Key result
A novel LC-MS/MS method for determining cAMP and cGMP concentrations in washed platelet supernatants demonstrated high precision with a lower limit of quantification of 10 pg/mL.
Why the study?
Cyclic nucleotides regulate platelet function and inhibit platelet aggregation, requiring improved methods to determine cAMP and cGMP concentrations in washed platelet supernatants.
A novel, highly sensitive LC-MS/MS method improves the quantification of cAMP and cGMP in washed platelet samples, facilitating research on antiplatelet pharmacological targets.
Provides sensitive assay for platelet signaling in animal models; leaves open translation to human antiplatelet therapies.
Cyclic Nucleotides are important in regulating platelet function. Increases in 3'5'-cyclic adenosine monophosphate (cAMP) and 3'5'-cyclic guanosine monophosphate (cGMP) inhibit platelet aggregation and are pharmacological targets for antiplatelet therapy. Here we report an improved method for determining cAMP and cGMP concentrations and, for the first time, in washed platelet supernatants by combining high-performance liquid chromatography and tandem mass spectrometry (LC-MS/MS). Characteristic peaks of the substrates, cGMP or cAMP and their internal standards were identified in negative-ion electrospray ionisation using multiple reaction monitoring. Compared with previously reported methods, the method presented here shows high precision with the lowest lower limit of quantification (LLoQ) to date (10 pg/mL). The effect of a novel catecholamine, 6-nitrodopamine, on cyclic nucleotide levels was quantified. Our results showed that this new method was fast, sensitive, and highly reproducible.
No takes yet. Share an insight, caveat, or question.
Nash et al. (2022) studied this question. LC-MS/MS method for determining cAMP and cGMP concentrations vs. previously reported methods was evaluated on Quantification of cAMP and cGMP levels. A novel LC-MS/MS method for determining cAMP and cGMP concentrations in washed platelet supernatants demonstrated high precision with a lower limit of quantification of 10 pg/mL.
Synapse has enriched 5 closely related papers on similar clinical questions. Consider them for comparative context: