Reliable quantification of bisphenol A and its chlorinated derivatives in human urine using UPLC–MS/MS method

Abstract

Bisphenol A (BPA), a widespread man-made chemical classified as an endocrine disruptor, is increasingly considered as a major cause of concern for human health. Chlorine present in drinking water may react with BPA to form chlorinated derivatives (ClxBPA), which have demonstrated a heightened level of estrogenic activity. If many epidemiological studies report that more than 90% of people have detectable BPA levels in their urine, then no such study has been undertaken regarding ClxBPA. The purpose of this work is to propose a highly sensitive and accurate analytical method adapted to large-scale biomonitoring studies aimed at assessing exposure to BPA and ClxBPA through the use of human urine. To achieve this, we have comprehensively validated a method using salting-out assisted liquid/liquid extraction (SALLE) coupled to UPLC-MS/MS and isotope dilution quantification, to measure unconjugated BPA and ClxBPA in human urine according to the accepted guidelines. Deutered BPA as well as deutered 2,2'-DCBPA was used as internal standards. The matrix calibration curve ranged from 0.05 to 1.60 ng mL(-1) and from 0.5 to 16.0 ng mL(-1) for ClxBPA and BPA respectively, and provided good linearity (r²>0.99). This method was precise (the intra- and inter-day coefficients of variation were <20% at three different concentrations: 0.05 ng mL(-1), 0.2 ng mL(-1), 0.8 ng mL(-1) and 0.5 ng mL(-1), 2 ng mL(-1), 8 ng mL(-1) for ClxBPA and BPA, respectively) and accurate (bias ranged from -13% to +12%). The limit of quantification, validated at 0.05 ng mL(-1) and 0.5 ng mL(-1) for ClxBPA and BPA respectively when using 300 µL of urine, was found to be suitable for the concentration existing in real samples. The matrix effect and the BPA cross-contamination were also investigated in this study. The analytical method developed in this study is in accordance with the requirements applicable to biomonitoring of BPA and ClxBPA in human urine.

Tutorial review on validation of liquid chromatography–mass spectrometry methods: Part I

“…In addition, there is a large number of articles demonstrating validation of LC-ESI-MS methods for practical analytical tasks with respect to certain validation guidelines such as FDA [68][69][70][71][72], ICH [73][74][75] Eurachem [76][77][78][79][80][81][82][83], EMA [84][85][86], IUPAC [87][88][89] or SANCO [90,91]. However, in most cases not all parameters are investigated.…”

Additional effect of drinking water endocrine-disrupting compound exposure on fetal growth compared to the effect of active tobacco exposure during pregnancy

“…EDC exposure assessment can be rendered more precise through exposure biomarkers such as blood and urine or cord blood, amniotic fluid and breast milk, with concentrations that could reflect fetal exposure (17,48). For example, BPA and its chlorinated derivatives have been successfully detected and reliably quantified in human urine and colostrum, and the methods applied can be used to estimate fetal exposure to these EDC in drinking water (49,50). Moreover, BPA and phthalate urinary levels have been combined with ultrasound measurements of fetal growth in the literature (51).…”

Exposition hydrique au bisphénol A et à ses dérivés chlorés et liens avec le cancer du sein : étude de faisabilité BREDI I (Breast and Endocrine Disrupter Investigation Part 1)

“…Leur analyse a été effectuée selon la méthode décrite par VENISSE et al (2014). Une gamme d'étalonnage associée à des points de contrôle a été préparée dans l'urine par surcharge via des solutions de BPA et de Clx-BPA de concentrations connues, de même qu'un blanc-urine et un blanc-eau.…”

“…Les analyses ont été adaptées de la méthode décrite par VENISSE et al (2014), mais préalablement développée et validée dans le tissu adipeux mammaire. Une gamme d'étalonnage a été réalisée par surcharge de solutions de BPA et de Clx-BPA de concentrations connues de même qu'un blanc-tissu adipeux et un blanc-eau.…”