Determination of Zearalanol and Its Analog Zearalanone in Muscle Tissues by Amplified Luminescent Proximity Homogeneous Assay (AlphaLISA)

A homogenous light-induced chemiluminescence immunoassay (AlphaLISA) method was established for the determination of residues of zearalanol and its analog zearalanone in muscle tissue samples. AlphaLISA is a bead-based proximity assay. When donor and acceptor beads proximity, a cascade of chemical reactions begin. The end result is a greatly amplified signal that contributes to the detection sensitivity down to the attomole level. Compared with other methods, the AlphaLISA has characteristics of homogeneity, being free of cleaning, high sensitivity. The method showed a linear relationship in the range of 0.01–4 ng/mL (R 2 > 0.99); the sensitivity of the assay was 0.066 ng/mL. Cross-reactivity rate of zearalanol was 100 % and zearalanone was 82.1 %, and other compounds were not more than 40 %. The average recovery rates of Zearalanol at spiked levels of 1–4 ng/mL were 96.3 to 105.0 and 91.7 to 100.5 % for pork and bovine muscles; the intra-day precision ranged from 2.6 to 9.8 %, and the inter-day precision ranged from 8.7 to 17.5 %. These results indicated that the proposed method was successfully applied in the analysis of zearalanol and its analog zearalanone in muscle tissues.