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Determination of pyridoxal, pyridoxal 5'-phosphate and 4-pyridoxic acid in plasma of selected groups of the patients with a view to proper sample preparation using HPLC with fluorescence detection
Year: 2009
Type of publication: článek v odborném periodiku
Name of source: Scientific Papers of the University of Pardubice, Series A, Faculty of Chemical Technology
Publisher name: Univerzita Pardubice
Place: Pardubice
Page from-to: 73-85
Titles:
Language Name Abstract Keywords
cze Stanovení pyridoxalu, pyridoxal 5'-fosfátu a 4-pyridoxové kyseliny v plazmě vybraných skupin pacientů pomocí HPLC s fluorescenční detekcí s ohledem na správnou přípravu vzorku Je zde popsána metoda pro stanovení pyridoxalu, pyridoxal 5'-fosfátu a 4-pyridoxové kyseliny pomocí kapalinové chromatografie s fluorescenční detekcí s ohledem na stabilitu a výtěžnost. Pro separaci byla použita kolona Purospher RP-18e, 5 um. Mobilní fází byla směs ethanolu a 25 mmol/l Na2HPO4 (7:93, v/v), pH 6,0. Analytické parametry metody jsou vyhovující: koeficienty přesnosti v sérii a mezi sériemi byly pod 10%. Výtěžnost se pohybovala v intervalu 93,0-101,8%, kromě pyridoxalu (82,5-98,8%). Detekční limit byl 1,0 nmol/l pro pyridoxal 5'-fosfát, 0,7 nmol/l pro 4-pyridoxovou kyselinu a 1,1 nmol/l pro pyridoxal. Orientační referenční hodnoty pro pyridoxal 5'-fosfát, pyridoxal a 4-pyridoxovou kyselinu u skupiny dobrovolných dárců krve jsou 42,0-75,6 nmol/l, 2,6-12,8 nmol/l a 16,2-33,0 nmol/l. HPLC s fluorescenční detekcí; lidská plazma; pyridoxal; pyridoxal 5'-fosfát, 4-pyridoxová kyselina
eng Determination of pyridoxal, pyridoxal 5'-phosphate and 4-pyridoxic acid in plasma of selected groups of the patients with a view to proper sample preparation using HPLC with fluorescence detection A method for the measurement of pyridoxal, pyridoxal 5'-phosphate and 4-pyridoxic acid using a high-performance liquid chromatography with fluorescence detection with regard to stability and recovery is described. For the separation, a reverse phase column LiChroCart 125-4, Purospher RP-18e, 5 um, was used. The mixture of ethanol and 25 mmol/L Na2HPO4 (7:93, v/v), pH 6.0 was used as a mobile phase. The analytical performance of this method is satisfactory: the intra-assay and inter-assay coefficients of variation were below 10%. Quantitative recoveries from spiked plasma samples were at intervals 93.0-101.8%, except for pyridoxal (82.5-98.8%). The limit of detection was 1.0 nmol/L for pyridoxal 5'-phosphate, 0.7 nmol/L for 4-pyridoxic acid and 1.1 nmol/L for pyridoxal. The preliminary reference range of pyridoxal 5'-phosphate, pyridoxal and 4-pyridoxic acid in a group of blood donors are 42.0-75.6 nmol/L, 2.6-12.8 nmol/L and 16.2-33.0 nmol/L, respectively. HPLC with fluorescence detection; human plasma; pyridoxal, pyridoxal 5'-phosphate, 4-pyridoxic acid

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