In this work, a new interface for continuous on-line MALDI-TOF MS is presented. The sample, mixed with a suitable matrix, was transported into the evacuated source chamber of the mass spectrometer at liquid flow rates of 100-400 nL/min. The liquid sample matrix was deposited on a rotating quartz wheel and transported to the repeller, where laser desorption took place. Rapid evaporation of the solvent (water or methanol) on the surface of the wheel resulted in formation of a thin, approximately 50-micron-wide, sample trace. Scanning electron microscopic photographs of the vacuum-dried trace revealed the deposited material to consist of an amorphous film. Furthermore, sample uniformity along the trace, in conjunction with its narrow width, resulted in excellent signal reproducibility, with detection limits in the attomole range. The interface permitted the on-line coupling of microcolumn separation techniques with MALDI MS, as demonstrated in the capillary electrophoresis MALDI-TOF MS analysis of a 12-peptide mixture. The approach offers the potential for rapid separation and trace analysis of complex mixtures.