Biological monitoring of 3-phenoxybenzoic acid in urine by an enzyme-linked immunosorbent assay.
An enzyme-linked immunosorbent assay (ELISA) method was employed for determination of the pyrethroid biomarker, 3-phenoxybenzoic acid (3-PBA) in human urine samples. The optimized coating antigen concentration was 0.5 ng/mL with a dilution of 1:4000 for the 3-PBA antibody and 1:6000 for the enzyme conjugate. Urine samples were hydrolyzed with concentrated hydrochloric acid; extracted with dichloromethane and solvent-exchanged into a methanol/buffer solution, prior to analysis in a 96-microwell plate immunoassay. Quantitative recoveries of 3-PBA were obtained for fortified urine samples by ELISA (92±18%) as well as by gas chromatography/mass spectrometry (GC/MS) (90±13%). The overall method precision of these samples was within ±20% for both the ELISA and GC/MS methods. Analytical results from over one hundred urine samples showed that the ELISA and GC/MS data were highly correlated, with a correlation coefficient of 0.95. At the 10 ng/mL comparative concentration level, the false positive rate was 0% and the false negative rate was 0.8% for ELISA when using GC/MS as the reference method. The ELISA method has a suitable low detection limit for 3-PBA to assess pyrethroid exposures in non-occupational settings.
Battelle, Columbus, OH 43201-2693, USA. chuangj@battelle.org
Jane C
JC
Chuang
Jeanette M
JM
Van Emon
Raquel M
RM
Trejo
Joyce
J
Durnford
D001565:Benzoates / Q000652:urine*; D015415:Biomarkers / Q000652:urine; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D007306:Insecticides / Q000737:chemistry / Q000652:urine; D057230:Limit of Detection; D011722:Pyrethrins / Q000737:chemistry / Q000652:urine
D001565:Benzoates; D015415:Biomarkers; D007306:Insecticides; D011722:Pyrethrins; C017618:3-phenoxybenzoic acid