Enzymatic methods for the determination of alpha-glycerophosphate and alpha-glycerophosphate oxidase with an automated FIA system
Φόρτωση...
Ημερομηνία
Συγγραφείς
Kiranas, E. R.
Karayannis, M. I.
Tzouwara-Karayanni, S. M.
Τίτλος Εφημερίδας
Περιοδικό ISSN
Τίτλος τόμου
Εκδότης
Περίληψη
Τύπος
Είδος δημοσίευσης σε συνέδριο
Είδος περιοδικού
peer reviewed
Είδος εκπαιδευτικού υλικού
Όνομα συνεδρίου
Όνομα περιοδικού
Talanta
Όνομα βιβλίου
Σειρά βιβλίου
Έκδοση βιβλίου
Συμπληρωματικός/δευτερεύων τίτλος
Περιγραφή
A procedure for the enzymatic determination of alpha-glycerophosphate (alpha-GP) has been developed, using an automated in-house FIA system, with immobilized glycerol-3-phosphate oxidase (GPO) on non-porous glass beads, following optimization of the immobilization and analytical parameters. Fabricated single bead string reactors (SBSR) were used in connection with the FIA system, following optimization of its parameters. The half-life of GPO-SBSR regarding reduction of the enzyme activity was found to be 110 days for its use in 20 triplicate measurements daily and storage at 4 degrees C in the appropriate buffer. The regression equation of the calibration graph for the determination of alpha-GP was: A(max) = (10 +/- 2) x 10(-4) + (22134 +/- 12) x 10(-4) (mmol l(-1) alpha-GP). The lower limit of quantitation was 0.74 mu mol l(-1) alpha-GP and the RSD of the method 0.05% (r = 0.9999). The same FIA system and procedure can be also used for the determination of the GPO activity, with the alpha-GP as substrate. The regression equation for this calibration graph was: A(max) = (23 +/- 18) x 10(-4) + (190 +/- 1) x 10(-4) (mu g ml(-1) GPO), the lower limit of quantitation was 0.782 x 10(-3) mg ml(-1) (0.782 ppm) GPO and the RSD of the method 0.53% (r = 0.9999). Serum samples obtained from hospitalized patients were deproteinized by gel filtration and analyzed under pseudo-first order conditions, at various concentrations of alpha-GP. A kinetic study of the reduction of alpha-GP in serum versus time is given and an observed reaction rate constant k(ob) = 106.5 x 10(-4) min(-1) was determined. (C) 1998 Elsevier Science B.V.
Περιγραφή
Λέξεις-κλειδιά
glycerol-3-phosphate oxidase, peroxidase, alpha-glycerophosphate, immobilization, fia, flow-injection analysis, glycerol, serum
Θεματική κατηγορία
Παραπομπή
Σύνδεσμος
<Go to ISI>://000072093700031
http://ac.els-cdn.com/S0039914097002105/1-s2.0-S0039914097002105-main.pdf?_tid=d15b401255620c8c709163c27350bff5&acdnat=1335370664_aebca99b541d91fb1f4f90367840fe69
http://ac.els-cdn.com/S0039914097002105/1-s2.0-S0039914097002105-main.pdf?_tid=d15b401255620c8c709163c27350bff5&acdnat=1335370664_aebca99b541d91fb1f4f90367840fe69
Γλώσσα
en
Εκδίδον τμήμα/τομέας
Όνομα επιβλέποντος
Εξεταστική επιτροπή
Γενική Περιγραφή / Σχόλια
Ίδρυμα και Σχολή/Τμήμα του υποβάλλοντος
Πανεπιστήμιο Ιωαννίνων. Σχολή Θετικών Επιστημών. Τμήμα Χημείας