Programa de Pós-Graduação em Ciência dos Alimentos, Laboratório de Bioquímica de Alimentos, Departamento de Ciência e Tecnologia de Alimentos, Universidade Federal de Santa Catarina, Rod. Admar Gonzaga, 1346, Itacorubi, CEP: 88034-001, Florianópolis, SC, Brazil.
Talanta. 2012 Sep 15;99:616-24. doi: 10.1016/j.talanta.2012.06.047. Epub 2012 Jun 27.
A simplified procedure for simultaneous quantification of ceftiofur (CEF), fluoroquinolone (FQ) and sulfonamide (SA) antibacterials in bovine milk was developed. The reverse-phase liquid chromatography (RP-LC) multiclass method for analysis of eleven distinct compounds, from three antibacterial classes, was validated in line with Commission Decision 2002/657/EC. Confirmation of the analytes identities was performed by electrospray mass spectrometry detection. The analytes were extracted from milk matrix by liquid-liquid extraction with acidified ultrapure water and directly analyzed in the chromatograph. The SA compounds were pre-column derivatized with fluorescamine for fluorescence detection. The method provided good results regarding the analytical parameters of linearity, selectivity, sensitivity, precision, recovery, decision limit (CCα), detection capability (CCβ), limit of detection (LOD), limit of quantification (LOQ), stability and robustness. Analytes were extracted by liquid-liquid extraction in the fortified matrix and the compounds identity was confirmed by their precursor ion and fragments through tandem mass spectrometry analysis. Additionally, milk samples from two state capitals in the South Region of Brazil were analyzed by both the quantitative and confirmatory methods. The validation process showed correlation coefficients (r(2)) greater than 0.98 for all the analytes, with recovery rates up to 98% for all the studied drugs. LOD and LOQ limits ranged from 8.0 to 20.0 ng mL(-1) and 10.0 to 32.0 ng mL(-1), demonstrating good specificity of the method. The intra-day and inter-day precisions for all the analytes were below or equal to 7.40 and 10.13, respectively. The studied antibacterials were not detected in milk samples. The developed method represents an efficient alternative for multi-residue analysis in milk, being suitable and especially viable for monitoring in developing countries.
建立了一种同时定量检测牛奶中头孢噻呋（CEF）、氟喹诺酮（FQ）和磺胺类抗菌药物的简化方法。该反相液相色谱（RP-LC）多类方法分析了来自三个抗菌类别的十一种不同化合物，符合委员会 2002/657/EC 号决定。通过电喷雾质谱检测确认了分析物的身份。用酸化超纯水进行液液萃取从牛奶基质中提取分析物，并直接在色谱仪中进行分析。磺胺类化合物用荧光胺柱前衍生化进行荧光检测。该方法在分析参数的线性、选择性、灵敏度、精密度、回收率、决策限（CCα）、检测能力（CCβ）、检测限（LOD）、定量限（LOQ）、稳定性和鲁棒性方面取得了良好的结果。在加标基质中通过液液萃取提取分析物，并通过串联质谱分析其母离子和碎片确认化合物的身份。此外，还通过定量和确证方法分析了来自巴西南地区两个首府的牛奶样品。验证过程表明，所有分析物的相关系数（r（2））均大于 0.98，所有研究药物的回收率高达 98%。LOD 和 LOQ 限值范围为 8.0 至 20.0ng/mL 和 10.0 至 32.0ng/mL，表明该方法具有良好的特异性。所有分析物的日内和日间精密度均低于或等于 7.40 和 10.13。研究中的抗菌药物未在牛奶样品中检出。所开发的方法代表了一种用于牛奶中多残留分析的有效替代方法，适用于特别是发展中国家的监测。
