HLB-MCX-Based Solid-Phase Extraction Combined with Liquid Chromatography–Tandem Mass Spectrometry for the Simultaneous Determination of Four Agricultural Antibiotics (Kasugamycin, Validamycin A, Ningnanmycin, and Polyoxin B) Residues in Plant-Origin Foods
Wenxi Li, Dai XueFang, Entang Pu, Haitao Bian, Zilei Chen, Xueyan Zhang, Zhixiang Guo, Peng Li, Huidong Li, Yanhua Yong, Chenchen Wang, Yan Zhang, Lijun Han
Abstract
An ultrahigh-performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS) method was established for the determination of four highly polar agricultural antibiotics kasugamycin, validamycin A, ningnanmycin, and polyoxin B in plant-derived foods. The samples were extracted with a 0.2% formic acid solution, purified by hydrophilic–lipophilic balance and mixed-mode cation-exchange solid-phase extraction, and then reconstituted for UPLC–MS/MS detection. The chromatographic analysis was performed on a BEH Amide column (100 mm × 2.1 mm, 1.7 μm) using gradient elution with a 0.1% formic acid solution and 0.1% formic acid acetonitrile as mobile phases. Method validation was performed on 15 matrices spiked at 0.02 (or 0.05), 0.5, and 2 mg/kg. The mean recovery rate ranged from 75 to 102% with relative standard deviations (RSD) was less than 20%. Good linearities (r > 0.99) in the range of 0.002–0.2 μg/mL were obtained. The limits of quantification (LOQs) were 0.02 and 0.05 mg/kg. Studies on the stability of the analytes in the stored kiwifruit samples showed that kasugamycin, validamycin A, and ningnanmycin were stable for at least 6 months, while polyoxin B was observed to be partially degraded (the degradation rate at 6 months was 31.3%). The method was demonstrated to be effective and reliable in real samples. In the kiwifruit samples treated after 7 days, no residues of ningnanmycin and polyoxin B were detected, while the residues of kasugamycin and validamycin A were 0.12 and 0.038 mg/kg, respectively.