Aflatoxin is a potent carcinogenic, curious metabolite mainly produced by many fungal species such as A. flavus and A. parasiticus. AFs are very toxic food contaminants that can acutely damage human health. There are four main groups of aflatoxins which include B1, B2, G1, and G2. A survey of red chili for aflatoxin contamination was performed in different varieties of chilies including Capsicum annum, L. Piper nigrum, L. C. frutescence, and fresh green chilies. In the present study detection of aflatoxin contamination in red chilies was analyzed through TLC (Thin Layer Chromatographic) technique. A total of 20 various red chili samples were collected from different local regions of Lahore, Pakistan. The occurrence of aflatoxin B1 was detected in 9 samples of chilies. In the case of red powdered chilies, 2 samples were contaminated with aflatoxin B1 ranging between 11.88ppb-11.89ppb. These two contaminated samples are unfit for human health and beyond the permissible limits. In the case of red round chilies, 3 samples were contaminated with aflatoxin B1 and its concentration ranged between 4.68ppb-13.37ppb. These two contaminated samples are unfit for consumption and outside the satisfactory limits. One contaminated sample is fit and below the permissible limit. While in P. nigrum four samples were contaminated with aflatoxin B1 ranging between 2.21ppb-13.47ppb. In which two contaminated samples were unfit for human health and outside the satisfactory limits. The other two contaminated samples were fit for consumption and below the satisfactory limits. In fresh green chilies, all samples were found to be free from contamination. While other types of aflatoxins B2, G1, and G2 were absent in all the samples. The relative percentage of aflatoxin contamination in C. annum (red chilli) was 40%, C. frutescence (round red chilli) was 60%, P. nigrum (black pepper) was 80% and fresh green chili was found to be free from contamination. The supply of red chilies in Lahore is extremely dangerous to human health because when exceeded beyond permissible level, aflatoxin can cause cancer. The main reason for aflatoxin contamination in chilies samples is the lack of implementation of qualitative methods.