[Objective] This study aimed to explore a Tricine-SDS-PAGE method for separation and purification of Periplaneta Americana small peptides. [Method] By comparing the separation effects of three types of gels with different ratios on P. Americana small peptides, an appropriate Tricine-SDS-PAGE method for separation of small peptides was established. [Result] Standard low molecular weight Marker was preliminarily separated to a certain extent with polyacrylamide gel electrophoresis, while the sample mixture was not separated due to the different levels of aggregation in separating gel, spacer gel and stacking gel. [Conclusion] More appropriate polyacrylamide gel electrophoresis separation method or other better separation methods were to be further explored for separating peptide mixture with smaller molecular weight. [Objective] This study aimed to explore a Tricine-SDS-PAGE method for separation and purification of Periplaneta Americana small peptides. [Method] By comparing the separation effects of three types of gels with different ratios on P. Americana small peptides, an appropriate Tricine-SDS-PAGE method for separation of small peptides was established. [Result] Standard low molecular weight Marker was preliminarily separated to a certain extent with polyacrylamide gel electrophoresis, while the sample mixture was not separated due to the different levels of aggregation in separating gel, spacer gel and stacking gel. [Conclusion] More appropriate polyacrylamide gel electrophoresis separation method or other better separation methods were to be further explored for separating peptide mixture with smaller molecular weight.