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Journal of Analytical Methods in Chemistry
Volume 2017, Article ID 5391832, 12 pages
https://doi.org/10.1155/2017/5391832
Research Article

Development of a LC-MS/MS Method for the Simultaneous Detection of Tricarboxylic Acid Cycle Intermediates in a Range of Biological Matrices

1Food and Health Programme, Quadram Institute Bioscience, Norwich NR4 7UA, UK
2Department of Urology, Norfolk and Norwich University Hospitals NHS Foundation Trust, Norwich NR4 7UY, UK

Correspondence should be addressed to Shikha Saha; ku.ca.mardauq@ahas.ahkihs

Received 27 February 2017; Accepted 5 June 2017; Published 18 September 2017

Academic Editor: Núria Fontanals

Copyright © 2017 Omar Al Kadhi et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

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 . 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.