Wet-chemical postcolumn reaction and fluorescence detection analysis of the reference interval of endogenous serum vitamin K1(20)
✍ Scribed by Willy E. Lambert; AndréP. De Leenheer; Eddy J. Baert
- Publisher
- Elsevier Science
- Year
- 1986
- Tongue
- English
- Weight
- 368 KB
- Volume
- 158
- Category
- Article
- ISSN
- 0003-2697
No coin nor oath required. For personal study only.
✦ Synopsis
The reference interval for serum vitamin K roO) 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 KroO) and vitamin K ro5), 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 [logx ? 2 SD -(logx)]. The method is linear over the entire physiological range and has a within-run precision of 3.6% (n = 5, X = 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 K,,20j levels as reported by other research groups are also discussed. o 1986 Academic Press. Inc.