1. The preparation, purification, chemical and spectral properties of potassium 4-methylumbelliferone sulphate are described. 2. The use of 4-methylumbelliferone sulphate as a substrate for the arylsulphatase of Patella vulgata is presented with specific reference to the fluorimetric assay procedure used with this substrate. 3. 4-Methylumbelliferone sulphate is compared with the previously used synthetic sulphatase substrates nitrocatechol sulphate and p-nitrophenyl sulphate with respect to K(m), V(max.) and sensitivity in the assay of arylsulphatase. 4. 4-Methylumbelliferone sulphate was strongly inhibited by phosphate. Sulphate, a less potent inhibitor, appeared to be of the competitive type with some anomalous characteristics.