Comprehensive Study of Mediterranean (Croatian) Propolis Peculiarity: Headspace, Volatiles, Anti-Varroa-Treatment Residue, Phenolics, and Antioxidant Properties.

Eight propolis samples from Croatia were analyzed in detail, to study the headspace, volatiles, anti-Varroa-treatment residue, phenolics, and antioxidant properties. The samples exhibited high qualitative/quantitative variability of the chemical profiles, total phenolic content (1,589.3-14,398.3 mg GAE (gallic acid equivalent)/l EtOH extract), and antioxidant activity (11.1-133.5 mmol Fe(2+) /l extract and 6.2-65.3 mmol TEAC (Trolox® equivalent antioxidant capacity)/l extract). The main phenolics quantified by HPLC-DAD at 280 and 360 nm were vanillin, p-coumaric acid, ferulic acid, chrysin, galangin, and caffeic acid phenethyl ester. The major compounds identified by headspace solid-phase microextraction (HS-SPME), simultaneous distillation extraction (SDE), and subsequent GC-FID and GC/MS analyses were α-eudesmol (up to 19.9%), β-eudesmol (up to 12.6%), γ-eudesmol (up to 10.5%), benzyl benzoate (up to 28.5%), and 4-vinyl-2-methoxyphenol (up to 18.1%). Vanillin was determined as minor constituent by SDE/GC-FID/MS and HPLC-DAD. The identified acaricide residue thymol was ca. three times more abundant by HS-SPME/GC-FID/MS than by SDE/GC-FID/MS and was not detected by HPLC-DAD.