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Development and validation of HPLC method for determination of DL-α-tocopherol in egg yolk


*Corresponding author
University of Novi Sad, Institute of Food Technology, Bulevar cara Lazara 1, 21000 Novi Sad, Serbia


The aim of the investigation was to develop and validate analytical method for DL-α-tocopherol determination in egg yolk by High Performance Liquid Chromatography. Obtained results indicated that this method has an acceptable level of accuracy and precision for DL-α-tocopherol. Recovery mean values were in the range of 99.2 percent to 100.3 percent with relative standard deviations between 0.2 percent and 0.8 percent. The limit of detection and limit of quantification were 4 ng/mL and 12ng/mL of DL-α-tocopherol, respectively. Testing the robustness of the method suggested that is not changeable with time or conditions. The results showed that this method is simple, accurate, precise, and applicable in laboratories for determination of vitamin DL-α-tocopherol in egg yolk.


Vitamin E is the family name for the group of eight natural compounds, including α-tocopherol, which is the most representative form of vitamin E. Vitamin E is essential nutrient of human and animal diet and plays an important role in the stimulation of synthesis and degradation of nutrients, improving immune function. In vitamin deficiency, humans and animals tend to develop diseases (1). Therefore, due to the inability of synthesizing vitamin E in human and animal cells, it must obtain it from exogenous sources (2).

The most widely accpeted biochemical function of the vitamin E is its antioxidant role. The vitamin E exhibited an antioxidant activity at low concentration and a prooxidant activity at high concentration (3, 4). Hence, vitamin E is very instable to light, oxygen, acid, heat and alkali (5).

Over the past few years, developing sensitive, selective, rapid and reliable method for the analysis of DL-α-tocopherol in egg yolk was the main goal of scientists. Determination by high performance liquid chromatography methods (HPLC), due to its rapid separation, high sensitivity, and accurate quantization, has become pop ...

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