Rapid determination of vitamin D3 in poultry eggs and their products byon-line column switching liquid chromatography
WANG Yin1, HUANG Baifen2, PAN Xiaodong2
1. Department of Toxicology, Zhuji Center for Disease Control and Prevention, Zhuji, Zhejiang 311800, China; 2. Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou, Zhejiang 310051, China
Abstract：Objective To establish an on-line column switching liquid chromatography for quantitative determination of vitamin D3 in poultry eggs and their products. Methods Antioxidant-protected samples were saponified, extracted with a mixture of ethyl acetate-n-hexane mixture (6︰4, v/v) and subjected for determination using two-dimensional liquid chromatography. One-dimensional chromatography was performed with the UniChiral® OD-5H column (4.6 mm × 100 mm, 5 μm), acetonitrile: methanol (75︰25, v/v) and water as a mobile phase for gradient elution at a flow rate of 1.00 mL/min to complete vitamin D purification, and two-dimensional chromatography was performed with the Agilent Eclipse PAH column (2.1 mm × 100 mm, 3.5 μm, column temperature of 35 ℃), detection wavelength of 264 nm for separation and determination of vitamin D3 and internal standard vitamin D2 with the internal standard method. Results Vitamin D3 had a good linear relationship within the range of 2.5 to 100 ng/mL, with r2 of >0.999. The mean spiked recovery rate of vitamin D3 was 101.23% to 102.08% in egg yolk powder, with relative standard deviation (RSD) of 3.91% to 5.85%, detection limit of 1.5 μg/100 g, and quantitative limit of 4.9 μg/100 g, and the limits of detection and quantitation of vitamin D3 were 0.15 μg/100 g and 0.49 μg/100 g in whole egg liquids. Conclusions The method is simple, rapid, highly accurate, sensitive and reproducible, which is suitable for rapid and quantitative determination of vitamin D3 in poultry eggs and their products.
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