单柱液相色谱-质谱联用法直接注入溶解细胞培养基测定PGE2

P. Araujo, Z. Du, Thu-Thao Nguyen, E. Holen
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引用次数: 5

摘要

通过液相或气相色谱联用质谱法来评估细胞培养物中的类二十烷类化合物的传统方法涉及耗时的程序,如衍生化、降解、固相萃取、液-液萃取、薄层色谱,有时在将样品注入色谱系统之前,需要将所有这些程序结合起来。评价了一种方法的性能,该方法允许将再溶解的细胞培养基直接注射到单柱液相色谱仪中,并与质谱相结合。Williams培养基E和L-15培养基的外标曲线在1 ~ 50 ng/ml之间呈线性关系。两种培养基的检测限和定量限分别为0.5和1 ng/ml。L-15培养基的回收率分别为98.6%和100.3%。由于响应因子的可变性,内标法未用于定量目的。所提出的方法在监测细胞培养物中的生物标记物方面具有广泛的应用潜力。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Direct Injection of Redissolved Cell Culture Media into a Single-Column Liquid Chromatography Coupled to Mass Spectrometry for the Measurement of PGE2
The traditional approach to assessing eicosanoids in cell cultures by liquid or gas chromatography coupled to mass spectrometry involves time-consuming procedures such as derivatisation, degradation, solid-phase extraction, liquid- liquid extraction, thin layer chromatography and sometimes, combination of all these procedures prior to the injection of the sample in the chromatography system. The performance of a method that allows the direct injection of redissolved cell culture media into a single-column liquid chromatography instrument coupled to mass spectrometry is evaluated. The ex- ternal standard calibration curves were linear between 1-50 ng/ml for Williams' medium E and L-15 medium. The limit of detection and quantification were 0.5 and 1 ng/ml in both media respectively. The recovery values were 98.6% for Wil- liams' medium E and 100.3% for L-15 medium. The internal standard method was not used for quantitative purposes due to the variability of the response factor. The proposed method has potential for broad implementation in monitoring bio- markers in cell cultures.
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