Quantification of Neuropeptide Y and Four of Its Metabolites in Human Plasma by Micro-UHPLC-MS/MS.

Details

Serval ID
serval:BIB_F64245F9B65C
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Quantification of Neuropeptide Y and Four of Its Metabolites in Human Plasma by Micro-UHPLC-MS/MS.
Journal
Analytical chemistry
Author(s)
Vocat C., Dunand M., Hubers S.A., Bourdillon N., Millet G.P., Brown N.J., Wuerzner G., Grouzmann E., Eugster P.J.
ISSN
1520-6882 (Electronic)
ISSN-L
0003-2700
Publication state
Published
Issued date
07/01/2020
Peer-reviewed
Oui
Volume
92
Number
1
Pages
859-866
Language
english
Notes
Publication types: Journal Article
Publication Status: ppublish
Abstract
Neuropeptide Y (NPY) is a 36-amino acid peptide circulating at a subpicomolar concentration participating in multiple physiological and pathological processes. NPY is prone to peptidolysis, generating metabolites with modified affinity for the five known receptors of NPY that mediate distinct effects. It is, therefore, crucial to distinguish each metabolite to understand the multiple functions of NPY. Since immunoassays are not able to distinguish NPY from its metabolites, we have validated a microliquid chromatography tandem mass spectrometry (micro-LC-MS/MS) assay for the quantification of endogenous NPY, NPY2-36, NPY3-36, NPY1-35, and NPY3-35 in human plasma. Sample preparation relies on immunoextraction in 96-well plates, followed by solid-phase extraction prior to micro-LC-MS/MS. The LLOQ ranged from 0.03 to 0.16 pM, intra- and inter-assay precision were <27% and trueness <22%. We determined reference intervals in 155 healthy volunteers and 40 hypertensive patients. We found that NPY3-36 is the main circulating peptide in resting conditions and that NPY and catecholamines are simultaneously increased during orthostasis. We also showed that the concentrations of NPY and its metabolites are similar in healthy volunteers and hypertensive patients. NPY is the prototype peptide that circulates in concentrations expected to be beyond instrumental capacities. We have been successful in developing a high-throughput specific and sensitive assay by including a deep knowledge of the physicochemical properties of these peptides to an efficient multistep sample preparation, and a micro-LC chromatography. We believe that our methodological approach opens the possibility to selectively quantify other endogenous peptides cleaved by peptidases whose concentrations are below 1 pM.
Pubmed
Web of science
Funding(s)
Other / Ligue suisse contre le cancer, KLS-4283-08-2017
Create date
21/01/2020 18:41
Last modification date
26/11/2020 7:24
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