Tear prealbumin was purified from crude tear prealbumin previously isolated from the saline soluble human ocular mucus. Purification was achieved by further column chromatographies on DEAE Sephadex A-25 and Sephadex G-75. Preliminary characterization included amino acid analysis, gel electrophoresis, and isoelectric focusing. Unlike serum prealbumin, the purified tear prealbumin showed a predominance of acidic residues and a trace amount of tryptophan. It exhibited polymorphic nature, with pi values of 4.8 and 4.9. The possibility of a tear prealbumin/retinol complex was also examined. The protein was found to incorporate with 3H retinol. The % retinol-incorporated tear prealbumin did not exhibit the characteristic UV spectrum of retinol; however, it did display emission and excitation fluorescence spectra at high concentrations similar to serum retinol-binding protein.