This dataset comprises field sensor physicochemical and optical/fluorescence measurements, as well as laboratory microbiological and chemical analysis, for urban surface water samples. Samples were collected at different locations throughout the urban area of Kolkata, with the latitude and longitude of all sample location provided within the spreadsheets. Samples/data were collected across three separate field surveys undertaken in June 2018 (file 1), March 2019 (file 2) and December 2019 (file 3). This dataset forms a case study of the water quality of three different types of urban surface freshwaters within the city of Kolkata, India. This case study was created to deploy a prototype multichannel fluorimeter and assess its ability to identify waters with a high bacterial load and biological contamination events through the use of Peak T fluorescence. Full details about this dataset can be found at https://doi.org/10.5285/9bc3dce7-7c2b-49dd-9b76-819267d7a352