Development and validation of an LC-MS/MS method for detection and quantification of in vivo derived metabolites of [Pyr1]apelin-13 in humans
| Autor: | Lutz Jermutus, Janet J. Maguire, Duuamene Nyimanu, Joseph Cheriyan, Richard G. Kay, Philip Ambery, Frank Reimann, Anthony P. Davenport, Rhoda E. Kuc, Fiona M. Gribble, Petra Sulentic |
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| Přispěvatelé: | Nyimanu, Duuamene [0000-0002-6212-9518], Reimann, Frank [0000-0001-9399-6377], Davenport, Anthony P [0000-0002-2096-3117], Apollo - University of Cambridge Repository |
| Rok vydání: | 2019 |
| Předmět: |
Adult
Male 0301 basic medicine Gene isoform Hydrochloride lcsh:Medicine Peptide 030204 cardiovascular system & hematology Cleavage (embryo) Plasma 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Tandem Mass Spectrometry In vivo Humans Protein Isoforms Guanidine lcsh:Science chemistry.chemical_classification Apelin Receptors Chromatography Multidisciplinary fungi lcsh:R Reproducibility of Results Metabolism Middle Aged Healthy Volunteers Apelin 3. Good health 030104 developmental biology chemistry Intercellular Signaling Peptides and Proteins Female lcsh:Q Peptides Chromatography Liquid |
| Zdroj: | Scientific Reports Scientific Reports, Vol 9, Iss 1, Pp 1-12 (2019) |
| Popis: | [Pyr1]apelin-13 is the predominant apelin peptide isoform in the human cardiovascular system and plasma. To date, few studies have investigated [Pyr1]apelin-13 metabolism in vivo in rats with no studies examining its stability in humans. We therefore aimed to develop an LC-MS/MS method for detection and quantification of intact [Pyr1]apelin-13 and have used this method to identify the metabolites generated in vivo in humans. [Pyr1]apelin-13 (135 nmol/min) was infused into six healthy human volunteers for 120 minutes and blood collected at time 0 and 120 minutes after infusion. Plasma was extracted in the presence of guanidine hydrochloride and analysed by LC-MS/MS. Here we report a highly sensitive, robust and reproducible method for quantification of intact [Pyr1]apelin-13 and its metabolites in human plasma. Using this method, we showed that the circulating concentration of intact peptide was 58.3 ± 10.5 ng/ml after 120 minutes infusion. We demonstrated for the first time that in humans, [Pyr1]apelin-13 was cleaved from both termini but the C-terminal was more susceptible to cleavage. Consequently, of the metabolites identified, [Pyr1]apelin-13(1–12), [Pyr1]apelin-13(1–10) and [Pyr1]apelin-13(1–6) were the most abundant. These data suggest that apelin peptides designed for use as cardiovascular therapeutics, should include modifications that minimise C-terminal cleavage. |
| Databáze: | OpenAIRE |
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