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Abstract
A modified cleanup procedure of 25-(OH)-vitamin D3 from a previous method based on the use of C18 and Si cartridges is proposed. The monitoring of the optimisation study was carried out by individual separation of the target analytes by HPLC and UV-detection. The improvement of the original procedure allows a dramatic reduction of plasma especies, which results in a more clean chromatogram in which other two hydroxymetabolites of vitamin D3 (namely, 24-R,25-(OH)2-vitamin D3 and 1-I,25-(OH)2-vitamin D3) can be determined. A closer to quantitativeness recovery of the target analytes (at least 4 times higher than in the original method) is achieved.