克仑特罗在猪肝脏中残留检测的ELISA法研究

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

Development of a Screening Assay of Clenbuterol Residue in Pig Liver by ELISA Kit

A kind of screening assay to detect the residue of clenbuterol in pi g liver was developed. 2g sample, added with appropriate amount of clenbuterol s tandard solution, was homogenized with 10mL of 0.1mol/L hydrochloric acid. It wa s centrifuged after frozen and unfrozen. The upper layer was purified with n-hex ane, then the pH was adjusted to 12.0 by 5 mol/L sodium hydroxide. 6mL iso-butan ol was used to extract clenbuterol, then iso-butanol was removed by N2 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 equatio n is y=-1.800x+2.285, r=0.997, linear arrange is 0.13～5.3ng/mL, the lowest detec table limit(LDL) is 0.13ng/mL, the lowest reliable quantitation is 0.25ng/g. Rec overies at 0.27, 0.53 and 1.06 ng/g were 68.9%, 72.7% and 77.4%, respectively. C oefficient of variations (CV) <20%. Maximum residue limit (MRL) of clenbuter ol I n animal liver is 0.5 ng/g, this method can be used to screen clenbuterol residu e in pig liver.