Allantoin as a marker of oxidative stress in human erythrocytes

Author(s):  
Roman Kand'ár ◽  
Pavla Žáková

Abstract: Uric acid is the final product of purine metabolism in humans. It was determined that this compound has important antioxidative properties and it may be oxidized to allantoin by various reactive oxygen species. Therefore, the measurement of allantoin may be useful for the determination of oxidative stress in humans.: We measured allantoin and uric acid in human plasma and erythrocytes obtained from patients with chronic renal failure before hemodialysis (n=30) and blood donors (n=30). We used a method based on selective isolation of allantoin from deproteinized plasma and erythrocyte lysate samples on AG 1-X8 resin and its derivatization to glyoxylate-2, 4-dinitrophenylhydrazone. Separation of glyoxylate-2, 4-dinitrophenylhydrazone from interfering substances was achieved on reversed phase HPLC with gradient elution and UV/VIS detection at 360 nm. Uric acid was determined by reversed phase HPLC with UV/VIS detection at 292 nm.: We found significant differences in allantoin and uric acid concentration between the patients with chronic renal failure and the control group both in plasma (20.5±6.5 μmol/L and 323.9±62.9 μmol/L vs. 2.1±1.1 μmol/L and 270.1±62.3 μmol/L, p<0.05) and erythrocytes [82.8±39.1 nmol/g hemoglobin (Hb) and 110.7±28.8 nmol/g Hb vs. 20.1±6.1 nmol/g Hb and 82.1±23.7 nmol/g Hb, p<0.05].: Significant higher levels of allantoin in both plasma and erythrocytes of patients with chronic renal failure indicate that allantoin may be used as a good marker of oxidative stress.Clin Chem Lab Med 2008;46:1270–4.

2000 ◽  
Vol 47 (3) ◽  
pp. 877-879 ◽  
Author(s):  
B Kochańska ◽  
R T Smoleński ◽  
N Knap

The profile and normal concentrations of nucleotide metabolites in human saliva and reproducibility of these determinations were analyzed. Samples of human saliva collected from healthy individuals at weekly intervals, were deproteinized and analysed for the content of adenine nucleotides and their metabolites by reversed-phase HPLC. Initial ATP, hypoxanthine and uric acid concentrations were 0.52 +/- 0.15 microM, 1.91 +/- 0.37 microM and 184 +/- 22 microM respectively. A substantial individual variation persisted within 3 weeks of sampling excepted hypoxanthine which showed some unrelated variations. Determination of nucleotides and their catabolites in saliva due to its simplicity and reproducibility, may be of clinical value in diagnosis of local or systemic disorders.


2020 ◽  
pp. 81-88
Author(s):  
Viktor Ivanovich Deineka ◽  
Elena Yur'yevna Oleinits ◽  
Aleksandr Alekseyevich Pavlov ◽  
Andrey Yur'yevich Mikheev ◽  
Ol'ga Vladimirovna Shelepova ◽  
...  

The anthocyanin composition of some species of gooseberries with colored fruits was studied in comparison with anthocyanins of currant fruits from the collection of the Tsitsin Main Botanical garden. In fruits of currants Ribes nigrum var. sibiricum E. Wolf, R. nigrum L., R. americanum Mill., as well as in dark-colored fruits R. aureum Pursh., 3-glucosides and 3-rutinosides of cyanidin and delphinidin were found as major components in species-dependent ratios. It was found that in the fruits of gooseberry species Ribes oxyacanthoides L., Ribes missouriensis (Nutt.) Coville & Britton, Ribes rusticum Jancz. and Ribes cynosbati (L.) Mill. and in the fruits of one of the currants (R. alpinum L.) there is practically no biosynthesis of delphinidin derivatives, and 3-glucoside and 3-rutinoside of cyanidin are detected. Meanwhile, the biosynthesis of acylated by substituted cinnamic acids (mainly – p-coumaric acid) anthocyanins were found in the fruits of Ribes oxyacanthoides and Ribes cynosbati (about 10%), Ribes rusticum (about 20%). The inheritance of characteristic for the fruit of some species of gooseberry biosynthesis of acylated mainly p-coumaric acid, was installed in the fruits of jostaberry - Ribes x nidigrolaria Rud. Bauer & A. Bauer (about 10%). The species composition of anthocyanins was established using traditional reversed-phase HPLC in gradient elution mode using diode-array and mass spectrometric detection options. In addition, a method for the separation of anthocyanins of the same objects in the isocratic mode under hydrophilic interaction chromatography on diol stationary phases, allowing the use of simpler equipment. The variants of sample preparation of extract samples for the subsequent HPLC determination of anthocyanins by two different methods to avoid the appearance of artifacts are proposed.


2019 ◽  
Vol 2019 ◽  
pp. 1-12
Author(s):  
Yansheng Wu ◽  
Qiang Wan ◽  
Liqiang Shi ◽  
Jiaoying Ou ◽  
YingQiao Li ◽  
...  

Objective. Renal anemia in patients with end-stage chronic kidney disease is closely related to the deterioration of cardiac function, renal function, and quality of life. This study involved adenine-induced renal anemic rat models and evaluated the treatment effect of Siwu granules and/or erythropoietin (EPO). Methods. Fifty SD rats were randomly divided into 5 groups: control, model, Siwu, EPO, and Siwu plus EPO groups. The expression levels of NO, MDA, SOD, CAT, IL-6, TNF-α, EPO, EPOR, α-SMA, and TGF-β1 were detected in rats after 8 weeks of treatment with Siwu granules and/or EPO. Results. After modeling, 47 rats entered the stage of treatment. Siwu plus EPO treatment significantly increased the rat hemoglobin content (p<0.05) and reduced blood urea nitrogen (p<0.05) and serum creatinine (p<0.001). Compared with the control group, the expression of EPO and EPOR in the kidney of rats with renal failure was significantly decreased (p<0.05). Moreover, the Siwu plus EPO group improved the level of oxidative stress in rats with chronic renal failure and reduced the expression of inflammatory factors. The expression of α-SMA and TGF-β1 in rats with renal failure was higher, but there was no expression in the control group. Conclusion. Combined treatment of Siwu granules with EPO increased the expression of EPO and EPOR in the renal tissues and inhibited oxidative stress and inflammatory factors, improving the renal function and anemia.


1987 ◽  
Vol 33 (8) ◽  
pp. 1427-1430 ◽  
Author(s):  
R Sakuma ◽  
T Nishina ◽  
M Kitamura

Abstract We evaluated six deproteinizing methods for determination of uric acid in serum by "high-performance" liquid chromatography with ultraviolet detection: those involving zinc hydroxide, sodium tungstate, trichloroacetic acid, perchloric acid, acetonitrile, and centrifugal ultrafiltration (with Amicon MPS-1 devices). We used a Toyosoda ODS-120A reversed-phase column. The mobile phase was sodium phosphate buffer (40 mmol/L, pH 2.2) containing 20 mL of methanol per liter. Absorbance of the eluate was monitored at 284 nm. The precipitation method with perchloric acid gave high recoveries of uric acid and good precision, and results agreed with those by the uricase-catalase method of Kageyama (Clin Chim Acta 1971;31:421-6).


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