Dermatological diseases of the scalp significantly affect the life quality of people. According tostatistical data, seborrheic dermatitis (SD) is the most common dermatological disease in morethan 50% of the population all over the world. There are many possible causes of SD includingsebum accumulation, less frequent shampooing, and sensitivity to hair care products.Maintaining personal cleanliness and reducing unwanted effects on scalp can be achieved byapplication of special shampoo. Most commercial antidandruff shampoos employ syntheticsubstances as active ingredients, but their regular use changes a biodiversity of scalp microfloraand determines subsequent antimicrobial resistance. Therefore, the aim of study was thedevelopment of medical shampoo with plant-based pharmaceutical substance for treatment ofSD. The inhibitory activity against S. epidermidis, S. aureus, C. albicans of the plant-basedsubstance based on M. alternifolia essential oil, 1,8-cineol (eucalyptol) and α-(-)-bisabolol andshampoo containing the substance were investigated by micro-broth dilution and broth dilutionmethod, respectively. The medical shampoo with 0.75% substance possessed high antibacterialactivity against S. epidermidis and S. aureus with log CFU>1.0. Interestingly, the antifungalactivity against C. albicans (model of Malassezia spp.) was compatible to that of ketoconazole,climbazole and piroctone olamine. Shampoo with developed combination of surfactants as laurylglucoside, sodium coco-sulfate, coco-glucoside and cocamidopropyl betaine had the bestformulation in terms of microbiology and pH-stability, density foam characteristics, hairsmoothness, soothing and scalp cleansing. Therefore, this medical shampoo with natural originingredients was suggested for use in treatment of SD and dandruff.