Fabric Phase Sorptive Extraction of Selected Steroid Hormone Residues in Commercial Raw Milk Followed by Ultra-High-Performance Liquid Chromatography-Tandem Mass Spectrometry.

Hormones in edible matrices, such as milk, are a subject of concern because of their adverse effects on the endocrine system and cell signaling and the consequent disruption of homeostasis in human consumers. Therefore, the assessment of the presence of hormones in milk as potential endocrine-disrupting compounds is warranted. However, the complexity of milk as a sample matrix and the ultra-low concentration of hormones pose significant analytical challenges. Fabric phase sorptive extraction (FPSE) has emerged as a powerful analytical technique for the extraction of emerging pollutants from complex aqueous matrices. FPSE allows for substantially simplified sample handling and short extraction and desorption times, as well as the decreased use of organic solvents. It is considered a green alternative to traditional extraction methodologies. In this work, the FPSE technique was evaluated to perform the simultaneous extraction of 15 steroid hormones from raw milk without employing any sample pretreatment steps. Clean and preconcentrated hormone solutions obtained from FPSE of raw milk were analyzed using ultra-high-performance liquid chromatography-tandem mass spectrometry to achieve low detection limits, which ranged from 0.047 to 1.242 ng·mL-1. Because of the presence of many interferents in milk, such as proteins, lipids, and sugar, the effect of fat content on the extraction procedure was also thoroughly studied. Additionally, for the first time, the effect of lactose on the extraction of steroid hormones was evaluated, and the results showed that the extraction efficiencies were enhanced in lactose-free samples. Finally, the optimized methodology was applied to commercial samples of cow and goat milk, and no measurable concentrations of the studied hormones were detected in these samples.

Pharmaceutical and personal care product residues in a macrophyte pond-constructed wetland treating wastewater from a university campus: Presence, removal and ecological risk assessment.

Pharmaceuticals and personal care products (PPCPs) constitute a group of chemicals of concern because of their potential toxicity when reaching aquatic environments. Wastewaters are one of the main pathways of introduction into the environment of the chemical compounds used in PPCPs because, in most cases, wastewater treatment facilities are not 100% efficient in their removal. This problem is accentuated in rural zones and isolated communities where conventional treatment systems are too expensive to build and operate. Waste-stabilization ponds and constructed wetlands (CWs) are natural wastewater treatment systems which are used to improve the quality of sewage from small communities because of their low cost and easy maintenance. There is growing interest in combining the two technologies to make a more robust system, taking into account their respective strengths and weaknesses. In this work, a combined macrophyte pond-CW system was evaluated for the presence at three sampling points (influent, pond effluent and CW effluent) of fifteen steroid hormones and six benzotriazole ultraviolet stabilizers (BUVSs). None of the targeted BUVS compounds were detected in either the influent or effluent, probably because of the particular characteristics of the population served by the wastewater system. In contrast, eight different steroid hormone compounds were detected at concentrations ranging from 17.3 to 247.7 ng·L-1 in influent samples and from 8.1 to 22.1 ng·L-1 in final effluent samples. The pond-CW system showed high elimination rates of steroid hormone residues with average removal efficiencies of over 77%. This efficacy was confirmed in the ecological risk assessment evaluation that was performed. Final effluents showed a low ecological risk associated with steroid hormones in contrast to the medium-high ecological risks found in the influent samples.

Study on the removal of hormones from domestic wastewaters with lab-scale constructed wetlands with different substrates and flow directions.

Eight wastewater samples from a university campus were analysed between May and July of 2014 to determine the concentration of 14 natural and synthetic steroid hormones. An on-line solid-phase extraction combined with ultra-high performance liquid chromatography coupled with mass spectrometry (on-line SPE-UHPLC-MS/MS) was used as extraction, pre-concentration and detection method. In the samples studied, three oestrogens (17ß-estradiol, estrone and estriol), two androgens (boldenone and testosterone), three progestogens (norgestrel, progesterone and norethisterone) and one glucocorticoid (prednisone) were detected. The removal of hormones was studied in primary and secondary constructed wetland mesocosms. The porous media of the primary constructed wetlands were palm tree mulch. These reactors were used to study the effect of water flow, i.e. horizontal (HF1) vs vertical (VF1). The latter was more efficient in the removal of 17ß-estradiol (HF1: 30%, VF1: 50%), estrone (HF1: 63%, VF1: 85%), estriol (100% both), testosterone (HF1: 45%, VF1: 73%), boldenone (HF1:-77%, VF1: 100%) and progesterone (HF1: 84%, VF1: 99%). The effluent of HF1 was used as influent of three secondary constructed wetland mesocosms: two double-stage vertical flow constructed wetlands, one with gravel (VF2gravel) and one with palm mulch (VF2mulch), and a mineral-based, horizontal flow constructed wetland (HFmineral). VF2mulch was the most efficient of the secondary reactors, since it achieved the complete removal of the hormones studied with the exception of 17ß-estradiol. The significantly better removal of BOD and ammonia attained by VF2mulch suggests that the better aeration of mulch favoured the more efficient removal of hormones.

Optimization and application of fabric phase sorptive extraction coupled to ultra-high performance liquid chromatography tandem mass spectrometry for the determination of cytostatic drug residues in environmental waters.

Every year, hundreds of tons of organic pollutants reach the environment through effluents released from wastewater treatment plants worldwide, and many of these compounds have harmful effects on the aquatic ecosystem. A new class of emerging pollutants of high concern is cytostatic drugs, which are designed to treat different types of cancers by attacking cells. Environmental concentrations of cytostatic drugs are known to be in the range of ngL-1, and for this reason, it is imperative to develop analytical methods of extraction and preconcentration to allow for subsequent instrumental analysis of these drugs. In this work, a rapid, simple and green method for the analysis of seven cytostatic drug compounds that are commonly used in anti-cancer therapies was developed using a novel extraction process based on a powerful miniaturized technique, fabric phase sorptive extraction (FPSE) coupled to ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS). The major parameters that affect the extraction process were optimized. The new method shows good linearity, with a relative standard deviation (RSD) of less than 12%. Relative recoveries higher than 40% were obtained for the studied compounds, and the detection limit of the method was within the values at which these compounds are usually found in environmental water (0.20ngL-1 to 80ngL-1). The Limit of Quantification ranged from 0.68 to 267ngL-1. Significant suppression of the signal due to the matrix effect, a common shortcoming attributed to interference from the extraction process as well as the use of ionization mode, was not observed. Subsequently, the method was applied to real wastewater samples from an effluent obtained from a hospital area and three wastewater treatment plants located in Gran Canaria Island, Spain.

Determination of steroid hormones in fish tissues by microwave-assisted extraction coupled to ultra-high performance liquid chromatography tandem mass spectrometry.

Steroid hormones produce adverse effects on biota as well as bioaccumulation in fish and seafood, making it necessary to develop methodologies to evaluate these compounds in samples related to the food chain. This work presents an analytical method for evaluating 15 steroid hormones in fish tissue. It is based on microwave-assisted extraction and solid-phase extraction coupled to ultra-high-performance liquid chromatography tandem mass spectrometry (MAE-SPE-UHPLC-MS/MS). The proposed method shows appropriate detection limits (0.14-49.0ngg-1), recoveries in the range of 50% and good repeatability. After optimization, the method was applied to different tissues from two small fishes of the Canary Islands that constitute an important level of the food web (Boops boops and Sphoeroides marmoratus) and were exposed to the outfall of the Las Palmas de Gran Canaria wastewater treatment plant. The concentrations of eight detected compounds ranged from below the quantification limits to 3.95µgg-1.

Application of microwave-assisted extraction and ultra-high performance liquid chromatography-tandem mass spectrometry for the analysis of sex hormones and corticosteroids in sewage sludge samples.

Hormonal compounds are a concern to the international community because they can affect the aquatic biota and are therefore considered to be endocrine-disrupting compounds. These compounds have lipophilic properties, so they tend to accumulate in solid matrices, such as sewage sludge. This work presents the optimization of a microwave-assisted extraction process combined with ultra-high performance liquid chromatography-tandem mass spectrometry for the determination of 15 hormonal compounds in sludge samples. The proposed method has relative standard deviations below 23 %, good recoveries (over 71 %) for all compounds, detection limits that ranged from 1.1 to 7.9 ng g(-1) and quantification limits which ranged from 3.7 to 26.3 ng g(-1). The method was used to analyse sludge samples from four different wastewater treatment plants of Gran Canaria (Spain) with different wastewater treatments. 17ß-estradiol, 17α-ethynylestradiol, norgestrel and cortisone were detected in sludge samples at concentrations that ranged from 17.3 to 1.44 × 10(3) ng g(-1). The developed method permits the use of small quantities of sample and organic solvents, presents short extractions times and is the first one based on microwave-assisted extraction for the analysis of both sex hormones and corticosteroids.

Determination of androgens and progestogens in environmental and biological samples using fabric phase sorptive extraction coupled to ultra-high performance liquid chromatography tandem mass spectrometry.

Androgens and progestogens are two important groups of endocrine disrupting compounds (EDCs) which are implicated to produce severe detrimental impact over aquatic biota, even at very low concentrations of ngL(-1). For this reason, one of the major challenges to analytical chemists is the development of sensitive and selective extraction processes which allow the rapid and green determination of these emerging pollutants at low concentrations in environmental samples. Fabric phase sorptive extraction is a new, highly sensitive, efficient and solvent minimized technique which combine the advantages of sol-gel derived microextraction sorbents and the rich surface chemistry of cellulose fabric substrate. This process has several advantages such as minimum usage of organic solvents, short extraction times, small sample volumes and high analyte preconcentration factors. In this study, an extraction method based on sorptive fabric phase coupled to ultra-high-performance liquid chromatography tandem mass spectrometry detection (FPSE-UHPLC-MS/MS) has been developed for the determination of four progestogens and six androgens in environmental and biological samples. All the parameters involved in the extraction, such as sample volume, extraction and desorption times, desorption solvent volume and sample pH values have been optimized. The developed method provides satisfactory limits of detection (between 1.7 and 264ngL(-1)), good recoveries and low relative standard deviations (below 10% in tap and osmosis water and below 20% in wastewater and urine). Subsequently, the method was used to analyse tap water, wastewater treated with different processing technologies and urine samples. The concentrations of the detected hormones ranged from 28.3 to 227.3 ngL(-1) in water samples and from 1.1 to 3.7µgL(-1) in urine samples.

Molecularly imprinted solid-phase extraction coupled with ultra high performance liquid chromatography and fluorescence detection for the determination of estrogens and their metabolites in wastewater.

Estrogens are an important class of endocrine-disrupting compounds, and their contamination of environmental waters through the effluents of wastewater treatment plants could have an important impact on aquatic biota, even at low concentrations. For this reason, the development of selective and sensitive extraction methodologies, which permit the identification and quantification of these compounds at trace level concentrations, is very important. In this study, a quantitative method based on molecularly imprinted solid phase extraction coupled to ultra high performance liquid chromatography with fluorescence detection has been developed. It has been used for the simultaneous determination of three estrogens and two of their metabolites in water samples from wastewater treatment plants. The method developed presents satisfactory limits of detection (between 0.18 and 0.45 ng·mL-1 ), good recoveries (higher than 60%) and low relative standard deviations (under 10%). The method was used to analyze wastewater from a veterinary hospital as well as influent and effluent samples of a wastewater treatment plant of Gran Canaria (Spain) The concentrations of the detected hormones ranged from 1.35 to 2.57 ng·mL-1 .

Simultaneous determination of hormonal residues in treated waters using ultrahigh performance liquid chromatography-tandem mass spectrometry.

In the last years, hormone consumption has increased exponentially. Because of that, hormone compounds are considered emerging pollutants since several studies have determinted their presence in water influents and effluents of wastewater treatment plants (WWTPs). In this study, a quantitative method for the simultaneous determination of oestrogens (estrone, 17 ß -estradiol, estriol, 17 α -ethinylestradiol, and diethylstilbestrol), androgens (testosterone), and progestogens (norgestrel and megestrol acetate) has been developed to determine these compounds in wastewater samples. Due to the very low concentrations of target compounds in the environment, a solid phase extraction procedure has been optimized and developed to extract and preconcentrate the analytes. Determination and quantification were performed by ultrahigh performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). The method developed presents satisfactory limits of detection (between 0.15 and 9.35 ng·L(-1)), good recoveries (between 73 and 90% for the most of compounds), and low relative standard deviations (under 8.4%). Samples from influents and effluents of two wastewater treatment plants of Gran Canaria (Spain) were analyzed using the proposed method, finding several hormones with concentrations ranged from 5 to 300 ng·L(-1).

An assessment of the concentrations of pharmaceutical compounds in wastewater treatment plants on the island of Gran Canaria (Spain).

An assessment of the concentrations of thirteen different therapeutic pharmaceutical compounds was conducted on water samples obtained from different wastewater treatment plants (WWTPs) using solid phase extraction and high- and ultra-high-performance liquid chromatography with mass spectrometry detection (HPLC-MS/MS and UHPLC-MS/MS), was carried out. The target compounds included ketoprofen and naproxen (anti-inflammatories), bezafibrate (lipid-regulating), carbamazepine (anticonvulsant), metamizole (analgesic), atenolol (ß-blocker), paraxanthine (stimulant), fluoxetine (antidepressant), and levofloxacin, norfloxacin, ciprofloxacin, enrofloxacin and sarafloxacin (fluoroquinolone antibiotics). The relative standard deviations obtained in method were below 11%, while the detection and quantification limits were in the range of 0.3 - 97.4 ng·L(-1) and 1.1 - 324.7 ng·L(-1), respectively. The water samples were collected from two different WWTPs located on the island of Gran Canaria in Spain over a period of one year. The first WWTP (denoted as WWTP1) used conventional activated sludge for the treatment of wastewater, while the other plant (WWTP2) employed a membrane bioreactor system for wastewater treatment. Most of the pharmaceutical compounds detected in this study during the sampling periods were found to have concentrations ranging between 0.02 and 34.81 µg·L(-1).

Determination and assessment of estradiol-mimicking compounds in the dissolved and particulate phases of wastewater treatment plant samples.

During the last two decades, a large number of publications have clearly shown that anthropogenic compounds that disrupt the endocrine system of wildlife species are a major cause for concern, and this concern has led to a demand for new screening methods. In this work, we have optimized and applied a new method to identify endocrine-disrupting chemicals (EDCs), such as nonylphenol, octylphenol, and their corresponding ethoxylates, 17alpha-ethynylestradiol, bisphenol-A, 17beta-estradiol, and estriol, in sewage samples. For the extraction and preconcentration of all analytes from the dissolved and particulate phases, we used SPE and ultrasonic assisted extraction, respectively. Identification and quantification were achieved by HPLC with fluorescence detection. Satisfactory LODs (between 0.5 and 7.6 ng/L in the dissolved phase and 12.3 and 21.4 ng/g in the particulate phase) and analyte recoveries (between 67 and 102%) were achieved for the target compounds. The optimized method was applied to the determination of EDCs in liquid sewage samples collected from July 2009 to July 2010 from a wastewater treatment plant in Las Palmas de G.C. (Spain). Concentrations of EDCs ranged from <10 to nearly 1300 ng/L in the dissolved phase, and from 0.1 to 7.7 microg/g in the suspended particulate matter.