The present invention belongs to the field of probe technology, and provides a trifunctional probe based on a photocrosslinking group, a preparation method thereof, and an application thereof. The method comprises the following steps: under a protective
atmosphere, compound 1, compound 2, 1-
hydroxybenzotriazole,
dimethylformamide,
carbodiimide, and N,N-diisopropylethylamine are mixed, and a
condensation reaction is performed to obtain a trifunctional probe based on a photocrosslinking group. The probe provided by the present invention can be bonded with
baicalein, TFAE, and
scutellaria baicalensis medicated serum to form a corresponding probe, and compete with its original compound. Under excitation of different
ultraviolet wavelengths, the probe can be cross-linked with small molecules and target proteins in
traditional Chinese medicine, and accurately reflect the level of
target protein labeling of the reactive group in the
proteome through a "reporter group", and can also selectively enrich the labeled
target protein, and achieve the identification of single compounds, compound combinations, and natural
drug extract targets through subsequent experiments.