Reid J A, MacNeil J D
Canadian Food Inspection Agency, Centre for Veterinary Drug Residues, Saskatoon, Saskatchewan, Canada.
J AOAC Int. 1999 Jan-Feb;82(1):61-7.
Tissue samples are digested under hot alkaline conditions after initial conditioning at room temperature with phosphate-buffered saline. The cooled digest is deproteinated with concentrated perchloric acid. After centrifugation and pH adjustment, the clear supernatant is applied to an ion-exchange cartridge, and after the cartridge is washed, the neomycin is eluted with dilute perchloric acid. This eluate is derivatized with 9-fluorenylmethyl chloroformate prior to liquid chromatography using a wide-pore spherical silica C4 column and fluorescence detection. Recovery and repeatability are calculated from tissue extract standard calibration curves produced from the same assay. Recoveries ranged from 80 to 120% for fortifications of 0.25-1.00 mg/kg for muscle tissue and from 80 to 100% for fortifications of 0.50-10.0 mg/kg for kidney tissue. Limits of quantitation were 0.25 and 0.50 mg/kg, respectively, for muscle and kidney tissues. Limits of detection were 0.125 and 0.20 mg/kg, respectively, for muscle and kidney tissues.
组织样本在室温下用磷酸盐缓冲盐水初步处理后，于热碱性条件下进行消化。冷却后的消化液用高氯酸进行脱蛋白处理。离心并调节pH值后，将澄清的上清液应用于离子交换柱，柱经洗涤后，用稀高氯酸洗脱新霉素。在使用宽孔径球形硅胶C4柱和荧光检测进行液相色谱分析之前，该洗脱液用9-芴基甲基氯甲酸酯进行衍生化。回收率和重复性根据同一分析产生的组织提取物标准校准曲线计算得出。肌肉组织中添加量为0.25 - 1.00 mg/kg时，回收率在80%至120%之间；肾脏组织中添加量为0.50 - 10.0 mg/kg时，回收率在80%至100%之间。肌肉和肾脏组织的定量限分别为0.25和0.50 mg/kg。肌肉和肾脏组织的检测限分别为0.125和0.20 mg/kg。
