Legumes constitute a significant protein source in the human diet. Among this category, the chickpea (Cicer arietinum L.) is recognized as an alternative crop due to its efficient water usage. This efficiency has generated a growing interest in its improvement, commercialization and generation of value-added products, particularly within the domain of functional foods. The literature contains a multitude of techniques for evaluating protein content in legumes, complicating the comparison of results among different research groups. The aim of this study was to assess the effect of four distinct extraction buffers on protein concentration in extracts of chickpea flour from two varieties, Costa 2004 and Blanoro, testing the effect of the extraction temperature at 25 and 37 °C. Additionally, the integrity of the samples was evaluated by analyzing the protein profile using SDS-PAGE. The results obtained showed a significant difference in protein concentration due to the choice of extraction buffer and extraction temperature. We propose the implementation of Tris buffer 500 mM pH 6.8 at 25°C to standardize the process of characterizing protein extracts from chickpea flour for comparative purposes. This standardized protocol serves as a valuable tool for agronomic characterization and offers a comprehensive framework for future research in this field.