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A new HPLC method for serum neopterin measurement and relationships with plasma thiols levels in healthy subjects
Author(s) -
Carru Ciriaco,
Zinellu Angelo,
Sotgia Salvatore,
Serra Roberta,
Usai Maria Franca,
Pintus Gian Franco,
Pes Giovanni Mario,
Deiana Luca
Publication year - 2004
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.325
Subject(s) - neopterin , chemistry , chromatography , high performance liquid chromatography , protein precipitation , elution , fluorescence , medicine , physics , quantum mechanics
Abstract Neopterin, a pyrazinopyrimidine compound, serves as a marker of cellular immune system activation, and it can be used as a prognostic predictor for certain types of diseases. We propose a new simple HPLC method to measure serum neopterin with highly sensitive uorimetric detection. After TCA serum protein precipitation, the supernatant was diluted ve times, injected into a C 18 reversed‐phase column and eluted at a ow rate of 1.5 mL/min by an isocratic water–acetonitrile (99:1) mobile phase. The natural uorescence of the molecule was detected at excitation wavelength 353 nm and emission 438 nm. In these conditions the neopterin retention time was about 4 min. Our proposed method was compared with a validated chromatographic separation, and the obtained data of the serum neopterin from 35 healthy volunteers were analysed by Passing–Bablok regression and Bland–Altman test. Neopterin measurement in healthy subjects was also employed to investigate on its potential relationships with plasma thiols levels. Copyright © 2004 John Wiley & Sons, Ltd.