Identification of Synthetic Steroids in River Water Downstream from Pharmaceutical Manufacture Discharges Based on a Bioanalytical Approach and Passive Sampling

Abstract

A bioanalytical approach was used to identify chemical contaminants at river sites located downstream from a pharmaceutical factory, where reproductive alterations in wild fish have been previously observed. By using polar organic compound integrative samplers (POCIS) at upstream and downstream sites, biological activity profiles based on in vitro bioassays revealed the occurrence of xenobiotic and steroid-like activities, including very high glucocorticoid, antimineralocorticoid, progestogenic and pregnane X receptor (PXR)-like activities (μg standard-EQ/g of sorbent range), and weak estrogenic activity (ng E2-EQ/g of sorbent range). Chemical analyses detected up to 60 out of 118 targeted steroid and pharmaceutical compounds in the extracts. In vitro profiling of occurring individual chemicals revealed the ability of several ones to act as agonist and/or antagonist of different steroids receptors. Mass balance calculation identified dexamethasone, spironolactone, and 6-alpha-methylprednisolone as major contributors to corticosteroid activities and levonorgestrel as the main contributor to progestogenic activities. Finally, RP-HPLC based fractionation of POCIS extracts and testing activity of fractions confirmed identified compounds and further revealed the presence of other unknown active chemicals. This study is one of the first to report environmental contamination by such chemicals; their possible contribution to in situ effects on fish at the same site is suggested.

Proposal to optimize ecotoxicological evaluation of wastewater treated by conventional biological and ozonation processes

“…To distinguish if a positive response comes from an activation of the androgen or glucocorticoid receptor, a co-exposure with specific AR (flutamide) or GR (RU486) antagonists was further realized. In complement to androgen receptor activity, information on glucocorticoid receptor activity is important since hospital and urban effluents are well-identified sources for glucocorticoid compounds in aquatic systems (Schriks et al 2010;Creusot et al 2014). …”

Additive effects of levonorgestrel and ethinylestradiol on brain aromatase ( cyp19a1b ) in zebrafish specific in vitro and in vivo bioassays

“…LNG has been detected in some effluents, sediments, ground water, tap water, but also in surface water of rivers at concentrations up to 38 ng/L (Vulliet et al 2008, Besse and Garric 2009, Al-Odaini et al 2010, Liu et al 2011, Vulliet and Cren-Olivé 2011, Fent 2015. LNG exerts biological activities that differ from the natural progestin (progesterone) since it has progestagenic and androgenic activities (Besse and Garric 2009) and also estrogenic activities both in vitro and in vivo (Jeng et al 1992, Zucchi et al 2012, Creusot et al 2014, Kroupova et al 2014. However, despite the joint exposure of aquatic wildlife to estrogens and progestins, very little information is available on their combined toxicity (Runnalls et al 2015, Säfholm et al 2015).…”

Passive sampling of pharmaceuticals and personal care products in aquatic environments

“…Alvarez et al (2013) and Jeffries et al (2011) address the analysis of steroid hormones in their studies, whereas Cernoch et al (2012) analyse only sulfonamides. A number of POCIS studies are also coupled to bioassays, evaluating endocrine-disruptive potential or dioxine-like activities (Jeffries et al 2011;Tapie et al 2011;Alvarez et al 2012;Jarosova et al 2012;Alvarez et al 2013;Jalova et al 2013;Creusot et al 2014).…”

“…Other extraction media, e.g. ACN (Amdany et al 2014a), acetone (Tanwar et al 2015), MeOH/water (Bailly et al 2013;Creusot et al 2014) or MeOH/dichloromethane (Vystavna et al 2012;Creusot et al 2013;Morin et al 2013;Vystavna et al 2013) are also suitable. Some authors adjust the pH in order to improve the extraction (Harman et al 2011;Cernoch et al 2012;Bayen et al 2014;Shi et al 2014).…”

“…In comparison, E-Screen and T47D-KBluc bioassays are more sensitive. Multi-receptor in vitro bioassays are often used to determine overall non-specific cytotoxicity, endocrine-disruptive potential and dioxin-like toxicity (Jeffries et al 2011;Tapie et al 2011;Jarosova et al 2012;Jalova et al 2013;Creusot et al 2014). Recombinant S. cerevisiae or MELN, MDA-kb2 and H4IIE-luc cell lines are the most commonly used.…”