Method based on GC-MS to study the influence of tricarboxylic acid cycle metabolites on cardiovascular risk factors

Metabolites involved in the tricarboxylic acid (TCA) cycle have previously been proposed as cardiovascular biomarkers. This cycle plays a key role in cell metabolism and the levels of the involved metabolites can also be affected by other physiological factors. The influence of three cardiovascular risk factors such as obesity, hypercholesterolemia, and smoking habit on serum levels of TCA-cycle metabolites has been studied in patients diagnosed with significant coronary lesion. For this purpose, a method based on GC-MS for determination of the target metabolites (viz. citric/isocitric, pyruvic, aconitic, oxaloacetic, malic, fumaric and succinic acids) in serum has been developed. The high accuracy and throughput analysis featuring the method have allowed application to a cohort of 223 patients, 172 of them with significant coronary lesion. Multifactor analysis of variance has revealed interactions between the occurrence or not of a coronary lesion and the risk factors considered in this study. These interactions were crucial to explain the levels of target TCA metabolites. Statistical evaluation by ROC curves allowed discrimination of the capability of significant metabolites with the occurrence of coronary lesions.