The purpose of this study is to find out the effect of the crystal water content on the crystal structure of lactitol monohydrate. Crystal water was removed by drying over silicagel at 40°C and by using phosphorus pentoxide as drying agent at 20°C. The amouts of water removals were identified by thermogravimetry, the melting points and the heat of fusions were calculated from the results of differential scanning calorimetry measurements and the structure of samples were identified by X-ray powder diffraction method. Over 23 w/w% of total water content could removed by gently drying until significant structural changes could be detected. The melting point of anhydrous lactitol obtained by drying lactitol monohydrate was 120°C and the melting enthalpy was 102 J g−1 when measured with heating rate 10°C min−1 by DSC.