Diagnostic potential of HPLC: experimental and clinical trials.
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Animals↗
Biomedical subjects
Tikhonov YuV
Publications and source records attributed to Tikhonov YuV.
Ion-pair high-performance liquid chromatography of purine compounds in the small intestinal mucosa of children with coeliac disease.
Isocratic methods are described for the separation of purine compounds in specimens of jejunal mucosa available by peroral biopsy from patients with coeliac disease. The first mode provided separation of both the ionic nucleoside monophosphates and the non-ionic nitrogen bases and nucleosides. The second mode was applicable to analyses of the nucleoside mono-, di- and triphosphates only. These chromatographic procedures were carried out with tetrabutylammonium phosphate as an ion-pair modifier and different concentrations of acetonitrile in the mobile phase. The modes are flexible in that they permit the optimization of the separation of these compounds according to the purpose of the investigation. The results of studies on purine metabolite patterns in coeliac mucosa are discussed.
Adolescent↗
Influence of perinatal hypoxia on purine contents in erythrocytes of newborn infants.
Alterations in the levels of hypoxanthine and some other purine derivatives occurring in the erythrocytes from umbilical cord blood of newborn infants with perinatal hypoxia were examined. Newborn babies were divided into two groups which included infants with favourable (Group 1) and complicated (Group 2) periods of early adaptation. Purine metabolites of erythrocytes were analyzed by isocratic reversed-phase HPLC. It was shown a reciprocal relation between concentrations of hypoxanthine and IMP in erythrocytes of infants from two groups that was expressed in elevation (Group 1) and a fall (Group 2) of hypoxanthine level while in Group 1 the IMP content had a tendency to decrease and Group 2 was characterized by an elevated level of this compound. The alterations in the concentrations of these interrelated metabolites may be due to the effects of hypoxia upon purine metabolism in erythrocytes.
Asphyxia Neonatorum↗
The adenine nucleotide catabolism in nonphosphorylating mitochondria of different tissues.
The degradation of endogenous adenine nucleotides was compared in mitochondria isolated from mouse and rat liver, rat renal cortex and solid hepatoma. Mitochondria were incubated for 30 min at 37 degrees C in the presence of carboxyatractyloside and oligomycin. In rat liver mitochondria ATP, ADP and AMP were degraded by about 25% each, whereas in kidney and hepatoma mitochondria a rapid decline of ATP and ADP but no change in the AMP contents were observed. Main products formed were adenosine, inosine and hypoxanthine in all mitochondria examined. Compartment studies revealed that the main route of intramitochondrial adenine nucleotide catabolism seems to proceed via AMP dephosphorylation and subsequent adenosine deamination.
Adenine Nucleotides↗
Purine compounds in mitochondria: a quantitative evaluation.
Applying a recently developed reverse-phase ion-pair HPLC technique we determined purine compounds in acid extracts of mitochondria isolated from rat and mouse liver, rat renal cortex, and hepatoma. Intramitochondrial purine nucleosides and bases were evaluated quantitatively for the first time. All mitochondria examined contain significant amounts of adenosine, guanosine, inosine, guanine, and hypoxanthine. Moreover, IMP was identified in all extracts. Adenine was detected in traces only. These results are in accordance with the concept of mitochondrial metabolism of purine compounds.
Animals↗
Simultaneous separation of ribonucleotides, nucleosides and nitrogen bases by ion-pair reversed-phase high-performance liquid chromatography on columns with radial compression.
A method is described for the simultaneous analysis of twelve major nucleotides, certain biochemically essential nucleosides and nitrogen bases. The separation was achieved using an ion-pair reversed-phase high-performance liquid chromatographic (HPLC) system with a radially compressed NovaPak C18 cartridge and tetrabutylammonium phosphate (TBA) as a mobile phase modifier under acetonitrile gradient conditions. The effects of TBA and acetonitrile on the retention of major nucleotides were studied. The retention of nucleotides increased significantly with an increase in the TBA concentration from 0 to 2 mM and remained constant up to 5 mM TBA. The logarithm of the capacity factor decreased in proportion to the concentration of acetonitrile in the mobile phase. The separation of nitrogen bases, nucleosides and some of their deoxy forms was performed on an isocratic ion-pair reversed-phase HPLC system with pentanesulphonic and heptanesulphonic acid using a mu Bondapak C18 column. The described chromatographic procedures were applied to the separation of various biological extracts.
Animals↗