甲醇-磷酸氢二钾-水双水相萃取-高效液相色谱检测肉类产品中四环素类残留  

Extraction and detection of tetracycline residues in meat products using methanol-K_2HPO_4aqueous two phase system coupled with liquid chromatography

作　　者：景超 文嘉慧 李晓通 邬旭然[1] 刘永明[1] JING Chao;WEN Jia-hui;LI Xiao-tong;WU Xu-ran;LIU Yong-ming(School of Chemistry and Chemical Engineering,Yantai University,Yantai 264005,China)

机构地区：[1]烟台大学化学化工学院,山东烟台264005

出　　处：《化学研究与应用》2018年第10期1733-1738,共6页Chemical Research and Application

基　　金：国家自然基金项目(21575122)资助

摘　　要：以甲醇-无机盐-水双水相体系,建立了一种高效的用于液相色谱法检测肉类产品中四环素类抗生素残留的样品前处理方法。实验考察了双水相的形成条件以及对四环素类药物的提取效率,用液相色谱法与瑞利散射法研究了蛋白质对甲醇-无机盐-水双水相体系萃取四环素的影响。结果表明,只有加入适量的磷酸氢二钾才能形成甲醇水双水相体系,该双水相体系对极性较强的四环素类药物具有较高的萃取效率;体积分数为70%的甲醇水溶液作为萃取液可以使肉类产品中的蛋白质基质发生缓慢而彻底地变性,有利于释放与蛋白质结合的药物。由于甲醇-K_2HPO_4-水双水相体系的上相萃取液中水溶性蛋白等杂质较少,样品净化步骤大大简化,在上相中加入少量PSA与C_(18)固体吸附剂,离心后取上清液即可进行HPLC分析。该方法用于肉类产品牛肉、鱼肉中四环素残留的萃取测定,回收率为76.8%~98.2%,检出限为0.039~0.084μg·g^(-1)。Based on the methanol–salt-H 2O aqueous two phase system(ATPS),an effective sample pretreatment method was developed for the analysis of 4 kinds of tetracyclines in meat by liquid chromatography(HPLC).The forming conditions of methanol-salt-H 2O system and the extrction efficiency of the ATPS for tetracyclines were detailedly investigated.Meanwhile,the influences of the protein on extracting tetracyclines with the methanol–salt-H 2O system were thoroughly studied by HPLC and Rayleigh scattering method.The results showed that the methanol-salt-H 2O ATPS could be formed only when the right amount of K 2HPO 4 was added.And this ATPS has high efficiency for extacting the tetracyclines with strong chemical polarity.In theφ(methanol)80%,a slow but full protein denaturation takes place,which causes the unfolding of protein and the release of tetracyclines which combined with protien.Furthermore,there were less water-soluble impurities in the upper phase of the methanol-K 2HPO 4-H 2O system.Therefore the purification steps were greatly simplified.The primary secondary amine(PSA)and C 18 were added in upper phase,and the supernatant was directly analyzed by HPLC after centrifugation.This method has been applied to the determination of tetracyclines residues in meat.The detection limits were in the range of 0.039~0.084μg·g-1,and the spiked recoveries were76.8%~98.2%.

关 键 词：四环素 甲醇水双水相体系 液相色谱 蛋白质 

分 类 号：O657.72[理学—分析化学]