Analytical method development for the determination of B-vitamins from foods
Nowadays there is an increasing attention on consuming healthy nutrition and vitamin rich foodstuff. A balanced diet should provide the necessary amount of vitamins, including vitamin B. B vitamins are waters soluble and absorbed from the gastro intestinal tract, where they have regulating role....
The aim of my thesis is the development of a chromatographic method for the determination of vitamin-B1 (thiamine), B2-(riboflavin) and B6-vitamins (vitamers: pyridoxin, pyridoxal, pyridoxamin) from various food samples. For the measurement of sole vitamins regulations MSZ EN 14122:2006, MSZ EN 14152:2004, MSZ EN 14663:2006, was taken as basis.
The measurement of all the three vitamins can be done in similar ways, but they have different physical and chemical properties. The regulations write different conditions. My purpose is to develop a common measurement method what is suitable for all vitamers from single injection.The determination of the whole vitamin content requires complex sample preparation, because B vitamins occur in nature (and in food) mostly in phosphorylated form. B6 vitamins occur as glycosides.The native forms can be freed up by acidic hydrolysis and enzymatic treatment.
The separation of the target components from the matrix can be achieved by reversed phase liquid chromatography. Detection can be done by UV and fluorescence detectors. The fluorescence allows for a more selective and sensitive detection, that is why it is necessary to derivatize the non-fluorescence active thiamine to thiochrome. The rest of the vitamins are fluorescence active in their native forms.
Author: Anita Klusovszki, Chemistry MSc student