Despite the fact that high-quality water is essential for existence on Earth, it is contaminated by a multitude of natural and human activities. This study compared the physicochemical and biological characteristics of water in the Kabul River with those of various water quality regulatory organizations over the course of six months at five distinct locations, in addition to calculating the water quality index (WQI). Color, turbidity, EC, alkalinity, pH, ammonia, phosphate, total hardness, Mn, Fe, and fecal coliform all exceeded the allowable threshold among the twenty parameters. The principal component analysis (PCA) study identified three primary factors that contribute to the variability in water quality: factor 1 (PC1) explains 46.57% of the variance; factor 2 (PC2) explains 39.27%; and factor 3 (PC3) explains 14.16%. It may contain an assortment of organic matter, metal concentrations, and nutrient content. According to the study, the contamination level in the river increases from upstream (at the city entrance) to downstream (at the city departure) as a result of solid refuse disposal, agricultural and industrial activities, and the discharge of domestic effluents. The assessment indicates that the water upstream of the Kabul River is unfit for potable use but appropriate for agricultural purposes, laundry, and aquatic life. In contrast, the water located midstream and downstream (specifically S3, S4, and S5) is deemed unsuitable for aquatic life.