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

Simultaneous determination of adenosine triphosphate and its metabolites in human whole blood by RP-HPLC and UV-detection

“…However, its main disadvantage compared to our method is that only ATP can be determined, whereas we can quantify the complete adenylate pool in one run. Ion-exchange methods often require more extensive pretreatment of samples and have difficulties in separating nucleobases, nucleosides and nucleotides in one run, given the charged nature of the nucleotides in the operating pH range (pH 2-7) [33,[36][37][38][39].…”

A Novel Method for Measuring the ATP-Related Compounds in Human Erythrocytes

“…μmol/L RBCs (Boulieu et al 1983;Werner et al 1987 (Tavazzi et al 2000). In addition, the present basal AEC suggest that the current process provided a stable metabolic state in RBCs because the decreased energy charge was associated with the degradation of nucleotides (Atkinson 1968;Matsumoto et al 1979;de Atauri et al 2006).…”

“…However, the ADP peak in blood samples might be eluted with peptides because purity factors were < 900, their spectra had an additional peak and the peptide bond typically absorbs from 180 to 240 nm in the UV region (Kelly and Price 2000). This impure ADP peak was not reported in other methods because this is the first time that the ATP-related compounds were evaluated using the 3D spectra and purity factors (Scholar et al 1973;Anderson and Murphy 1976;Schweinsberg and Loo 1980;Harmsen et al 1981;Ericson et al 1983;Crescentini and Stocchi 1984;de Korte et al 1985;Stocchi et al 1985Stocchi et al , 1987Werner et al 1987;Tekkanat and Fox 1988;Maessen et al 1988;Smolenski et al 1990;Formato et al 1990;Nishikawa et al 1991;Guieu et al 1994;Caruso et al 2004;Taniai et al 2006;Coolen et al 2008;Yeung et al 2008;Contreras-Sanz et al 2012).…”

“…Moreover, the ranges in Table 3 were established using the normal values from human samples of whole blood and RBCs (Scholar et al 1973;Rapoport et al 1976;Harmsen et al 1981;Ericson et al 1983;Crescentini and Stocchi 1984;de Korte et al 1985;Stocchi et al 1985Stocchi et al , 1987Werner et al 1987;Trulock 1990;Özer et al 2000;Van den Bossche et al 2002;Coolen et al 2008;Dudzinska et al 2010;Díaz et al 2012); however, some normal amounts of nucleotides were not included in these ranges because these values were not reported per volume of RBCs (Beutler et al 1983;Crescentini and Stocchi 1984;de Korte et al 1985;Tekkanat and Fox 1988;Coolen et al 2008) or the units did not agree with the amounts (Schweinsberg and Loo 1980;Formato et al 1990;Buoncristiani et al 1996;Bolzonella et al 2001).…”

“…The complete interpretation of the energetic homeostasis of RBCs under stressful conditions might be performed with a liquid chromatographic (LC) method using ultraviolet (UV) detection that measures all the compounds involved in the degradation pathway for adenine nucleotides alongside the uric acid (Anderson and Murphy 1976;Schweinsberg and Loo 1980;Harmsen et al 1981;Crescentini and Stocchi 1984;Stocchi et al 1985Stocchi et al , 1987Bontemps et al 1986;Werner et al 1987;Maessen et al 1988;Tekkanat and Fox 1988;Smolenski et al 1990;Nishikawa et al 1991;Guieu et al 1994;Smoleńska et al 1999;Caruso et al 2004;Taniai et al 2006;Coolen et al 2008;Yeung et al 2008;Contreras-Sanz et al 2012). The commercial kits (ab65313, abcam; K255-200, BioVision; TB288, Promega; FL-AA, Sigma-Aldrich) and LC methods using fluorometric detection or mass spectrometry (Levitt et al 1984;Ramos-Salazar and Baines 1985;Fujimori et al 1990; Kawamoto et al 1998;Katayama et al 2001;Tuytten et al 2002;Xing et al 2004;Klawitter et al 2007;Wang et al 2009;Birkler et al 2010;Pabst et al 2010;Jiang et al 2012) can only detect some of these compounds simultaneously.…”