Ultra-high performance hydrophilic interaction liquid chromatography - Triple quadrupole tandem mass spectrometry method for determination of cysteine, homocysteine, cysteinyl-glycine and glutathione in rat plasma
Ultra-high performance hydrophilic interaction liquid chromatography - Triple quadrupole tandem mass spectrometry method for determination of cysteine, homocysteine, cysteinyl-glycine and glutathione in rat plasma
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DOI：
10.1016/j.jpba.2018.10.053
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发表时间：
2019-02-05
影响因子：
3.4
通讯作者：
Kovac, Andrej
中科院分区：
文献类型：
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作者：
Forgacsova, Andrea;Galba, Jaroslav;Kovac, Andrej
建立了超高效亲水作用液相色谱-三重四极杆串联质谱法测定大鼠血浆中具有重要生物活性的巯基化合物半胱氨酸、同型半胱氨酸、半胱氨酰甘氨酸和谷胱甘肽的方法。对样品制备程序和分析方法进行了全面优化，并随后进行了验证。一个最佳的样品制备方案是基于简单和快速的衍生化的硫醇与新的衍生试剂，N-苯基马来酰亚胺，使血浆基质中的高选择性和灵敏度的定量。该方法的特征在于有利的性能参数，并满足FDA的生物医学分析标准，成功地应用于监测从tau蛋白病的转基因大鼠模型的样品中的选定的硫醇的浓度水平。该研究揭示了与tau蛋白病相关的同型半胱氨酸和谷胱甘肽水平的显著变化，而其他硫醇没有表明这种关系。事实上，这些发现可以在进一步了解大脑中的tau蛋白病变过程中发挥重要作用。此外，所提出的高效，可靠和稳健的分析方案可以很容易地适用于其他硫醇化合物，扩大其在生物医学领域的应用范围。(C)2018 Elsevier B. V.版权所有。
An ultra-high performance hydrophilic interaction liquid chromatography - triple quadrupole tandem mass spectrometry method was developed for the determination of biologically important thiols, namely cysteine, homocysteine, cysteinyl-glycine, glutathione, in rat plasma. The sample preparation procedure as well as the analytical method were comprehensively optimized and subsequently validated. An optimum sample preparation protocol was based on the simple and fast derivatization of the thiols with new derivatization reagent, N-phenylmaleimide, enabling highly selective and sensitive quantification in plasma matrices. The method, characterized by favourable performance parameters and meeting the FDA criteria for biomedical analysis, was successfully applied for monitoring the concentration levels of the selected thiols in the samples from transgenic rat model for tauopathy. The study revealed significant changes in homocysteine and glutathione levels related to tauopathy while other thiols did not indicate such relationship. Indeed, these findings could play an important role in further understanding of tauopathy process in the brain. Moreover, the proposed highly effective, reliable and robust analytical protocol can be easily adapted for other thiol compounds, spreading its application range in this biomedical field. (C) 2018 Elsevier B.V. All rights reserved.