Diatom indices developed in certain geographic regions are frequently used elsewhere, despite the strong evidence that such metrics are less useful when applied in regions other than that where species-environment relationships were originally assessed, showing that species have particular autoecological requirements in different geographic areas. The goal of this study was to develop a new metric, the Duero Diatom Index (DDI), aimed at monitoring water quality in Duero basin watercourses (NW Spain). In summer 2008 and 2009, a total of 355 epilithic diatom samples were collected following standard protocols. The 2008 samples were used to develop the DDI, whereas the samples collected during 2009 were used in the index testing. Weighted averages method was used to derive the autoecological profiles of diatoms with respect to pH, conductivity, biological oxygen demand, ammonia, nitrates, and phosphates. The optimum and tolerance values for the measured environmental variables were determined for 137 taxa with abundances and frequencies of occurrence above 1%, and subsequent trophic indicator and sensitivity values were defined for the DDI. The correlation between the observed and the diatom-inferred nutrient concentrations was highest for phosphates (ρ (S) = 0.72). Significant statistical relationship were observed between DDI values and the chemistry-based General Quality Index values (p = 0.006) and the specific pollution index (SPI) diatom metric (p = 0.04). DDI has demonstrated a better correlation with water chemistry than SPI diatom metric.