Profiling phospholipid elution in reversed-phase LC-MS/MS bioanalytical methods in order to avoid matrix effects.
Profiling phospholipid elution in reversed-phase LC-MS/MS bioanalytical methods in order to avoid matrix effects.
复制标题
在反相 LC-MS/MS 生物分析方法中分析磷脂洗脱以避免基质效应。
DOI：
10.4155/bio.12.33
复制
发表时间：
2012
期刊：
影响因子：
1.8
通讯作者：
Luke Smith
中科院分区：
文献类型：
--
作者：
Steve Silvester;Luke Smith
背景
内源性磷脂在生物分析LC-MS方法中具有深刻的基质效应，并且在通过在样品处理期间去除或在分析运行期间进行色谱分离来最小化其影响的策略上投入了相当大的努力。本文提出的研究的目的是调查后一种方法，在反相条件下。
结果
甘油磷酸胆碱（GPC）在移动的乙腈流动相中的保留与有机物的百分比呈复杂的“U形”关系。相反，在采用甲醇的移动的相中，保留和有机物百分比之间的关系完全是预测性的，不受移动的相pH值变化的影响。GPC洗脱曲线也是定性等效的，与血浆来源的物质无关。
结论
GPC保留的预测性质，在反相色谱法和甲醇作为有机改性剂，是一个重要的发现，应该允许一个更精简和简化的策略，在生物分析测定的发展。
BACKGROUND
Endogenous phospholipids have a profound matrix effect in bioanalytical LC-MS methods and considerable effort is invested in strategies to minimize their impact either by removal during sample processing or chromatographic separation during the analytical run. The aim of the research presented in this article was to investigate the latter approach, under reversed-phase conditions.
RESULTS
The retention of glycerophosphocholines (GPCs) in mobile phases employing acetonitrile demonstrated a complex 'U-shaped' relationship with the percentage of organic. Conversely, in mobile phases employing methanol, the relationship between retention and percentage of organic was entirely predictive and unaffected by changes in the mobile phase pH. The GPC elution profile was also qualitatively equivalent irrespective of the species from which the plasma was derived.
CONCLUSION
The predictive nature of GPC retention, under reversed-phase chromatography and with MeOH as organic modifier, is an important finding that should allow for a more streamlined and simplified strategy in the development of bioanalytical assays.