Quantitative detection method for steroid hormone in human/animal body fluid and application

The invention discloses a quantitative detection method for 24 steroid hormones in human/animal plasma, serum, urine and other body fluids, the method is based on ultra-high performance liquid chromatography-tandem mass spectrometry combined with online enzymolysis-solid phase extraction, the total amount of the 24 steroid hormones in the free state and the combined state can be detected at a time, the detection limit of the method can reach pg/mL, and the detection time is greatly shortened. A methodological verification result meets a 9101-analysis method verification guidance principle. The detection method provided by the invention can accurately and quantitatively detect 24 steroid hormones in human/animal plasma, serum, urine and other body fluids at the same time, has the advantages of high sensitivity, strong specificity, small required sample size, short detection time, improvement of the sensitivity, precision, detection efficiency and detection flux of the method, low cost, avoidance of resource wa