Abstract

4-Hydroxybenzoate is a phenolic derivative of alkyl benzoates and is a widely used preservative in cosmetic and pharmaceutical products. The presence of 4-hydroxybenzoates in the human body may result from the use of pharmaceutical and personal care products. These compounds are also known to exhibit estrogenic and genotoxic activities. The potential adverse effects of these compounds include endocrine disruption, oxidative and DNA damage, contact dermatitis, and allergic reactions. This study used two mass spectrometry methods that are applicable when using a derivatization-enhanced detection strategy (DEDS) to screen 4-hydroxybenzoates and their metabolites. Chemical derivatization was used to enhance the detection of these compounds. To evaluate the metabolic process triggered by UV radiation, human keratinocyte HaCaT cells treated with these 4-hydroxybenzoates were further exposed to UVA, UVB and UVC radiation. Metabolites transformed by human keratinocytes in the chemical derivatization procedure were identified by a nano ultra-performance liquid chromatographic system (nanoUPLC) coupled with LTQ Orbitrap. The experiments confirmed the feasibility of this method for identifying 4-hydroxybenzoate metabolites and for high-throughput screening of 4-hydroxybenzoate in commercial products (50 samples) by the DEDS.

Highlights

  • Esters of 4-hydroxybenzoic acid (4-hydroxybenzoate), commonly known as parabens, are used as antimicrobial preservatives in cosmetics and pharmaceuticals

  • Cell line samples were composed of complicated components, and these biological samples were difficult to analyze by mass spectrometry (MS) without sample preparation

  • After implementing the derivatization-enhanced detection strategy (DEDS), all paraben metabolites transformed by human keratinocytes were identified by liquid chromatography (LC) coupled with LTQ Orbitrap

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Summary

Introduction

Esters of 4-hydroxybenzoic acid (4-hydroxybenzoate), commonly known as parabens, are used as antimicrobial preservatives in cosmetics and pharmaceuticals. Parabens are known to have estrogenic and genotoxic activities[2,3] Because of their widespread use as preservatives in various personal care products, cosmetics, pharmaceuticals, and food, parabens may be introduced to humans via many different environmental sources (including water, soil, sediment and sludge, air and dust, and biota)[3,4]. Because these compounds are ubiquitous in the environment, their safety and toxicity should be clearly determined. Experiments showed that the simple and rapidly performed method developed in this study is applicable for identifying paraben metabolites in human keratinocyte cells exposed to UV radiation and for screening for the presence of parabens in commercial pharmaceutical and cosmetic products

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