Rough lemon (C. jambhiri Lush.), widely employed as a rootstock in India as well as worldwide, is widely consumed in Assam. Its fruits are highly nutritious and in high local demand, often priced higher than Assam lemon during the harvest season. As a heterozygous species indigenous to the state, this crop displays extensive variation. These variations can be observed in the morphology of the plant and its fruits. However, limited research on its biochemical constituents has hindered efforts to promote this fruit for both processing and fresh consumption nationwide. This paper aims to provide a gist of the work conducted at Assam Agricultural University, based on biochemical characteristics and group them to assess their biochemical similarity. This study classifies the available germplasm of rough lemon collected across the state of Assam in three main groups based on biochemical constituents of fruit. This may be attributed to the genetic makeup of the crop or the influence of soil properties and nutrient availability in the study areas. Sensory evaluations were conducted to assess consumer preference for fresh consumption. Consequently, this research offers a distinct understanding of the specific type of rough lemon that can be chosen for fresh consumption as well as processing purposes.