accurate quantitation, a matrix-matched curve was used to eliminate the matrix effects. The results showed that the calibration curves are linear (R²>0.99) over a concentration range of 1.0-200 μg/L for 26 antibiotics. The limits of quantification are 1.2 μg/kg for all antibiotics. At three spiked levels, the average recoveries ranged from 60.4% to 113% with the relative standard deviations (RSDs) of 1.5%-13.6%. This method has high sensitivity, good reproducibility and wide applicability, which is suitable for rapid screening and detection of 26 antibiotics. Sulfonamides, quinolones and macrolides antibiotics were determined in 20 sediment samples collected from aquaculture ponds in southern Fujian Province. Sulfamerazine, sulfamethazine, sulfamethoxazole, sulfadiazine, erythromycin, roxithromycin and clarithromycin were detected, and the detection rates of sulfadiazine, sulfamethazine and sulfamethoxazole were the highest.