Ultrasound-assisted extraction coupling to high performance liquid chromatography for enantiomerically quantitative analysis of two preservatives in cosmetics and the potentially cytotoxic study

Abstract

A simple and efficient high performance liquid chromatography method based on a home-made 3, 5-dichloro-phenylcarbamated mono-6-ethylenediamine-β-cyclodextrin chiral column for simultaneously enantiomeric analysis of two preservatives in cosmetics, including chlorphenesin and climbazole was successfully developed. Ultrasound-assisted extraction in combination with a sample clean-up procedure allowed to provide satisfactory enrichment and purification of the target analytes from the cosmetics. Response surface methodology using a Box-Behnken design was applied to screen experimental variables that affected the extraction efficiency. The proposed method was fully validated in terms of system suitability test, specificity, limits of detection and quantitation, linearity, accuracy, and precision, showing a reliable performance. Good linearity (R2 ≥ 0.9974), accuracy (% recovery ≥ 87.5), and intra-/inter-day precision (% RSD ≤ 9.3) with limits of quantitation from 0.5 to 1.0 μg mL−1 were obtained. This method was further applied to a broad range of cosmetics, in which the target analytes were detected in seven out of twenty-four analyzed samples at concentrations ranging from 0.32 to 1.20 mg g−1. Additionally, the cytotoxicity of the enantiomers of chlorphenesin to HaCAT cells was measured by in vitro MTT assay. The enantiomers of chlorphenesin inhibited the proliferation of cells in a dose dependent manner with significant enantioselectivity.