高效液相色谱-高分辨质谱法定性筛查牛奶中49种β-内酰胺类抗生素残留

Qualitative screening of 49 kinds of β-lactam antibiotic residues in milk by high performance liquid chromatography coupled with high resolution mass spectrometry

中文关键词:  β-内酰胺类抗生素  高分辨质谱法  目标物筛查  牛奶

英文关键词:β-lactam antibiotic  high resolution mass spectrometry  targeted screening  milk

中文摘要:

      目的 开发和验证定性筛查牛奶中49种β-内酰胺类抗生素残留的高效液相色谱-高分辨质谱法(high performance liquid chromatography coupled with high resolution mass spectrometry, HPLC-HRMS)。方法 牛奶经磷酸盐缓冲液及乙腈提取后, 加入硫酸钠(Na2SO4)和氯化钠(NaCl)进行盐析, 离心后取部分上清液经Na2SO4和C18混合的分散式固相萃取填料(dispersive solid phase extraction, d-SPE)净化, 净化液浓缩定容后, 使用HPLC-HRMS分别以全扫描-数据依赖型二级扫描(full scan-data dependent secondary acquisition, Full MS/dd-MS2)和全扫描-平行反应监测二级扫描(full scan-selected ion monitoring/parallel reaction monitoring, Full MS/PRM)两种模式进行数据采集。结果 在Full MS/PRM模式下, 45种抗生素通过方法验证, 假阳性率和假阴性率都小于5%; 在Full MS/dd-MS2模式下, 43种抗生素通过方法验证, 假阳性率和假阴性率在规定的目标浓度下均低于5%, Full MS/PRM数据采集模式更适用于多兽药残留的定性筛查。结论 此方法适用于牛奶样品中β-内酰胺类抗生素的定性筛查, 并且高分辨质谱的Full MS/PRM采集模式相较于Full MS/dd-MS2采集模式, 抗干扰性更好, 更适用于复杂基质中目标物的定性筛查。

英文摘要:

      Objective To developed and validate a qualitative screening method for the determination of 49 kinds of β-lactam antibiotic residues in milk by high performance liquid chromatography coupled with high resolution mass spectrometry (HPLC-HRMS). Method Milk was extracted with a mixture of a phosphate buffer solution and acetonitrile followed by a salt partition using sodium sulphate (Na2SO4) and sodium chloride (NaCl). After centrifugation, an aliquot of the supernatant was cleaned by dispersive solid phase extraction (d-SPE) which consisted of a mixture of Na2SO4 and octadecyl-silica C18 sorbents. After concentration and constant volume of the purification solution, HPLC-HRMS were used for data acquisition in 2 modes: Full scan-data dependent secondary acquisition (Full MS/dd-MS2) and full scan-selected ion monitoring/parallel reaction monitoring (Full MS/PRM). Result The 45 kinds of antibiotic passed through the validation with false positive and negative rate less than 5% in Full MS/PRM mode; 43 kinds of antibiotics passed through the validation with false positive and negative rate less than 5% at the specified target concentration in Full MS/dd-MS2 mode, Full MS/PRM was more suitable for the qualitative screening of multiple veterinary drug residues. Conclusion This method is suitable for the qualitative screening of β-lactam antibiotics residues in milk. Comparing to Full MS/dd-MS2, Full MS/PRM is more robust, and it is more suitable for the qualitative screening of veterinary drug residues in complex matrix.