Zhao Ke-Meng, Zhong Shu-Shu, Zhang Jun, Zhang Cun-Sheng, Dang Zhi, Liu Ze-Hua
School of Environment and Energy, South China University of Technology, Guangzhou, China.
Key Lab Pollution Control and Ecosystem Restoration in Industry Cluster, Ministry of Education, Guangzhou, China.
Water Environ Res. 2022 Apr;94(4):e10704. doi: 10.1002/wer.10704.
Arylsulfatase and β-glucuronidase are two important enzymes in wastewater and surface water, which play important roles on cleavage of sulfate/glucuronide estrogens. In this work, a high-performance liquid chromatography (HPLC)-based new method was firstly established for arylsulfatase/β-glucuronidase with determination of p-nitrophenyl sulfate (pNPS)/p-nitrophenyl-β-D-glucuronide (pNPG). The limits of detections (LODs) of the developed method for pNPS and pNPG were 0.164 and 0.098 μM, respectively. Intraday and interday reproducibility expressed as relative standard deviation (RSD) values of retention times and peak areas was 0.39%-3.68% and 0.23%-4.74%, respectively. The respective recovery efficiencies of this HPLC-based method spiking at three different concentrations for p-nitrophenol (pNP), pNPS, and pNPG in activated sludge were 76.5%-88.1%, 79.2%-93.1%, and 84.2%-96.1%, with RSD below 3.9%. The HPLC-based method was finally applied to estimate the enzyme activity of arylsulfatase/β-glucuronidase in one activated sludge system and along which the classical spectrophotometric method was also evaluated. Compared with the classic spectrophotometric analytical method, the HPLC-based new method could simultaneously measure arylsulfatase/β-glucuronidase one time, which was convenient and time-saving. Moreover, the developed method could effectively avoid possible underestimation that the spectrophotometric method might encounter. PRACTITIONER POINTS: A new HPLC-based method for activity estimation of arylsulfatase and β-glucuronidase was developed. The HPLC-based method can simultaneously estimate enzyme activity of both arylsulfatase and β-glucuronidase. The HPLC-based method can avoid possible underestimation that spectrophotometric method may encounter.
芳基硫酸酯酶和β-葡萄糖醛酸酶是废水和地表水中的两种重要酶，它们在硫酸酯/葡萄糖醛酸酯雌激素的裂解中起重要作用。在本研究中，首次建立了一种基于高效液相色谱（HPLC）的新方法，用于测定芳基硫酸酯酶/β-葡萄糖醛酸酶以及对硝基苯硫酸酯（pNPS）/对硝基苯-β-D-葡萄糖醛酸苷（pNPG）。该方法对pNPS和pNPG的检测限分别为0.164和0.098μM。以保留时间和峰面积的相对标准偏差（RSD）值表示的日内和日间重现性分别为0.39%-3.68%和0.23%-4.74%。该基于HPLC的方法对活性污泥中三种不同浓度的对硝基苯酚（pNP）、pNPS和pNPG加标的回收率分别为76.5%-88.1%、79.2%-93.1%和84.2%-96.1%，RSD低于3.9%。该基于HPLC的方法最终应用于评估一个活性污泥系统中芳基硫酸酯酶/β-葡萄糖醛酸酶的酶活性，同时还对经典分光光度法进行了评估。与经典分光光度分析方法相比，基于HPLC的新方法可以一次性同时测定芳基硫酸酯酶/β-葡萄糖醛酸酶，方便省时。此外，所开发的方法可以有效避免分光光度法可能遇到的低估问题。从业者要点：开发了一种基于HPLC的新方法用于芳基硫酸酯酶和β-葡萄糖醛酸酶活性的评估。基于HPLC的方法可以同时评估芳基硫酸酯酶和β-葡萄糖醛酸酶的酶活性。基于HPLC的方法可以避免分光光度法可能遇到的低估问题。
