Cham B E, Roeser H P, Kamst T W
Department of Medicine, University of Queensland, Royal Brisbane Hospital, Brisbane, Australia.
Clin Chem. 1989 Dec;35(12):2285-9.
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.
我们描述了一种用于同时测定人血清中维生素K1和维生素E的高效液相色谱法。使用脱脂人血清（不含维生素K1和维生素E）制备这些维生素的标准溶液，十六烷基萘酸酯和α-生育酚乙酸酯分别作为维生素K1和维生素E的内标。一种利用液-液分配色谱的简单新颖分离方法用作制备性“净化”程序。十六烷基萘酸酯和维生素K1（柱后还原后）通过荧光检测，α-生育酚乙酸酯和维生素E通过紫外吸收检测。该测定方法的灵敏度（检测限）为每次进样维生素K1 30 pg，维生素E 5 ng。该方法具有特异性、精确性，且比先前描述的方法更快。维生素K1的批内和批间变异系数分别为8.1%和12.9%；维生素E分别为3.5%和6.0%。维生素K1和维生素E从血清和脱脂血清（标准品）中的分析回收率分别为80%和93%。新生儿血清中维生素K1的平均浓度为83 ng/L，维生素E为2.5 mg/L；正常血脂成年人的值分别为343 ng/L和7.9 mg/L，高脂血症成年人分别为541 ng/L和11.1 mg/L。
