An HPLC method for determination of inosine and hypoxanthine in human plasma from healthy volunteers and patients presenting with potential acute cardiac ischemia

Farthing, Don; Sica, Domenic A.; Gehr, Todd W.B.; Wilson, Bill; Fakhry, Itaf; Larus, Terri; Farthing, Christine; Karnes, H. Thomas

doi:10.1016/j.jchromb.2007.04.013

Cited by 59 publications

(42 citation statements)

References 27 publications

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“…Our research group utilized blood samples acquired from non-traumatic chest pain patients admitted by a local hospital ER. 28 These patients had chest pain with potential acute cardiac ischemia for several hours and we found elevated plasma concentrations of inosine and/or hypoxanthine in all of the samples.…”

Section: Plasma Componentsmentioning

confidence: 59%

“…28 In this study, residual plasma samples (heparinized) from ER non-traumatic chest pain patients were obtained and evaluated using a validated quantitative HPLC-UV method. Briefly, the plasma samples were prepared using 10 K MWCO filters, with 15 mL of the filtrate injected for HPLC-UV analysis.…”

Section: Introductionmentioning

confidence: 99%

“…Figure 2 depicts an overlay of HPLC-UV chromatograms representing a calibration standard, a healthy individual (Control), two positive cardiac troponin T (cTnT) samples (cTnT 6.64 and cTnT 0.55 mg/L), and two chest pain patient samples (CP-294 and CP-307). 28 The representative chromatogram of a control subject had a typical quantifiable concentration of hypoxanthine, with no detectable amount of inosine. The hypoxanthine concentrations for normal subjects were consistent with those previously reported in literature.…”

Section: Introductionmentioning

confidence: 99%

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Inosine and hypoxanthine as novel biomarkers for cardiac ischemia: From bench to point-of-care

Farthing

2015

Exp Biol Med (Maywood)

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Cardiac ischemia associated with acute coronary syndrome and myocardial infarction is a leading cause of mortality and morbidity in the world. A rapid detection of the ischemic events is critically important for achieving timely diagnosis, treatment and improving the patient's survival and functional recovery. This minireview provides an overview on the current biomarker research for detection of acute cardiac ischemia. We primarily focus on inosine and hypoxanthine, two by-products of ATP catabolism. Based on our published findings of elevated plasma concentrations of inosine/hypoxanthine in animal laboratory and clinical settings, since 2006 we have originally proposed that these two purine molecules can be used as rapid and sensitive biomarkers for acute cardiac ischemia at its very early onset (within 15 min), hours prior to the release of heart tissue necrosis biomarkers such as cardiac troponins. We further developed a chemiluminescence technology, one of the most affordable and sensitive analytical techniques, and we were able to reproducibly quantify and differentiate total hypoxanthine concentrations in the plasma samples from healthy individuals versus patients suffering from ischemic heart disease. Additional rigorous clinical studies are needed to validate the plasma inosine/hypoxanthine concentrations, in conjunction with other current cardiac biomarkers, for a better revelation of their diagnostic potentials for early detection of acute cardiac ischemia.

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Section: Plasma Componentsmentioning

confidence: 59%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Inosine and hypoxanthine as novel biomarkers for cardiac ischemia: From bench to point-of-care

Farthing

2015

Exp Biol Med (Maywood)

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“…The HPLC results from healthy individuals (ProMedDx) and non-traumatic chest pain patients (Chippenham Hospital) from our previous research (14,16) were used to estimate plasma levels of inosine and hypoxanthine for development of the chemiluminescence method. Since the luminometer is a detection device, and does not separate a mixture of components (as does HPLC), it was necessary to utilize enzyme PNP to convert inosine to hypoxanthine, and then measure the resulting total plasma hypoxanthine level (i.e.…”

Section: Methods Development and Optimizationmentioning

confidence: 99%

“…Current test methods for measurement of selected ATP catabolic by-products inosine, hypoxanthine, xanthine and uric acid utilize high-performance liquid chromatography ultraviolet (HPLC-UV) detection with sample preparation techniques including solid-phase extraction (15), centrifugal fi ltration (16), protein precipitations (e.g. ethanol or TCA), as well as other methods requiring use of internal standards (17,18).…”

Section: Introductionmentioning

confidence: 99%

A rapid and simple chemiluminescence method for screening levels of inosine and hypoxanthine in non‐traumatic chest pain patients

et al. 2011

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A rapid and simple chemiluminescence method was developed for detection of inosine and hypoxanthine in human plasma. The method utilized a microplate luminometer with direct injectors to automatically dispense reagents during sample analysis. Enzymatic conversions of inosine to hypoxanthine, followed by hypoxanthine to xanthine to uric acid, generated superoxide anion radicals as a useful metabolic by-product. The free radicals react with Pholasin(®) , a sensitive photoprotein used for chemiluminescence detection, to produce measurable blue-green light. The use of Pholasin(®) and a chemiluminescence signal enhancer, Adjuvant-K™, eliminated the need for plasma clean-up steps prior to analysis. The method used 20 μL of heparinized plasma, with complete analysis of total hypoxanthine levels (inosine is metabolized to hypoxanthine using purine nucleoside phosphorylase) in approximately 3.7 min. The rapid chemiluminescence method demonstrated the capability of differentiating total hypoxanthine levels between healthy individuals, and patients presenting with non-traumatic chest pain and potential acute cardiac ischemia. The results support the potential use of chemiluminescence methodology as a diagnostic tool to rapidly screen for elevated levels of inosine and hypoxanthine in human plasma, potential biomarkers of acute cardiac ischemia.

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SERS Analysis of Bacteria, Human Blood, and Cancer Cells: a Metabolomic and Diagnostic Tool

Premasiri

Lemler

Chen

et al. 2014

Frontiers of Surface‐Enhanced Raman Scattering

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An HPLC method for determination of inosine and hypoxanthine in human plasma from healthy volunteers and patients presenting with potential acute cardiac ischemia

Cited by 59 publications

References 27 publications

Inosine and hypoxanthine as novel biomarkers for cardiac ischemia: From bench to point-of-care

Inosine and hypoxanthine as novel biomarkers for cardiac ischemia: From bench to point-of-care

A rapid and simple chemiluminescence method for screening levels of inosine and hypoxanthine in non‐traumatic chest pain patients

SERS Analysis of Bacteria, Human Blood, and Cancer Cells: a Metabolomic and Diagnostic Tool

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