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Rapid high‐performance liquid chromatographic method for the estimation of toxicological levels of 25‐hydroxycholecalciferol (25‐OH D 3 ) in human serum
Author(s) -
Łukaszkiewicz Jacek,
Kozłowski Kazimierz,
Lorenc Roman
Publication year - 1983
Publication title -
journal of applied toxicology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.784
H-Index - 87
eISSN - 1099-1263
pISSN - 0260-437X
DOI - 10.1002/jat.2550030602
Subject(s) - chromatography , acetonitrile , high performance liquid chromatography , chemistry , extraction (chemistry) , solvent , hexane , vitamin d and neurology , medicine , endocrinology , organic chemistry
A reversed‐phase HPLC method for determination of 25‐hydroxycholecalciferol (25‐OH D 3 ) in serum samples is described. The method involves extraction of 25‐OH D 3 with hexane, followed by differential resolubilization in acetonitrile. The sample is then applied directly on the C 18 bonded phase column of the liquid chromatograph, and developed in a acetonitrile‐water solvent system. In the range 12.5–400 ng, a linear relationship was observed between detector response and the amount of 25‐OH D 3 placed on the column. Also, a very good correlation was observed between the amounts of 25‐OH D 3 added to serum samples, and the calculated amounts. The method can be used for screening for toxicological levels of 25‐OH D 3 in patients treated with high doses of vitamin D 3 .