Abstract: Surface hydrophobicity of leghemoglobin (Lb), myoglobin (Mb) ,cytochrome c (Cyt C) and hemoglobin (Hb) under heated (80 °C for 30 min.) and unheated conditions and at varying pH values (3.0, 5.0, 7.0, and 9.0) were measured using ANS, CPA, and PRODAN. Hydrophobicity was lower at pH 9.0, 5.0 and 7.0 for all proteins measured by PRODAN, for ANS and for CPA respectively. Lb shows maximum hydrophobic nature with ANS and CPA may and less with PRODAN compared to other heme proteins, indicating that Lb favor less ionic interaction. These results suggest that the presence of a permanent charge of fluorescent probes can affect protein hydrophobicity values measured under various pH conditions for Lb.