Abstract This study investigated the influence of vitamin B7 (biotin) at different concentrations (0.1, 0.3 and 0.5) mM on the micellisation behaviour of three different ionic liquids with different cationic headgroups, namely N-methylmorpholinium bromide [Mor1,12][Br], N-dodecyl-N-methylpyrrolidinium bromide [Pyrr1,12][Br], and 1-dodecyl-3-methylimidazolium bromide [C12mim][Br], at four different temperatures (298.15, 303.15, 308.15, and 313.15) K using electrical conductivity and surface tension measurements. The evaluated CMC values for the studied system were used to obtain information about the colloidal behavior of the ionic liquids in the presence of biotin. The different thermodynamic parameters were studied. Tensiometry was used to determine the interfacial properties. The parameters obtained were used to study the different types of interactions for micelle formation. The main objective of studying these interactions is to further utilize this type of mixture in pharmaceutical industry, medicinal chemistry, and bio industry.