A new HPLC method for serum neopterin measurement and relationships with plasma thiols levels in healthy subjects.
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.
Chair of Clinical Biochemistry, University of Sassari, Sassari, Italy. carru@uniss.it
Ciriaco
C
Carru
Angelo
A
Zinellu
Salvatore
S
Sotgia
Roberta
R
Serra
Maria Franca
MF
Usai
Gian Franco
GF
Pintus
Giovanni Mario
GM
Pes
Luca
L
Deiana
D000328:Adult; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D019075:Electrophoresis, Capillary / Q000379:methods; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D019798:Neopterin / Q000097:blood*; D012016:Reference Values; D013050:Spectrometry, Fluorescence; D013438:Sulfhydryl Compounds / Q000097:blood*
D013438:Sulfhydryl Compounds; D019798:Neopterin