Activity measurement of arylsulfatase and β-glucuronidase in activated sludge: HPLC-based versus classical spectrophotometric method,Water Environment Research

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Activity measurement of arylsulfatase and β-glucuronidase in activated sludge: HPLC-based versus classical spectrophotometric method
Water Environment Research Pub Date : 2022-03-13 , DOI: 10.1002/wer.10704
Ke-Meng Zhao ₁ , Shu-Shu Zhong ₁ , Jun Zhang _{1,

2,

3} , Cun-Sheng Zhang ₄ , Zhi Dang ₁ , Ze-Hua Liu _{1,

2,

3}

Affiliation

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.

中文翻译：

活性污泥中芳基硫酸酯酶和 β-葡萄糖醛酸酶的活性测量：基于 HPLC 与经典分光光度法的比较

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，在硫酸盐/葡萄糖醛酸雌激素的裂解中发挥重要作用。本工作首次建立了基于高效液相色谱（HPLC）的芳基硫酸酯酶/β-葡萄糖醛酸酶测定对硝基苯硫酸盐（ p NPS）/对硝基苯基-β-D-葡萄糖醛酸苷（ p NPG）的新方法。）。所开发的 pNPS 和 pNPG 方法的检测限 (LOD) 分别为 0.164 和 0.098 μM。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 的方法在活性污泥中添加三种不同浓度的对硝基苯酚 ( p NP)、 p NPS 和p NPG 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%。最终应用基于 HPLC 的方法来估计活性污泥系统中芳基硫酸酯酶/β-葡萄糖醛酸酶的酶活性，并同时对经典分光光度法进行了评估。与经典的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量芳基硫酸酯酶/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。

更新日期：2022-03-13

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