To find the target protein and its characteristic peptides suitable for the determination of the whole scorpion of medicinal materials by means of proteomics，and to use this characteristic peptide to form a quantitative method.Methods:After the whole scorpion was crushed，it was ultrasonically extracted with PBS buffer solution and filtered to obtain the protein extract solution of the medicinal material.The above solution was analyzed by high-resolution mass spectrometry after trypsin digestion to determine the target protein to be tested(β-actin)and the corresponding characteristic peptide(IIAPPER)，and the characteristic peptide and its isotope internal standard synthesis(IIA*(13C3，15N)PPER).The obtained characteristic peptide was used as a reference substance，and the isotopic internal standard was used as an internal standard for content determination.The column was Waters ACQUITY UPLC HSS T3(2.1 mm×100 mm，1.8 μm)，the mobile phase was 0.1% formic acid solution(V/V)-water and acetonitrile，and the gradient eluted.Mass spectrometry used the electrospray source ion(ESI)positive ion mode and the multi-reaction monitoring mode(MRM)for signal acquisition.The quantitative method was the internal standard method.Results:The characteristic peptide of β-actin was well linear in the concentration range of 0.1-40 nmol/L，and the correlation coefficient(r2)was above 0.999.The detection limit of the method was 0.04 mg/kg，and the limit of quantification was 0.13 mg/kg.The recovery rate of spiked standard was 87%-93%，and the repeatable RSD is 5.2%.The intermediate precision RSD was 4.8%，and the test solution was stable within 4 h.Using this method，the content of β-actin in 37 batches of whole scorpion medicinal materials from different origins was determined，and the result was between 4.66-100.26 mg/kg.Conclusion:The method has high sensitivity，good accuracy and stable repeatability，and can provide an effective method for the quality control of the whole scorpion medicinal materials.