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alliedacademies
September 20-22, 2017 | Toronto, Canada
10 TH AMERICAN PEDIATRICS HEALTHCARE &
PEDIATRIC INFECTIOUS DISEASES CONGRESS
Pediatric Healthcare & Pediatric Infections 2017
In humans, genetic defects of the synthesis or regeneration
of tetrahydrobiopteridin (BH4), an essential cofactor
in hydroxylation reactions, are associated with severe
neurological disorders. The diagnosis of these conditions
relies on the determination of BH4, and its metabolites
and precursors in biological fluids. This paper describes a
reversed-phase high-performance liquid chromatographic
(HPLC), new method with fluorometric detection (ERRECI,
Milano) for precise and sensitive quantification in urine
of the pterins after oxidation: neopterin, monapterin,
isoxanthopterin, pterin, biopterin and primapterin. The HPLC
method employs a C18, 5-µm particle size analytical column
(250 mm×4.6 mm), 10-µL injection volume, column at room
temperature, excitation at 350 nm and emission at 450 nm,
and an acetate buffer mobile phase at a flow rate of 1.1 mL/
min. These conditions resolve the six molecules as well as
diluent peaks within 20 min. The method is linear for all the
six molecules (0.2-34.0 µmol/L). The detection limits were
<0.08 µmol/L at a signal-to-noise ratio of two. The relative
standard deviation was <5% (<10% only for monapterin)
for the within-assay imprecision (n=10) and <9% for the
between-assay imprecision (n=6). The recovery of different
amount (1.3-5.5 µ mol/L) of the six molecules added to
an urine sample was 92-108%. The specific and sensitive
method described may offer a means for determining
BH4 and five metabolites. The method is characterized by
high recovery and good reproducibility; it is well suited for
routine operation in every newborn with even slight but
persistent hyperphenylalaninemia unresponsive to a low-
phenylalanine diet.
Speaker Biography
A Barassi has completed her PhD from Insubria University School of Medicine (Varese,
Italy) and Post-doctoral studies from Insubria University School of Medicine (Varese,
Italy). She is working as an Assistant Professor of Clinical Biochemistry and Clinical
Molecular Biology, University of Milan from 2006 and Assistant of Laboratory of Clinical
Chemistry, San Paolo University Hospital, Milan, Italy from 2007. She has published
67 papers in reputed journals and has been serving as an Editorial Board Member of
repute.
e:
Determination of pterines as markers of hyperphenylalaninemias
Alessandra Barassi1, C A L Damele2, R Stefanelli2, M Di Dario2, L Massaccesi1 and G V Melzi d’Eril1
1Università degli Studi di Milano, Italy
2ASST Santi Paolo e, Italy