Simultaneous determination of deuterated and non-deuterated alpha-tocopherol in human plasma by high-performance liquid chromatography.
Labelled tocopherol is used to evaluate its absorption by biodiscriminating the dietary intake from the endogenous tocopherol pool of subject. A normal-phase high-performance liquid chromatographic method is described for the easy separation and quantification of deuterated (d(6)) and non-deuterated alpha-tocopherol. The alpha-tocopherol isotopomers were extracted from plasma triacylglycerol-rich lipoproteins in hexane, separated by two EC Nucleosil columns in series with a mobile phase of hexane-isopropanol (659.34:0.786, w/w) running isocratically. The detection of d(6)-alpha-tocopherol was performed by its UV absorbance at 297 nm with a limit of detection of 34 pmol/ml, a limit of quantification of 83 pmol/ml and a range of determination of 34-9905 pmol/ml. Between- and within-assay RSDs were 2.4% (n=10) and 2.7% (n=5), respectively.
Richelle
Myriam
M
Department of Nutrition, Nestlé Research Center, PO Box 44, CH-1000 26, Lausanne, Switzerland. myriam.richelle@rdls.nestle.com
Tavazzi
Isabelle
I
Fay
Laurent Bernard
LB
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003903:Deuterium; D006801:Humans; D008297:Male; D012016:Reference Values; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet; D024502:alpha-Tocopherol / Q000097:blood*
D003903:Deuterium; D024502:alpha-Tocopherol