Trace Analysis of 5 Androgens in Environmental Waters by optimization of Enzymolysis and Solid-Phase Extraction Ultra-Performance Liquid Chromatography-Mass Spectrometry and its Risk Assessment.

Steroid hormones (SHs), have received widespread attention in recent years. However, current studies of SHs have primarily focused on estrogenic substances and androgen-related studies are still very limited. In this study, the solid-phase extraction (SPE) pre-treatment method, as well as the enzymolysis conditions of 5 androgenic materials, such as Androstenedione, Boldenone, Methandienone, Nandrolone, and Testosterone, were optimized to simultaneously determine their concentrations in the effluent from wastewater treatment plants (WWTP) and surface water samples. Then, the ecological risks of the 5 androgens in the effluent and Pearl River basin of Guangzhou were evaluated with the risk quotient RQ method. The results showed that the recovery rates of the targets are 90%-99% in water samples when digested with β-glucosidase for 90 mins before solid-phase extraction, extracted with a Poly-Sery HLB column, and washed with 15% methanol aqueous solution and 2% ammonia. The established instrument's limit of detection (LOD) was between 0.02 μg/L and 0.39 μg/L, and the limit of quantification (LOQ) was between 0.05 μg/L and 1.29 μg/L. Androstenedione, Boldenone, Methandienone, Nandrolone, and Testosterone were detected in all samples from the 2018 and 2022 wastewater influent and 2018 surface water, with concentrations of 3.06×10 1 ng/L-1.33×10 3 ng/L, 1.03×10 2 ng/L-8.15×10 2 ng/L, and 0.93×10 1 ng/L-5.50×10 2 ng/L, respectively. The ecological risk of androgens in wastewater influent and surface water were medium to high and low to medium respectively. Moreover, the bio-toxicity of androgens was predicted by the ECOSAR model, with Methandienone and Androstenedione having the highest and lowest acute and chronic toxicity, respectively. This study suggests that the risk of environmental androgens should not be ignored, and further research should be carried out.