A total of 120 highly diverse cowpea [Vigna unguiculata (L.) Walp] genotypes, including indigenous and exotic lines, were evaluated for different biochemical traits using AOAC official methods of analysis and other standard methods. The results exhibited wide variability in the content of proteins (ranging from 19.4 to 27.9%), starch (from 27.5 to 42.7 g 100 g−1), amylose (from 9.65 to 21.7 g 100 g−1), TDF (from 13.7 to 21.1 g 100 g−1), and TSS (from 1.30 to 8.73 g 100 g−1). The concentration of anti-nutritional compounds like phenols and phytic acid ranged from 0.026 to 0.832 g 100 g−1 and 0.690 to 1.88 g 100 g−1, respectively. The correlation coefficient between the traits was calculated to understand the inter-trait relationship. Multivariate analysis (PCA and HCA) was performed to identify the major traits contributing to variability and group accessions with a similar profile. The first three principal components, i.e., PC1, PC2, and PC3, contributed to 62.7% of the variation, where maximum loadings were from starch, followed by protein, phytic acid, and dietary fiber. HCA formed six distinct clusters at a squared Euclidean distance of 5. Accessions in cluster I had high TDF and low TSS content, while cluster II was characterized by low amylose content. Accessions in cluster III had high starch, low protein, and phytic acid, whereas accessions in cluster IV contained high TSS, phenol, and low phytic acid. Cluster V was characterized by high protein, phytic acid, TSS, and phenol content and low starch content, and cluster VI had a high amount of amylose and low phenol content. Some nutri-dense accessions were identified from the above-mentioned clusters, such as EC170579 and EC201086 with high protein (>27%), TSS, amylose, and TDF content. These compositions are promising to provide practical support for developing high-value food and feed varieties using effective breeding strategies with a higher economic value.