scholarly journals Differences in Nucleotide Hydrolysis Contribute to the Differences between Erythrocyte 6-Thioguanine Nucleotide Concentrations Determined by Two Widely Used Methods

2003 ◽  
Vol 49 (2) ◽  
pp. 260-268 ◽  
Author(s):  
Maria Shipkova ◽  
Victor William Armstrong ◽  
Eberhard Wieland ◽  
Michael Oellerich

Abstract Background: Measurement of 6-thioguanine nucleotide (6-TGN) concentrations in erythrocytes is widely accepted for use in optimization of thiopurine therapy. Various chromatographic methods have been developed for this purpose. In preliminary experiments we observed a considerable difference between 6-TGN concentrations determined with two widely used methods published by Lennard (Lennard L. J Chromatogr 1987;423:169–78) and by Dervieux and Boulieu (Dervieux T, Boulieu R. Clin Chem 1998;44:551–5). We therefore investigated methodologic differences between the two procedures with respect to hydrolysis of 6-TGNs to 6-thioguanine (6-TG) in more detail. Methods: We analyzed 6-TGNs in erythrocyte preparations (n = 50) from patients on azathioprine therapy by both methods, using the original protocols. In one set of experiments, we replaced the 0.5 mol/L sulfuric acid in the Lennard method with the 1 mol/L perchloric acid used by Dervieux and Boulieu. In a second set of experiments, we investigated the effect of various dithiothreitol (DTT) concentrations on 6-TG recovery with both methods. In a third set of experiments, we determined the effect of hydrolysis time on both protocols. Results: Direct comparison of both methods showed that 6-TGN concentrations were, on average, 2.6-fold higher in the Dervieux–Boulieu method over the concentration range tested, although the correlation (r = 0.99; P <0.001) was good. Replacement of sulfuric acid by perchloric acid reduced this difference to ∼1.4-fold (r = 0.99; P <0.001). Increasing the DTT concentration enhanced 6-TG recovery. The hydrolysis time used in the Lennard method (1 h) was not sufficient to achieve complete hydrolysis. Conclusions: The difference between 6-TGN concentrations measured by the two methods is attributable, at least in part, to differences in the extent of nucleotide hydrolysis. For optimization of thiopurine therapy, method-dependent therapeutic ranges are necessary, which precludes comparison of results from clinical studies derived with these methods. Efforts must therefore be made to standardize the analytical procedures for the determination of 6-TGN.

1984 ◽  
Vol 39 (3-4) ◽  
pp. 249-251 ◽  
Author(s):  
Vassilios M. Kapoulas ◽  
Sofia K. Mastronicolis ◽  
Ibrahim C. Nakhel ◽  
Helen J. Stavrakakis

A rapid method for initial quantitative estimation of the phosphate present in compound containing a carbon-phosphorus bond is described. Two phosphorus assays are employed. One assay is for total phosphorus, which can be determined by digesting with perchloric acid and the other assay is for total non phosphonate phosphorus which can be determined by digesting with sulfuric acid simultaneously. The difference between total phosphorus and the non phosphonate phosphorus determined represents the amount of phosphorus present in a carbon-phosphorus linkage in a crude phospholipids sample


1964 ◽  
Vol 10 (9) ◽  
pp. 799-823 ◽  
Author(s):  
H Hoch ◽  
S L Sinnett ◽  
T H McGavack

Abstract The chloric acid digestion of serum, followed by the ceric sulfate reduction test for iodine has been shown to give results reproducible to within ± 0.00035 µg. iodine per test sample (1) if the entire digestion tubes are kept hot during the combustion step so that products of incomplete oxidation are vaporized, (2) if the perchloric acid and sulfuric acid concentrations in the final digest are adjusted to 1.64 normal (16.5%) and 0.37 normal, respectively, (3) if attention is paid to the order of addition of the reagents, and (4) if the arsenite reagent is added within a fraction of a second. The method gave values for total serum iodine in 20 healthy subjects between 5 and 10.5 µg./100 ml.


1970 ◽  
Vol 53 (6) ◽  
pp. 1172-1175 ◽  
Author(s):  
M T Jeffus ◽  
J S Elkins ◽  
C T Kenner

Abstract Mercury in biological samples is determined by digestion in nitric acid, sulfuric acid, and permanganate, followed by reduction and aeration for measurement by atomic absorption at room temperature. The average recovery is 95.8% with a standard deviation of 13.3%. The standard deviation, calculated from the difference between duplicates of 23 samples, is 0.063 μg mercury, which represents 9.4% of the average value of the samples. The method is simple and requires approximately 4 hr for completion. Mercury can be confirmed by adsorption on gold foil after maximum absorbance has been obtained during aeration.


1966 ◽  
Vol 49 (2) ◽  
pp. 307-309
Author(s):  
W Glover ◽  
H Kirschenbaum ◽  
A Caldwell

Abstract A method has been developed for determining starch in meat food products in which simple sugars are extracted from meat food products with hot 80% ethanol while starch remains. The starch residue is solublized with dilute perchloric acid, readily forms a colored derivative when heated with anthrone-sulfuric acid reagent, and is measured at 630 mμ. It was not necessary to prepare a standard dextrose curve daily. The anthrone-sulfuric acid reagent is good for 3–4 days if kept near 0.0°C.


1978 ◽  
Vol 56 (7) ◽  
pp. 935-940 ◽  
Author(s):  
John T. Edward ◽  
Graeme Welch ◽  
Sin Cheong Wong

The rates of hydrolysis of thioacetic, thiobenzoic, and three substituted thiobenzoic acids increase with concentration of solvent sulfuric or perchloric acid to a maximum in 30–40% acid and then decrease. Yates–McClelland r, Bunnett–Olsen [Formula: see text], and Hammett ρ parameters, and entropies of activation indicate an AAC2 mechanism over this range of acid concentrations. In acid concentrations above 50–60% the rates increase sharply and the same mechanistic criteria now indicate an AAc1 mechanism. The difference between the rate–acidity profile of thiobenzoic acid and that of ethyl thiolbenzoate can be explained by the different response of the activity coefficients of their transition states to increase in sulfuric acid concentration.


1990 ◽  
Vol 68 (10) ◽  
pp. 1653-1656 ◽  
Author(s):  
Y. Chiang ◽  
A. J. Kresge ◽  
R. A. More O'ferrall ◽  
B. A. Murray ◽  
N. P. Schepp ◽  
...  

Rates of ketonization of the enol of acetophenone, generated by flash photolytic photohydration of phenylacetylene, were measured in aqueous sulfuric and perchloric acid solutions over the concentration range 1–50 wt.% acid; rates of enolization of acetophenone, monitored by bromine scavenging, were also measured in aqueous perchloric acid solutions over the same concentration range. The results suggest that the curvature observed in a previous X acidity function correlation of the rate of enolization in sulfuric acid solutions was an artifact produced by insufficiently efficient scavenging, and that introduction of the activity of water in the correlating expression, used previously to eliminate the curvature and believed to reflect covalent involvement of water in the enolization reaction, is unnecessary. The present results also show that the keto–enol equilibrium constant for acetophenone decreases with increasing acidity in these concentrated sulfuric and perchloric acid solutions. Key words: acetophenone, enolization, ketonization, keto–enol equilibrium, concentrated acid solutions.


2008 ◽  
Vol 73 (8-9) ◽  
pp. 871-878 ◽  
Author(s):  
Radmila Dzudovic ◽  
Ljiljana Jaksic

A coulometric-potentiometric procedure for the determination of relative acidity scales of acetone, methylethyl ketone, methyl-isobutyl ketone and propylene carbonate is described. The range of the relative acidity scale of a solvent was determined from the difference between the half-neutralization potential of perchloric acid and that of tetrabutylammonium hydroxide. The perchloric acid was generated in situ from a hydrogen-palladium electrode in presence of sodium perchlorate or tetrabutylammonium perchlorate as the supporting electrolyte. The electrode pairs glass-SCE and (H2/Pd)ind-SCE were applied for the measurement of the half-neutralization potentials of the acid and base. A wider range of relative acidity scale of the solvents was obtained with the glass-SCE electrode pair and tetrabutylammonium perchlorate as the supporting electrolyte.


1962 ◽  
Vol 41 (1) ◽  
pp. 123-128 ◽  
Author(s):  
Pentti A. Järvinen ◽  
Sykkö Pesonen ◽  
Pirkko Väänänen

ABSTRACT The fractional determination of 17-ketosteroids in the daily urine was performed in nine cases of hyperemesis gravidarum and in four control cases, in the first trimester of pregnancy both before and after corticotrophin administration. The excretion of total 17-KS is similar in the two groups. Only in the hyperemesis group does the excretion of total 17-KS increase significantly after corticotrophin administration. The fractional determination reveals no difference between the two groups of patients with regard to the values of the fractions U (unidentified 17-KS), A (androsterone) and Rest (11-oxygenated 17-KS). The excretion of dehydroepiandrosterone is significantly higher in the hyperemesis group than in the control group. The excretion of androstanolone seems to be lower in the hyperemesis group than in the control group, but the difference is not statistically significant. The differences in the correlation between dehydroepiandrosterone and androstanolone in the two groups is significant. The high excretion of dehydroepiandrosterone and low excretion of androstanolone in cases of hyperemesis gravidarum is a sign of adrenal dysfunction.


Author(s):  
Irina Mordous

The development of modern civilization attests to its decisive role in the progressive development of institutions. They identified the difference between Western civilization and the rest of the world. Confirmation of the institutional advantages of the West was its early industrialization. The genesis and formation of institutionalism in its ideological and conceptualmethodological orientation occurs as a process alternative to neoclassic in the context of world heterodoxia, which quickly spread in social science. Highlighting institutional education as a separate area of sociocultural activity is determined by the factor of differentiation of institutional theory as a whole. A feature of institutional education is its orientation toward the individual and his/her transformation into a personality. The content of institutional education is revealed through the analysis of the institution, which includes a set of established customs, traditions, ways of thinking, behavioral stereotypes of individuals and social groups. The dynamics of socio-political, economic transformations in Ukraine requires a review of the foundations of national education and determination of the prospects for its development in the 21st century in the context of institutionalism.


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