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PubMed1980

Current state of the art in the HPLC analyses of free nucleotides, nucleosides, and bases in biological fluids.

Because of the use of HPLC for the determination of free nucleotides, nucleosides, and bases in samples of biological origin, investigations that were very difficult or impossible to perform only a decade ago are now possible. Microparticulate, totally porous, chhemically bonded particles appear to be the column packings that will be used routinely in the future because analyses are achieved with high sensitivity, selectivity, efficiency, and speed. These packings are stable, the results are reproducible, and the data are quantitative. In addition, the reversed-phase mode of HPLC offers improvements over the ion-exchange mode for the analysis of nucleosides, bases, and other UV-absorbing compounds which are not ionic. However, the ion-exchange mode may be used as a complementary method for very polar or ionic compounds which are eluted too rapidly with reversed phase. A sample can be chromatographed with each mode and a profile--a complete picture of nucleotide, nucleoside, and base levels in the sample--obtained. Alternatively, ion pairing can be used with the reversed-phase mode; thus both the ionic and lipophilic compounds can be determined in one separation.

Chromatography, High Pressure Liquid↗
PubMed1987

Determination of nucleotides, nucleosides and nucleobases in cells of different complexity by reversed-phase and ion-pair high-performance liquid chromatography.

Procedures are presented for the analysis of profiles of purine and pyridine compounds in human and rabbit red blood cells by reversed-phase high-performance liquid chromatography and in Ehrlich ascites tumour cells of mouse by ion-pair high-performance liquid chromatography. These compounds are present in rabbit erythrocytes in higher concentrations than in human blood cells, and in rabbit reticulocytes the concentration of purine compounds is still higher. During glucose-free incubation, human red cells accumulate adenosine and adenine in the presence of coformycin owing to the inhibition of adenosine and AMP deamination. Ehrlich ascites tumour cells lose major portions of purine mono-, di- and triphosphates between the seventh and eleventh day after inoculation into mouse peritoneal cavities.

Animals↗
PubMed1995

Quantitation of nucleotides, nucleosides and bases in antemortem and postmortem bloodstains by high-performance liquid chromatography.

Ante- and post-mortem bloodstains prepared from the blood of volunteers and corpses were analysed for ATP and its related compounds by reversed-phase high-performance liquid chromatography (HPLC). The results showed that (1) ATP was present in a large amount in antemortem bloodstains but not in postmortem stains, (2) AMP, adenosine, inosine, hypoxanthine, xanthine and uracil either were not detected or were detected in smaller amounts in antemortem than in postmortem bloodstains, and (3) ADP was present in both ante- and post-mortem bloodstains. These differences suggest that quantitation of these compounds may be useful in identifying whether bloodstains are ante- or post-mortem.

Blood Stains↗
PubMed2003

Neuron-specific enolase, nucleotides, nucleosides, purine bases, oxypurines and uric acid concentrations in cerebrospinal fluid of children with meningitis.

To determine the effects of meningitis on cerebral energy metabolism, cerebrospinal fluid concentrations of adenosine monophosphate, inosine monophosphate, inosine, adenosine, guanosine, adenine, guanine, hypoxanthine, xanthine and urate were determined by high-performance liquid chromatography, and neuron-specific enolase by an enzyme immunoassay method, in 100 children with meningitis (45 bacterial, 46 viral and nine tuberculous), aged between 1 month and 13 years, and in 160 age-matched controls. Compared with controls, patients with bacterial meningitis showed high concentrations of hypoxanthine, xanthine and urate; patients with viral meningitis showed high concentrations of inosine, guanosine, xanthine, urate and neuron-specific enolase; and patients with tuberculous meningitis showed very high concentrations of inosine, xanthine and urate. Xanthine and urate concentrations were significantly higher in patients with tuberculous meningitis than in patients with viral or bacterial meningitis. These results suggest that in the acute stage of bacterial, viral and tuberculous meningitis, neuronal energy metabolism may be altered. The measurement of cerebrospinal xanthine and uric acid concentrations may be useful for the early diagnosis of a tuberculous origin.

Child↗
PubMed1997

1H and (31)P NMR and EPR of Pentaammineruthenium(III) Complexes of Endocyclically Coordinated Nucleotides, Nucleosides, and Related Heterocyclic Bases. Autoxidation of [(Guokappa(N7))(NH(3))(5)Ru(III)] (Guo = Guanosine). Crystal Structure of [7MeGuakappa(N9)(NH(3))(5)Ru]Cl(3).3H(2)O.

The (1)H-NMR spectra of complexes involving the paramagnetic metal center [(NH(3))(5)Ru(III)] coordinated at ring nitrogens have been examined with pyridine, purine, nucleoside, and nucleotide ligands along with (31)P-NMR of the nucleotide complexes and EPR of representative complexes. Variations in the spectra have been investigated as a function of the coordination site and pH. Pseudocontact and contact shifts have been calculated for various protons, and an attempt has been made to correlate sugar conformations in coordinated 5'GMP, 5'IMP, Guo, and Ino with paramagnetically induced shifts. The compound [(7MeGuakappa(N9))(NH(3))(5)Ru]Cl(3).3H(2)O crystallizes in the orthorhombic space group Pna2(1) with cell parameters a = 25.375(4) Å, b = 11.803(4) Å, c = 6.958(2) Å, Z = 4, and R = 0.042. The autoxidation of [L(NH(3))(5)Ru(III)], where L = Guo, dGuo, and 1MeGuo, to the corresponding 8-oxo complexes under atmospheric oxygen is first order in the complex and [OH(-)]. For L = Guo, k = 6.6 x 10(-5) M(-1) s(-1), DeltaH = 58 kJ/mol, and DeltaS = -124 J/(mol K).

Journal Article↗