Enzymatic methods for the determination of alpha-glycerophosphate and alpha-glycerophosphate oxidase with an automated FIA system

dc.contributor.authorKiranas, E. R.en
dc.contributor.authorKarayannis, M. I.en
dc.contributor.authorTzouwara-Karayanni, S. M.en
dc.date.accessioned2015-11-24T16:57:47Z
dc.date.available2015-11-24T16:57:47Z
dc.identifier.issn0039-9140-
dc.identifier.urihttps://olympias.lib.uoi.gr/jspui/handle/123456789/10619
dc.rightsDefault Licence-
dc.subjectglycerol-3-phosphate oxidaseen
dc.subjectperoxidaseen
dc.subjectalpha-glycerophosphateen
dc.subjectimmobilizationen
dc.subjectfiaen
dc.subjectflow-injection analysisen
dc.subjectglycerolen
dc.subjectserumen
dc.titleEnzymatic methods for the determination of alpha-glycerophosphate and alpha-glycerophosphate oxidase with an automated FIA systemen
heal.abstractA 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.en
heal.accesscampus-
heal.fullTextAvailabilityTRUE-
heal.identifier.secondary<Go to ISI>://000072093700031-
heal.identifier.secondaryhttp://ac.els-cdn.com/S0039914097002105/1-s2.0-S0039914097002105-main.pdf?_tid=d15b401255620c8c709163c27350bff5&acdnat=1335370664_aebca99b541d91fb1f4f90367840fe69-
heal.journalNameTalantaen
heal.journalTypepeer reviewed-
heal.languageen-
heal.publicationDate1998-
heal.recordProviderΠανεπιστήμιο Ιωαννίνων. Σχολή Θετικών Επιστημών. Τμήμα Χημείαςel
heal.typejournalArticle-
heal.type.elΆρθρο Περιοδικούel
heal.type.enJournal articleen

Αρχεία

Φάκελος/Πακέτο αδειών

Προβολή: 1 - 1 of 1
Φόρτωση...
Μικρογραφία εικόνας
Ονομα:
license.txt
Μέγεθος:
1.74 KB
Μορφότυπο:
Item-specific license agreed upon to submission
Περιγραφή: