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

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Data
2004-06-30
Autores
Castro, M. R. de
Di Marco, G. S.
Arita, D. Y.
Teixeira, L. C.
Pereira, A. B.
Casarini, D. E.
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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.
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Journal of Biochemical and Biophysical Methods. Amsterdam: Elsevier B.V., v. 59, n. 3, p. 275-283, 2004.