期刊
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
卷 402, 期 4, 页码 1635-1641出版社
SPRINGER HEIDELBERG
DOI: 10.1007/s00216-011-5619-6
关键词
Homocitrulline; Carbamylation; Plasma; LC-MS/MS
资金
- Committee of American Memorial Hospital of Reims, France
Homocitrulline (HCit), an amino acid formed by the carbamylation of epsilon-amino groups of lysine residues, is considered a promising biomarker for monitoring diseases such as chronic renal failure and atherosclerosis. This paper describes a tandem mass spectrometric method for total, protein-bound and free HCit measurement in plasma samples. HCit was separated from other plasma components by hydrophilic interaction liquid chromatography. Detection was achieved by monitoring transitions of 190.1 > 127.1 and 190.1 > 173.1 for HCit, and 183.1 > 120.2 for d(7)-citrulline used as internal standard. This method allowed HCit quantification within 5.2 min and was precise (inter-assay CV < 5.85%), accurate (mean recoveries ranging from 97% to 106%), and exhibited a good linearity from 10 nmol/L to 1.6 mu mol/L. Plasma samples from control and uremic mice (n=10) were analyzed. In control mice, mean total plasma HCit concentration was 0.78 +/- 0.12 mu mol/mol amino acids, whereas it was increased 2.7-fold in uremic mice plasma, reaching 2.10 +/- 0.50 mu mol/mol amino acids (p < 0.001). In conclusion, this method exhibits good analytical performances and meets the criteria of sensitivity suitable for HCit concentration assessment in plasma samples.
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