Method for detecting endogenous steroid hormone testosterone in animal serum

The invention relates to the technical field of chemical analysis and quantitative detection, in particular to a method for detecting endogenous steroid hormone testosterone in animal serum. The invention relates to a sample pretreatment method for detecting endogenous steroid hormone in animal serum, the endogenous steroid hormone is testosterone, the sample pretreatment method comprises the following steps: adding a stable isotope internal standard solution into a to-be-detected serum sample, mixing, adding formic acid and ammonium acetate, and reacting to obtain a pretreated to-be-detected serum sample; the invention discloses a method for detecting endogenous steroid hormones in animal serum by using an ultra-high performance liquid chromatography-triple quadrupole tandem mass spectrometry (UPLC-MS/MS) technology, only a small amount of animal serum needs to be collected, and the method has the advantages of no need of derivatization, high selectivity, high flux, high sensitivity, short operation time and