High-performance liquid chromatographic determination of hypoxanthine and xanthine in biological fluids

Boulieu, Roselyne; Bory, C.; Baltassat, P; Gonnet, C.

doi:10.1016/s0378-4347(00)81739-9

Cited by 48 publications

(10 citation statements)

References 17 publications

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“…56–58 Subsequently, techniques based on high-performance liquid chromatography following various procedures for deproteinization, have been employed for determinations of hypoxanthine levels in whole blood or blood plasma. 59–65 The ability to accurately measure levels of hypoxanthine in blood or plasma is important for purine metabolism disorders such as xanthine oxidase deficiency, 59, 66 hyperuricemia (gout) 67 and fetal hypoxia. 68 Elevated hypoxanthine has also been used in the early detection of acute cardiac ischemia 62, 65 and may be a marker of pre-labor fetal development problems.…”

Section: Resultsmentioning

confidence: 99%

Surface-Enhanced Raman Scattering of Whole Human Blood, Blood Plasma, and Red Blood Cells: Cellular Processes and Bioanalytical Sensing

2012

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SERS spectra of whole human blood, blood plasma, and red blood cells on Au nanoparticle SiO 2 substrates excited at 785 nm have been observed. For the sample preparation procedure employed here, the SERS spectrum of whole blood arises from the blood plasma component only. This is in contrast to the normal Raman spectrum of whole blood excited at 785 nm and open to ambient air, which is exclusively due to the scattering of oxyhemoglobin. The SERS spectrum of whole blood shows a storage time dependence that is not evident in the non-SERS Raman spectrum of whole blood. Hypoxanthine, a product of purine degradation, dominates the SERS spectrum of blood after ∼10−20 h of storage at 8 °C. The corresponding SERS spectrum of plasma isolated from the stored blood shows the same temporal release of hypoxanthine. Thus, blood cellular components (red blood cells, white blood cells, and/or platelets) are releasing hypoxanthine into the plasma over this time interval. The SERS spectrum of red blood cells (RBCs) excited at 785 nm is reported for the first time and exhibits well-known heme group marker bands as well as other bands that may be attributed to cell membrane components or protein denaturation contributions. SERS, as well as normal Raman spectra, of oxy-and met-RBCs are reported and compared. These SERS results can have significant impact in the area of clinical diagnostics, blood supply management, and forensics.

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Section: Resultsmentioning

confidence: 99%

Surface-Enhanced Raman Scattering of Whole Human Blood, Blood Plasma, and Red Blood Cells: Cellular Processes and Bioanalytical Sensing

2012

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“…Fluorimetry (30, 3 1) and high-performance liquid chromatography (HPLC) (28,(32)(33)(34)(35) provide methods with greater sensitivity. Although several other methods have been described, pOz, fluorimetric, or HPLC methods are the most generally used.…”

Section: Methodsmentioning

confidence: 99%

Hypoxanthine as an Indicator of Hypoxia: Its Role in Health and Disease through Free Radical Production

Saugstad¹

1988

Pediatr Res

374

206

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Hypoxia is a common insult during the perinatal and neonatal period. New and better ways to evaluate hypoxia are needed. In 1975 we demonstrated high concentrations of the purine metabolite hypoxanthine in umbilical cord plasma after intrauterine hypoxia and proposed that hypoxanthine could be used as an indicator of hypoxia (1). Since then a large number of studies have been published dealing with different aspects of hypoxanthine in hypoxia. However, investigators in this field have encountered several methodologic problems: 1) Hypoxanthine leaks rapidly from erythrocytes (2, 3); if plasma is not separated promptly from red cells, falsely elevated plasma hypoxanthine concentrations will be found. 2) There are large variations in purine metabolism among species. 3) No clear definition of clinical hypoxia is available. Many authors do not distinguish between the terms hypoxemia and hypoxia, which adds to the confusion. We define tissue hypoxia as: "oxygen deficiency resulting in altered or interrupted energy metabolism" (4). It is important to be aware that hypoxia has two stages. In the first stage it is compensated because the cells are able to meet energy demands through anaerobic metabolism and other mechanisms. (We are not dealing with physiologic adaptation to hypoxia.) In the second stage of hypoxia or uncompensated hypoxia, energy demands are not met and cell injury ensues. At present there are no techniques for distinguishing between these two stages in clinical medicine, and such a distinction would be useful. Theoretically hypoxanthine should only be elevated in uncompensated hypoxia, while pH and lactate changes occur in compensated hypoxia. It seems, however, that hypoxanthine is also elevated to some extent in uncompensated hypoxia.Renewed interest in hypoxanthine developed when it was realized that hypoxanthine is a potential free radical generator (5, 6). Hypoxanthine seems to play a role in posthypoxic reoxygenation cell injury through oxygen radical production (7) and is therefore involved in the pathogenesis of a number of diseases. Hypoxanthine also modulates a number of other processes because it reacts with benzodiazepine receptors (8) and inhibits phosphodiesterase in the brain (9). Hypoxanthine inhibits the effect of several cytotoxic drugs and may therefore influence treatment with such drugs (10).Herein we summarize the extensive literature on hypoxanthine published during the last decade and try to answer the question: How useful in clinical medicine are hypoxanthine measurements in plasma and other body fluids? Is hypoxanthine a better marker of hypoxia than lactate or pH? Further, we discuss the significance of hypoxanthine as a potential oxygen radical generator and put forward a hypothesis for the pathogenesis of an "oxygen radical disease in neonatology". In hypoxia there is an accelerated breakdown of AMP to hypoxanthine. Figure 1 illustrates several important aspects of hypoxanthine metabolism in hypoxia. Normally about 90% of the hypoxanthine formed is reutilized through the sa...

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“…In these measurements, we chose 1.9 lM for the initial concentration of xanthine, according to the reported values of xanthine in human blood plasma [29,30]. Under this condition, the XO reaction was 95% complete at the reaction time of 30 s, which was confirmed by the amperometry of urate at E = 0.2 V with a Pt disk electrode.…”

Section: å Oh Generation Induced By Xanthine-xo Systemmentioning

confidence: 91%

Sensitive electrochemical measurement of hydroxyl radical generation induced by the xanthine–xanthine oxidase system

Tatsumi

Tsuchiya

Sakamoto

2014

Analytical Biochemistry

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High-performance liquid chromatographic determination of hypoxanthine and xanthine in biological fluids

Cited by 48 publications

References 17 publications

Surface-Enhanced Raman Scattering of Whole Human Blood, Blood Plasma, and Red Blood Cells: Cellular Processes and Bioanalytical Sensing

Surface-Enhanced Raman Scattering of Whole Human Blood, Blood Plasma, and Red Blood Cells: Cellular Processes and Bioanalytical Sensing

Hypoxanthine as an Indicator of Hypoxia: Its Role in Health and Disease through Free Radical Production

Sensitive electrochemical measurement of hydroxyl radical generation induced by the xanthine–xanthine oxidase system

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