High sensitivity and specificity Enzyme-Linked immunosorbent assay for detection of Acetaminophen in herbal tea and liquor

Abstract

The acidified structure of acetaminophen (ACA) and 3-Oxo-3-(phenylamino) propanoic acid (3-O-3) were used to couple bovine serum albumin (BSA) to prepare haptens, respectively. A highly sensitive and specificive competitive indirect enzyme-linked immunosorbent assay (ciELISA) method was developed for the detection of acetaminophen using 3-O-3-BSA for New Zealand white rabbit immunization to produce antibodies. The IC50 was 6.66 ng/mL, the detection limit was 0.32 ng/mL, and the quantitative detection range was 0.91-48.54 ng/mL. Comparing the performance of ACA-BSA antibody and 3-O-3-BSA antibody recognition of acetaminophen, it was found that the carbonyl site of 3-O-3 plays a major role in antigen-antibody recognition, while para-phenolic hydroxyl does not play an important role in the recognition process. The study provided theoretical guidance for the preparation of acetaminophen recognition antibodies, and established a rapid detection method for acetaminophen.