Water Hormone Research Articles

On-Site Monitoring of Steroid Hormones in Environmental Waters with a Low-Cost, Integrated Polymer Lab-on-Chip Device

Routine analysis of steroid hormones in environmental water samples demands for cost-effective tools that can detect multiple targets simultaneously. This study reports a high-throughput polymer platform integrated to polymer optical sensors for on-site monitoring of hormones in water. This opto-microfluidic device concept is fully compatible to low-cost fabrication methods. A competitive chemiluminescence immunoassay was performed onto gold thin film coated chambers, and a detection resolution of roughly 0.2 ng/mL was obtained using 17β-estradiol as the model target. Furthermore, the integrated polymer platform showed good recovery for the estradiol target when spiked in surface water samples.

Enrichment of steroid hormones in water with porous and hydrophobic polymer‐based SPE followed by HPLC–UV determination

There have been great concerns about the persistence of steroid hormones in surface water. Since the concentrations of these compounds in water samples are usually at a trace level, the efficient enrichment of steroid hormones is vital for further analysis. In this work, a porous and hydrophobic polymer was synthesized and characterized. The composition of solvent used as porogen in the synthetic process was shown to have an effect on the morphology of the polymer, which was successfully used as an SPE sorbent for simultaneously enriching steroid hormones in surface water samples. The recoveries of the steroid hormones on the custom-made polymer ranged from 93.4 to 106.2%, whereas those on commercialized ENVI-18, LC-18, and Oasis HLB ranged from 54.8 to 104.9, 66 to 93.6, and 77.2 to 106%, respectively. Five types of steroid hormones were simultaneously measured using HPLC-UV after they were enriched by the custom-made sorbent. Based on these findings, the SPE-HPLC method was developed. The LODs of this method for estriol, estradiol, estrone, androstenedione, progesterone were 0.07, 0.43, 0.61, 0.27, and 0.42 μg/L, respectively, while precision and reproducibility RSDs were <6.40 and 7.49%, respectively.

Veterinary antibiotics and hormones in water from application of pig slurry to soil

Although the use of liquid waste from piggeries as agricultural fertilizer is common practice, problems arise from contamination of water by chemical compounds. This paper reports on the presence of three hormones and six veterinary antibiotics in water of the surface runoff and drainage flow from volumetric lysimeters to which pig slurry was applied, and which had been planted with cereals, horticultural crops and permanent pasture. High-efficiency liquid chromatography was used for chemical analysis of soil water following two applications of liquid pig slurry. Concentrations of the hormones and antibiotics found were between 4.6 and 1350.8ngL−1, and were detectable up to the ninth day after the second application of manure. The presence of different crops did not influence the occurrence of the analytes.

Online molecularly imprinted solid-phase extraction coupled to liquid chromatography-tandem mass spectrometry for the determination of hormones in water and sediment samples

The molecularly imprinted SPE directly coupled to RP LC-MS/MS method has been developed and successfully validated for the determination of six hormones in water and sediment samples. The method is based on the use the home-made column filled with a molecularly imprinted sorbent (imprinted against estrogens) that was used under nonaqueous conditions. Thus, its high selectivity could be utilized resulting in low matrix components' coextraction. The method showed excellent recovery (92-105%) and satisfactory sensitivity (LOQs water: 1.9-4.0 ng/L; LOQs sediment: 0.2-0.5 ng/g). The intra- and interprecision for water and sediment was in the range of 4.0-6.0% and 4.4-7.6%, respectively. Finally, 20 water and sediment samples collected from the Svratka river were analyzed. Only estrone was quantified in eight water samples (4.4-7.1 ng/L); no analytes were found in sediment samples.

Simultaneous analysis of non-steroidal anti-inflammatory drugs and estrogenic hormones in water and wastewater samples using gas chromatography–mass spectrometry and gas chromatography with electron capture detection

Non-steroidal anti-inflammatory drugs are the group of pharmaceuticals that is most often found in the environment, whereas estrogenic hormones are considered to be potent endocrine disruptors. However, the fate and persistence of these compounds in the environment are still unclear. In this study we propose two approaches for determining these compounds in environmental water samples: GC–MS using time windows and operating in selected ion-monitoring mode (SIM) and, for the first time, gas chromatography with electron capture detection (GC–ECD). The identification criteria of both methods fulfilled the requirements of Directive 2002/657/EC. The use of time windows improved the sensitivity of GC–MS measurements. In GC–MS analysis the pharmaceuticals were determined as trimethylsilyl, in GC–ECD as pentafluoropropionyl derivatives. The influence of such parameters as the type of reagent, type of solvent, reaction time, reaction temperature and microwave irradiation in a household microwave oven on the efficacy of silylation was investigated. Derivatization using N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA) and 1% trimethylchlorosilane (TMCS) in pyridine (1:1, v/v) for 30min in 60°C was found to be optimal. Optimization of the solid phase extraction procedure (SPE) confirmed that the application of Oasis HLB cartridges, the acidification of loading samples to pH2 and the use of methanol as eluent gave the best absolute recoveries (ARs) of the target compounds. The following ARs of all the compounds were achieved: 58.2–106.8% in influent wastewater, 77.8–103.4% in effluent wastewater and 81.2–101.9% in surface water samples. Validation of the SPE–GC–MS method enables 13 pharmaceuticals to be determined with MDLs between 3.3 and 343.6ng/L, depending on the analytes and matrices. GC–ECD analysis enables the determination of 6 pharmaceuticals in surface water samples with MDLs between 0.7 and 5.4ng/L. The proposed methods were successfully used for analyzing selected pharmaceuticals in wastewaters and river waters in Poland.

Using a novel sol–gel stir bar sorptive extraction method for the analysis of steroid hormones in water by laser diode thermal desorption/atmospheric chemical ionization tandem mass spectrometry

A new coating material was used for a stir bar sorptive extraction (SBSE) method coupled to a high throughput sample analysis technique. This allowed for a simple procedure for fast determinations of eight steroid hormones (estriol, estradiol, ethynylestradiol, estrone, progesterone, medroxyprogesterone, levonorgestrel, northindrone) in water. Sample pre-treatment was performed using an in-house SBSE method based on a polydimethylsiloxane/phenyltrimethylsiloxane/β-cyclodextrin sol–gel material. The analytes were desorbed by liquid extraction prior to their analysis by laser diode thermal desorption/atmospheric pressure chemical ionization coupled to tandem mass spectrometry (LDTD–APCI–MS/MS). Several parameters, including ionic strength, volume and time of extraction as well as volume and time of desorption, were investigated to maximize extraction efficiency by SBSE in aqueous solutions. The in-house stir bar showed good reproducibility and could be used for at least 50 extractions without affecting analytical performance. The recoveries of the spiked steroid hormones ranged from 55% to 96% in all water matrices studied (HPLC grade water, tap water and raw wastewater). Only one compound showed poor recovery values (<2% for estriol) in all matrices. The method detection limits (MDLs) in real matrices were within the range of 0.1–0.3μgL−1 except for estriol at 48μgL−1. The extraction performance of the in-house SBSE for the eight selected hormones was also compared with that of a commercially-available stir bar coated with polydimethylsiloxane (PDMS). This novel stir bar coating could prove to be useful method for the detection and quantification of trace levels of steroid hormones.

Simultaneous determination of estrogenic and androgenic hormones in water by isotope dilution gas chromatography–tandem mass spectrometry

A rapid gas chromatography–tandem mass spectrometry (GC–MS/MS) analytical method was developed for the simultaneous analysis of 7 estrogenic hormones (17α-estradiol, 17β-estradiol, estrone, mestranol, 17α-ethynylestradiol, levonorgestrel, estriol) and 5 androgenic hormones (testosterone, androsterone, etiocholanolone, dihydrotestosterone, androstenedione) in aqueous matrices. This method is unique in its inclusion of all 12 of these estrogens and androgens and is of particular value due to its very short chromatographic run time of 15 min. The use of isotope dilution for all analytes ensures the accurate quantification, accounting for analytical variabilities that may be introduced during sample processing and instrumental analysis. Direct isotopically labelled analogues were used for 8 of the 12 hormones and satisfactory isotope standards were identified for the remaining 4 hormones. Method detection levels (MDLs) were determined to describe analyte concentrations sufficient to provide a signal with 99% certainty of detection. The established MDLs for most analytes were 1–5 ng L −1 in a variety of aqueous matrices. However, slightly higher MDLs were observed for etiocholanolone, androstenedione, testosterone, levonorgestrel and dihydrotestosterone in some aqueous matrices. Sample matrices were observed to have only a minor impact on MDLs and the method validation confirmed satisfactory method stability over intra-day and inter-day analyses of surface water and tertiary treated effluent samples.

Levels of Human Steroid Hormones in Water and Sediment From 4 Major Rivers in Korea

PP-30-122 Background/Aims: Steroid hormones remained in river water or soil may influence endocrine disrupting effects to ecological system even though low level of concentration. This study was performed to assess the residual level of 4 steroid hormones at river water and sediment. Methods: Estrone (E1), 17-β-estradiol (E2), estriol (E3), and 17-α-ethinylestradiol (EE2) were analyzed at water (n = 80) and sediment (n = 8) from 4 major river in Korea by LC-MS/MS technique. Water and sediment samples were prepared by continuous solid phase extraction method subsequent by liquid-liquid extraction. Results: The concentration of steroid hormone ranged ND ∼59.4 ng/L for E1, ND ∼52.8 ng/L for E2, ND ∼8.6 ng/L for E3 in river water and ND ∼5.75 ng/g for E1, ND ∼1.29 ng/g for E2 in sediment samples, respectively. EE2 was not found at any of the samples. The occurrence rate of E1 was most high; 21.3% for river water samples and 75% for sediment samples. Conclusion: We detected low concentration of steroid hormones in river water sediments in Korea.

Presence of Pharmaceuticals and Hormones in Waters from Sewage Treatment Plants

This paper describes the presence of 33 pharmaceuticals and hormones in waters from two sewage treatment plants (STPs) situated in Catalonia, in northeastern Spain. The target compounds were one psychoactive stimulant, one antiepileptic, four analgesics and non-steroidal anti-inflammatories, one lipid regulators, two anti-ulcer agents, nine antibiotics (sulfonamides and macrolides), two beta-blockers, two metabolites, and 11 hormones (free and conjugates). The determination was performed using liquid chromatography coupled to tandem mass spectrometry after enrichment by solid-phase extraction with Oasis HLB sorbent. Most of the pharmaceuticals were found in both influent and effluent samples from the two STPs. The most frequently detected were caffeine, acetaminophen, carbamazepine, diclofenac, ibuprofen, naproxen, sulfamethoxazole, sulfapyridine, sulfathiazole, ranitidine, omeprazole, estrone 3-sulfate, and estradiol 17-glucuronide. Specifically, the highest concentrations found in influents were 19,850 ng/L (acetaminophen), 9,945 ng/L (caffeine), 4,215 ng/L (ibuprofen), 5,695 ng/L (sulfamethoxazole), and 5,140 ng/L (sulfathiazole). Most of the pharmaceuticals present in influent waters were found in effluents at lower concentrations. The highest concentrations in effluents were 970 ng/L (caffeine), 670 ng/L (sulfamethoxazole), 510 ng/L (bezafibrate), and 1,032 ng/L (diclofenac).

Determination of steroid sex hormones in water and urine matrices by stir bar sorptive extraction and liquid chromatography with diode array detection

In this study, stir bar sorptive extraction and liquid desorption followed by high performance liquid chromatography with diode array detection (SBSE–LD–HPLC/DAD) were combined for the simultaneous determination of nine steroid sex hormones (estrone, 17α-estradiol, 17β-estradiol, 17α-ethynylestradiol, diethylstilbestrol, mestranol, progesterone, 19-norethisterone and norgestrel) in water and urine matrices. During the method development, it has been demonstrated that equilibrium time, ionic strength and back extraction solvents are the most important parameters to control, for determining the nine-hormones in water matrices, in which stir bars coated with 126 μl of polydimethylsiloxane were used. Assays performed on 30 ml water samples spiked at 10 μg/l levels under optimised experimental conditions, yielded recoveries ranging from 11.1 ± 4.9% (17β-estradiol) to 100.2 ± 10.4% (mestranol), showed that the methodology is well described by the octanol–water partition coefficients ( K PDMS/W ≈ K O/W) for the latter, while pronounced deviations to the theoretical efficiency ( K PDMS/W ≠ K O/W) were observed for the remaining hormones. From calibration studies, a good analytical performance for all hormones was attained, including a suitable precision (2.1–17.1%), low limits of detection (0.3–1.0 μg/l) and an excellent linear dynamic range (1.25–50.0 μg/l). Assays on environmental water and urine matrices showed recovery yields in worthy good agreement with the spiking level (10 μg/l), and suitability for profiling low μg/l levels of natural hormones in urine samples taken from pregnant women. The present methodology is easy, reliable and sensitive at the trace level, only requiring a low sample volume, showing to be a good analytical alternative to routine quality control for environmental and biomedical laboratories.

Concentration of Hydrophobic Organic Compounds by Polymer-Mediated Extraction

A water-soluble polymer, poly(N-isopropylacrylamide), becomes water-insoluble above a lower critical solution temperature (∼32 °C) and forms gumlike precipitates (polymer phase) having a very small volume fraction. The features in the polymer precipitation were dependent on the molecular weight of the polymer and/or the concentration of salts or buffer components. Accompanying the formation of the polymer phase, hydrophobic organic compounds including polycyclic aromatic hydrocarbons, alkylphenols, chlorobenzenes, chlorophenols, phthalic esters, pesticides, and steroid hormones in water were efficiently incorporated into the polymer phase. On the other hand, hydrophilic components such as inorganic ions or polysaccharides remained in the bulk aqueous solution. The extractabilities of the compounds increased with increasing hydrophobic properties. The distribution coefficients, based on the volume fractions of their aqueous and polymer phases, between the bulk aqueous and polymer phases were correlated with the water−octanol distribution coefficients. The linear relationship between the two equilibrium coefficients indicated that the hydrophobic interaction is the predominating force for incorporating solutes. The introduction of the concentrated polymer solution into the hygroorganic mobile phase of the reversed-phase HPLC rarely interfered with the chromatographic operation. The application of the polymer-mediated extraction to HPLC significantly enhanced the peak intensities of the hydrophobic analyte varieties by reducing the interferences of the hydrophobic matrix components.

Cytoplasmic dilution induces antidiuretic hormone water channel retrieval in toad urinary bladder.

Antidiuretic hormone (ADH) increases the osmotic water permeability (Pf) of the toad urinary bladder by insertion of water channels into the apical cell membrane. Transepithelial water flow (Jv) reduces Pf by inducing endocytosis of apical water channels despite continuous ADH stimulation. This phenomenon is termed flux inhibition. We wished to determine whether cytoplasmic dilution or transcellular Jv causes flux inhibition because both have been proposed previously as a primary regulatory mechanism for this process. Apical membrane endocytosis was quantified by monitoring the uptake of the fluid phase marker fluorescein isothiocyanate dextran (FITC-dextran). FITC-dextran fluorescence was monitored in Triton X-100 extracts of epithelial cells as the ratio of total tissue fluorescence compared with background fluorescence. The background was defined as cellular autofluorescence and nonspecific tissue staining due to the presence of small amounts of free fluorescein contaminating the FITC-dextran. FITC-dextran uptake measured under symmetric isotonic (220 mosmol/kgH2O) conditions in either the absence (1.0 +/- 0.4 SD; n = 14) or presence (1.3 +/- 0.3; n = 4) of ADH was not statistically different from that of background. In contrast, flux inhibition induced by a 180 mosmol/kgH2O apical-to-basolateral osmotic gradient increased FITC-dextran uptake to 3.4 +/- 1.3 (n = 7). FITC-dextran uptake was identical in bladders exposed to symmetric hypotonic (150 mosmol/kgH2O) solutions during ADH (3.6 +/- 0.9; n = 6) or adenosine 3',5'-cyclic monophosphate (3.1 +/- 0.4 fold; n = 3) stimulation.(ABSTRACT TRUNCATED AT 250 WORDS)

High proton flux through membranes containing antidiuretic hormone water channels

Antidiuretic hormone (ADH) stimulation of toad urinary bladder granular cells causes simultaneous increases in transepithelial water and H+ permeabilities (PF and PH+, respectively), suggesting that ADH-elicited water channels inserted into granular cell apical membranes might be permeable to both water and H+. We have previously used self-quenching fluorophores entrapped within endocytic vesicles selectively retrieved from water-permeable apical membranes to measure vesicle PF. The membranes of these vesicles possess an extremely high PF such that our measurements provide only minimum estimates of vesicle PF and have limited our ability to quantitate the properties of ADH water channels. We therefore quantitated vesicle PH+ using similar rapid mixing techniques. Vesicle PH+ was 5.1 +/- 0.5 x 10(-3) cm/s. Activation energy of this process was 3.6 +/- 0.6 kcal/mol, indicative of H+ flux through an aqueous channel. The mercurial reagent, para-chloromercuribenzenesulfonate (PCMBS), which inhibits ADH-stimulated transepithelial PF in intact bladders by 50-60%, inhibited vesicle PH+ by 55%. N-Ethylmaleimide and phloretin, which do not alter ADH-stimulated PF, did not affect vesicle PH+. We conclude that membranes containing ADH water channels possess substantial PH+ that likely reflects proton flux through water channels. The apparent high PH+ of the ADH water channel may have important implications for intracellular trafficking of these water channels in ADH-responsive epithelial cells.

A Raman spectroscopic investigation of the disulfide conformation in oxytocin and lysine vasopressin.

The conformation of the CCSSCC moiety in oxytocin and lysine vasopressin is investigated using laser Raman spectroscopy. The Raman spectra of solutions of these hormones in water and in dimethyl sulfoxide show an intense band at 508 cm-1 which is assigned to the S-S stretching mode. The presence of shoulders on this band between 490 and 525 cm-1 shows that there is an equilibrium among several conformations for the disulfide unit of these hormones in solution. Most of the CS-SC dihedral angles are within 30 degrees of +/-90 degrees, but some of the molecules have CS-SC dihedral angles strained away from this value by more than 30 degrees. The previously published circular dichroism spectra of these hormones are reinterpreted, and it is shown that the circular dichroism spectra indicate the presence of more than one conformation for the disulfide unit, in agreement with the Raman results.

EFFECT OF A SYNTHETIC ANALOGUE OF VASOPRESSIN IN ANIMALS AND IN PATIENTS WITH DIABETES INSIPIDUS

A synthetic analogue of vasopressin, in which the L-arginine in position 8 has been replaced by D-arginine and the free amino group in position 1 has been removed by replacing the hemicystine component at position 1 by β-mercaptopropionic acid, has been tested in rats and dogs, and in patients with diabetes insipidus. Compared with the natural hormone, the antidiuretic potency of the analogue is higher, but pressor activity is decreased, so that the compound is more specifically a "water hormone" than is the mother substance. Not only can it be administered intranasally, as well as by injection, but it also has a prolonged antidiuretic action even as pure substance in aqueous solution.

Use of desoxycorticosterone acetate in dehydration and malnutrition in infancy.

THE CASES to be reported cannot be considered proved cases of adrenal cortical hypoplasia or deficiency. The patients did, however, have many of the symptoms of adrenal cortical deficiency, and they did respond to desoxycorticosterone acetate and salt therapy usually after other forms of treatment had failed. Unfortunately, there was a lack of sufficient laboratory data, especially of serum potassium, sodium, and chloride levels and of electrocardiograms. At present three clinical syndromes due to disturbances in adrenal cortical secretions are recognized, according to Deamer and Silver, 1 namely, (1) hypofunction of the adrenal cortex causing disturbance of the salt-controlling and water-controlling hormone (Addison's disease), (2) hyperfunction of the adrenal cortex leading to masculinization and precocious maturation (excess androgen secretion), and (3) the combination of an excess of one hormone—androgen—and the simultaneous deficiency of the other—the salt and water hormone. Cushing's syndrome may be considered a fourth possibility. Here there is