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Originally published In Press as doi:10.1194/jlr.D700006-JLR200 on June 19, 2007

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Journal of Lipid Research, Vol. 48, 2079-2085, September 2007
Copyright © 2007 by American Society for Biochemistry and Molecular Biology

Methods

Specific and rapid analysis of ubiquinones using Craven's reaction and HPLC with postcolumn derivatization

Haruo Shimada1,*, David Kodjabachian{dagger} and Masami Ishida*

* Frontier Science Laboratories, H&BC Development Center, Shiseido Co., Ltd., Kanazawa, Yokohama 236-8643, Japan
{dagger} Ecole Nationale Supérieure de Chimie de Paris, 75231 Paris Cedex 05, France

Published, JLR Papers in Press, June 20, 2007.

1 To whom correspondence should be addressed. e-mail: haruo.shimada{at}to.shiseido.co.jp

A new method for the analysis of ubiquinones in various samples was developed using an HPLC system with postcolumn derivatization. Craven's reaction, a specific color reaction for the analysis of ubiquinones, was used in the system. Because the reaction progressed in organic solvents that contained ubiquinones and ethylcyanoacetate under an alkaline condition, the selectivity for ubiquinone detection was higher than that for ubiquinone detection using the nonderivatized ultraviolet detection system at 275 nm, a system widely used for the analysis of ubiquinones. The new detection system can avoid the adverse effects of impurities. Furthermore, it can confirm specificity by stopping the color reaction under a neutral condition. The detection limit for ubiquinone-10 was 1 ng (1.2 pmol). A good linearity for the calibration curve was observed in the range of 11.7 pmol to 11.7 nmol. To investigate the possible application of this method, various samples, such as soybean capsules used as a dietary supplement and biological materials (rice as well as bovine plasma and liver samples), were applied to the system and their ubiquinone contents were quantified. This method is thought to be widely and conveniently applicable for determining the level of ubiquinones because of its high selectivity for ubiquinone detection.

Supplementary key words quantification • high-performance liquid chromatography • coenzyme Q9 and Q10

Abbreviations: RSD, relative standard deviation


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