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Determination of ascorbic acid in human plasma with a view to stability using HPLC with UV detection
Year: 2008
Type of publication: článek v odborném periodiku
Name of source: Journal of Separation Science
Publisher name: Wiley-VCH
Place: Weinheim
Page from-to: 3503-3508
Titles:
Language Name Abstract Keywords
cze Stanovení kyseliny askorbové v lidské plazmě pomocí HPLC-UV s ohledem na stabilitu Je zde popsána metoda stanovení kyseliny askorbové pomocí HPLC s UV detekcí, kde je kladen důraz na vhodnou precipitaci proteinů s ohledem na stabilitu a výtěžnost. Byla testována různá precipitační činidla. Byla zjišťována stabilita kyseliny askorbové ve vzorcích přes více jak 10 hodin. Při precipitaci pomocí kyseliny metafosforečné byla kyselina askorbová při 4 °C stabilní po dobu deseti hodin (s poklesem 1,8% pro standardní roztok a 2,8% pro plazmu). Kyselina chloristá byla jako precipitační činidlo nevhodná (s poklesem 36,0% pro standardní roztok a 7,3% pro plazmu). Analytické parametry metody byly vyhovující. Koeficienty variace pro přesnost v sérii a mezi sériemi byly 2,1% (n = 10) a 5,8% (n = 12). Kalibrační křivka byla lineární v rozsahu 2,0 ? 250,0 umol/l. Výtěžnost metody byla 96,1% s CV = 4,8% (n = 6) a LOD byla 3 umol/l. Referenční hodnoty se pohybovaly v rozmezí 50,8?22,4 umol/l. Prezentovaná metoda je vysoce citlivá a reprodukovatelná. Kyselina askorbová; Dithiothreitol; HPLC s UV detekcí; Deproteinační činidla; Krátkodobá a dlouhodobá stabilita
eng Determination of ascorbic acid in human plasma with a view to stability using HPLC with UV detection A method for the measurement of ascorbic acid using HPLC with UV detection and investigation into the protein precipitation techniques with regard to stability and recovery are described. The effectiveness of various protein precipitants was tested. Stability of ascorbic acid samples for analysis was investigated over 10 h. Ascorbic acid samples extracted with metaphosphoric acid were stable on a cooled autosampler (4 °C) for at least 10 h (with a decline of 1.8% for ascorbic acid solution and 2.8% for plasma). Perchloric acid as protein precipitant for ascorbic acid was unsuitable (with a decline of 36.0% for ascorbic acid solution and 7.3% for plasma). Analytical performance of this method is satisfactory. The intra- and interassay coefficients of variation were 2.1% (n = 10) and 5.8% (n = 12), respectively. The calibration curve was linear with the tested range of 2.0?250.0 umol/L. The recovery was 96.1% with CV = 4.8% (n = 6) and the LOD was 3 umol/L. The preliminary reference ranges of ascorbic acid in a group of blood donors are 50.8?22.4 umol/L. This assay is a highly sensitive and reproducible HPLC method for the determination of ascorbic acid in human plasma. Ascorbic acid; Dithiothreitol; HPLC with ultraviolet detection; Protein precipitants; Short-term and long-term stability

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