Simultaneous liquid-chromatographic determination of vitamin K1 and vitamin E in serum

• Published in: Clinical chemistry (Baltimore, Md.) Vol. 35; no. 12; pp. 2285 - 2289
• Main Authors: Cham, BE, Roeser, HP, Kamst, TW
• Format: Journal Article
• Language: English
• Published: Washington, DC Am Assoc Clin Chem 01.12.1989; American Association for Clinical Chemistry
• Subjects: Adult; Analysis of complex biological substances; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Blood and biological fluids; Chromatography, High Pressure Liquid; Female; Fetal Blood - analysis; Fundamental and applied biological sciences. Psychology; Health Status; Humans; Hyperlipidemias - blood; Infant, Newborn; Reference Standards; Temperature; Vitamin E - blood; Vitamin E - standards; Vitamin K 1 - blood; Vitamin K 1 - standards; Human; Biological fluid; Newborn; Clinical biology; Simultaneous measurement; HPLC chromatography; Adult; Lipids; Serum; Tocopherol; Quantitative analysis; Vitamin K
• ISSN: 0009-9147; 1530-8561
• DOI: 10.1093/clinchem/35.12.2285

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.