title: An immunoassay for the detection of triclosan-O-glucuronide, a primary human urinary metabolite of triclosan.
abstract: Triclosan-O-glucuronide (TCSG) is one of the primary urinary metabolites of the antibacterial compound triclosan or TCS that is found in many personal care products and consumer goods. We have developed a competitive, indirect heterologous ELISA for the detection of the target TCSG in urine. Such an ELISA for TCSG could be developed as a useful tool to measure this important biomarker of human exposure to TCS. Immunogens were prepared by conjugating TCSG to thyroglobulin, via heterobifunctional cross-linkers AEDP or 3-[(2-aminoethyl)dithio] propionic acid•hydrochloride and TFCS or N-[ε-trifluoroacetylcaproyloxy]succinimide ester. The coating antigen was prepared by the direct conjugation of TCSG to bovine serum albumin. Antibodies raised in rabbits 2619, 2621 (immunogen TCSG-AEDP-Thy), and 2623 (immunogen TCSG-TFCS-Thy), and the coating antigen were screened and characterized to determine their optimal concentrations. The optimized ELISA, developed with antibody 2621, gave an IC50 value of 2.85 ng/mL, with the linear range (IC20-IC80) determined to be 2.6-24.8 ng/mL. Selectivity of the assay was assessed by measuring cross-reactivity of antibody 2621 to related congeners such as the aglycone TCS, triclosan-O-sulfate, triclocarban, a polybrominated diphenyl ether derivative, and 3-phenoxybenzyl alcohol glucuronide. There was virtually no recognition by antibody 2621 to any of these cross-reactants. Graphical Abstract Urinary biomarker analysis of triclosan glucuronide.
authors.affiliation: Department of Entomology and UCD Comprehensive Cancer Center, University of California Davis, Davis, CA, 95616, USA.
authors.forename: Anupama
authors.initials: A
authors.lastname: Ranganathan
authors.affiliation: Department of Entomology and UCD Comprehensive Cancer Center, University of California Davis, Davis, CA, 95616, USA.
authors.forename: Shirley J
authors.initials: SJ
authors.lastname: Gee
authors.affiliation: Department of Entomology and UCD Comprehensive Cancer Center, University of California Davis, Davis, CA, 95616, USA. bdhammock@ucdavis.edu.
authors.forename: Bruce D
authors.initials: BD
authors.lastname: Hammock
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000378:metabolism / Q000652:urine*; D020719:Glucuronides / Q000378:metabolism / Q000652:urine*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007106:Immune Sera; D007118:Immunoassay / Q000379:methods*; D009994:Osmolar Concentration; D011817:Rabbits; D014260:Triclosan / Q000378:metabolism / Q000652:urine*
chemical_list: D000900:Anti-Bacterial Agents; D020719:Glucuronides; D007106:Immune Sera; D014260:Triclosan
keywords: Biomarker; ELISA; Polyclonal antibodies; Triclosan-O-glucuronide; Urinary metabolite