Simultaneous liquid-chromatographic determination of vitamin K1 and vitamin E in serum.
We describe a high-performance liquid chromatographic procedure for the simultaneous measurement of vitamins K1 and E in human serum. Delipidated human serum (free of vitamins K1 and E) was used to make standard solutions of these vitamins, and cetyl naphthoate and alpha-tocopheryl acetate were the internal standards for vitamin K1 and vitamin E, respectively. A simple, novel separation method utilizing liquid-liquid partition chromatography was used as a preparative "clean-up" procedure. Cetyl naphthoate and vitamin K1 (after post-column reduction) were detected by fluorescence, alpha-tocopheryl acetate and vitamin E by ultraviolet absorption. Sensitivity (detection limit) of the assay was 30 pg for vitamin K1 and 5 ng for vitamin E per injection. The method is specific, precise, and more rapid than previously described procedures. Within- and between-assay CVs were 8.1% and 12.9%, respectively, for vitamin K1; 3.5% and 6.0%, respectively, for vitamin E. Analytical recoveries of vitamins K1 and E were 80% and 93%, respectively, from serum and from delipidated serum (standards). The average neonatal serum concentration of vitamin K1 was 83 ng/L, 2.5 mg/L for vitamin E; for normolipidemic adults, the values were 343 ng/L and 7.9 mg/L, respectively, and for hyperlipidemic adults, 541 ng/L and 11.1 mg/L, respectively.
Cham
B E
BE
Department of Medicine, University of Queensland, Royal Brisbane Hospital, Brisbane, Australia.
Roeser
H P
HP
Kamst
T W
TW
D000328:Adult; D002851:Chromatography, High Pressure Liquid; D005260:Female; D005312:Fetal Blood / Q000032:analysis; D006304:Health Status; D006801:Humans; D006949:Hyperlipidemias / Q000097:blood; D007231:Infant, Newborn; D012015:Reference Standards; D013696:Temperature; D014810:Vitamin E / Q000097:blood* / Q000592:standards; D010837:Vitamin K 1 / Q000097:blood* / Q000592:standards
D014810:Vitamin E; D010837:Vitamin K 1