Background and Objective Alcohol, tobacco, and cannabis use are more prevalent in sexual minority females than heterosexual females, and their use is associated with adverse consequences. Identifying disparities in substance use patterns by sexual identity may inform interventions targeting this vulnerable group. This study examined differences between heterosexual and sexual minority females on patterns of past 30-day tobacco, alcohol, and cannabis use. Methods N = 3020 females (18.8% sexual minority) completed an online survey (September 2020-October 2021) that queried about past 30-day tobacco/nicotine (cigarettes, e-cigarettes, large cigar/LCCs, and other products), alcohol, and cannabis use. Participants were classified into one of eight patterns: no use, tobacco/nicotine-only, alcohol-only, cannabis-only, alcohol and tobacco/nicotine, tobacco/nicotine and cannabis, alcohol and cannabis, and polysubstance use. A multinomial logistic regression model examined the association between sexual identity and each substance use group, controlling for demographics. Results Across both groups, no substance use was the most common pattern. Polysubstance use was the most common substance use pattern among sexual minority females. In adjusted regression models, sexual minority females were more likely to report cannabis-only (adjusted odds ratio [AOR] = 2.54), tobacco/nicotine and cannabis co-use (AOR = 1.73), alcohol and cannabis co-use (AOR = 2.49), and polysubstance use (AOR = 2.58), compared to heterosexual females. Discussion and Conclusions Substance use patterns that involve cannabis are more common among sexual minority females. Scientific Significance This study extends prior research by using a large sample of females to examine differences based on sexual identity in patterns of tobacco/nicotine, alcohol, and cannabis use beyond single substance use and considers co-use and polysubstance use.