Background:Genital tobacco powder, locally known as "tabaa," is a mixture of powdered tobacco and other substances that women use vaginally for sexual enhancement. Through a media report, the Ministry of Health issued a warning for women to stop the practice. However, literature search showed that no any scientific study documenting the burden of this cultural practice beyond media reports. The aim of this study was to determine the prevalence and factors associated with genital tobacco powder use among women in The Gambia. Methods: A sequential exploratory mixed approach was used. The research was carried out in two areas of The Gambia: one rural (Farafenni) and one urban (Banjul). Focus group discussions and in-depth interviews were used to recruit (30) participants in the qualitative study. The quantitative study interviewed (400) women using structured questionnaire that was developed from the qualitative results. SPSS version 24 and STATA version 7 were used to analyze the data. Descriptive, Bivariate and multivariate statistics were used to summarize the data. Results: Most of the study participants were between 30-39 years old who were married. The prevalence (17.0%; n=68) of genital tobacco powder was high among women aged 40 years and above (OR = 2.88; 95% CI = 1.03–8.05; p = 0.04), from Farafenni and satellite villages (OR = 2.01; 95% CI = 1.02-3.96; p = 0.04), had non-formal education (OR = 3.09; 95% CI 0.85–11.26; p = 0.02) and had husbands who traveled for 6 (OR = 6.67; 95% CI = 1.71–26.01; p= 0.006) or 12 or more months (OR = 9.64; 95% CI 3.19-29.18; p<0.001). The perception of women and peer influence were associated with genital tobacco powder use. Conclusion: Genital tobacco powder is practiced all over the Gambia and highest in the rural areas. Socio-demographic variables such age, residence, marital status, education status were significantly associated with the practice. According to the findings, women with low level of education and have poor perception can easily be convinced by close friends/family members to use genital tobacco powder. Therefore, awareness creation is the most important intervention to mitigate this practice.