Critical evaluation of extracellular polymeric substances extraction methods: Extraction efficiency, molecular characteristics, and heavy metals binding properties.

Abstract

Extracellular polymeric substances (EPS) significantly influence the properties and performance of waste activated sludge. Various pretreatment protocols with different extraction efficiency and characteristics of EPS have been reported, which markedly impact subsequent treatment and disposal of sewage sludge. This study systematically assesses the EPS properties from twelve extraction pretreatment methods. The organic and inorganic matters content, cell lysis, basic physicochemical property, molecular weight distribution, and fluorescence properties of extracted EPS were determined. Physical extraction methods (Centrifugation, Heat, and Ultrasound) were relatively mild, resulting in lower extraction of organic matter contents from EPS (< 6 mg total organic carbon /g volatile solids). Biological extraction methods (Enzyme and Enzyme-NaOH) exhibited high EPS extraction efficiency however led to significant cell lysis, thereby contaminating the extracted EPS with intracellular substances. In addition, heating and biological extraction methods excessively degraded EPS, resulting in a large amount of small molecular weight matters (≤ 10³ Da) generation. Chemical extraction methods offered efficient EPS extraction with the different degrees of cell lysis, but it would introduce chemical reagent residues in EPS. Among those extraction methods, EPS extracted by cation exchange resin (CER) method had uniform and abundant molecular weight matters and fluorescence matters distribution. After dialysis, the residual Na and other metal elements could be significantly removed. Then, the fluorescence quenching effect of Cu(II) and Zn(II) at 400 μM on the Bound-EPS after dialysis increased up to the maximum value of 65 % and 30 %, respectively, compared to that without dialysis. It indicates that dialysis coupled with CER extracted EPS has a well extraction efficiency and can maintain the original state of EPS for the subsequent investigation.