The aim of this study is to determine the endogenous hormone activities of abscisic acid (ABA), Jasmonic acid (JA) and Salicylic acid (SA) in compatible (Intraspecific = ISP) and incompatible pollination (Intergeneric = IGP) in stigmatic ovaries during orchid post-pollination process. It was designed to be Himantoglossum robertianum in ISP experiments and Himantoglossum robertianum and Orchis italica in IGP experiments. Polynariums taken from O. italica have been applied with needles to the flower stigmas of H. robertianum. Ovaries with stigma from both pollination types were taken separately for each day for a total of 10 days and quantitative endogenous hormone analyzes have been performed by LC-MS/MS. After all, SA hormone has been detected for the first time in both ISP and IGP, but only ISP values were significant. The statistical analysis revealed that while SA is significant in only ISP, ABA values were found to be highly significant in both ISP and IGP. ISP9 also showed high significance. Additionally, ABA values were generally higher than SA values, especially in IGP experiments. The results also emphasized the importance of ovarian endogenous hormone activities for the first time. JA was not detected in the analysis.