超高压液相色谱-串联四级杆飞行时间质谱法同时筛查猪肉中54种兽药

Screening Method for 54 Drugs in Pork by Ultra Performance Liquid Chromatography- Quadrupole Time-of-Flight Mass Spectrometry

关键词:  超高效液相色谱-飞行时间质谱  兽药  筛查  猪肉

Key words:  ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF)  drugs  screening  pork

摘要:

应用超高压液相色谱串联四级杆飞行时间质谱建立了筛查猪肉中54种兽药的新方法。样品经过乙腈：水=5:1(V:V)和5%三氯乙酸两次提取,旋蒸后经Waters Oasis HLB柱净化后上机测定,样品在正负离子模式下进行检测,在50 ~ 1500 Da质量范围内进行一级全扫描,以保留时间、化合物精确分子离子质量和特征碎片离子质量对药物进行定性分析。结果表明,在低中高三个添加浓度下测定了各种药物的回收率,54种兽药在4 ~ 1.5×104 μg/L范围内线性关系良好,相关系数r2≥0.98。54种药物的质量数误差绝对值小于7.71 ppm,不同兽药在猪肉中回收率为10% ~ 128%。猪肉中检出限为2 ~ 10 μg/kg。该方法适用于猪肉中54种兽药的检测需要,该方法操作简单,准确,杂质干扰少。

Abstract:

A method based on ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF) has been developed for quantitative analysis of 54 kinds of drugs in pork. The conditions of sample preparation and analysis were optimized in this thesis. The drugs were fristly extracted with 25 mL acetonoitrile and 5 mL deionized water, after adding 2 g NaCl, the extract was evaporated to 5 mL under nitrogen at 40 ℃. The second extraction of residues was carried out by 20 mL 5% trichloroacetic. After adjusting the second extracting solution to pH=6.5 with 6 mol/L NaOH, all of the extarcting solutions were purified with a 500 mg Waters Oasis HLB SPE cartridge. The separation of analytes were carried on a BEH C18 column(2.1 mm×100 mm, 1.7 μm)by a gradient elution using acetonitrile(A) - water(added 0.1% formic acid) (B) as mobile phase with a flow rate of 0.4 mL/min. 54 drugs were detected under both positive electrospray ionization (ESI) and negtive electrospray ionization (ESI) based on retention time, accurate mass and fragment ions information. The method was quantified by external standard method and the linearity of respond ranged from 4 μg/L to 1.5×10₄ μg/L and the correlation coefficient was greater than 0.98. The average recoveries ranged from 10% to 128% of 54 kinds of drugs in pork at three different levels. The limits of detection were ranged from 2 μg/kg to 10 μg/kg. The results indicated that the method develop is highly sensitive and selective in screening for 54 kinds of drugs in pork.