Abstract
BackgroundThe increasing prevalence of male infertility and the declining trend in sperm quality has been associated to compounds known as “endocrine-disruptors”. The proven endocrine-disrupting effects of atrazine and propazine herbicides led us to conduct long-term research based on highly accurate specific analytical methods with a view to confirming the suspected association. Among the proposed developments was a sensitive analytical method for the simultaneous determination of three metabolites of atrazine and propazine.ResultsIn this work, the method was for first time used for the chromatographic separation and determination of deethyl- and deisopropyl-atrazine (DEA and DIA, respectively) and propazine-2-hydroxy (PP-2OH) in human seminal plasma by LC–ESI-MS/MS using deuterated atrazine (d5-AT) as internal standard (IS). Chromatographic and mass spectrometric conditions such as the mobile phase composition and flow-rate, injected volume, dry gas source temperature and flow-rate, nebulizer pressure and capillary voltage were all carefully optimized. Analytes were identified and quantified by using the multiple reaction monitoring (MRM) mode as applied to positive ions ([M + H]+). Transitions at three different m/z values for each analyte were selected from precursor ions, and the 212.1 → [128]+, 188.1 → [146]+ and 174.1 → [68.1]+ transitions for PP-2OH, DEA and DIA, respectively, were found to be quantitative. The proposed method was validated in terms of precision (repeatability and reproducibility), linear range (10–240 ng mL–1), limit of detection (150–210 pg mL–1), and quantification (500–700 pg mL–1), recovery, accuracy and matrix effects on extracts from variably treated seminal plasma samples. The overall analytical method was successfully applied to human seminal plasma samples from volunteers. PP-2OH was found at concentrations from 1.10 to 11.3 ng mL–1 in four of the six samples, and so was DIA at 9.60 ng mL–1 in one.ConclusionsThese results are suggestive of bioaccumulation of the target analytes in humans. Untargeted analytes including suspected parent molecules (atrazine and propazine) and other ions [viz., deethyldeisopropyl-atrazine (DD) and diamino-s-chlorotriazine (DACT)] were also detected under the working conditions used. These results may open up new prospects for as yet very incipient research into the bioaccumulation of endocrine disruptors in seminal plasma.Graphical
Highlights
Defective sperm function can arise from a wide range of primary genetic, lifestyle and environmental factors acting alone or, more frequently, in combination
We developed the first accurate, sensitive, selective method for the simultaneous determination of two active intermediate metabolites of atrazine (DIA and DEA), and the degradation product of propazine PP-2OH, which US EPA has deemed endocrine disruptors
The samples were previously subjected to enzymatic hydrolysis (EH) and solid-phase extraction (SPE) in order to isolate the analytes, clean-up and preconcentrate the final extract
Summary
Defective sperm function can arise from a wide range of primary genetic, lifestyle and environmental factors acting alone or, more frequently, in combination. Selvaraju et al (2020) reviewed the effects of environmental contaminants on male reproductive function and their potential mechanisms of action. EDCs, which have been deemed a global threat to public health by the World Health Organization (WHO), are arousing increasing interest [4] These compounds are highly heterogeneous in nature and have been classified according a number of factors [2] including physicochemical properties, environmental persistence and bioaccumulation in humans [5]. The production and use of various chemicals as additives in agriculture has grown substantially in recent decades Some such chemicals can have harmful effects on health, so their release and/ or accumulation in the environment or in food chains is a cause for concern. Among the proposed developments was a sensitive analytical method for the simultaneous determination of three metabolites of atrazine and propazine
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