Citation: WU Dan, HAN Mei-Lin, ZOU De-Xun, WANG Xu-Ming, GAO Min, QIU Tian-Lei. Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry Analysis of 16 Kinds of Residual Antibiotics in Chicken Manure. Chinese Journal of Analytical Chemistry, 2017, 45(9): 1389-1396. doi: 10.11895/j.issn.0253-3820.170173 [复制]
Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry Analysis of 16 Kinds of Residual Antibiotics in Chicken Manure
建立了固相萃取-超高效液相色谱-串联质谱（UPLC-MS/MS）同时检测畜禽粪便中四环素类、磺胺类、氟喹诺酮类和大环内酯类16种抗生素的分析方法。针对目标物化学性质和样品杂质情况，对质谱条件、提取液种类、超声功率等参数进行了优化。最终以50%乙腈（V/V）的磷酸盐缓冲溶液（pH=4）提取3次，经过超声、离心、旋蒸、稀释后，SAX-HLB串联小柱净化富集，用10 mL甲醇-丙酮混合液（80：20，V/V）洗脱，35℃氮吹近干后，用含0.1%甲酸-甲醇（1：1，V/V）定容，在UPLC-MS/MS多反应检测模式下进行定性及定量分析。结果表明，粪便中四环素类、磺胺类、氟喹诺酮类和大环内酯类抗生素的平均加标回收率为56.4%-94.6%，相对标准偏差（RSD）在2.6%-19.8%之间，方法检出限（LOD，S/N=3）和定量限（LOQ，S/N=10）分别为0.01-2.50 μg/kg和0.05-7.90 μg/kg。本方法简便、稳定性好、灵敏度高、重现性好，适用于畜禽粪便中多种抗生素的同时检测。
A sensitive and effective method for determination of 16 kinds of antibiotics, including tetracycline, sulfonamide, fluoroquinolone and macrolide, in livestock and poultry manure using solid phase extraction-ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was established. Aiming at the chemical properties and sample impurities of the target, the parameters such as mass spectrum conditions, types of extraction and ultrasonic power were optimized. Finally, the samples were extracted with 50% acetonitrile in phosphate buffer solution (pH=4) for three times, followed by ultrasonic steaming, centrifugal and rotary, dilution, and purified by SAX-HLB. After sample loading, the solid phase was washed with 10 mL of methanol-acetone (80:20, V/V), evaporated to near dryness at 35℃, and then re-dissolved and vortex mixed in 1 mL of 0.1% formic acid:methanol (1:1, V/V). The extracts were analyzed with UPLC-MS/MS and calculated by external standard method based on the monitored product ion. The results indicated that the average spiked recoveries of tetracycline, sulfonamide, fluoroquinolone and macrolide in manure were 56.4%-94.6% with relative standard deviations (RSDs) of 2.6%-19.8%, the LODs (S/N=3) were 0.01-2.50 μg/kg, and the LOQs (S/N=10) were 0.05-7.90 μg/kg. The method was simple with high stability, high sensitivity and good reproducibility, and suitable for the simultaneously determination of many antibiotics in animal and poultry manure.
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