Detection of 17β-Estradiol in River Water and Human Urine by Highly Sensitive Chemiluminescence Enzyme Immunoassay

A highly sensitive chemiluminescence enzyme immunoassay (CLEIA) was developed for the determination of 17β-estradiol (E2) in river water and human urine using the FITC-anti-FITC system as solid phase, the avidin-biotin system as signal amplification system, and the highly sensitive 4-methoxy-4-(3-phosphatephenyl)-spiro-(1,2-dioxetane-3,2′-adamantane) (AMPPD)-alkaline phosphatase (ALP) system as chemiluminescence detection. After optimizing the conditions of enzyme reaction, the linear range of E2 is 2.5–2000 pg ml−1 and detection limit is 1.1 pg ml−1. The coefficient of variation (CVs) of intra- and inter-assay was below 10%. The percent recoveries are in the range of 92–112% in human urine. This method has been successfully applied to detect E2 in river water and human urine. The concentration of E2 in river water is in the range of 0.1–0.57 pg ml−1, and in human urine is in the range of 0.75–135 μg ml−1