改进的液相色谱法测定牛肾中新霉素B的含量

Improved liquid chromatographic determination of neomycin B in bovine kidney.

作者信息

Shaikh B, Jackson J

机构信息

U.S. Food and Drug Administration, BARC-East, MD 20705.

出版信息

J AOAC Int. 1993 May-Jun;76(3):543-8.

PMID:8318846

Abstract

An improved liquid chromatographic (LC) method was developed for the determination of neomycin B in bovine kidney tissue. The tissue is homogenized twice in phosphate buffer; homogenate is centrifuged, and the supernatant is deproteinated by heat. The extract is acidified and mixed with ion-pair concentrate, and neomycin B is determined by an LC system consisting of an ion-pairing mobile phase, a reversed-phase ODS column, postcolumn derivatization with o-phthalaldehyde reagent, and fluorometric detection. Average recoveries of neomycin B from kidney tissues spiked at 3, 6, and 12 ppm were 103, 99, and 104%, with 9.7, 7.9, and 3.7% intralaboratory coefficients of variation, respectively, using a standard curve prepared in buffer. The method was used to determine neomycin B in kidney tissue obtained from a calf killed 14 days after intramuscular dosing with neomycin (5 mg/lb). The tissue was found to contain about 3 ppm neomycin B. The LC conditions were also used to assay control samples of muscle, liver, milk, plasma, and urine. No interfering peaks were noted at the elution position of neomycin B.

摘要

开发了一种改进的液相色谱（LC）方法，用于测定牛肾组织中的新霉素B。将组织在磷酸盐缓冲液中匀浆两次；匀浆液离心，上清液通过加热进行脱蛋白处理。提取物酸化并与离子对浓缩物混合，然后用由离子对流动相、反相ODS柱、邻苯二甲醛试剂柱后衍生化和荧光检测组成的LC系统测定新霉素B。使用在缓冲液中制备的标准曲线，新霉素B在添加浓度为3、6和12 ppm的肾组织中的平均回收率分别为103%、99%和104%，实验室内变异系数分别为9.7%、7.9%和3.7%。该方法用于测定一只犊牛在肌肉注射新霉素（5 mg/磅）14天后宰杀获取的肾组织中的新霉素B。发现该组织中含有约3 ppm的新霉素B。LC条件还用于分析肌肉、肝脏、牛奶、血浆和尿液的对照样品。在新霉素B的洗脱位置未发现干扰峰。