Microbial biofilms may form on wide variety of surfaces and are problematic in medical, industrial and environmental settings. Natural antibiofilm agents are promising candidates for treatment of biofilm-associated infections. One such natural products used for therapeutic purposes is Alum (Sphatika). Besides, Ayurvedic Bhasmas are used for treatment of various ailments in India. In the present in vitro study, anti-Biofilm potential of Alum as well as its Bhasma- Sphatika Bhasma was evaluated against total 8 bacteria, viz., 4 gram negative and 4 gram positive using crystal violet staining method. Antibiofilm activity of Sphatika Bhasma was noted in the order of P. aeruginosa > S. lutea > E. coli > K. pneumoniae > S. typhi > MRSA > B. subtilis > E. faecalis, whereas, Antibiofilm effect of Alum was found to be in the order of S. lutea > P. aeruginosa > E. coli > MRSA > S. typhi > B. subtilis > K. pneumoniae > E. faecalis. In general, Sphatika Bhasma exhibited better Anti-Biofilm activity than Alum. Furthermore, four different combinations prepared from Alum (AL), Sphatika Bhasma (SB) and Ciprofloxacin (CP)- standard antibiotic were evaluated for their combinatorial effect against 8 bacterial biofilms. In general, the combination (AL + SB) displayed the highest Anti-biofilm activity against all the 8 bacterial biofilms. Antibiofilm effects of Alum and Sphatika Bhasma could be attributed to their Flavonoid content. Moreover, to best of our knowledge, this is the first study revealing the anti-biofilm potential of Sphatika Bhasma.