Piperine, the bioactive alkaloid compound of the spice black pepper (Piper nigrum) exhibits a wide range of beneficial physiological and pharmacological activities. Being essentially water-insoluble, piperine is presumed to be assisted by serum albumin for its transport in blood. In this study, the binding of piperine to serum albumin was examined by employing steady state and time resolved fluorescence techniques. Binding constant for the interaction of piperine with human serum albumin, which was invariant with temperature in the range of 17-47 degrees C, was found to be 0.5 x 10(5)M(-1), having stoichiometry of 1:1. At 27 degrees C, the van't Hoff enthalpy DeltaH degrees was zero; DeltaS degrees and DeltaG degrees were found to be 21.4 cal mol(-1) K(-1) and -6.42 kcal mol(-1). The binding constant increased with the increase of ionic strength from 0.1 to 1.0M of sodium chloride. The decrease of Stern-Volmer constant with increase of temperature suggested that the fluorescence quenching is static. Piperine fluorescence showed a blue shift upon binding to serum albumin, which reverted with the addition of ligands -triiodobenzoic acid and hemin. The distance between piperine and tryptophan after binding was found to be 2.79 nm by Förster type resonance energy transfer calculations. The steady state and time resolved fluorescence measurements suggest the binding of piperine to the subdomain IB of serum albumin. These observations are significant in understanding the transport of piperine in blood under physiological conditions.