Development of a highly sensitive chemiluminescent assay for hydrogen peroxide under neutral conditions using acridinium ester and its application to an enzyme immunoassay.

We developed a highly sensitive chemiluminescent (CL) assay for hydrogen peroxide using 10-methyl-9-(phenoxycarbonyl) acridinium fluorosulfonate (PMAC) that produced chemiluminescence under neutral conditions and applied it to an enzyme immunoassay (EIA). One picomole of hydrogen peroxide could be detected using the optimized PMAC-CL method and 6.2 x 10(-20) mol beta-d-galactosidase (beta-gal) could be detected by combining an indoxyl derivative substrate and the proposed PMAC-CL method. This highly sensitive CL beta-gal assay was applied to an EIA for thyroid-stimulating hormone (TSH) using beta-gal as a label enzyme; 0.02-100.0 mu U/mL TSH in human serum could be assayed directly and with high reproducibility. Copyright (c) 2013 John Wiley & Sons, Ltd.