Simple solvatochromic spectroscopic quantification of long-chain fatty acids for biological toxicity assay in biogas plants

Oily organic waste is a promising feedstock for anaerobic co-digestion. Free long-chain fatty acids (LCFAs) produced from lipids can inhibit methanogenic consortia, so optimal control of LCFA concentration is the key to successful operation of co-digestion. Most LCFAs are present in the solid phase, making them difficult to be detected and monitored. This study proposes a simple and easy method for detecting LCFAs in both the liquid and solid phases of anaerobic digestate by combining liquid–liquid extraction followed by solid-phase extraction (SPE) and spectrophotometric analysis. The extraction procedure successfully removed impurities that interfere with the absorbance spectrum and ensured high recovery rates of LCFAs. The utility of the pretreatment used for the extraction was discussed using thermodynamic analysis and calculations of phase equilibrium for the solvent extraction system. The absorbance spectrum shift of pyridinium N-phenolate betaine (PNPB) dye-stained solution showed a good correlation with LCFA concentration and enabled highly sensitive measurements. Good quantification was demonstrated in experiments using various digestate samples obtained from the laboratory, pilot, and full-scale reactors.