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In the present study, a new extraction method based on acid digestion and SPE clean-up (Oasis Wax) was developed for measuring four PFR metabolites (i.e. dibutyl phosphate (DBP), diphenyl phosphate (DPhP), bis(1,3-dichloro-2-propyl)phosphate (BDCPP) and bis(2-butoxy ethyl) phosphate (BBEP)) in hair and nails. The method optimization was done according to a combinatorial design (Taguchi) where several parameters were efficiently optimized. Precision was lower for hair than for nails (RSD % 18 and 28%). Recoveries were >74%. High DBP levels in procedural blanks were traced back to the use of SPE cartridges. Therefore a new SPE pre-treatment was tested, reducing significantly DBP levels (<1 ng).

Levels of the PFR metabolites were measured in scalp hair, finger, and toe nails collected over two months in two volunteers (female and male). DPhP levels were extremely high (in mu g/g range) in both finger and toe nails in the female. BDCPP and BEEP were the minor metabolites detected in nails (average levels of 28-64 ng/g and <2.2-4.1 ng/g, respectively). DPhP was the only metabolite detected in hair (0.23-0.25 ng/g). Results showed that there is a possible contribution from both an external (via deposition) and an internal exposure, however it was not possible to fully understand their extent. Since there were no records of lifestyle and due to the small sample size, the major exposure source could not be addressed here. Nevertheless, there is evidence that hair and nails (finger and toe) might be good indicators of human exposure to PFRs, especially to TPhP. (C) 2016 Elsevier Ltd. All rights reserved.