Urinary neopterin quantification by reverse-phase high-performance liquid ultraviolet chromatography with detection

Urinary neopterin quantification by reverse-phase high-performance liquid ultraviolet chromatography with detection

Author Castro, M. R. de Google Scholar
Di Marco, G. S. Google Scholar
Arita, D. Y. Google Scholar
Teixeira, L. C. Google Scholar
Pereira, A. B. Google Scholar
Casarini, D. E. Google Scholar
Institution Universidade Federal de São Paulo (UNIFESP)
Abstract Neopterin plays an important role in the malignant disease diagnostics. However, the methods employed in neopterin determination are generally difficult and/or time consuming. the aim of this work was to standardize a practical method to quantify neopterin using high-performance liquid chromatography-ultraviolet (HPLC-UV) and quantify it in patients with systemic lupus erythematosus (SLE). Urine was collected from healthy subjects (n=49), patients with inactive (n = 15), active (n = 28), and highly active SLE (it = 6). the HPLC was performed using two coupled reverse-phase columns eluted with 150 mM sodium phosphate, pH 4.0, under a flow rate of 0.8 ml/ min, with UV detector set at 353 nm and 100-fold diluted urines. the inter- and intra-assay studies presented an imprecision of 12.5% and 12.9% for quality controls of 3.94 and 1.1 mumol/ml, respectively. Recovery from 79.5% to 82% was observed throughout the assay's linear range. Subjects with active (874.2 +/- 165.3 8 mumol/mol creatinin) and highly active SLE (1753.8 +/- 453.9 mumol/mol creatinin) showed three- and sixfold increased neopterin levels, respectively, compared to subjects with inactive SLE (314.3 mu 121.3 mumol/mol creatinin) and healthy subjects (294.6 +/- 178.6 mumol/mol creatinin) (P > 0.05). Briefly, the proposed method was precise, specific, and reproducible, not invasive and allows the urinary neopterin quantification only with UV detection, (C) 2004 Elsevier B.V. All rights reserved.
Keywords neopterin
HPLC
UV detection
reverse phase
urine
systemic lupus erythematosus
Language English
Date 2004-06-30
Published in Journal of Biochemical and Biophysical Methods. Amsterdam: Elsevier B.V., v. 59, n. 3, p. 275-283, 2004.
ISSN 0165-022X (Sherpa/Romeo, impact factor)
Publisher Elsevier B.V.
Extent 275-283
Origin http://dx.doi.org/10.1016/j.jbbm.2004.03.004
Access rights Closed access
Type Article
Web of Science ID WOS:000222144700008
URI http://repositorio.unifesp.br/handle/11600/27796

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