Department of Pharmacology, School of Medical Sciences, Universiti Sains Malaysia, 16150, Kubang Kerian, Kelantan, Malaysia.
Toxicol Mech Methods. 2010 Mar;20(3):137-42. doi: 10.3109/15376511003602112.
Many previous published methods for the quantitative determination of propranolol (PRN) in human plasma have poor recoveries and were not validated according to the FDA guideline. The aim of this study is to develop a simple HPLC method for detecting PRN in human plasma and to validate it so that it can be applied to a clinical study. Chromatographic separation was achieved using a mixture of a mobile phase consisting of 160 ml water, 180 ml methanol, 70 ml acetonitrile, 2.5 ml acetic acid, and 125 microl triethylamine (v/v). The pH of the whole mixture was adjusted to 3.4. A flow rate of 0.5 ml/min was employed throughout with a 15 microl injection volume. Detection was done using a UV detector at 291 nm. The validated method was linear for concentrations ranging from 15-180 ng/ ml with a good separation and specificity for both PRN and its internal standard, oxprenolol (OXP), with excellent recoveries, precision, and accuracies. The limit of detection (LOD) and limit of quantification (LOQ) were 1 and 10 ng/ml, respectively. The stability studies demonstrated that PRN is stable in the autosampler vials and also up to 3.5 months. To the authors' knowledge, the recovery, that ranged between 97.9-102.7%, is the highest among all previously reported methods that used HPLC with UV detection. The developed and validated method for PRN analysis is excellent and applicable to a clinical study.
许多先前发表的用于定量测定人血浆中普萘洛尔（PRN）的方法回收率较差，并且不符合 FDA 指南的要求。本研究旨在开发一种简单的 HPLC 方法来检测人血浆中的 PRN，并对其进行验证，以便能够应用于临床研究。色谱分离采用由 160ml 水、180ml 甲醇、70ml 乙腈、2.5ml 乙酸和 125 微升三乙胺（v/v）组成的流动相混合物实现。整个混合物的 pH 值调整为 3.4。整个过程采用 0.5ml/min 的流速，进样量为 15 微升。检测采用 291nm 的紫外检测器进行。验证的方法对于 15-180ng/ml 的浓度范围呈线性，对于 PRN 和其内标物奥普洛尔（OXP）具有良好的分离度和特异性，回收率、精密度和准确度均很好。检测限（LOD）和定量限（LOQ）分别为 1 和 10ng/ml。稳定性研究表明，PRN 在自动进样器小瓶中稳定，并且在 3.5 个月内稳定。据作者所知，回收率在 97.9-102.7%之间，在所有使用 HPLC 与紫外检测的先前报道的方法中是最高的。开发和验证的 PRN 分析方法非常出色，适用于临床研究。
