Determination of adenosine, inosine and hypoxanthine in human plasma by high performance liquid chromatography

KLEPÁRNÍK, Martin and Josef TOMANDL. Determination of adenosine, inosine and hypoxanthine in human plasma by high performance liquid chromatography. In VITAMINS, NUTRITION, DIAGNOSTICS 2009, The Abstract Book. Zlín: Univerzita Tomáše Bati ve Zlíně, 2009, p. 149-356. ISBN 978-80-7318-809-2.

Basic information

• Original name: Determination of adenosine, inosine and hypoxanthine in human plasma by high performance liquid chromatography
• Name in Czech: Stanovení adenosinu, inosinu a hypoxanthinu v plazmě pomocí HPLC
• Authors: KLEPÁRNÍK, Martin and Josef TOMANDL.
• Edition: Zlín, VITAMINS, NUTRITION, DIAGNOSTICS 2009, The Abstract Book, p. 149-356, 2009.
• Publisher: Univerzita Tomáše Bati ve Zlíně

Other information

• Original language: English
• Type of outcome: Proceedings paper
• Field of Study: 10406 Analytical chemistry
• Country of publisher: Czech Republic
• Confidentiality degree: is not subject to a state or trade secret
• Organization unit: Faculty of Medicine
• ISBN: 978-80-7318-809-2
• Keywords (in Czech): inosin; hypoxanthin; adenosin; HPLC
• Keywords in English: inosine; hypoxanthine; adenosine; HPLC
• Tags: International impact

Abstract

Inosine and hypoxanthine are endogenous low molecular plasma constituents normally found at low concetrations (200-400 ng/mL) in human plasma resulting from dietary and endogenous purine metabolism. Using the mouse model, inosine levels increased from cardiac tissue subjected to cardiac oxidative stress and may serve as potential biomarker indicative of early cardiac ischaemia. We report here development of a new HPLC method for the determination of adenosine, inosine and hypoxanthine using ultra-violet (UV) detection. Before analysis, plasma samples were deproteinized by centrifugal filtration through 10-kDa cut-off membranes. The protein-free ultrafiltrates were analysed on a monolithic reversed phase column Chromolith Performance RP-18e using gradient elution and UV detection at 250nm. Mobile phase consisted of 25 mM acetate buffer and methanol, the flow rate was 1mL/min. Optimal wavelength were confirmed by absorption spectra of inosine and hypoxanthine. Plasma levels of analytes were quantified on the basis of peak area using external standardization.

Abstract (in Czech)

Inosine and hypoxanthine are endogenous low molecular plasma constituents normally found at low concetrations (200-400 ng/mL) in human plasma resulting from dietary and endogenous purine metabolism. Using the mouse model, inosine levels increased from cardiac tissue subjected to cardiac oxidative stress and may serve as potential biomarker indicative of early cardiac ischaemia. We report here development of a new HPLC method for the determination of adenosine, inosine and hypoxanthine using ultra-violet (UV) detection. Before analysis, plasma samples were deproteinized by centrifugal filtration through 10-kDa cut-off membranes. The protein-free ultrafiltrates were analysed on a monolithic reversed phase column Chromolith Performance RP-18e using gradient elution and UV detection at 250nm. Mobile phase consisted of 25 mM acetate buffer and methanol, the flow rate was 1mL/min. Optimal wavelength were confirmed by absorption spectra of inosine and hypoxanthine. Plasma levels of analytes were quantified on the basis of peak area using external standardization.

Links

• LC06023, research and development project: Name: Integrované bioanalytické technologie pro mikroanalýzy a diagnostiku s využitím LIF a hmotnostní spektrometrie

Investor: Ministry of Education, Youth and Sports of the CR

• MSM0021622402, plan (intention): Name: Časná diagnostika a léčba kardiovaskulárních chorob

Investor: Ministry of Education, Youth and Sports of the CR, Early diagnostics and treatment of cardiovascular diseases