Implementations herein relates to effect-directed identification of targeted and non-targeted androgen disruptors. The implementations include primary separation, androgenic activity testing, high throughput separation and preparation of toxicants, active component scanning based on high performance liquid chromatography-time of flight mass spectrometry, targeted screening of suspicious androgenic substances, non-target identification of androgenic compounds combining mass spectrum, chromatography and toxicity characteristics and toxicity confirmation. The implementations perform separation using SPE and preparative separation in series to obtain high throughput separation fractions, utilize DMSO as a protective agent to optimize concentration of second fractions, utilize target databases to achieve target identification of key toxicants. In addition, assess mass spectrum characteristic identification based on TOF-MS library spectrum, assess chromatography characteristic identification methods based on relationship between retention times and characteristics of compounds, assess toxicity characteristic identification methods based on molecular dynamics simulation techniques, and further assess non-targeted identification and determination of key androgenic substances in the fractions.