Detection of acrylamide in food based on MIL-glucose oxidase cascade colorimetric aptasensor

Abstract

Background: Maillard reaction involves the polymerization, condensation, and other reactions between compounds containing free amino groups and reducing sugars or carbonyl compounds during heat processing. This process endows unique flavors and colors to food, while it can also produce numerous hazards. Acrylamide (AAm) is one of Maillard's hazards with neurotoxicity and carcinogenicity, these effects can trigger mutations and alternations in gene expression in human cells and accelerate organ aging. An accurate and reliable acrylamide detection method with high sensitivity and specificity for future regulatory activities is urgently needed.

Results: Herein, we constructed a colorimetric aptasensor with the hybridization of MIL-glucose oxidase (MGzyme)-cDNA and magnetic nanoparticle-aptamer (MNP-Apt) to specifically detect AAm. The incorporation of MB-Apt and AAm released MGzyme-cDNA in the supernatant, took the supernatant out, with the addition of glucose and TMB, MGzyme would oxidize glucose, the resulting •OH facilitated the oxidation of colorless TMB to blue ox-TMB. The absorbance value at 652 nm, which indicates the characteristic absorption peak of ox-TMB, exhibited a proportion to the concentration of AAm. MGzyme avoided the addition of harmful intermediate H₂O₂ and created an acid microenvironment for the catalytic reaction. MNP-Apt possessed the advantages of high specificity and simplified separation. Under optimal conditions, this method displayed a linear range of 0.01-100 μM with the limit of detection of 1.53 nM. With the spiked analysis data cross-verified by ELISA kit, this aptasensor was proven to specifically detect AAm at low concentrations.

Significance: This colorimetric aptasensor was the integration of aptamer and the enzyme-cascade system, which could broaden the applicable range of enzyme-cascade system, break the limits of specific detection of substrates, eliminate the need for harmful intermediates, improve the reaction efficiency, implement the specific detection, whilst enabling the accurate detection of AAm. Given these remarkable performances, this method has shown significant potential in the field of food safety inspection.

Keywords: Acrylamide; Aptamer; Cascade; Glucose oxidase; Metal-organic frameworks.