ORIGINAL RESEARCH article
Front. Anal. Sci.
Sec. Biomedical Analysis and Diagnostics
Volume 5 - 2025 | doi: 10.3389/frans.2025.1600781
Comprehensive Measurement of Purines in Biological Samples
Provisionally accepted
- 1HPLC Bioanalytical Core, School of Medicine, Emory University, Atlanta, United States
- 2Department of Neuroglogy, Human Genetics, and Pediatrics, Emory University, Atlanta, United States
- 3Department of Pharmacology and Chemical Biology, School of Medicine, Emory University, Atlanta, Georgia, United States
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Defects in numerous aspects of purine metabolism are well-recognized causes for human diseases. The applicability of ultra performance liquid chromatography (UPLC) with photodiode-array (PDA) detection for analysis of the most abundant biological relevant purines metabolites is described. This method was optimized to resolve and quantify 15 purine metabolites including ATP, ADP, AMP, adenosine, adenine, GTP, GDP, GMP, IMP, ZMP, guanosine, hypoxanthine, inosine, xanthine and uric acid in 33 min with a 5 μL injection volume. With purified standards, the detection was linear in a range from 0.1-100 μM. The within-run and between-run variances were <2% overall, indicating excellent reproducibility and reliability.Samples from cultured human cells were prepared to assess the applicability of the method in biological samples. When compared to normal cell lines, mutant cell lines in which purine salvage was absent showed small or no changes for most intracellular purines. Conditioned medium contained no detectable purines, except for hypoxanthine, which was elevated in the mutant lines as expected. Compared to previous methods, this new UPLC-PDA method provides better resolution of key purine metabolites, higher sensitivity with a smaller sample size and half the run time. Similar to prior methods, the new method appears well suited to the simultaneous analysis of the most abundant biologically relevant purines in biological samples.
Keywords: purine, UPLC-PDA, quantification, Biological sample, human diseases
Received: 26 Mar 2025; Accepted: 18 Jul 2025.
Copyright: © 2025 Fu, Sutcliffe, Dinasarapu, Hess and Jinnah. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence:
Rong Fu, HPLC Bioanalytical Core, School of Medicine, Emory University, Atlanta, United States
H. A. Jinnah, Department of Neuroglogy, Human Genetics, and Pediatrics, Emory University, Atlanta, United States
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