This study investigates gender-specific bile acid (BA) profiles in non-alcoholic fatty liver disease (NAFLD) and alcohol-associated liver disease (ALD). The research involved 205 participants, including 45 NAFLD patients, 103 healthy controls, and 57 ALD patients. Bile acid levels were measured using targeted mass spectrometry, revealing significant gender differences in BA profiles. Women with NAFLD had higher concentrations of total BA, primary BA, and specific secondary BA subtypes, including glycocholic and glycochenodeoxycholic acids. In contrast, men with NAFLD showed elevated levels of several secondary BA subtypes, such as glycodeoxycholic, glycolithocholic, and lithocholic acids, while cholic acid levels were significantly lower. These differences suggest that men may better compensate for bile acid dynamics, as indicated by higher levels of taurine-conjugated BA, which are associated with beneficial metabolic functions. In ALD, only male patients showed significant correlations between BA levels and the Enhanced Liver Fibrosis (ELF) score. The study also found that higher ELF scores were associated with increased BA levels in both NAFLD and ALD. The results highlight the importance of considering gender differences in BA profiles for the diagnosis and management of NAFLD and ALD. The findings suggest that gender-specific BA profiles may provide insights into the pathogenesis of these diseases and could be useful for developing targeted therapies.This study investigates gender-specific bile acid (BA) profiles in non-alcoholic fatty liver disease (NAFLD) and alcohol-associated liver disease (ALD). The research involved 205 participants, including 45 NAFLD patients, 103 healthy controls, and 57 ALD patients. Bile acid levels were measured using targeted mass spectrometry, revealing significant gender differences in BA profiles. Women with NAFLD had higher concentrations of total BA, primary BA, and specific secondary BA subtypes, including glycocholic and glycochenodeoxycholic acids. In contrast, men with NAFLD showed elevated levels of several secondary BA subtypes, such as glycodeoxycholic, glycolithocholic, and lithocholic acids, while cholic acid levels were significantly lower. These differences suggest that men may better compensate for bile acid dynamics, as indicated by higher levels of taurine-conjugated BA, which are associated with beneficial metabolic functions. In ALD, only male patients showed significant correlations between BA levels and the Enhanced Liver Fibrosis (ELF) score. The study also found that higher ELF scores were associated with increased BA levels in both NAFLD and ALD. The results highlight the importance of considering gender differences in BA profiles for the diagnosis and management of NAFLD and ALD. The findings suggest that gender-specific BA profiles may provide insights into the pathogenesis of these diseases and could be useful for developing targeted therapies.