| A high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) method and an ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method for simultaneous determination of residues of eight coccidiostats (Robenidine, Halofuginone, Lasalocid, Monensin, Nigericin, Salinmoycin, Narasin and Maduramicin) in egg, chicken muscle and beef were established and validated, using Jinghai yellow chicken and Chinese Holstein as experimental materials in this study. The main resultsare as follows:1. A HPLC-MS/MS method was developed for simultaneous extraction and confirmation of residues of in albumen, egg yolk, whole egg, chicken muscle and beef. At the fortified levels within the range of the limit of quantitation (LOQs) to 100.00Oμg/kg for eight coccidiostats, the peak area ratio of qutantificational ion showed a linear correlation with the concentration(R2= 0.99934,0.99933,0.99951,0.99975,0.99966,0.99974, 0.99947,0.99957, respectively. At the fortified levels of LOQs for eight coccidiostats in specified tissues, the recovery values of four analytes in egg, egg yolk, whole egg, chicken muscle and beef were all higher than 75.38%,73.57%,71.24%,75.39%,72.49%,72.81%,71.62% and 74.21%, respectively. The relative standard deviations (RSD) were all less than 16.78%,12.65%,14.33%,14.68%,14.85%,18.79%,14.92% and 12.68%. respectively. At spiked levels of 0.5,1, and 2 fold of max residue limit (MRL), recoveries of eight coccidiostats simultaneously were 81.16%～88.30%,79.38%～89.24%,81.10%～88.25%,86.33%～97.25%, 81.26%～90.89%,82.46%～92.56%,80.27%～89.21% and 81.66%-89.37%; with relative standard deviations(RSD)of 9.89%,9.13%,4.84%,5.47%,4.76%,7.54%,5.19%,5.72%, respectively. Under current experimental conditions, intra-day RSD were all less than 9.96%,9.82%,5.96%,8.19%,9.84%, 7.32%,8.36%,8.81% and 8.81%, respectively. Inter-day RSD were no higher than 14.65%,14.83%, 9.65%,9.27%,9.89%,10.12%,9.97% and 9.49%, respectively. limits of detection (LOD) and the highest values were 0.48,0.58,0.44,0.46,0.52,0.56,0.58 and 0.61 μg/kg (S/N≥3), respectively, limit of quantitation (LOQ)and the highest values were 1.60,1.92,1.28,1.56,1.78,1.85,1.89 and 1.97μg/kg(S/N ≥10), respectively. The CCa and CCp were all near the MRLs in different tissues (SD≤5.17% and SD SD≤ 10.34%).2. A UPLC-MS/MS method was developed for simultaneous extraction and detection of eight coccidiostats simultaneously in albumen, egg yolk, whole egg, chicken muscle, and beef. At fortified levels within the range of the limit of quantitation(LOQs) to 100.00μg/kg for eight coccidiostats simultaneously, the peak area ratio of qutantificational ions showed a linear correlation with the concentration. The coefficients of determination(R2) of eight analytes in different tissues are all higher than 0.99947,0.99947,0.99961 0.99959,0.99969,0.99979,0.99947,0.99957. At the fortified levels of LOQs for eight coccidiostats in specified tissues, the recovery values of four analytes in egg, chicken muscle and beef were all higher than 75.24%,72.26%,72.34%,73.28%,74.20%,80.92%,72.32% and 71.96%, respectively. The relative standard deviations (RSD) were all less than 12.06%,11.73%,10.36%,12.38%,13.25%,12.16%,11.28% and 11.39%, respectively. At the fortified level of 0.5MRL, 1MRLand 2MRL, the recovery were 84.07%～88.19%,77.57%～89.07%,82.35%～91.22%,87.62%～96.80%,86.846%～99.52%, 88.99%～102.74%,82.16%～88.71% and 81.98%～89.15%. The relative standard deviations (RSD) were all less than 7.31%,6.40%,7.40%,8.75%,8.67%,7.98%,8.61% and 8.52%. The corresponding intra-day and intra-day variation were found to be less than 9.96%,7.89%,8.98%,9.87%,9.77%,8.55%,9.21% and 9.87%. The corresponding inter-day variation were found to be less than 13.37%,10.46%,10.79%,11.39%, 10.80%,12.71%,11.51% and 10.91%. In HPLC-MS/MS detection under the condition of detection limit and the highest value were 0.33,0.34,0.38,0.32,0.29,0.31,0.28 and 0.35μg/kg (S/N≥3), limit of quantitation and the highest values were 1.10,1.13,1.25,1.08,0.99,0.95,1.02 and 1.21μg/kg (S/N>10). The CCa and CCβ were all near the MRLs in different tissues (SD<5.09% and SD<10.18%).Comparing the two methods developed in this study, the UPLC-MS/MS method was proved to possess higher recovery and precision and also had faster analyzing speed, better separating effect of the peaks, as well as economy of the mobile phase. In this study, the HPLC-MS/MS and UPLC-MS/MS method were successfully applied in practice of the determination of eight coccidiostats residue in albumen, egg yolk, whole egg, chicken muscle, and beef. Both of the two methods meet the requirements of the Chinese Ministry of Agriculture, the Food and Drug Administration of the United States, and the European Union and was proved to be suitable for detection in practice. |
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