Quantification of succinic acid levels, linked to succinate dehydrogenase (SDH) dysfunctions, by an automated and fully validated liquid chromatography tandem mass spectrometry method suitable for multi-matrix applications.

Abstract

The hallmarks of cancer include metabolism with deregulating cellular energetics. Dysfunctions in succinate dehydrogenase (SDH) metabolic enzyme activity, leading to an abnormal accumulation of succinic acid has been described in solid tumors but also in inflammation and ischemia reperfusion injury. Succinic acid is a potential biomarker of SDH related pathologies for diagnostic, evaluation of treatment response and follow-up of the disease. We developed a liquid chromatography tandem mass spectrometry (LC-MS/MS) method allowing a rapid, accurate and precise quantification of succinic acid levels in clinical (serum, urine) and preclinical (cellular pellets, supernatants) samples. 13C4 succinic acid disodium salt was used as internal standard and added to samples before a solid phase extraction (SPE) on Phenomenex STRATATM XL-A (200 mg - 3 mL) 33 µm cartridges. This method is automated by a Freedom EVO® platform from TECAN and succinic acid is separated on a C18 column combined to a Xevo® TQ-S micro Waters mass spectrometer with electrospray ionization (ESI) source. This biomedical analysis allows standard curves to be linear over the range 1.0-135.5 µM with r2 values > 0.999 and low matrix effects (<9.1 %). This method, which is validated according updated European Medicine Agency (EMA) guidelines, is accurate between-run (<11.0 %) and within-run (<7.8 %), precise between-run (<14.4 CV %) and within-run (<3.7 CV %), and is suitable for clinical and preclinical applications.

• Authors: Constance Lamy, Clémence Mansard, Louis Blondel, Lionel Mercier, Angelo Paci, Sophie Broutin
• Journal: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences (J Chromatogr B Analyt Technol Biomed Life Sci) Vol. 1189 Pg. 123085 (Jan 15 2022) ISSN: 1873-376X [Electronic] Netherlands
• PMID: 34974318 (Publication Type: Journal Article)
• Copyright: Copyright © 2021. Published by Elsevier B.V.

Chemical References

• Biomarkers, Tumor
• SDHD protein, human
• Succinic Acid
• Succinate Dehydrogenase

Topics

• Biomarkers, Tumor (analysis, metabolism)
• Chromatography, Liquid (methods)
• Humans
• Limit of Detection
• Linear Models
• Neoplasms (diagnosis, metabolism, physiopathology)
• Reproducibility of Results
• Solid Phase Extraction
• Succinate Dehydrogenase (metabolism)
• Succinic Acid (analysis, metabolism)
• Tandem Mass Spectrometry (methods)