# Simultaneous Detection and Quantification of Adenine Nucleotides in Mammalian Cultured Cells by HPLC

## TL;DR

This paper introduces a new HPLC method to accurately measure adenine nucleotides in mammalian cells, which is important for understanding cellular metabolism and signaling.

## Contribution

A novel RP-HPLC method with UV–vis detection for simultaneous quantification of multiple adenine nucleotides in cultured cells.

## Key findings

- The developed RP-HPLC method is sensitive and capable of detecting physiological concentrations of adenine nucleotides.
- The method successfully detects changes in ATP, AMP, and NAD+ levels caused by pharmacological treatments.
- The approach provides a reliable and accessible alternative for quantifying adenine nucleotides in biological samples.

## Abstract

Adenine nucleotides,
including ATP, ADP, ADP-ribose, AMP, NAD+, and NADH, play
central roles in cellular homeostasis and
are involved in multiple metabolic and signaling pathways. Owing to
their broad functional relevance in cell biology, the accurate quantification
of these metabolites is essential for diverse research areas such
as bioenergetics, cell signaling, and cancer biology. Several analytical
methods have been described for the measurement of adenine nucleotides,
ranging from enzymatic assays to mass spectrometry-based approaches.
In this study, we developed a reverse-phase high-performance liquid
chromatography (RP-HPLC) method with UV–vis detection that
enables the simultaneous quantification of ATP, ADP, ADP-ribose, AMP,
NAD+, and NADH. This method is simple, sensitive within
the physiological concentration range of all analytes, and capable
of detecting biologically relevant changes in ATP, AMP, and NAD+ levels induced by pharmacological treatments. Therefore,
it presents an accessible and reliable alternative for the quantification
of these nucleotides in biological samples.