Abstract: Organophosphorus pesticides are one of the most widely used pesticides at present, which can enter the human body through various pathways including the respiratory tract, digestive tract, skin and mucous membrane. Long term exposure will cause damage to the cardiovascular system, reproductive system, nervous system of human, and even lead to cancer. Human biomonitoring is the “gold standard” for assessing the exposure level of human environmental pollutants. Through the analysis of environmental chemicals and their metabolites in human tissues and body fluids (blood, urine, and breast milk, etc.), the total amount of environmental chemicals entering the human body through different exposure routes can be accurately detected. In order to better understand and evaluate the population exposure dose of organophosphorus pesticides and its effect on health, the common pretreatment methods (liquid-liquid extraction, solid phase extraction, QuEChERS and other methods) and detection methods (gas chromatography, gas chromatography-mass spectrometry, gas chromatography-tandem mass spectrometry, and liquid chromatography-tandem mass spectrometry) for organophosphorus pesticides in human biological samples were reviewed mainly (68 ref. cited).