Suitability of Visualized Fluorescent Zebrafish Model for Estrogen Detection in Cosmetics

To address the limitation of single-target detection in existing estrogen analysis techniques, a biological effect-oriented method based on in vivo imaging was developed to systematically evaluate the performance of fluorescent zebrafish model for estrogen detection in cosmetic matrices. The estrogen response element-driven green fluorescent protein (Tg(5×ERE:eGFP)cpu110) transgenic zebrafish was constructed by Tol2 transposase, and the gene regulatory network of 17α-ethinylestradiol (EE2) was resolved by a combination of reverse transcription quantitative polymerase chain reaction (RT-qPCR) and transcriptomics to determine its sensitive window period (48 h). The model could simultaneously detect 9 kinds of estrogenic active substances, including typical endocrine disruptors such as estrone (E1) and estradiol (E2), which were banned and restricted by the Technical Code for Cosmetic Safety (2015 version), and the limits of detection (LOD, 0.04 μg/g) of hexestrol, ethinylestradiol and dienestrol were comparable to those of the high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method (0.03 μg/g). The qualitative results of the blinded experiments were in 100% agreement with those of HPLC-MS/MS. With the triple validation of “gene regulation-fluorescence quantification-instrument calibration”, the model showed a sensitivity level of μg/g, strong specificity and matrix interference resistance, which provided an innovative methodology for high-throughput screening and risk assessment of estrogenic endocrine disruptors in cosmetics and raw materials, and also provided a high-throughput screening tool for regulation of estrogenic safety in cosmetic products.