Online coupling of molecularly imprinted solid-phase extraction to HPLC for determination of trace tetracycline antibiotic residues in egg samples.

An automated system has been developed for the determination of trace tetracycline antibiotics (TCs) in egg samples, based on online molecularly imprinted solid-phase extraction (MISPE) coupling with high-performance liquid chromatography (HPLC). Oxytetracycline and chlortetracycline were chosen as mixed templates to synthesize highly selective molecularly imprinted polymers for online extraction. Under the optimal online MISPE-HPLC condition, 10 mL egg samples were injected into the MISPE column and then the matrix was washed out. By rotating the switching valve, TCs were transferred to the analytical column and then separated by HPLC. Because sample pretreatment and chromatographic separation were carried out simultaneously, the whole analytical time (18 min) was significantly shortened compared with conventional offline techniques. The detection limits ranged from 0.8 to 1.3 ng/g. The enhancement factors were in the range of 159-410. The spiked recoveries of TCs in real egg samples ranged from 91.6 to 107.6% and the relative standard deviations (RSDs) were not higher than 4.0%.