A liquid chromatography and tandem mass spectrometry method for the determination of potential biomarkers of cardiovascular disease
2013 (English)In: Journal of chromatography. B, ISSN 1570-0232, E-ISSN 1873-376X, Vol. 919, 20-29 p.Article in journal (Refereed) Published
A simple, accurate and sensitive liquid chromatography tandem mass spectrometry (LC-MS/MS) method was developed and validated for the quantitation of alpha-ketoglutaric acid (alpha-KG), L-carnitine (L-CAR) and acetyl-L-carnitine (acetyl-L-CAR) in human urine as potential biomarkers of cardiovascular disease. The separation was performed using an isocratic elution of 0.1% formic acid in water and acetonitrile (97:3, v/v) on an Acclaim 120 C8 column (150 mm x 4.6 mm, 3.0 mu m). The flow rate of the mobile phase was 1.2 mL/min and the total assay run time was 3 min. Detection was performed on a triple-quadrupole mass spectrometer in selected reaction monitoring (SRM) mode via an electrospray ionization (ESI) source in positive and negative ion modes. This method covered a linearity range of 0.1-500 ng/mL for L-CAR and acetyl-L-CAR and 1-1000 ng/mL for alpha-KG with lower limits of quantification (LLOQ) of 0.08 ng/mL for L-CAR, 0.04 ng/mL for acetyl-L-CAR and 0.8 ng/mL for alpha-KG. The intra-day and inter-day precision and accuracy of the quality control samples exhibited relative standard deviations of less than 5.54% and relative error values from -5.95% to 3.11%. Analyte stability was evaluated under various sample preparation, analysis and storage conditions and varied from -9.89% to -0.47%. A two-step solid-phase extraction (SPE) procedure using silica gel and quaternary amine cartridges was used for urine sample cleanup. The average recoveries for all analyzed compounds were better than 86.64% at three concentrations. The method was successfully applied for the quantitation of alpha-KG, L-CAR and acetyl-L-CAR in human urine samples.
Place, publisher, year, edition, pages
Elsevier, 2013. Vol. 919, 20-29 p.
LC-MS/MS, Biomarkers, alpha-Ketoglutaric acid, L-Carnitine, Acetyl-L-carnitine
Engineering and Technology
IdentifiersURN: urn:nbn:se:liu:diva-90758DOI: 10.1016/j.jchromb.2012.12.015ISI: 000316043300003OAI: oai:DiVA.org:liu-90758DiVA: diva2:617212
Funding Agencies|National Science Center (Cracow, Poland)|N N204 355840|2013-04-222013-04-052013-04-29