Biochemical assays of TCA cycle and β-oxidation metabolites

Abstract

This chapter focuses on the methods to measure unique metabolites, specific enzymes, and metabolic flux in fatty acid β-oxidation, and on biochemical assays of tricarboxylic acid (TCA) cycle enzymes and the pyruvate dehydrogenase complex. These assays play an important role in the diagnosis of genetic diseases, newborn screening, and in cancer and metabolism research. The rationale, protocol, pros and cons, and alternative methods are discussed. Nevertheless, each laboratory should adapt the preferred method optimizing sample preparation and assay conditions for linearity and a low signal-to-noise ratio. The reader is also referred to the additional literature citing methods and clinical descriptions of the various diseases.

Keywords: Acylcarnitine; Acylglycines; Carnitine; Fatty acid oxidation; Krebs cycle; Mitochondria; Organic acids; Pyruvate dehydrogenase; TCA cycle.