Korean J Physiol Pharmacol.  2012 Jun;16(3):205-209. 10.4196/kjpp.2012.16.3.205.

Development of Estimation Methods of Skin Oxidation and Evaluation of Anti-Oxidative Effects of Genistein in Topical Formulations

• Kim SY
• Na YJ
• Kim D
• Kim Y
• Kim HM
• Hwang SH
• Kwak J
• Kuh HJ
• Lee J

• KMID: 1768015
• DOI: http://doi.org/10.4196/kjpp.2012.16.3.205

Abstract

The objective of the present study was to establish the method of measurement of hydrogen peroxide and to estimate the anti-oxidative effect of genistein in the skin. UVB induced skin oxidation and anti-oxidative effect of genistein formulations were evaluated by determining levels of hydrogen peroxide. The mechanism involved in the determination of hydrogen peroxide is based on a color reaction between ferric ion (Fe3+) and xylenol orange, often called FOX assay and subsequent monitoring of absorbance values of the reactant at 540 nm. The reaction was to some extent pH-dependent and detection sensitivity was greatest at pH 1.75. Genistein liposomal gel demonstrated better anti-oxidative effect with regard to lowering hydrogen peroxide levels elevated by UVB irradiation compared to genistein-suspended gel. A linear relationship has been observed between anti-oxidative effect of genistein and drug deposition in the skin tissue. Genistein liposomal gel resulting in the localization of the drug in the deeper skin led to improved anti-oxidative effect compared to genistein gel. The suggested method for evaluation of oxidation of the skin can be used as a tool to screen effective anti-oxidative agents and their delivery systems acting on the skin.

Keyword

Oxidation; Skin; Hydrogen peroxide; Genistein

MeSH Terms

Citrus sinensis
Genistein
Hydrogen Peroxide
Hydrogen-Ion Concentration
Phenols
Skin
Sulfoxides
Genistein
Hydrogen Peroxide
Phenols
Sulfoxides

Figure

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  Fig. 1 Influence of deproteinization solvents in absorbance of reaction solution at 540 nm. Mean±SD (n=3).

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  Fig. 2 Calibration curve of hydrogen peroxide constructed in different pH conditions. •, pH 1.5; ♦, pH 2.5; ▴, pH 1.75.

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  Fig. 3 Levels of hydrogen peroxide in the hairless mouse skin. Genistein from genistein-suspended gel and liposomal gel significantly reduced the increased concentration of hydrogen peroxide caused by UVB irradiation (p<0.05). Mean±SD (n=3).

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  Fig. 4 Percentages of genistein retained in the hairless mouse dorsal skin. Statistically more genistein was retained in the skin from liposomal gel, compared to genistein-suspended gel (p<0.05). Mean±SD (n=3).

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  Fig. 5 Correlation between skin deposition and anti-oxidative activity of genistein from formulations. Activity values are differences between levels of hydrogen peroxide measured after UVB irradiation without drug treatment and levels of hydrogen peroxide measured after drug treatment three times. •, Genistein-suspended gel; ▪, Genistein liposomal gel. Mean±SD (n=3).

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