Development of a LC-MS/MS Method for the Simultaneous Detection of Tricarboxylic Acid Cycle Intermediates in a Range of Biological Matrices.
Development of a LC-MS/MS Method for the Simultaneous Detection of Tricarboxylic Acid Cycle Intermediates in a Range of Biological Matrices.
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DOI：
10.1155/2017/5391832
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发表时间：
2017
影响因子：
2.6
通讯作者：
Saha S
中科院分区：
文献类型：
--
作者：
Al Kadhi O;Melchini A;Mithen R;Saha S
现在已经确定，三羧酸（TCA）循环的扰动在癌症（包括前列腺癌）中发生的代谢转变中发挥着重要作用。通过 LC-MS/MS 技术，使用常见的 C18 反相色谱柱，开发了一种对人体体液、组织和培养细胞系中的 TCA 循环中间体进行同步定性和定量分析的方法。这种用于分析 TCA 循环中间体的 LC-MS/MS 方法具有显着的优势，包括简单快速地制备各种人类生物样品。分析方法根据英国皇家化学学会分析方法委员会的指导方针进行验证。除乳酸和富马酸外，大多数 TCA 中间体的检测限均低于 60nM。所有 TCA 分析物的校准曲线均呈线性，相关系数 r2 > 0.9998。所有 TCA 分析物的回收率 >95%。该方法是考虑到现有技术的问题和局限性而建立的。我们设想，将其应用于不同的生物基质将有助于通过体外、离体和体内研究更深入地了解 TCA 循环中的代谢变化。
It is now well-established that perturbations in the tricarboxylic acid (TCA) cycle play an important role in the metabolic transformation occurring in cancer including that of the prostate. A method for simultaneous qualitative and quantitative analysis of TCA cycle intermediates in body fluids, tissues, and cultured cell lines of human origin was developed using a common C18 reversed-phase column by LC-MS/MS technique. This LC-MS/MS method for profiling TCA cycle intermediates offers significant advantages including simple and fast preparation of a wide range of human biological samples. The analytical method was validated according to the guideline of the Royal Society of Chemistry Analytical Methods Committee. The limits of detection were below 60 nM for most of the TCA intermediates with the exception of lactic and fumaric acids. The calibration curves of all TCA analytes showed linearity with correlation coefficients r2 > 0.9998. Recoveries were >95% for all TCA analytes. This method was established taking into consideration problems and limitations of existing techniques. We envisage that its application to different biological matrices will facilitate deeper understanding of the metabolic changes in the TCA cycle from in vitro, ex vivo, and in vivo studies.
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影响因子：
2.2
作者：
Keevil, BG;Owen, L;Kavanagh, J
通讯作者：
Kavanagh, J
影响因子：
6.1
作者：
Mariano, Filippo;Morselli, Maurizio;Triolo, Giorgio
通讯作者：
Triolo, Giorgio
影响因子：
4.3
作者：
Hempen, Christel;Wanschers, Harry;Veer, Gertjan van der Sluijs
通讯作者：
Veer, Gertjan van der Sluijs
影响因子：
6.6
作者：
Kline, Eric E.;Treat, Eric G.;Sillerud, Laurel O.
通讯作者：
Sillerud, Laurel O.
影响因子：
4.1
作者：
Luo, Bing;Groenke, Karsten;Oldiges, Marco
通讯作者：
Oldiges, Marco