Lambert W E, De Leenheer A P, Baert E J
Anal Biochem. 1986 Nov 1;158(2):257-61. doi: 10.1016/0003-2697(86)90546-4.
The reference interval for serum vitamin K1(20) levels was assayed in healthy fasting adults by a method based on high-performance liquid chromatography (HPLC). The isolation procedure involves a solvent extraction of the plasma lipids followed by two chromatographic steps, consisting of a purification of the extract on a semipreparative adsorption column and a final quantitation on a reverse-phase column. Vitamin K1(20) and vitamin K1(25), the internal standard, are monitored by fluorescence detection after postcolumn reduction with a methanolic solution of tetramethylammonium octahydridotriborate. This reaction is performed in an open tubular reaction coil at elevated temperature. The median plasma concentration in 50 healthy fasting adults was 247 pg/ml. The levels showed a skewed distribution with a range of 62 to 980 pg/ml [log x +/- 2 SD (log x)]. The method is linear over the entire physiological range and has a within-run precision of 3.6% (n = 5, mean = 311 pg/ml). The minimum detectable amount in serum is 50 pg/ml. Other extraction procedures resulted in lower recoveries or in interferences in the final measurement. The vitamin K1(20) levels as reported by other research groups are also discussed.
采用基于高效液相色谱法（HPLC）的方法，对健康空腹成年人的血清维生素K1(20)水平的参考区间进行了测定。分离过程包括对血浆脂质进行溶剂萃取，随后进行两个色谱步骤，即在半制备吸附柱上对提取物进行纯化，并在反相柱上进行最终定量。维生素K1(20)和内标维生素K1(25)在用八氢硼酸四甲铵甲醇溶液进行柱后还原后，通过荧光检测进行监测。该反应在开放管式反应盘管中于高温下进行。50名健康空腹成年人的血浆中位浓度为247 pg/ml。这些水平呈偏态分布，范围为62至980 pg/ml [log x +/- 2 SD (log x)]。该方法在整个生理范围内呈线性，批内精密度为3.6%（n = 5，均值 = 311 pg/ml）。血清中的最低检测量为50 pg/ml。其他提取程序导致回收率较低或在最终测量中产生干扰。还讨论了其他研究小组报告的维生素K1(20)水平。
