采用离子对液相色谱-荧光检测法测定动物组织中的新霉素。

Determination of neomycin in animal tissues, using ion-pair liquid chromatography with fluorometric detection.

作者信息

Shaikh B, Allen E H, Gridley J C

出版信息

J Assoc Off Anal Chem. 1985 Jan-Feb;68(1):29-36.

Abstract

A liquid chromatographic (LC) method is described for the determination of neomycin in animal tissues. Tissues are homogenized in 0.2M potassium phosphate buffer (pH 8.0); the homogenate is centrifuged, and the supernate is heated to precipitate the protein. The heat-deproteinated extract is acidified to pH 3.5-4 and directly analyzed by LC. The LC method consists of an ion-pairing mobile phase, a reverse phase ODS column, post-column derivatization with o-phthalaldehyde reagent, and fluorometric detection. The LC method uses paromomycin as an internal standard, and separates neomycin from streptomycin or dihydrostreptomycin because they have different retention times. The LC column separates neomycin in 25 min; the detection limit is about 3.5 ng neomycin. The overall recovery of neomycin from kidney tissues spiked at 1-30 ppm was 96% with a 9.0% coefficient of variation. The method was also applied to muscle tissue.

摘要

本文描述了一种用于测定动物组织中新霉素的液相色谱（LC）方法。将组织在0.2M磷酸钾缓冲液（pH 8.0）中匀浆；匀浆液离心，上清液加热使蛋白质沉淀。经热脱蛋白的提取物酸化至pH 3.5 - 4，然后直接用LC分析。该LC方法由离子对流动相、反相ODS柱、邻苯二甲醛试剂柱后衍生化和荧光检测组成。该LC方法使用巴龙霉素作为内标，由于新霉素与链霉素或二氢链霉素的保留时间不同，可将它们分离。LC柱在25分钟内分离出新霉素；检测限约为3.5 ng新霉素。在肾脏组织中添加1 - 30 ppm新霉素时，新霉素的总回收率为96%，变异系数为9.0%。该方法也应用于肌肉组织。