[Background] Protein sample preparation is a central step in a 2-DE proteomics as pure protein samples are necessary. There is a need to establish a routine protocol for the application of proteomics analysis in insects.[Method] This study focused on the specific protein extraction problems in insect tissued and evaluated two protocols to bypass them. The methods of phenol extraction (PE) and TCA/acetone precipitation (TCA) were evaluated for proteins isolation from adult rice water weevil, Lissorhoptrus oryzophilus Kuschel.[Result] TCA method yielded a greater protein extraction rate (14.55 μg·mg^-1) than the PE protocols (9.30 μg·mg^-1). The two methods got similar SDS-PAGE results but the expression of protein was higher in TCA than PE; For 2-DE, the PE protocol was optimal, resulting in good background and clear protein spots and TCA had an indistinct background and occurred a few bands were present.[Conclusion and significance] Among the applied methods, PE precipitation extraction was found to be more suitable than TCA for rice water weevil proteomic analysis. These protocols should also be applicable to other insects. Different insects can vary considerably in the amounts and types of interfering compounds they produce, so the results of protein extraction rate and purity in different insects should be assessed. The study proves that various protein extraction methods result in different protein rates and 2-DE products. It provides basic protocols for proteomic analysis of adult rice water weevil.