Abstract

Benzophenone (BP)-type UV (ultraviolet) filters, especially 2-hydroxy-4-methoxybenzophenone (2OH–4MeO–BP), are widely used in the U.S., to protect the skin and hair from UV irradiation. Despite human exposures to UV filters through the dermal application of products containing sunscreen agents, few studies have examined the occurrence of UV filters in humans. Thus far, few sensitive methods are available for the determination of 2OH–4MeO–BP in human urine. Furthermore, methods for the determination of other BP derivatives, including 2,4-dihydroxybenzophenone (2,4OH–BP), which is formed from 2OH–4MeO–BP via metabolic activities of the cytochrome P450 enzymes, have not been available. In this study, we have developed a method for the analysis of five BP derivatives: 2OH–4MeO–BP, 2,4OH–BP, 2,2′-dihydroxy-4-methoxybenzophenone (2,2′OH–4MeO–BP), 2,2′,4,4′-tetrahydroxybenzophenone (2,2′,4,4′OH–BP), and 4-hydroxybenzophenone (4OH–BP) in human urine, using liquid–liquid extraction and liquid chromatograph (LC)-tandem mass spectrometer (MS/MS) analysis. The instrumental calibration range for each of the BP derivatives ranged from 0.05 to 100 ng ml−1, and showed excellent linearity (r > 0.99). The respective limits of detection (LODs) and limits of quantification (LOQs) were determined to be 0.082 and 0.28 ng ml−1 for both 2,4OH–BP and 4OH–BP; 0.13 and 0.44 ng ml−1 for 2,2′OH–4MeO–BP; and 0.28 and 0.90 ng ml−1 for both 2OH–4MeO–BP and 2,2′,4,4′OH–BP. Recoveries of BP derivatives through the entire analytical procedure were between 85.2 and 99.6%. The coefficients of variation (CVs) of five replicate analyses within 1 day, and across 5 days, were respectively 1.4 and 3.7% for 2,4OH–BP; 1.7 and 3.0% for 2OH–4MeO–BP; and 2.8 and 4.5% for 4OH–BP. When BP derivatives were determined in urine samples from 23 U.S. (Albany, New York) and 22 Japanese (Matsuyama, Ehime) volunteers, higher concentrations of 2,4OH–BP, 2OH–4MeO–BP, and 4OH–BP were found in samples collected from females in the Albany cohort, probably reflecting great usage by U.S. females. The urine sample from a known sunscreen user contained very high concentrations of 2,4OH–BP and 2OH–4MeO–BP. 2,2′,4,4′OH–BP and 2,2′OH–4MeO–BP were not detected in any of the urine samples analyzed. Our results indicate considerable exposure to highly estrogenic 2,4OH–BP and 2OH–4MeO–BP by females in the U.S., and suggest the need for further studies on potential health effects.

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