Targeted and untargeted LC-MS polyphenolic profiling and chemometric analysis of propolis from different regions of Croatia.

Propolis is a complex biological matrix consisting mostly of plant resins and waxes, and in a small proportion of the herbal secondary metabolites, phenols. Phenols are components that are responsible for biological activities of propolis, however, their qualitative and quantitative composition is strongly influenced by climate and vegetation. Although studies on profiling of propolis samples from different countries have been carried out for some time propolis from Croatia is still not characterized till now. Targeted liquid chromatography coupled to triple quadrupole (LC-QQQ), untargeted liquid chromatography coupled to quadrupole time-of-flight (LC-QTOF) and direct injection QTOF methods were developed and 56 propolis samples from different geographical regions of Croatia were analyzed. Results revealed that there is not only one expected type of propolis in the territory of Croatia; i.e. beside expected European "poplar" propolis another type can be distinguished. Principal component analysis (PCA) and Partial least squares Discriminant Analysis (PLS-DA) indicated that the phenolic content of propolis samples significantly changes under the influence of the Mediterranean, so the "European" propolis type mixes with the Mediterranean type on the Croatian coast, especially on the islands. For fast screening of propolis type, direct injection QTOF analysis demonstrated to be fast and reliable method, but for unambiguous identification of phenolic compounds, chromatographic separation is indispensable. This paper presents the findings from the first research on phenolic profiling of propolis from Croatia.