The formation of the inclusion complex of p-sulfonatocalix[6]arene (SCX6) with different forms of vitamin B 6 (VB 6 ) was studied by using fluorescence spectroscopy. VB 6 can exist in one of three forms (the acidic form, neutral zwitterionic form and basic form) depending on pH. The fluorescence intensities of acidic and basic forms of VB 6 remarkably decreased in presence of SCX6. SCX6 preferred to form 1:1 inclusion complexes with acidic and basic forms of VB 6 but hardly form inclusion complex with neutral zwitterionic form. According to the nonlinear curve fitting method, the inclusion constant (K) for the formation of inclusion complexes of acidic and basic forms of VB 6 with SCX6 were evaluated to be 1.4 9 10 4 and 9 9 10 3 L/mol, respectively. The binding affinity of SCX6 towards acidic form is attributed to hydrogen bonds and hydrophobic interaction, furthermore, additional electrostatic interaction also plays a crucial role. The possible inclusion mode was given by 1 H NMR technique.