Complete separation of adenine nucleotides for ATPase activity assay by ion-pair reversed-phase high-performance liquid chromatography

Determination of adenosine nucleotides in cultured cells by ion-pairing liquid chromatography–electrospray ionization mass spectrometry

“…Perchloric acid has been used for metabolites extraction due to its ability to precipitate proteins. In a recent publication, Ushimaru and Fukushima [27] suggested that sulfuric and/or hydrochloric acids might be a better choice for the extraction procedure as perchloric ions might affect subsequent analysis. In the present and previous studies [26], the perchloric ion was effectively removed by precipitation with equal molar KOH.…”

Liquid chromatographic methods for the determination of endogenous nucleotides and nucleotide analogs used in cancer therapy: A review

“…As above described for monophosphate nucleosides, the retention achieved for nucleoside triphosphates was satisfactory using either IP chromatography [39,40,45,[49][50][51][52]54,55,57,58,66,67,[72][73][74][75]77,78,84,86,[88][89][90] or PGC column [47,82,87]. The complete resolution between the eight intracellular nucleoside triphosphates (ATP, CTP, guanosine triphosphate (GTP), uridine triphosphate (UTP), dATP, dCTP, deoxyguanosine triphosphate (dGTP), TTP) was achieved using an IP-HPLC with high salts concentrations in the mobile phase (10 mM tetrabutylammonium hydroxide, 10-50 mM K 2 HPO 4 ) [54].…”

A highly sensitive high-performance liquid chromatography–mass spectrometry method for quantification of fludarabine triphosphate in leukemic cells

“…The response of F-ara-ADP observed resulting from such fragmentation is routinely 9% of the F-ara-ATP peak area; that of F-ara-AMP is less than 1%. Although recent work [18] has suggested that residual PCA in samples can interfere with the separation of nucleotide phosphates, we have circumvented the problem by neutralizing the sample with potassium bicarbonate. The resulting sample has low ion strength at pH ∼7.5, ideal for subsequent chromatography.…”