一种使用内标物分析两性霉素B的新型高效液相色谱法。

New high-performance liquid chromatographic method for amphotericin B analysis using an internal standard.

作者信息

Lambros M P, Abbas S A, Bourne D W

机构信息

University of Oklahoma Health Science Center, College of Pharmacy, Oklahoma City 73190, USA.

出版信息

J Chromatogr B Biomed Appl. 1996 Oct 11;685(1):135-40. doi: 10.1016/0378-4347(96)00162-4.

DOI:10.1016/0378-4347(96)00162-4

PMID:9026302

Abstract

A simple and reproducible HPLC method for the analysis of amphotericin B (AmB) in serum, lung and liver using natamycin as the internal standard was developed. AmB and natamycin were extracted from serum, lung and liver and were separated using an isocratic elution from C18 reversed-phase column. The mobile phase consisted of acetonitrile-10 mM acetate buffer pH 4.0 (37:63, v/v). The HPLC system had two detectors in series. One was set at 303 nm and the other at 383 nm for the detection of natamycin and AmB, respectively. The retention times of AmB and natamycin were 15 and 6 min, respectively. The recovery efficiency was 96%-70%. The limit of quantification was 0.1 microgram/ml. The assay was reproducible, the within-day coefficient of variation (n = 6) was < 8% for serum, lungs and liver. The between-day variability (n = 6) was < 7.7% for serum, liver and lungs at 1 microgram/ml or 1 microgram/g tissue concentration. The assay was linear within the range 1-40 micrograms/ml (r2 = 0.99).

摘要

建立了一种简单且可重复的高效液相色谱法，以游霉素作为内标物，用于分析血清、肺和肝脏中的两性霉素B（AmB）。从血清、肺和肝脏中提取AmB和游霉素，并使用C18反相柱等度洗脱进行分离。流动相由乙腈-10 mM醋酸盐缓冲液pH 4.0（37:63，v/v）组成。高效液相色谱系统串联有两个检测器。一个设置在303 nm用于检测游霉素，另一个设置在383 nm用于检测AmB。AmB和游霉素的保留时间分别为15分钟和6分钟。回收率为96%-70%。定量限为0.1微克/毫升。该测定方法具有可重复性，血清、肺和肝脏的日内变异系数（n = 6）< 8%。在1微克/毫升或1微克/克组织浓度下，血清、肝脏和肺的日间变异性（n = 6）< 7.7%。该测定方法在1-40微克/毫升范围内呈线性（r2 = 0.99）。