Measuring NAD(+) levels in mouse blood and tissue samples via a surrogate matrix approach using LC-MS/MS.
Bioanalysis
confidence
Key findings
Describes an LC-MS/MS method and sample collection procedure for quantifying NAD(+) in mouse blood and tissue; no clinical/biological endpoints reported.
View source on PubMed (PMID 25046046) ↗
- Sample size
- Not reported
- Population
- Mouse blood and tissue samples
- Dosing
- Oral administration of GNE-617 (nicotinamide phosphoribosyltransferase inhibitor)
- Duration
- Not reported
- Route
- Oral
- Blinding
- not_reported
- Controls
- none
- Drug class
- coenzyme
Full abstract
NAD(+) is an endogenous analyte and is unstable during blood sample collection, both of which present obstacles for quantitation. Moreover, current procedures for NAD(+) sample collection require onsite treatment with strong acid to stabilize the NAD(+) in mouse blood cells. NAD(+) can be stabilized by addition of acid before the frozen mouse blood sample was thawed. A simple sample collection procedure was proposed to facilitate the analysis of NAD(+) in mouse blood and tissue samples. A LC-MS/MS method was developed for quantifying NAD(+) in mouse blood and various tissue samples. The described method was used to measure endogenous NAD(+) levels in mouse blood following oral administration of the nicotinamide phosphoribosyltransferase inhibitor GNE-617. This study presents a suitable assay and sample collection procedure for high throughput screening of NAD(+) samples in preclinical discovery studies.