Black aspergilli are, the most causes of otomycosis and Aspergillus niger and A. tubingensis are two more frequently isolates. Although, amphotericin B was a gold standard for the treatment of invasive fungal infection for several decades, it replaced by fluconazole and /or voriconazole. Luliconazole, appears to offer the potential for in vitro activity against black aspergilli. The aim of the present study was to compare the in vitro activity of a novel antifungal agent, luliconazole, with commonly used antifungals against clinical and environmental strains of black aspergilli. Sixty seven strains of black aspergilli were identified using morphological and molecular tests (β-Tubulin gene). Aspergillus niger was the common isolate followed by, A. tubingensis and 54.1% (clinical) and 30% (environmental) of isolates were resistant to caspofungin. The highest resistant rate was found in amphotericin B for both clinical (86.5%) and environmental (96.7%) strains. Clinical strains of Aspergillus were more sensitive to voriconazole (86.7%) than environmental strains (70.3%). On the other hand, 83.8% of clinical and 70% of environmental isolates were resistant to posaconazole, respectively. In conclusion, luliconazole compare to routine antifungals is a potent antifungal for A. niger complex in vitro. The MIC range, MIC 50 , MIC 90 and MIC GM of luliconazole against black aspergilli were the lowest among the representative tested antifungals.