https://doi.org/10.15255/KUI.2018.016
Published: Kem. Ind. 67 (13) (2018) P107–P114
Paper reference number: KUI-16/2018
Paper type: Conference paper
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Reproducibility of the Selected Methods for Determination of Phenol in Urine by High Performance Liquid Chromatography

E. Horozić, A. Begić, Z. Ademović and S. Brekalo Lazarević

Abstract

Determination of phenol in urine is considered as a suitable biomarker of benzene exposure, which is classified as Group A carcinogen by the International Agency for Cancer Research (IARC) and the Environmental Protection Agency (EPA). The aim of this work was to test reproducibility of several published high performance liquid chromatography (HPLC) methods for phenol determination in urine samples. Previous methods mostly included additional equipment (e.g. fluorescence detector) or an additional step in a sample preparation protocol (e.g. derivatization or steam distillation) which is time consuming and can also cause the loss of analyte. The method that was chosen in this work showed good reproducibility with certain modifications in chromatographic conditions and sample preparation protocol. Optimisation of the method included changes in wavelength, constituents, and the flow rate of mobile phase, and excluded the addition of internal standard in sample preparation protocol. Mobile phase consisted of distilled water: methanol: glacial acetic acid (65 : 34 : 1 v/v %), and Selectra C18 (4.6 × 250 mm, 5 μm) column was used. Chromatographic conditions included the flow rate of 1 ml min^–1, room temperature, wavelength at 218 nm and 270 nm, and an injection volume of 20 µl. At 270 nm, linearity was shown in the range of concentrations of 1–30 mg l^–1 (y = 10290x + 15138; R² = 0.9932), while the achieved percentage of recovery was between 73.8 % and 111.2 % for the tested concentrations.

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Keywords

phenol, extraction, HPLC, urine