Groundwater is an essential and indispensable resource that meets the critical needs for drinking and irrigation purposes. Different sources such as industrial, municipal, urban, and agricultural runoff are the main contributors to water pollution. Nitrate, an inorganic form of nitrogen, is a significant water pollutant that can degrade water quality both ecologically and clinically by causing eutrophication in aquatic bodies and methemoglobinemia (blue baby syndrome) in human infants, and various health complications in animals. According to the Nitrate Pollution Index (NPI), it is essential to regularly monitor well sites to prevent nitrate contamination resulting from human activities, including waste disposal in open areas and sewage infiltration. By testing groundwater samples through spectrophotometry, this study aims to understand the extent of nitrate contamination and develop a practical and sustainable solution for its removal. This study performs a comparative analysis of filtration efficiency using two treatment combinations: activated charcoal with Chlorella vulgaris and activated charcoal with duckweed. These combinations are layered in a column chromatography setup to enhance the filtration process. This study aims to determine the most effective and environmentally sustainable method for nitrate removal, contributing to the development of scalable water treatment solutions for regions facing similar contamination challenges.