Journal of Food, Agriculture and Environment

Vol 10, Issue 1,2012
Online ISSN: 1459-0263
Print ISSN: 1459-0255

High-performance liquid chromatography (HPLC) analysis of the α-tocopherol deposition in the egg yolk: Effect of dl-α-tocopheryl acetate application in feed of breeding hens


Hongxin Zhang 1, 2, Renlu Huang 1*, Qiuxia Yang 1, Xiaohu Guo 1, Hui Chen 1

Recieved Date: 2011-09-12, Accepted Date: 2012-01-06


A method to quickly measure the content of alpha-tocopherol (α-T) in egg yolk was studied by high-performance liquid chromatography and then to further analyze effects of dl-α-tocopheryl acetate (α-TAC) of different doses in breeding hens’ feed on the deposition of α-T in the egg yolk.  A total of 288 Hyline brown breeding hens, 39 weeks old, were used and divided into six groups. Diets were based on standard layer rations with 0, 20, 50, 80, 160 or 320 mg of α-TAC/kg. At the forth weekend, the eggs were selected to measure the content of α-T of egg yolk. The following three methods were adopted: dealing with samples of the egg yolk by traditional saponification, ultrasonic processing after methanol extraction and keeping them for 12 hours over night after methanol extraction; methanol as mobile phase at flow rate of 1 ml/min, and using UV-detector to analyze the content of α-T in the egg yolk by HPLC at 292 nm wavelength. The method of ultrasonic processing 20 min after methanol extraction was much better than the other two methods in terms of extraction efficiency, with simplicity of operation and less loss of α-T. By using this method, a good correlation was shown (r = 0.9999) in the observed concentration range. The egg yolk should be extracted at the temperature of 20ºC for 10 hours, with a recovery rate of samples from 100.8% to 109%. The deposition of α-T in the egg yolk increased significantly when the amount of α-TAC was more than 40 mg/kg (P<0.01) and the highest concentration was reached when 320 mg/kg was added. In conclusion, the HPLC method of ultrasonic processing 20 min after methanol extraction is simple, fast and quick-response. It can be used effectively to measure the content of α-T in the egg yolk; through adding  a heavy dose of α-TAC into the daily ration, α-T egg yolk concentration will be increased and hatching egg rich in α-T will be produced, thus improving the hatching efficiency of breeding hens. 


High-performance liquid chromatography (HPLC), egg yolk, dl-α-tocopheryl acetate, alpha-tocopherol

Journal: Journal of Food, Agriculture and Environment
Year: 2012
Volume: 10
Issue: 1
Category: Agriculture
Pages: 385-390

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