In Malaysia, air pollution still measured using instrumental approach. Hence, this study used lichen as biological indicator to assess air pollution. Study has been conducted in four different locations which are Rumah Tumbuhan, UKM (Control station), Section 51 Industrial Area, Petaling Jaya, Spring Crest Industrial Area, Batu Caves and Nilai Industrial Area, Negeri Sembilan. This study used lichen Dirinaria sp.; an endemic species of tropical country; as the biological indicator. Heavy metal concentrations in lichen samples been analyzed using ICP-OES and number of motor vehicles were recorded for every sampling location. One way ANOVA and Pearson’s correlation used to test the relationship between heavy metals and sampling locations also relationship between heavy metals and motor vehicles. Result shows that heavy metals such as Cr, Fe, Cu, Ni, Zn and Pb have been recorded. One way ANOVA test shows there is significant relationship between heavy metals and sampling locations where F is 2.7728 and P value is 0.0001 (99% significant level). Pearson’s correlation also shows the relationships between all recorded heavy metals with number of motor vehicles where all the P value is < 0.05. This study found that lichen can be used as the alternative approach in determining the heavy metals content in the environment and it also cheaper and time saving rather than using instrumental approach. Heavy metals; Lichen; Air pollution; Environment; Urban ecosystem