Determination of pyridinium crosslinks in plasma and serum by high-performance liquid chromatography

Gianalfredo Abbiati, Massimo Rigoldi, Serena Frignani, Laura Colombo, Emilio Mussini

Research output: Contribution to journalArticle

Abstract

A chromatographic method for the determination of pyridinoline (Pyr) and deoxypyridinoline (Dpyr) in serum and plasma is described. The analytical procedure involved plasma or serum purification by ultrafiltration (20 000 relative molecular mass cut-off) under centrifugation at 2500 g for 4 h, as an innovative step. Analysis was done by isocratic high-performance liquid chromatography with fluorescence detection. The linearity of the method was tested from 0.6 to 15 pmol/ml and 0.12 to 3 pmol/ml for Pyr and Dpyr, respectively. The detection limit was 60 fmol/ml for both crosslinks. Except for Dpyr in plasma (coefficient of variation 19.9%), intra-assay variation was always below 10% in serum and plasma. The method has been applied to the quantification of crosslinks in serum and plasma of healthy volunteers and also in mouse and rat plasma. Serum proved to be the most suitable biological fluid for the systemic measurement of these compounds in humans and under the experimental conditions used, contained an average of 3.62 ± 0.65 and 0.7 ± 0.18 pmol/ml Pyr and Dpyr, respectively.

Original languageEnglish
Pages (from-to)303-310
Number of pages8
JournalJournal of Chromatography B: Biomedical Sciences and Applications
Volume656
Issue number2
DOIs
Publication statusPublished - Jun 17 1994

ASJC Scopus subject areas

  • Chemistry(all)

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