作　　者： ; ; ;

机构地区： 华中农业大学畜牧兽医学院兽药研究所

出　　处： 《华中农业大学学报》 2000年第3期252-255,

摘　　要： 本研究旨在用ELISA试剂盒建立一种简便、可靠的猪肝脏中克仑特罗残留的检测方法。添加了适量克仑特罗标准工作液的肝脏样品同 5倍 0 .1mol/L的盐酸溶液匀浆 ,冻藏、解冻后离心。上清液用正已烷萃取脂溶性杂质 ,再用氢氧化钠溶液调节pH至 1 2。用异丁醇提取待测药物 ,将提取液在 6 0℃用氮气吹干。用 1mL试剂盒中的稀释液将残渣溶解后 ,用ELISA法在 4 50nm处检测克仑特罗的含量。该方法的线性范围为 0 .1 3～ 5.3ng/mL ,回归方程为y=- 1 .80 0x + 2 .2 85,相关系数r=0 .997,最低检测限为 0 .1 3ng/mL ,最低可定量限为 0 .2 5ng/g,浓度为 0 .2 7、0 .53和 1 .0 6ng/g时的回收率分别为 6 8.9%、72 .7%和 77.4 % ,CV <2 0 %。本法的可靠定量限低于克仑特罗在动物肝脏中的最高残留限量 ,且回收率和变异系数均能满足残留检测的要求 ,是一种较简便、可靠的克仑特罗在猪肝脏中残留的快速检测方法 A kind of screening assay to detect the residue of clenbuterol in pig liver was developed. 2g sample, added with appropriate amount of clenbuterol standard solution, was homogenized with 10mL of 0.1mol/L hydrochloric acid. It was centrifuged after frozen and unfrozen. The upper layer was purified with n-hexane, then the pH was adjusted to 12.0 by 5 mol/L sodium hydroxide. 6mL iso-butanol was used to extract clenbuterol, then iso-butanol was removed by N\-2 at 60℃. The residue was dissolved with 1mL diluted diluent/wash buffer of the ELISA kit. The amount of clenbuterol was detected by ELISA kit at 450nm.The linear equation is y =-1.800 x +2.285, r =0.997, linear arrange is 0.13～5.3ng/mL, the lowest detectable limit(LDL) is 0.13ng/mL, the lowest reliable quantitation is 0.25ng/g. Recoveries at 0.27, 0.53 and 1.06 ng/g were 68.9%, 72.7% and 77.4%, respectively. Coefficient of variations ( CV ) <20%. Maximum residue limit (MRL) of clenbuterol in animal liver is 0.5 ng/g, this method can be used to screen clenbuterol residue in pig liver.

关 键 词： 克仑特罗 残留 酶联免疫吸附测定 猪 肝脏

领　　域： [农业科学]