Honey is a complex natural product that has been used since ancient times because of its beneficial effects on human health. The composition of honey depends on the botanical and geographical origin, climate and storage conditions. Honey is rich in phenolic compounds that can act as natural antioxidants and have been of particular interest in recent years due to their potential positive effects on human health. In addition, phenolic compounds can be used as biomarkers of honey's plant and geographical origin. The aim of this thesis is the development of spectrophotometric methods for the analysis of phenolic compounds in honey and determination of their antioxidant activity. Additionally, methods based on mass spectrometry for absolute quantification of individual phenols were developed. Each method was tested on six different honey samples. The ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) and DPPH (2,2-diphenyl-1-picrylhydrazyl) methods were used to test the antioxidant activity of honey. A positive correlation of total phenols (R2=0,8643) and flavonoids (R2=0,7630) with the antioxidant activity determined using the ABTS method was observed. Furthermore, the methods for extraction of phenolic compounds from honey samples using solid phase extraction (SPE) and LC-QQQ for absolute quantification of twelve phenolic compounds were developed. Matrix effect and partial loss of phenolic compounds due to the selected extraction procedure were minimized with the standard addition method in which a mixture of phenolic standards was added before the extraction procedure. The developed methods proved to be effective regardless of the type of honey and showed significant progress in the quantitative analysis of phenolic compounds in honey.