Dandruff is a scalp disorder that is affected by three factors, namely the fungus Malassezia globosa, sebaceous gland activity, and individual susceptibility. Selenium sulfide (SeS2) and Zinc Pyrithion (ZPTO) is used as an antidandruff shampoo active ingredient because it has anti-fungal properties. The study was conducted to determine the potentiality of shampoo with active ingredient SeS2 1% and ZPTO 1% to M.globosa in vitro. The colonies of M.globosa (CBS 7966 ATCC 96 807) were exposed to a non-antifungal base shampoo, shampoo containing 1% SeS2, shampoo containing 1% ZPTO and shampoo containing combination of 1% SeS2 and 1% ZPTO. Fungal contact time with shampoo is 3 and 5 minutes. Fungal colonies are grown on Sabourraud Dextrose agar (SDA) medium coated with olive oil. Incubation was carried out at 37° C for 5 days. After fiveday, assessment was performed to observe whether the fungal culture was grown, and then the colony growth is calculated. The research was conducted at Clinical Parasitology Laboratory, Parasitology Department, Faculty of Medicine, University of Indonesia from November 2011 until April 2012. Data were analyzed using ANOVA and LSD Fisher test. Shampoo containing 1% SeS2 inhibited fungal growth in 5 min contact time (p = 0,000 <0.05), whereas 1% ZPTO shampoo fungal growth was inhibited either in 3 min or 5 min contact time (p = 0,000 <0,05), and combination of 1% SeS2 and 1% ZPTO shampoo in both contact times (p = 0,000 <0.05). In conclusion, combination of 1% SeS2 and 1% ZPTO shampoo has the most potency in inhibiting the growth of M.globosa colonies in vitro.