Background: The chewing of Khat leaves, a natural psychoactive substance is widely chewed in countries of East Africa and the southern Arabian Peninsula, and is reported to be associated with a range of unfavorable health outcomes including khat dependence. The impact of Khat chewing on Health Related Quality of Life is yet to be explored. Aims: to measure and compare the quality of life of the khat chewers and non-khat chewers using a short form health survey (SF36), and to assess factors associated with Khat chewing using SF36 in a sample of adult population in Jazan region, Kingdom of Saudi Arabia.Methods: A total of 630 participants from two independent male populations of khat chewers and non-khat chewers were recruited into a cross-sectional survey study. A self administrative survey based on the SF-36 questionnaire was used to collect data on measures of health-related quality of life (HRQoL). Socioeconomic data of the respondents were also collected for detailed analysis. Data analysis include: descriptive statistics, reliability tests (Cronbach's alpha and intraclass correlation coefficient), and bivariate analysis (Chi square and Mann-Whitney U-test) to compare HRQoL of Khat chewers and non-Khat chewers.Results: The odds of being a khat chewer were higher in respondents with a lower socioeconomic status. The SF-36 scores were significantly lower in all domains for respondents with khat chewing, indicating that non-khat chewers had higher health perceptions compared with those chewing khat. The overall mean score of HRQoL for non-khat chewers was 92.7% (SD 5.53) compared with 63.5% (SD 21.73) for the khat chewing group. The study had shown good internal consistency and reproducibility across the eight subscales of SF-36 questionnaire (α 0.74-0.95). The Mann-Whitney U-test showed a significant difference between khat chewers and non-khat chewers (P < 0.001).Conclusions: This study measured and compared the quality of life of khat chewers and non-khat chewers using a generic health survey (SF36). The study had shown that khat chewing is associated with lower quality of life (HRQoL) and lower socioeconomic status. However in future a more refined SF36 developed especially for Khat chewers can provide more useful information.