Human biomonitoring of toxic trace elements is of critical importance for public health protection. The current study aims to assess the levels of selected trace metals (Cd, Co, Cr, Cu, Mn, Ni, Pb, and Zn) into paired human nail and hair samples (n = 180 each) from different altitudinal setting along the Indus River, and which were measured by using inductively coupled plasma mass spectrometry (ICP-MS). The human samples (hair and nail) were collected from four different ecological zones of Pakistan which include frozen mountain zone (FMZ), wet mountain zone (WMZ), riverine delta zone (RDZ), and low-lying southern areas (LLZ). Our results showed the following occurrence trends into studied hair samples: higher values (ppm) of Zn (281), Co (0.136), and Mn (5.65) at FMZ; Cr (1.37), Mn (7.83), and Ni (1.22) at WMZ; Co (0.15), Mn (11.89), and Ni (0.99) at RDZ; and Mn (8.99) and Ni (0.90) at LLZ. While in the case of nails, the levels (ppm) of Mn (9.91) at FMZ and Mn (9.38, 24.1, and 12.5), Cr (1.84, 3.87, and 2.33), and Ni (10.69, 8.89, and 12.6) at WMZ, RDZ and LLZ, respectively, showed higher concentration. In general, among the studied trace elements, Mn and Ni in hair/nail samples were consistently higher and exceeded the WHO threshold/published reference values in most of the studied samples (> 50-60%) throughout the Indus basin. Similarly, hair/nail Pb values were also higher in few cases (2-10%) at all studied zones and exceeded the WHO threshold/published reference values. Our area-wise comparisons of studied metals exhibited altitudinal trends for Cd, Cr, Zn, and Mn (p < 0.05), and surprisingly, the values were increasing from south to north (at higher altitudes) and indicative of geogenic sources of the studied toxic elements, except Mn, which was higher at lower floodplain areas. Estimated daily intake (EDI) values showed that food and drinking water had the highest contribution towards Zn, Cu, Mn, and Ni and accumulation at all studied zones. Whereas, dust also acts as the main exposure route for Mn, Co, Cr, and Cd followed by the food, and water.

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