Serrano Juan Manuel, Silva Manuel
Department of Analytical Chemistry, University of Cordoba, Cordoba, Spain.
Electrophoresis. 2006 Dec;27(23):4703-10. doi: 10.1002/elps.200600184.
This work describes a straightforward and sensitive method for the multi-residue analysis of aminoglycoside antibiotics (kanamycin B, amikacin, neomycin B and paromomycin I) in bovine milk samples. The method involves the pre-capillary derivatization of antibiotics with sulfoindocyanine succinimidyl ester (Cy5) and their separation and determination by MEKC with LIF detection. The optimum procedure includes a derivatization step of the antibiotics at 25 degrees C for 30 min and direct injection for MEKC analysis, which is performed in about 20 min by using borate buffer (35 mM; pH 9.2) with 55 mM SDS as an anionic surfactant and 20% ACN as the organic modifier. Under these conditions, dynamic ranges of 10-500 microg/L and RSDs (within-day precision) from 3.8 to 5.3% were obtained. These results indicate that the proposed MEKC-LIF method is useful as a selective and sensitive tool for the determination of these antibiotics and surpasses other reported electrophoretic alternatives. Finally, the method was successfully applied to bovine milk samples after a simple solid-phase extraction clean-up and preconcentration procedure. The aminoglycosides were readily detected at 0.5-1.5 microg/kg levels with average recoveries ranging from 89.4 to 93.3%.
本研究描述了一种直接且灵敏的方法,用于分析牛奶样品中的氨基糖苷类抗生素(卡那霉素B、阿米卡星、新霉素B和巴龙霉素I)的多残留。该方法包括用磺基吲哚菁琥珀酰亚胺酯(Cy5)对抗生素进行毛细管前衍生化,然后通过MEKC结合LIF检测进行分离和测定。最佳步骤包括在25℃下对抗生素进行30分钟的衍生化步骤,并直接进样进行MEKC分析,使用硼酸盐缓冲液(35 mM;pH 9.2)、55 mM SDS作为阴离子表面活性剂和20% ACN作为有机改性剂,约20分钟内完成分析。在这些条件下,获得了10 - 500 μg/L的动态范围和3.8%至5.3%的RSD(日内精密度)。这些结果表明,所提出的MEKC-LIF方法作为测定这些抗生素的选择性和灵敏工具是有用的,并且优于其他报道的电泳方法。最后,该方法在经过简单的固相萃取净化和预浓缩程序后成功应用于牛奶样品。氨基糖苷类抗生素在0.5 - 1.5 μg/kg水平下易于检测,平均回收率为89.4%至93.3%。