建立超高效液相色谱-串联质谱法(ultra performance liquid chromatography-tandem mass spectrometry, UPLC-MS/MS)同时测定蜂蜜中9种氨基糖苷类药物(新霉素、阿米卡星、双氢链霉素、链霉素、潮霉素B、卡那霉素、庆大霉素、妥布霉素、大观霉素)残留量的方法。蜂蜜样品中的氨基糖苷类药物采用三氯乙酸的缓冲盐溶液提取,经HLB固相萃取柱富集净化,选择水性洗脱液洗脱,在最终提取液中添加离子对试剂优化色谱行为,选取乙腈-水体系作为流动相进行梯度洗脱,采用反相色谱柱进行分离,以串联质谱法正离子模式扫描,外标法定量。实验结果表明,该方法在20~1 000 ng/mL线性关系良好,新霉素、双氢链霉素、链霉素、卡那霉素、妥布霉素的定量限为5 μg/kg,阿米卡星、潮霉素B、庆大霉素、大观霉素的定量限为10 μg/kg,在线性范围内,分别在空白蜂蜜样品中进行3个浓度的添加实验,样品的回收率在72.6%~109.4%,相对标准偏差为2.13%~8.23%。该方法操作简单,灵敏度高,净化效果好,在流动相中避免使用离子对试剂,有效减少了对质谱仪的污染,能够较好地满足蜂蜜中氨基糖苷类药物残留量的检测需要。
Ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed for the simultaneous detection of aminoglycosides (neomycin, amikacin, dihydrostreptomycin, streptomycin, hygromyxin B, kananmycin, gendamicin, tobramycin, spectinomycin). The residues of aminoglycosides in the honey were extracted with TCA buffer, enriched and cleaned up with HLB solid phase extraction column, and eluted with aqueous elution. And the ion-pairing reagent was added to the final extract and used to optimize the chromatography conditions. Moreover, the extracts were separated on reversed-phase chromatographic column using acetonitrile-water as mobile phase by gradient elution, and detected by liquid chromatography-mass spectrometry under multiple reaction monitoring mode via positive ion mode. All the analytes were calibrated by the external method. The results showed that the method has a good linear relationship in the range of 20-1 000 ng/mL. Moreover, the quantification limit of neomycin, dihydrostreptomycin, streptomycin, kanamycin, and tobramycin is 5 μg/kg; the quantification limit of amikacin, hygromycin B, gentamicin, and spectinomycin is 10 μg/kg. Within the linear range, three concentrations of addition experiments were carried out in the blank honey samples, the sample recovery rates ranged from 72.6%-109.4%, and the relative standard deviations were 2.13%-8.23%. The method developed here has the advantages of easy operation, high sensitivity and good filter effect. By avoiding the use of ion-pairing reagent in the mobile phase, the method could effectively reduce the pollution of mass spectrometer and better meet the detection requirements of the aminoglycoside residues in honey.
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