In this study, the optimisation of horse chestnut seed extraction was performed to determine the best extraction solvent in the recovery of pharmacologically active escin from raw material. All extracts were obtained by accelerated solvent extraction and were analysed for their antioxidant potential (DPPH test), phenolic content (Folin-Ciocalteu (FC) assay) and escin content (ESI-TOF-MS extracts profiling). Methanol was found to recover the highest amount of escin from the plant matrix as well as to produce the most active antiradical solutions rich in polyphenols. Radical scavenging properties of methanol extracts were calculated as 2 GAE in DPPH test and 23 GAE (mg/g) in FC assay. Quantitative analysis of extracts performed by ESI-TOF-MS measurements in ion-positive mode confirmed the presence of four major escin isomers in the extracts and showed that the high antioxidant potential of methanolic extracts went hand in hand with the highest content of escin (8.92%).