CONCLUSION

This study provides the first comprehensive integration of untargeted LC–HRMS metabolomic profiling with standardized CLSI-guided antimicrobial susceptibility testing and antioxidant evaluation of ethanol extracts of Miana leaves against the clinically important genital pathogens Neisseria gonorrhoeae and C. albicans. Untargeted LC–HRMS identified 243 annotated metabolites, including 20 major bioactive compounds from flavonoids, phenolic acids, terpenoids, diterpenoids, coumarins, unsaturated fatty acids, and alkaloids, thereby establishing a comprehensive phytochemical profile of the extract. The extract demonstrated fungicidal activity against C. albicans (MIC/MFC, 62,500 ppm [62.5 mg/mL]) and bactericidal activity against N. gonorrhoeae (MIC/MBC, 100,000 ppm [100 mg/mL]), with MFC/MIC and MBC/MIC ratios of 1.0. In addition, the extract exhibited very strong antioxidant activity in the DPPH assay (IC₅₀, 43.68 ppm) and moderate antioxidant activity in the ABTS assay (IC₅₀, 128.7 ppm), indicating its dual antimicrobial and antioxidant potential.

The principal strength of this study lies in the combination of high-resolution untargeted metabolomic characterization and standardized antimicrobial susceptibility testing, providing quantitative microbiological data alongside detailed phytochemical information. The identification of several bioactive metabolites not previously reported in Miana further expands the current understanding of the phytochemical diversity of this medicinal plant and provides valuable reference data for future phytopharmaceutical standardization.

Despite these promising findings, the antimicrobial potency of the crude ethanol extract remained relatively low compared with conventional antimicrobial agents, indicating that therapeutic application of the unfractionated extract is currently limited. Furthermore, the absence of cytotoxicity evaluation, determination of the selectivity index, and in vivo efficacy studies precludes definitive conclusions regarding its clinical applicability.

Future investigations should prioritize bioassay-guided fractionation to isolate the principal active constituents, evaluation of synergistic interactions with conventional antimicrobial agents, comprehensive cytotoxicity and pharmacokinetic studies, and validation using appropriate in vivo infection models. Standardization of extracts through LC–HRMS chemical fingerprinting and development of optimized pharmaceutical formulations will also be essential steps toward translation into clinical applications.

Overall, the findings demonstrate that ethanol extracts of Miana leaves possess measurable dual antimicrobial and antioxidant activities supported by a diverse metabolomic profile. Although further optimization is required to improve antimicrobial potency and establish safety, this study provides a robust scientific foundation for the future development of standardized Miana-based phytopharmaceutical products as complementary therapeutic candidates for the management of gonorrhea, candidiasis, and mixed genital infections.