高效液相色谱-串联质谱法快速测定兽药制剂中 喹诺酮类、磺胺类等26种药物

Rapid Determination of 26 Compounds in Veterinary Pharmaceutical Powder by Liquid Chromatography-Tandem Mass Spectrometry

关键词:  兽药制剂  喹诺酮类  磺胺类  孔雀石绿  金刚烷胺  高效液相色谱-串联质谱（LC-MS/MS）

Key words:  vterinary parmaceutical pwder  qinolones  sulfonamides  malachite green  amantadine  HPLC-MS/MS

摘要:

建立了兽药制剂中喹诺酮类、磺胺类、孔雀石绿、金刚烷胺等26种药物的高效液相色谱-串联质谱法的快速筛查与定量方法。样品经0.2%甲酸乙腈超声提取，甲醇水(1∶1，V∶V)稀释10000倍后经LC-MS/MS测定。采用C18色谱柱分离，以5 mmol/L甲酸-甲酸铵与甲醇-乙腈(1∶1, V∶V)为流动相，梯度洗脱方式；质谱采用电喷雾正离子模式离子化，多反应监测模式(MRM)，外标法标准曲线定量。26种药物在0.1 ~10.0 ng/mL浓度范围内线性关系良好，相关系数r2均大于0.99。该方法检出限为0.01 mg/g，定量限为0.02 mg/g。兽药制剂中26种药物在001 mg/g~0.1 mg/g添加范围内，平均回收率为70.0%~101%，相对标准偏差(RSD) 为1.9%~8.8%。该方法简便快捷、准确度和灵敏度高，能满足兽药制剂中常规添加药物及禁用药物进行快速筛查和准确定量。

Abstract:

A rapid screening and quantification method for 26 compounds such as quinolones, sulfonamides, malachite green and amantadine et al,in veterinary pharmaceutical powder was established by high performance liquid chromatography-tandem mass spectrometry.The samples were ultrasonically extracted with 0.2% formic acid-acetonitrile,and determined by LC-MS/MS after diluted with methanol-water (1∶1, V∶V) for 10000 times.The separation was performed on a C18 column using 5 mmol/L formic-ammonium formate and methanol-acetonitrile (1∶1, V∶V) as mobile phases in a gradient elution mode.The mass spectrometry was performed by electrospray ionization in positive ion mode, multiple reaction monitoring mode (MRM) and standard curve external standard method.The calibration curves of the 26 compounds were linear in the range of 01~10.0 ng/mL,with correlation coefficients more than 0.99.The limit of detections(LODs) were 0.01 mg/g, and the limit of quantification(LOQs) were 0.02 mg/g.The method was validated at spiked levels of 0.01,0.02,0.1 mg/g,and the overall recoveries were in a range of 70.0%~101%,with relative standards deviations(RSDs) of 1.9%~8.8%. The method was simple and convenient,efficient,accurate and sensitive,which would meet the requirements of screening and accurate quantification for the routine addition of drugs and banned drugs in veterinary pharmaceutical powder.