Development and validation of a novel quantification approach for gradient elution reversed phase high-performance liquid chromatography coupled to tandem ICP-mass spectrometry (RP-HPLC-ICP-MS/MS) and its application to diclofenac and its related compounds
Development and validation of a novel quantification approach for gradient elution reversed phase high-performance liquid chromatography coupled to tandem ICP-mass spectrometry (RP-HPLC-ICP-MS/MS) and its application to diclofenac and its related compounds
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DOI：
10.1016/j.aca.2017.04.030
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发表时间：
2017-06-29
影响因子：
6.2
通讯作者：
Vanhaecke, Frank
中科院分区：
文献类型：
--
作者：
Klencsar, Balazs;Balcaen, Lieve;Vanhaecke, Frank
一种新的定量方法，在反相高效液相色谱-电感耦合等离子体串联质谱分析中，补偿梯度洗脱对仪器响应的影响，已被开发和验证。作为概念验证，已在人血浆基质中定量测定了双氯芬酸及其相关化合物，包括其主要代谢产物，即4 '-羟基-双氯芬酸。本文采用电感耦合等离子体-串联质谱仪，在碰撞/反应池中，以（ClH ~（2+））-Cl ~（35）为反应气体，无干扰地测定了Cl ~（2+）。在反相高效液相色谱法中，最常见的有机溶剂，即甲醇和乙腈的洗脱液组合物对仪器响应氯的效果进行了彻底的研究。一个适当的数学函数描述的影响，洗脱液组成的灵敏度为Cl，监测（ClH 2+）-Cl-35，允许充分校正的梯度洗脱的两种溶剂的其他不利影响。使用加标其主要代谢产物4 '-羟基-双氯芬酸的合成降解双氯芬酸样品进行验证，证明了适当的准确度（4'-羟基-双氯芬酸的回收率在95 - 105%之间），使用乙腈和甲醇时Cl的回收率分别>90%和>80%。当应用于加标的人血浆样品（药物代谢研究中最重要的基质）时，令人满意的准确度（回收率为92-98%）和精密度（
A novel quantification approach, compensating for the effect of gradient elution on the instrumental response during reversed phase high-performance liquid chromatography - inductively coupled plasma tandem mass spectrometry analysis, has been developed and validated. As a proof of concept, diclofenac and its related compounds, including its major metabolite i.e. 4'-hydroxy-diclofenac, have been quantitatively determined in human plasma matrix. An inductively coupled plasma - tandem mass spectrometer has been applied for the interference-free determination of Cl as (ClH2+)-Cl-35 using H-2 as a reaction gas in the collision/reaction cell. The effect of the eluent composition on the instrumental response for Cl has been thoroughly investigated for the most common organic solvents in reversed phase high-performance liquid chromatography, i.e. methanol and acetonitrile. A proper mathematical function describing the effect of the eluent composition on the sensitivity for Cl, monitored as (ClH2+)-Cl-35, permitted adequate correction for the otherwise detrimental effect of gradient elution for both solvents. Validation using synthetically degraded diclofenac samples spiked with its major metabolite, 4'-hydroxy-diclofenac, demonstrated appropriate accuracy (recovery for 4'-hydroxy-diclofenac between 95 and 105%) and >90% and >80% recovery for Cl using acetonitrile and methanol, respectively. When applied to spiked human plasma samples (the most important matrix in drug metabolism studies), a satisfactory accuracy (recovery of 92-98%) and precision (