Abstract
A rapid, sensitive and inexpensive method has been developed for the simultaneous determination of fat-soluble vitamins A and E in infant formula (IF) samples. The isolation of fat-soluble vitamins includes a saponification step and an extraction step with petroleum ether. The qualitative and quantitative determination of vitamins was performed by High Performance Liquid Chromatography (HPLC) coupled to Diode Array Detector (DAD). More specifically, method development involved optimization of saponification time, testing of antioxidants, and number of extractions along with the extraction time applied. The method proved to be reliable as results agreed with the sample’s labels and the permitted variation according to the EE law. The relative standard deviation decreased significantly after the optimization (%RSD 1.8 % for retinol and 3.1% for a-tocopherol).
Highlights
Vitamins are organic compounds which are essential for human life, and are differentiated into fat and water soluble compounds
Vitamin A and E are two of the fat soluble vitamins, which must be supplied in human nutrition, because they cannot be composed in the body and have many important biological functions
The isolation of fat-soluble vitamins was divided into three main parts: sample preparation, saponification and extraction
Summary
Vitamins are organic compounds which are essential for human life, and are differentiated into fat and water soluble compounds. They are needed in relatively small amounts to sustain life and good health. Vitamin A is essential for normal vision, growth, cellular differentiation, development and proper functioning of skin and mucous membranes. It is defined as sum of trans-retinol or other retinoids as carotenoids. It prevents oxidative damage to neonates exposed to higher oxygen levels than those in the intrauterine environment8,9 It is defined as a collective group of related lipids called tocopherols (a- b-, g- and d-) and tocotrienols. A developed, optimized and validated HPLC-DAD method for simultaneous determination of retinol acetate as well as a-tocopherol acetate is presented
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