Carru, Ciriaco and Zinellu, Angelo and Sotgia, Salvatore and Serra, Roberta and Usai, Maria Franca and Pintus, Gianfranco and Pes, Giovanni Mario and Deiana, Luca (2004) A new HPLC method for serum neopterin measurement and relationships with plasma thiols levels in healthy subjects. Biomedical Chromatography, Vol. 18 (6), p. 360-366. eISSN 1099-0801. Article.
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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 fluorimetric detection. After TCA serum protein precipitation, the supernatant was diluted five times, injected into a C18 reversed-phase column and eluted at a flow rate of 1.5 mL/min by an isocratic water-acetonitrile (99:1) mobile phase. The natural fluorescence 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.
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