Propylparaben-induced disruption of energy metabolism in human HepG2 cell line leads to increased synthesis of superoxide anions and apoptosis.
We developed a method using on-line solid phase extraction (SPE) coupled to high performance liquid chromatography-isotope dilution tandem mass spectrometry (HPLC-MS/MS) to measure the serum concentrations of seven environmental phenols and five parabens: bisphenol A; ortho-phenylphenol; 2,4-dichlorophenol; 2,5-dichlorophenol; 2,4,5-trichlorophenol; benzophenone-3; triclosan; and methyl-, ethyl-, propyl-, butyl-, and benzyl-parabens. The phenols and parabens present in serum were retained and concentrated on a C18 reversed-phase size-exclusion SPE column, back-eluted from the SPE column while the eluate was diluted through a mixing Tee (analyte peak focusing), separated using a pair of monolithic HPLC columns, and detected by isotope dilution-MS/MS. Sample preparation did not require protein precipitation, only dilution of the serum with 0.1M formic acid. This method, which combines an on-line SPE with analyte peak focusing feature and the selective atmospheric pressure photoionization MS detection, resulted in limits of detection ranging from 0.1 to 0.5 ng/mL for most of the analytes. The high throughput and adequate sensitivity with yet a relative low serum volume used (100 microL) confirm that analytically it is possible to measure simultaneously these phenols and parabens with the precision and accuracy at sub-parts-per-billion levels required for biomonitoring. However, important additional factors, including validated sample collecting, handling, and storing protocols, as well as toxicokinetic data, are required if these measures are used for exposure assessment.