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Analysis of neuropeptide Y and its metabolites by high-performance liquid chromatography–electrospray ionization mass spectrometry and integrated sample clean-up with a novel restricted-access sulphonic acid cation exchanger
Kaunas University of Technology, Department of Organic Technology, Kaunas, Lithuania.
Bioanalytical Chemistry, AstraZeneca R&D Mölndal, Mölndal, Sweden.
Institut für Anorganische Chemie und Analytische Chemie, Johannes Gutenberg Universität, Mainz, Germany.
Merck KGaA, Specialty Laboratory Products, Darmstadt, Germany.
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2000 (English)In: Journal of Chromatography A, ISSN 0021-9673, E-ISSN 1873-3778, Vol. 890, no 1, p. 135-144Article in journal (Refereed) Published
Abstract [en]

A novel restricted access cation exchanger with sulphonic acid groups at the internal surface was proven to be highly suitable in the sample clean up of peptides on-line coupled to HPLC–electrospray ionization (ESI)-MS. Neuropeptide Y (NPY) and several of its fragments in plasma were subjected to the sample clean-up procedure. The peptides were eluted by a step gradient from the restricted access column, applying 10 mM phosphate buffer pH 3.5 from 5 to 20% (v/v) of acetonitrile with 1 M NaCl and transferred to a Micra ODS II column (33×4.6 mm). The separation of the peptides and their fragments was performed by a linear gradient from 20 to 60% (v/v) acetonitrile in water with 0.1% formic acid and 0.01% trifluoroacetic acid in 4 min at a flow-rate of 0.75 ml/min. An integrated and completely automated system composed of sample clean up–HPLC–ESI-MS was used to analyze real life samples. The sample volumes ranged between 20 and 100 μl. Peaks due to the fragments NPY 1–36, 3–36 and 13–36 in porcine plasma were identified by ESI-MS. The limit of detection was in the 5 nmol/ml range. The total analysis required 21 min and allowed the direct injection of plasma. © 2000 Elsevier Science B.V. All rights

Place, publisher, year, edition, pages
Amsterdam: Elsevier B.V , 2000. Vol. 890, no 1, p. 135-144
Keywords [en]
Restricted-access media, Sample handling, Column switching, Neuropeptides, Peptides
National Category
Analytical Chemistry
Identifiers
URN: urn:nbn:se:hh:diva-24121DOI: 10.1016/S0021-9673(00)00639-7ISI: 000088736900014PubMedID: 10976801Scopus ID: 2-s2.0-0343962626OAI: oai:DiVA.org:hh-24121DiVA, id: diva2:679543
Available from: 2013-12-16 Created: 2013-12-09 Last updated: 2018-03-22Bibliographically approved

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Lutz, Mareike

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