Coprecipitation of ATP with potassium perchlorate: The effect on the firefly enzyme assay of ATP in tissue and blood

Wiener, Stanley L.; Wiener, Roy S.; Urivetzky, Morton; Meilman, Edward

doi:10.1016/0003-2697(74)90302-9

Cited by 32 publications

(8 citation statements)

References 7 publications

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“…To restore a neutral pH to samples, the HClO 4 was neutralized with KOH, producing a precipitate that needed to be removed from the samples by centrifugation. It is likely that, despite our efforts to completely remove these precipitates, they continued to form over time as compounds adsorbed to the remaining perchlorate [36]. It was noted that, for some analytes, this treatment increased the area under the peak and modified the retention times, and hence, we normalized for these effects by also treating the standards with HClO 4 /KOH.…”

Section: Resultsmentioning

confidence: 99%

Simultaneous quantification of 12 different nucleotides and nucleosides released from renal epithelium and in human urine samples using ion-pair reversed-phase HPLC

Contreras-Sanz

Scott-Ward

Gill

et al. 2012

Purinergic Signalling

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Nucleotides and nucleosides are not only involved in cellular metabolism but also act extracellularly via P1 and P2 receptors, to elicit a wide variety of physiological and pathophysiological responses through paracrine and autocrine signalling pathways. For the first time, we have used an ion-pair reversed-phase high-performance liquid chromatography ultraviolet (UV)-coupled method to rapidly and simultaneously quantify 12 different nucleotides and nucleosides (adenosine triphosphate, adenosine diphosphate, adenosine monophosphate, adenosine, uridine triphosphate, uridine diphosphate, uridine monophosphate, uridine, guanosine triphosphate, guanosine diphosphate, guanosine monophosphate, guanosine): (1) released from a mouse renal cell line (M1 cortical collecting duct) and (2) in human biological samples (i.e., urine). To facilitate analysis of urine samples, a solid-phase extraction step was incorporated (overall recovery rate ≥ 98 %). All samples were analyzed following injection (100 μl) into a Synergi Polar-RP 80 Å (250 × 4.6 mm) reversed-phase column with a particle size of 10 μm, protected with a guard column. A gradient elution profile was run with a mobile phase (phosphate buffer plus ion-pairing agent tetrabutylammonium hydrogen sulfate; pH 6) in 2–30 % acetonitrile ( v/v ) for 35 min (including equilibration time) at 1 ml min −1 flow rate. Eluted compounds were detected by UV absorbance at 254 nm and quantified using standard curves for nucleotide and nucleoside mixtures of known concentration. Following validation (specificity, linearity, limits of detection and quantitation, system precision, accuracy, and intermediate precision parameters), this protocol was successfully and reproducibly used to quantify picomolar to nanomolar concentrations of nucleosides and nucleotides in isotonic and hypotonic cell buffers that transiently bathed M1 cells, and urine samples from normal subjects and overactive bladder patients. Electronic supplementary material The online version of this article (doi:10.1007/s11302-012-9321-8) contains supplementary material, which is available to authorized users.

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Section: Resultsmentioning

confidence: 99%

Simultaneous quantification of 12 different nucleotides and nucleosides released from renal epithelium and in human urine samples using ion-pair reversed-phase HPLC

Contreras-Sanz

Scott-Ward

Gill

et al. 2012

Purinergic Signalling

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“…This point was emphasized by Langercrantz and Stjarne (1974), who reported recoveries of 59-73% for ATP, using internal standards. This potential error was discussed by Wiener et al (1974).…”

Section: Discussionmentioning

confidence: 99%

“…The values derived by luciferin-luciferase analysis were uncorrected for losses of nucle otide which might be attributed to the potassium carbonate neutralization step, as discussed by Wiener et al (1974). Values obtained by luciferin-luciferase analysis were provided in order to verify, using an entirely different method, those values obtained from the high pressure liquid chromatographic separations and ultraviolet analysis.…”

Section: Analysis O F Nucleotides By Luciferin-luciferasementioning

confidence: 99%

An Analysis of Nucleotides and Catecholamines in Bovine Medullary Granules by Anion Exchange High Pressure Liquid Chromatography and Fluorescence

et al. 1977

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The catecholamine and nucleotide content of chromaffin granules from bovine adrenal medulla was analyzed both qualitatively and quantitatively. Nucleotides were examined by separation on anion exchange high pressure liquid chromatography and ultraviolet absorption, as well as by anion exchange thin-layer chromatography and luciferin-luciferase light emission analysis. Catecholamines were quantified via automated continuous-flow analysis using fluorescence. We found the average catecholamine/ATP ratio to be 8.2:1 with an SD of 1.98 and an SE of 0.50 for 16 separate glands. Relative percentages of epinephrine and norepinephrine were found to be 79.8 and 20.2, respectively. The granules contained 80.9% ATP, 12.3% GTP, and 6.8% UTP, as well as detectable amounts of ADP, AMP, GDP, and UDP. If we accept the catecholamine/ATP ratio of 4:1 proposed by Hillarp earlier, then those catecholamines constituting the remainder of our ratio of 8.2:1 should represent a nucleotide-unassociated pool. In view of recent evidence of a direct agonistic function of nucleotides, we propose that the granule-associated nucleotides may act locally as coagonists with certain biogenic amines, and may additionally provide a circulating pool of purines and pyrimidines for use by the heart and lungs.

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“…The loss was most pronounced at high concentrations of red blood cells [12]. A sim ilar problem of inadequate extraction has been recognized previously using perchloric acid [19,20], Supernatant must be used in spectrophotometric measurements since tides during the last week in RBC-Hct 79 and RBC-Hct 62 ( fig. 2d, e).…”

Section: Discussionmentioning

confidence: 90%

Preservation of Red Blood Cells: Studies of Erythrocyte Adenine Nucleotides Using Luminescence Analysis

Gulliksson¹,

Olsson²,

Palmeborn³

et al. 1983

Vox Sanguinis

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The levels of ATP and total adenine nucleotides (ATP + ADP + AMP) were determined by firefly luciferase assay in red blood cells during storage for 5 weeks at 4°C. With few exceptions, no significant differences in nucleotide levels were found between whole blood stored in CPD-adenine and various preparations of red blood cells in CPD-adenine or CPD with saline-adenine-glucose (SAG) as additive. The levels of ATP and total adenine nucleotides during storage are discussed in relation to glucose levels, extracellular pH and shelf life of the red blood cells.

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Coprecipitation of ATP with potassium perchlorate: The effect on the firefly enzyme assay of ATP in tissue and blood

Cited by 32 publications

References 7 publications

Simultaneous quantification of 12 different nucleotides and nucleosides released from renal epithelium and in human urine samples using ion-pair reversed-phase HPLC

Simultaneous quantification of 12 different nucleotides and nucleosides released from renal epithelium and in human urine samples using ion-pair reversed-phase HPLC

An Analysis of Nucleotides and Catecholamines in Bovine Medullary Granules by Anion Exchange High Pressure Liquid Chromatography and Fluorescence

Preservation of Red Blood Cells: Studies of Erythrocyte Adenine Nucleotides Using Luminescence Analysis

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