Determination of egg yolk xanthophylls by isocratic high-performance liquid chromatography
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- 作者：Lu膷ka Brulc ; Breda Simonovska ; Irena Vovk ; Vesna Glavnik
- 关键词：Xanthophylls ; Egg yolk ; HPLC ; Triethylammonium acetate ; C30 column
- 刊名：Journal of Chromatography A
- 出版年：29 November, 2013
- 年：2013
- 卷：1318
- 期：Complete
- 页码：134-141
- 全文大小：1109 K
文摘
An isocratic HPLC method was developed for the determination of eight xanthophylls (lutein, capsanthin, zeaxanthin, canthaxanthin, 尾-apo-8鈥?carotenal, ethyl-8鈥?apo-尾-carotene-8鈥?oate, citranaxanthin and 尾-cryptoxanthin; registered as additives in poultry feeding) in egg yolks. Optimum separation of all-E-isomers of these xanthophylls was achieved in less than 18 min on a ProntoSIL C30 column at 27 掳C using acetone-methanol-0.5 M triethylammonium acetate buffer pH 7 14:5:1 (v/v) as the mobile phase with a flow rate of 1 mL/min using spectrophotometric detection at 450 nm. Other mobile phases were also found suitable, including acetone-water 93:7 (v/v) and acetone-methanol 1:4 (v/v) and the influences of column temperature on the separation and addition of triethylammonium acetate buffer pH 7 to the mobile phase on enhancement of the peak areas were evaluated. Preparation of test solution from yolks included a short vortexing of 0.5 g of yolk in 10 mL of acetone, followed by 15 min magnetic stirring under nitrogen and centrifugation. The method was validated for 5 analytes. The calibration range was between 0.04 and 2 渭g/mL and the mean recovery of the analytes (95%) and the intra-day precision of the method (RSD less than 5%) on three levels in triplicates were obtained. Analyses of eggs from four husbandry classes showed the presence of up to four xanthophylls (lutein, zeaxanthin, canthaxanthin and ethyl-8鈥?apo-尾-carotene-8鈥?oate) and traces of 尾-cryptoxanthin.