title: HPLC determination of serum pteridine pattern as biomarkers.
abstract: Pteridinic derivatives are important biomolecules considered as biomarkers for several diseases, especially in cancer and infectious pathologies. A new fluorimetric-HPLC method for the analysis of nine pteridines in human serum has been reported. Two analytical columns composed by C18 porous and fused core particles were assayed and the results compared. Fused core particle column allows us adequate separation, in only one run and in 15 min. Acid precipitation step of the proteins and clean-up process with an Isolute ENV+ (hydroxylated polystyrene-divinylbenzene copolymer) cartridge of the serum samples have been optimized. Analytes were determined by fluorimetric detection, exciting at 272 nm and measuring the fluorescence emission at 410 nm for isoxanthopterin, at 465 nm for xanthopterin, and at 445 nm for the analysis of the other pteridines. Detection limits between 0.07 and 0.61 ng mL(-1) were calculated according to Clayton criterium. Intraday precision varied from 1.2 to 5.3 and interday precision between 1.2 and 7.4, both expressed as RSD (%). External standard and standard addition calibrations were compared in the analysis of serum samples. The pteridine amounts in serum (expressed as ng mL(-1) ± confidence interval) were 3.69 ± 1.78; 1.35 ± 0.24; 0.46 ± 0.14; 0.54 ± 0.24; 0.84 ± 0.55; 2.10 ± 0.51 and 0.23 ± 0.11 for XAN, NEO, MON, ISO, BIO and 6HMPT, respectively, using the external standard method. Comparable results were obtained by the standard addition method. It is noticeable that 7BIO was not detected in the healthy serum samples analyzed.
authors: Department of Analytical Chemistry, University of Extremadura, Avd. de Elvas, s/n, 06006 Badajoz, Spain.
authors: E
authors: E
authors: Martín Tornero
authors: Department of Analytical Chemistry, University of Extremadura, Avd. de Elvas, s/n, 06006 Badajoz, Spain.
authors: I
authors: I
authors: Durán Merás
authors: Department of Analytical Chemistry, University of Extremadura, Avd. de Elvas, s/n, 06006 Badajoz, Spain. Electronic address: nuncy@unex.es.
authors: A
authors: A
authors: Espinosa-Mansilla
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D015415:Biomarkers / Q000097:blood*; D002138:Calibration; D002648:Child; D002675:Child, Preschool; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005470:Fluorometry / Q000379:methods*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007223:Infant; D007231:Infant, Newborn; D008875:Middle Aged; D011621:Pteridines / Q000097:blood*; D012016:Reference Values; D015203:Reproducibility of Results; D055815:Young Adult
chemical_list: D015415:Biomarkers; D011621:Pteridines
keywords: Fluorescence detection; Fused core reverse column; HPLC; Human serum; Pteridines