Inhalation Bioaccessibility, Pollution Assessment, and Speciation of Heavy Metals in PM2.5RD Road Dust from Urban and Semi-Urban Areas of Pakistan

Ambient exposure to fine particulate matter (PM2.5) remains a critical global public health concern; however, the speciation and bioaccessibility mechanisms of particle-bound heavy metals within the lung environment are still insufficiently understood. In particular, the role of soluble and insoluble metal species in simulated lung fluids requires further clarification. Road dust, a major contributor to PM2.5, contains heavy metals that pose significant inhalation risks, leading to oxidative stress, DNA damage, and systemic toxicity. This study investigated the contamination characteristics, bioaccessibility, pollution level, and speciation of heavy metals in PM2.5-fractionated road dust (PM2.5RD) collected from Karachi (a megacity with >27 million inhabitants) and Shikarpur (a medium-sized city of ~0.3 million) in Pakistan. A grid-based sampling approach was applied across four functional zones (residential, school, hospital, and office), yielding 48 samples. Inhalation bioaccessibility was evaluated using artificial lysosomal fluid (ALF; pH 4.5, 37 °C, 24 h), revealing moderate bioaccessibility of Cr, As, and Pb. Pollution status was assessed using contamination factor (CF), integrated pollution index (IPI), pollution load index (PLI), and potential ecological risk index. Geochemical modeling with PHREEQC 3.0 and Visual MINTEQ 3.1 was employed to simulate metal speciation, release behavior, and mineral saturation states. Overall, this study provides mechanistic insights into speciation-driven toxicity of As, Pb, and Cr in simulated lung environments, offering a scientific basis for improved risk assessment and regulatory strategies for PM2.5-associated metal exposure.