Acne vulgaris (AV) is the most common skin disease, including in Indonesia. This research aims to determine the effectivity of preovulation phase autologous serum application in patients with moderate-severe AV. This research was conducted in Makassar, South Sulawesi. Thirteen patient samples with moderate-severe AV who have met the inclusion criteria were taken from Dr. Wahidin Sudirohusodo General Hospital, Public Senior High School, and Medical Faculty of Hasanuddin University Laboratory. The method used is clinical trial. The examination was done by observing the clinical manifestations, sebum level using 815 sebumeter, and estradiol level examination. All patiens were given the same autologous serum treatment which is applied once a day every night and followed-up on day 21 and day 42. The data were processed using paired t test and Fisher's exact test. The result showed that there were improvements in clinical manifestation and decreased sebum level after autologous serum application (p < 0.05) on day 21 compared to day 1, on day 42 compared to day 21, and day 42 compared to day 1. Preovulation estradiol level were higher than the estradiol level on menstruation or in lutheal phase (p < 0.05). There were no significant relationship between routine activity, familial history, peanuts, chocolate, spicy and oily food consumption, wearing cosmetics, stress, and squeezing pimples (trauma) with number of acne lesion and sebum level (p < 0.05).