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Bioactivity guided isolation of dichloromethane extract of <em>Alternaria alternata</em> identified tenuazonic acid (1) as potentially active against <em>Mycobacterium tuberculosis</em> H37Rv, MIC at 250 μg/mL concentration. This active metabolite 1, was also evaluated for osmotic hemolysis using the erythrocyte as a model system. It was observed that this fungal metabolite showing antitubercular activity exhibited concentration dependent toxicity to human erythrocytes.