Separation of vitamin d2 and d3 by hplc from milk sample
Answers
Therefore, an on-line two-dimensional HPLC (2D-HPLC) method, that is similar to the reported design for the separation of vitamin D2 and D3 in feedstuff,9 was developed to measure vitamin D3 as well as vitamins A and E in milk-based samples.
Explanation:
The methods used for the determination of vitamin D in some dairy products (milk and infant formulas) by high performance liquid chromatography (HPLC). The low vitamin D contents and the presence of interfering compounds require sample treatment and purification of the extract. The advantages and drawbacks of hot and cold saponification, direct extraction and different types of extract purification are also discussed, taking into account the lack of vitamin D stability by heating, exposure to light and oxidation. With respect to chromatographic determination, the mode (normal or reverse phase), type of column (microcolumn or conventional) and detection system (UV, electrochemical and mass spectrometry) are overviewed, considering the vitamers to be determined