Application of high-performance liquid chromatography to determine the concentration of α-tocopherol in the blood of cattle

The purpose of the work was to develop within the laboratory method of analysis of alpha-tocopherol in the blood of animals corresponding to the laboratory equipment and the practical needs of the laboratory. The main objective of the study is to develop a method for the extraction and analysis of alpha-tocopherol in the blood serum of cattle, which could be used in studies related to the assessment of animal health and their productivity. Since most of the techniques are somehow related to the use of mass spectrometry (due to the low concentration of alpha-tocopherol in the blood), and the content of alpha-tocopherol is an important indicator in the system of evaluation criteria, an attempt was made to adapt the existing methods of extraction and analysis to the conditions and instrument base of the laboratory. In the process of work, the extraction of alpha-tocopherol in various variations was performed, the methods already described in scientific publications were tested. The result was an adapted extraction technique, as well as chromatographic analysis using fluorimetric detection, the separation conditions were selected. In particular, the selection of the composition of the rolling phase for the most efficient separation was carried out, which made it possible to reduce the analysis time from 23 minutes to 18 minutes (relative to the previously used technique).