Wet-chemical postcolumn reaction and fluorescence detection analysis of the reference interval of endogenous serum vitamin K1(20)
| Autor: | Willy E. Lambert, AndréP. De Leenheer, Eddy J. Baert |
|---|---|
| Rok vydání: | 1986 |
| Předmět: |
Vitamin
Tetramethylammonium Chromatography Vitamin K Extraction (chemistry) Biophysics Fluorescence spectrometry Endogeny Cell Biology Biochemistry Fluorescence High-performance liquid chromatography chemistry.chemical_compound Adsorption Spectrometry Fluorescence chemistry Humans Molecular Biology Chromatography High Pressure Liquid |
| Zdroj: | Analytical biochemistry. 158(2) |
| ISSN: | 0003-2697 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|