The application relates to a method for constructing a target
proteome spectrum
library and application in micro target
proteome analysis, and the method comprises the following steps: performing multiple times of
mass spectrum scanning on a target
proteome in a data-dependent acquisition mode without dynamic exclusion through different scanning ranges, so as to obtain
mass spectrum data of the target proteome, wherein the
mass spectrum data of the target proteome comprises primary spectra of multiple scanning ranges of the target proteome and secondary spectra of parent ions in each scanning range, which meet a preset threshold; and performing
library searching based on the
mass spectrum data of the target proteome to construct the target proteome spectrum
library. The application constructs a spectrum library through
mass spectrum data in a data-dependent acquisition mode without dynamic exclusion in multiple scanning ranges, performs
library searching analysis on DIA data of a micro proteome, improves the success rate of DIA
data matching, and further improves the accuracy and sensitivity of DIA
data analysis, and improves the accuracy and sensitivity of micro
protein identification.