牛奶中七种氟喹诺酮类药物残留检测高效液相色谱-串联质谱法研究

Determination of Fluoroquinolones Residue in Milk by HPLC-MS/MS

关键词:  牛奶  氟喹诺酮类药物  高效液相色谱-串联质谱

Key words:  milk  Fluoroquinolones  HPLC-MS/MS

摘要:

建立了牛奶中氧氟沙星、诺氟沙星、环丙沙星、恩诺沙星、达氟沙星、二氟沙星和沙拉沙星等七种氟喹诺酮类药物（FQs）残留检测的高效液相色谱-串联质谱（HPLC-MS/MS）方法。液相色谱条件为：色谱柱为Waters XBridge C18柱（150×2.1mm,5µm）；流动相为0.1％甲酸甲醇溶液＋0.1％甲酸水溶液，梯度洗脱；柱温为35℃；流速为0.2mL/min；进样量为20µL。质谱条件为：电喷雾离子源（ESI＋），多反应监测（MRM）方式进行采集；外标法定量。结果表明：七种FQs在5～250ng/mL浓度范围内呈现良好的线性关系，相关系数R2均大于0.997；方法检测限为2ng/mL，定量限为5ng/mL；从10、20和100ng/mL三个添加浓度检测结果可以看出，方法平均回收率为70.2%～119%，批内RSD为1.8％～16％，批间RSD为9.1％～18％。

Abstract:

A HPLC-MS/MS method was established for the determination of Ofloxacin, Norfloxacin,Ciprofloxacin,Enrofloxacin,Danofloxacin,Difloxacin and Sarafloxacin residue in milk. HPLC condition: the chromatography column is XBridge C18 column of 150×2.1mm,5µm；mobile phase is methanol(0.1% formic acid) and water(0.1% formic acid)；column temperature is 35℃；flow rate is 0.2 mL/min；injection volume is 20µL. Mass Spectrometry condition: ESI+ and MRM mode. The calibration curve was good linear between the peak areas and concentrations of Fluoroquinolones from 5 to 250 ng/mL，the correlation coefficient R2＞0.997. The limit of detection of the method was 2ng/mL, and the limit of quantification was 5ng/mL. The average recoveries from spiked milk at the three concentrations of 10,20 and 100ng/mL ranged from 70.2%～119%（n＝6）, and intra- and inter-batch variation coefficients were 1.8％～16％ and 9.1％～18％,respectively.