超高效液相色谱串联高分辨四极杆飞行时间质谱法快速筛选鉴别猪血浆中的大环内酯类抗生素

Rapid screening, indentification of macrolides antibiotics in swine plasma using ultra-performance liquid chromatography-tandem high resolution quadrupole-time-of-flight mass spectrometry

关键词:  超高效液相色谱  高分辨四极杆飞行时间质谱  猪血浆  大环内酯类抗生素  筛选  鉴别

Key words:  ultra-performance liquid chromatography  high resolution quadrupole-time-of-flight mass spectrometry  swine plasma  macrolides antibiotics  screening  indentification

摘要:

采用超高效液相色谱串联高分辨四极杆飞行时间质谱法(UPLC-Q-TOP-MS)结合大环内酯类抗生素数据库,建立快速筛查鉴别猪血浆中常见大环内酯类抗生素的方法。猪血浆经甲醇提取,通过UPLC-Q-TOP-MS正离子模式全扫描分析,采用Agilent Extend-C18色谱柱(50×2.1 mm,1.8μm)分离,以乙腈-2mmol/L乙酸铵水溶液(含0.1%甲酸)为流动相梯度洗脱,流速0.4mL/min,柱温40℃,得到猪血浆中大环内酯类抗生素准分子离子峰精确质量数、主要碎片峰的精确质量数和保留时间后,结合大环内酯类抗生素数据库筛查,实现猪血浆中大环内酯类抗生素的快速定性识别。所建数据库包含26种常见猪血浆中大环内酯类抗生素的准分子离子峰精确质量数、主要碎片峰的精确质量数和保留时间,其中13种常见大环内酯类抗生素的检测限在10-80ng/mL范围内。该方法分离度高、检测灵敏,可以作为快速筛查鉴别猪血浆中的大环内酯类抗生素的有效方法。

Abstract:

A method was developed for rapid screening and identification of common macrolide antibiotics in swine plasma by ultra-performance liquid chromatography-tandem high resolution quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOP-MS) combined with macrolide antibiotics database. The swine plasma sample was extracted with methanol, and then the extracts were analyzed by UPLC-Q-TOP-MS in positive full scan modes. The best detection conditions were as follows: Agilent Extend-C18 chromatographic column(50×2.1 mm, 1.8μm) was used for separation, with acetonitrile and 2mmol/L ammonium acetate aqueous solution (containing 0.1% formic acid) as mobile phases. The flow rate was 0.4 mL/min and the temperature of column was set at 40℃. The accurate-mass of quasi-molecular ion peak, accurate-mass of main fragment ion peaks and retention time of macrolide antibiotics in swine plasma were acquired and the macrolide antibiotics in swine plasma could be successfully qualitative identification by database screening. The created database includes accurate-mass of quasi-molecular ion peak, accurate-mass of main fragment ion peaks and retention time of 26 common macrolide antibiotics. The limits of detection (LOD) of 13 macrolide antibiotics were in the range of 10-80ng/mL. The results showed that the proposed UPLC-Q-TOP-MS method was sensitive and had the high separation efficiency. The method was a useful tool for the rapid screening and qualitative identification of common macrolide antibiotics in swine plasma.