A new HPLC method for serum neopterin measurement and relationships with plasma thiols levels in healthy subjects |
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Authors: | Carru Ciriaco Zinellu Angelo Sotgia Salvatore Serra Roberta Usai Maria Franca Pintus Gian Franco Pes Giovanni Mario Deiana Luca |
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Institution: | Chair of Clinical Biochemistry, University of Sassari, Sassari, Italy. carru@uniss.it |
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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 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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Keywords: | high‐performance liquid chromatography neopterin thiols homocysteine cysteine cardiovascular risk |
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