Stability, preservation, and quantification of hormones and estrogenic and androgenic activities in surface water runoff

Abstract

Degradation of hormones that may occur during storage of surface water samples can lead to underestimations in estrogenic and androgenic activities and inaccuracies in hormone concentrations. The current study investigated the use of sodium azide, hydrochloric acid (HCl), and sulfuric acid (H₂SO₄) to inhibit the degradation of hormones and estrogenic and androgenic activities in samples of surface water runoff from cattle manure-amended fields during storage at 4°C. Hormones and hormone metabolites were extracted using solid-phase extraction and analyzed using high-performance liquid chromatography (HPLC) with tandem MS. Estrogenic and androgenic activities were assessed by E-screen and A-screen, respectively. Results of the current study indicate significant degradation of estrogenic, androgenic, and progestogenic hormones and activities, which is likely attributable to microbial activity, within hours of sample collection. The inclusion of internal standards provides a means to account for hormone losses caused by extraction inefficiency and to some extent degradation. However, internal standards are unable to adequately account for significant losses and are not available for E-screen and A-screen. Sodium azide did not adequately inhibit androgen degradation at the concentration used (1 g/L). Acid preservation (HCl or H₂SO₄, pH 2) stabilized the estrogenic and androgenic activities, and coupling acid preservation with the use of internal standards resulted in reliable and accurate recovery of a suite of androgens, estrogens, and progestogens for up to 14 d of storage at 4°C.

Effects of progesterone on reproduction and embryonic development in the fathead minnow (Pimephales promelas)

“…4). In addition, during a prolonged rain event, P4 could continuously be washed into aquatic environments from concentrated animal feeding operations at concentrations as high as 375 ng/L [7]. 4).…”

“…With regard to the potential for chronic exposure, it has been reported that sustained levels of P4 can enter aquatic environments from wastewater treatment plants with concentrations in the range of 5 to 199 ng/L [8]. In addition, during a prolonged rain event, P4 could continuously be washed into aquatic environments from concentrated animal feeding operations at concentrations as high as 375 ng/L [7]. Given these high levels and the ubiquitous presence of P4 in the environment and its effects on fish reproduction, we conclude that P4 is a significant EDC that warrants more attention from investigators and managers.…”

“…Target P4 concentrations were confirmed in a preliminary one-week trial by measuring the P4 concentrations in three randomly sampled exposure tanks within each treatment using high-performance liquid chromatography mass spectrometry/ mass spectrometry (Wisconsin State Laboratory of Hygiene ESS Organic Chemistry Method 1690; MDL ¼ 1.0 ng/L [7]). The measured concentrations of P4 in the 0, 10, 100, and 1,000 ng/L tanks were <MDL, 15.4 AE 7.1, 111.3 AE 23.3, and 987 AE 10.5 ng/L, respectively (mean AE SEM, n ¼ 3).…”

“…In a recent field investigation of the steroid hormones associated with bovine dairy and beef operations in Wisconsin, researchers from the Wisconsin State Laboratory of Hygiene detected progesterone (P4) in spring runoff events at concentrations as high as 375 ng/L [7]. Progesterone and other natural and synthetic progestins (norethindrone, megesterol acetate, and others) have previously been measured in surface waters at concentrations ranging from 5 to 199 ng/L and originate primarily from human wastewater treatment plants and concentrated animal-feeding operations (CAFOs) [8][9][10].…”

Pollution by endocrine disrupting estrogens in aquatic ecosystems in Morogoro urban and peri-urban areas in Tanzania

“…The pH of water samples was adjusted to about 3 by adding hydrochloric acid so as to fix the estrogens. The added acid suppressed microbial activity which could degrade the estrogens to some extent before analysis (Havens et al, 2010). Thereafter, the samples were carried in cool box packed with ice packs to the Ecotoxicology and Natural Products research Laboratory in the Faculty of Veterinary Medicine at Sokoine University of Agriculture, for pretreatment and solid phase extraction of estrogens that was done within 12 h after sample collection.…”