IQUIR   05412
INSTITUTO DE QUIMICA ROSARIO
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
DETERMINATION OF MARKER PTERIDINS IN URINE BY HPLC WITH FLUORIMETRIC DETECTION AND SECOND-ORDER MULTIVARIATE CALIBRATION USING MCR-ALS.
Autor/es:
A. MUÑOZ DE LA PEÑA; A. ESPINOSA; F. CAÑADA; A. MANCHA; H. GOICOECHEA; M. DE ZAN; J. CULZONI
Reunión:
Congreso; VII Colloquium Chemiometricum Mediterraneum; 2010
Resumen:
A liquid chromatographic method has been developed, in combination with the MCR-ALS algorithm, for the simultaneous determination of marker pteridins in urine samples. A central composite design was applied to optimize the factors influencing the separation. A set of 15 calibration samples were randomly prepared, in a concentration range of 0.5-10.5 ng mL-1 for NEO, BIO and PT; 4.0-8.0 ng mL-1 for XAN and 0.5-4.5 ng mL-1 for ISO. The validation was carried out with urine samples from healthy adults (dilution 1:250). The optimized conditions were a mobile phase composition of citric buffer 10 mM at pH=5.44-acetonitrile (94.5/5.5, v/v). The flow rate was 1.0 mL min-1 and the oven temperature was maintained at 25 ºC. The detection system consisted of a fast-scanning spectrofluorimeter, which allows one to obtain second-order data matrices containing the fluorescence intensity as a function of retention time and emission wavelength. In this work, MCR-ALS was used to cope with coeluting interferences, on account of the second order advantage inherent to this algorithm, which, in addition, is able to handle data sets deviating from trilinearity. The developed approach enabled us to determine five pteridins, some of them with overlapped profiles, reducing the experimental time and reagent consumption. Ratio values for pteridins/CREA in urine, for infected children with different pathologies, are reported in this communication.