# Quantitative Analysis of Labile Metabolites in Biological Samples

> **NIH NIH R01** · UNIVERSITY OF WASHINGTON · 2024 · $370,650

## Abstract

PROJECT SUMMARY
Coenzymes and antioxidants mediate hundreds of biochemical reactions and are fundamental to cellular and
mitochondrial function. The redox coenzymes undergo reversible oxidation and reduction reactions, while the
balance between oxidized and reduced forms drive important cellular functions including regulation of ion
channels, cell signaling, cell survival and death. Cellular dysfunction associated with these coenzymes has been
implicated in many diseases including cancer, heart failure, diabetes, obesity and aging. Given the importance
of NAD metabolites and their declining levels in aging, numerous clinical trials of nutritional supplementation of
the coenzymes' precursors are currently underway. Despite the immense interest and the need to determine
cellular and subcellular levels of these metabolites, no reliable method exists currently for their simultaneous and
comprehensive measurement. The major challenge is that these molecules are unstable and their levels are
sensitive to sample harvesting, extraction and measurement conditions. As a result, errors involved with their
measurement using conventional methods often outweigh biological variations and potentially lead to incorrect
inferences. To overcome these challenges, and building on our preliminary studies of methodological
developments using nuclear magnetic resonance (NMR) spectroscopy, in this proposal, we seek to develop
methods to reliably measure the coenzymes and antioxidants in blood, cells, tissue as well as two subcellular
components: mitochondria and cytoplasm. Further, we seek to develop methods to measure these coenzymes
in live cells and mitochondria in real time. We will also translate the protocols and measurements to the widely
used mass spectrometry platform for broader dissemination. The proposed project has three main aims: (1)
Develop methods to reliably measure the coenzymes and antioxidants in blood, tissue, cells and two subcellular
components: mitochondria and cytoplasm using NMR spectroscopy; (2) Develop methods to measure the
coenzymes and antioxidants in live cells and mitochondria in real time using NMR spectroscopy; and (3) use
NMR spectroscopy to guide the translation of the methods for analysis of the unstable (coenzymes and
antioxidants) as well stable metabolites to mass spectrometry for wider applications in the metabolomics field.
The outcome will provide robust methods to analyze important coenzymes and antioxidants in a broad range of
biological sample types that can be used by many researchers. The new methods will also provide novel avenues
for investigation of live cells and mitochondrial metabolism in real time. These developments will impact
numerous areas of biomedical science.

## Key facts

- **NIH application ID:** 10844620
- **Project number:** 5R01GM138465-04
- **Recipient organization:** UNIVERSITY OF WASHINGTON
- **Principal Investigator:** G. A. Nagana Gowda
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $370,650
- **Award type:** 5
- **Project period:** 2021-06-01 → 2026-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10844620

## Citation

> US National Institutes of Health, RePORTER application 10844620, Quantitative Analysis of Labile Metabolites in Biological Samples (5R01GM138465-04). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10844620. Licensed CC0.

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