Glucosinolates are secondary metabolites that are synthesized from amino acid. They were found in 16 families of Brassicales, among the most important Brassicaceae family, which includes well-known plants of everyday nutrition such as cabbage, broccoli, cauliflower and others. Chemically these are β-thioglucoside-N-hydroxysulfates with a variable side chain. Glucosinolates are chemically and bioloyically inactice compounds, and with their degradation occur volatile sulfur compound which have a number of medicinal effects. They can be broken down by enzymatic, thermal and chemical ways. In our study we used different methods of degradation which includes hydrodistillation in a modified apparatus by Clevenger (100 °C, 150 min), microwave assisted distillation and extraction (500 W, 98 °C, 15 - 30 min), which are an example of thermal degradation, and autolysis followed by extraction with dichloromethane which is an example of enzymatic degradation. The aim of this study was to isolate and identify sulfur compounds from the horseradish (Armoracia rusticana P. Gaertn., B. Mey. et Scherb.). The roots and the leaves of horseradish were collected from the wild-growing population in Konavle (near Dubrovnik, Croatia) in March 2019. GC-MS and HPLC-DAD techniques were used to identify compounds. Analysis of the volatile compounds hasidentified the presence of allyl isothiocyanate, as the main glucosinolate degradation product originating form sinigrin, along with phenylethyl isothiocyanate and benzenpropanonitrile, originating from glucosinolate gluconasturtiin. Only sinigrin was present in the leaf of horseradish, while at the root, along with sinigrin, the presence of gluconasturtiin was also confirmed. It was later confirmed with HPLC-DAD technique acros desulfated (desulfoglucosinolates) products.