Sensitive Estrogens Determination in Wastewater Samples by HPLC and Fluorescence Detection

Abstract

A sensitive high-performance liquid chromatography-fluorescence detection method (HPLC-FLD) was developed and validated for 17β-estradiol (E2) and 17α-ethinylestradiol (EE2) determination in wastewater samples. Sample preparation was performed by solid-phase extraction (SPE) on Hypersep C18 cartridges. Conditioning and elution solvents and sample volume were investigated by a 2 3 factorial design for both analytes. The pre-concentration factor was 2500 times, allowing to quantify 5.0 ng L -1 for both hormones. The SPE-HPLC-FLD method was successfully applied to detect and quantify trace amount of target compounds in raw and treated wastewater samples. E2 was found below limit of quantification in influent and effluent samples. Nevertheless, the recoveries of E2 in standard addition experiments were between 41 and 72%, with relative standard deviations between 4 and 16%. An important matrix effect was observed in the determination of EE2, with fluorescence signal suppression. Keywords: estrogens, SPE, HPLC, fluorescence, wastewater IntroductionContamination of water resources represents a worldwide concern. Excessive population growth with the inefficient wastewater collection, particularly in developing countries, low treatment efficiency and inadequate sewage disposal, are the common causes of water contamination.1 Endocrine disrupting compounds (EDCs), especially estrogens, are chemical species that have negative activity on the endocrine system in humans 2,3 and animals. 4,5 Estrone (E1), 17β-estradiol (E2) and estriol (E3) are estrogens naturally produced by mammals and excreted mainly by pregnant and menstrual women via urine and feces.1 Otherwise, 17α-ethinylestradiol (EE2) is a synthetic estrogen widely used as contraceptive and for therapeutic purposes such menopausal syndrome and prostate cancer control. 6 Despite of being unregulated contaminants in environmental waters, they have already been included in the New Contaminant Candidate List-3 of the U.S.EPA and in the list of priority substances of the EU Water Framework Directive. 7,8 These compounds have been detected in wastewater treatment plants (WWTP), [9][10][11][12] in natural waters 9,13 and, even in water supplies.14 Since 1990, estrogenic activity has been detected in many countries. For instance, E1, E2, and EE2 were found in concentrations about ng L -1 in the effluent from a sewage treatment plant in the UK. 15 In Brazil, monitoring studies of natural estrogens in domestic sewage also were initiated in the 90s. 16 This study revealed estrogen concentrations between 20 and 40 ng L -1 and only partial removal by conventional treatments based on biological filter and activated sludge systems. Since then, other monitoring works reported the presence of endocrine disruptors in surface waters in Brazil. Sodré et al. 17 The estrogens monitoring in sewage and natural waters is a challenge due to the low concentration of the analytes and the complexity of the environmental matrices. Most of the reported methods to estrogens determination in aque...

Development of Electrochemical Platform Based on Molecularly Imprinted Poly(methacrylic acid) Grafted on Iniferter‐modified Carbon Nanotubes for 17β‐Estradiol Determination in Water Samples

“…The t-value obtained experimentally (t = 1.49) was lower than the critical value of t tab = 2.47, which indicates no significant differences between the proposed method and HPLC-DAD. Although there is no national and international legislation regarding the maximum allowed levels of E2 in natural waters, the LOQ obtained with this procedure allows the monitoring of this molecule considering the levels already detected in water bodies [66,67].…”

High Performance Liquid Chromatography (HPLC) with Fluorescence Detection for Quantification of Steroids in Clinical, Pharmaceutical, and Environmental Samples: A Review

“…Steroids can be found in various environmental samples, including water, plant, and animal samples. Water is the transporter medium and reservoir for various synthetic steroids, whereas plants were found to contain many different natural steroids [ 17 , 52 , 93 , 94 , 95 ], which can be classified according to their biological relevance. For example, plant physiological steroids can be present as hormones (e.g., brassinolide) and pheromones (e.g., antheridiol), whereas plant allelochemical substances are biologically related to animal hormones such as vertebrate hormones (progesterone) and insect hormones (ecdysone).…”

“…Many studies used HPLC-FLD for the analysis of steroids in environmental samples due to its sensitivity and selectivity. Studies investigated the steroid content in samples collected from drinking water, wastewater, river water, tap surface water, and mineral and underground river water [ 17 , 20 , 37 , 52 , 92 , 93 , 94 , 95 ]. Steroids were also analyzed in samples taken from effluent, influent, and sediment wastewater in treatment plant [ 41 ].…”

“…Levels of steroids are required to be continuously monitored since any increase or decrease may be associated with major consequences on health and environment. Therefore, steroid quantification has been gaining increasing importance in disease diagnosis (e.g., increased levels of cortisol in stress and Cushing’s syndrome) [ 15 ], contamination assessment of water and food samples [ 16 , 17 , 18 , 19 , 20 , 21 ], detection of anabolic steroids among sportsmen due to doping [ 22 , 23 , 24 , 25 , 26 ], and detection of growth promoting steroids in animal tissue [ 18 , 25 , 27 , 28 , 29 , 30 , 31 , 32 , 33 ]. While there are several reviews on steroid quantification by HPLC methods coupled to MS and UV [ 8 , 34 ], to the best of our knowledge no similar reports summarizing data on HPLC-FLD methods exist.…”

Research developments in carbon materials based sensors for determination of hormones

“…It is used in menopausal hormone therapy and prostate cancer control, and is excreted through urine and feces. EN is an endocrinedisrupting chemical that affects endocrine systems and can lead to carcinogenic effects, birth faults, and other growth issues [135][136][137][138][139][140]. So, its detection is important for human health and environmental concern.…”