Integrated recombinant gene yeast bioassay and HPLC-MS analysis for detection of low-dose multi-component residue of hormone-like compounds in environment.

Illegal addition of low-dose multi-component residue of hormone-like compounds have sprung up in human activities and production as a cunning strategy to escape from sanitation monitoring, sowing a horrible seed for accidents in food security. Hence, combined strategies with accuracy and efficiency are urgently needed to overcome current circumstance and practical hurdles. Herein, an integrated analysis that compromises recombinant gene yeast bioassay (V400E-YES) and high-performance liquid chromatography-mass spectrometry (HPLC-MS) determination was developed promptly with distinguishing advantage as optimized V400E-YES provides a sum of hormone-like compounds in sample, and subsequent HPLC-MS analysis can accurately quantify the concentration of compounds. In terms of V400E-YES, large-scale random mutagenesis was performed to obtain higher sensitivity hormone receptors. Excitingly, single mutation of estrogen receptor (ER) at V400E and amphimutation of androgen receptor (AR) at G581R/D831E conferred the highest detection sensitivity with a more than 10-fold increase. Subsequently, sensitive HPLC-MS methods for simultaneous detection of eleven estrogens and nine androgens, respectively, were well-constructed. Utilizing this integrated and validated methods, we successfully investigated the hormone residue in environmental samples from farms in Jiangsu, China. Collectively, with multiple estrogens and androgens residue are being widely detected in soil and sewage samples, the feasibility of this method is highlighted, as well as the increasing lurking peril of hormone-like substance residues in environment.