Cimadevilla H.M.,University of Oviedo |
Hevia D.,University of Oviedo |
Hevia D.,Instituto Universitario Oncologico Del Principado Of Asturias |
Miar A.,University of Oviedo |
And 7 more authors.
Biomedical Chromatography | Year: 2015
A straightforward and common analytical method for α-tocopherol (αT) determination in various biological samples, including plasma, red blood cells (RBC), tissues and cultured cell lines, was developed and validated, using a reverse phase-chromatographic method (RP-HPLC). Even though many chromatographic methods for αT determination have been reported, most of them require readjustment when applied to different types of samples. Thus, an effective and simple method for αT determination in different biological matrices is still necessary, specifically for translational research. This method was applied using a C18 column (250×4.6mm, 5μm particle size) under isocratic elution with MeOH:ACN:H2O (90:9:1v/v/v) at a flow rate of 1mL/min and detected using photodiode array at 293nm. Linearity (r >0.9997) was observed for standard calibration with inter- and intraday variation of standard <4%. Lower limits of detection and quantification for αT in this assay were 0.091 and 0.305μg/mL respectively. Validation proved the method to be selective, linear, accurate and precise. The method was successfully applied in great variety of biological samples, that is, human and mouse plasma, RBCs, murine tissues and human/mouse/rat cultured cell lines. More importantly, a single protocol of extraction and detection can be applied, making this method very convenient for standardization of different types of samples. © 2014 John Wiley & Sons, Ltd.