HPLC‐MS/MS analysis of steroid hormones in environmental water samples

Abstract

Today, freshwaters, such as lakes and rivers, are subject to controlled pollution. Steroid hormones are chemically very stable highly lipophilic molecules. Their biological properties have a strong impact on the endocrine regulation of species. Steroids have estrogenic, androgenic, thyroidogenic or progestogenic effects and based on them, they could disturb the physiological mechanisms of freshwater species. We focused on progestins as they are the main active ingredients of contraceptive pharmaceuticals. Progestins have been shown to impair reproduction in fish, amphibians, and mollusks at low ng/L concentrations. Certain progestins, such as levonorgestrel (LNG) have androgenic properties also. We selected the most used active substances drospirenone (DRO), LNG, and progesterone (PRG) and then developed and optimized a liquid chromatographic-mass spectrometric method with solid-phase extraction to measure them. Using our sensitive method (LOQ 0.03-0.11 ng/L) we could measure steroids even between 0.1 and 1 ng/L. Analyzing freshwater samples from the Lake Balaton catchment area, we found influents where the concentration of these hormones was 0.26-4.30 (DRO), 0.85-3.40 (LNG), and 0.23-13.67 (PRG) ng/L. Out of 53 collecting places, 21 contained measurable progestin levels, which clearly demonstrates the applicability of our method, legitimates toxicology experiments with effected species, and indicates monitoring efforts.

β-Estradiol and ethinyl-estradiol contamination in the rivers of the Carpathian Basin

“…The rest of the samples were not filtrated. 2.4 Solid phase extraction Solid phase extraction was carried out according to our previous method for progestogens with small modifications (Avar et al 2015). SPE cartridges were conditioned with 15 mL methanol and equilibrated with 20 mL LC-MS grade water.…”

Pond Snail Reproduction as Model in the Environmental Risk Assessment: Reality and Doubts

“…SPE), finally quantitative measured by using LC-MS/MS. Generally, around 0.1 ng/L limit of quantification (LOQ) value are achieved [33,34,71,72]. The detection of steroid EDCs from various solid samples are complicated because more sample preparation steps are required (drying, homogenization, destruction, extraction and purification).…”

“…progesterone, estradiol, testosterone [33][34][35] and synthetic (e.g. drospirenone, levonorgestrel, ethinylestradiol, cyproterone acetate (CPA), t-methyltestosterone [23,[33][34][35]) hormones, phytosterols (e.g.…”

“…L. stagnalis, Anodonta cygnaea, Dreissena polymorpha) and fish species (e.g. Rutilus rutilus, Alburnus alburnus, Abramis brama, Carassius carassius, Cyprinus carpio, Perca fluviatilis), where the estrogen and progestogen concentrations were found between 0.07-0.68 ng/L and 0.23-13.67 ng/L, respectively [33,34]. The presence of steroid hormones has also been found in the drinking water, which is a warning sign that the current handling of pharmaceuticals may lead to future global human health problems [52][53][54][55][56].…”

“…ethinylestradiol, estradiol, estrone and estriol), and the risk is now well documented [47,49,75,76]. Several studies have also been conducted on the risk related to anti-androgens [77], but contrast to these, surprisingly, relatively few data are published about the occurrence of progestogens in different waters [34,41,49] and mainly their neuroendocrine effects on non-target freshwater organisms, including particularly invertebrates [49].…”