采用反相高效液相色谱法测定血浆新生霉素水平。

Determination of plasma novobiocin levels by a reversed-phase high-performance liquid chromatographic assay.

作者信息

Chen T L, Kennedy M J, Dunlap V M, Colvin O M

机构信息

Division of Pharmacology and Experimental Therapeutics, Johns Hopkins Oncology Center, Baltimore, MD 21287.

出版信息

J Chromatogr. 1994 Jan 14;652(1):109-13. doi: 10.1016/0378-4347(94)80113-4.

DOI:10.1016/0378-4347(94)80113-4

PMID:8014221

Abstract

A simple and specific reversed-phase high-performance liquid chromatographic (HPLC) assay for the determination of novobiocin levels in human plasma has been developed. The sample preparation was performed by deproteinization with methanol. Prednisone was used as an internal standard. Both novobiocin and prednisone were separated on a C8 column with a gradient elution of acidic water (pH 3.0)-methanol. The recovery of novobiocin from plasma was nearly complete. The linear range was 5-1000 microM in 0.5 ml of plasma with a minimum limit of determination of 2.25 fmol of novobiocin at 254 nm. The method has been implemented and validated in an ongoing clinical trial.

摘要

已开发出一种简单且特异的反相高效液相色谱（HPLC）分析法，用于测定人血浆中的新生霉素水平。样品制备通过用甲醇进行脱蛋白处理来完成。泼尼松用作内标。新生霉素和泼尼松均在C8柱上通过酸性水（pH 3.0）-甲醇梯度洗脱进行分离。新生霉素从血浆中的回收率几乎是完全的。在0.5 ml血浆中的线性范围为5 - 1000 μM，在254 nm处新生霉素的最低检测限为2.25 fmol。该方法已在一项正在进行的临床试验中实施并验证。