Previous research on specialized metabolites of olive leaves has focused on the phenolic components and their biological role. The research in this article focuses on the metabolites that form free volatile compounds (FVCs). The composition of FVCs is divided into compounds isolated in the oil phase (essential oils; EO) and in the aqueous phase (hydrosols; Hy) from leaves of Olea europaea L. cultivar Oblica. Plant material was collected from the same olive tree over a six-month period, from December to May, and analyzed by gas chromatography–mass spectrometry (GC–MS). The compounds β-caryophyllene, α-humulene, allo-aromadendrene, docosane, hexadecanoic acid and oleic acid were identified in all EO study periods. In the Hy in all studied periods, the major compounds are α-pinene, β-ionone, myristicin, docosane, 1-hexanol, oleic acid and (E)-β-damascenone. The differences in the qualitative composition of FVC are directly related to the phenological development of the leaves. Antioxidant capacity of the EOs and hydrosols was measured with two methods, ORAC and DPPH. Hydrosol extracts showed higher capacity than the EOs in all methods.

Chemical profile and antioxidant activity of the species Veronica saturejoides Vis. ssp. saturejoides (Plantaginaceae)—which is endemic to Croatia, Bosnia and Herzegovina and Montenegro —were investigated. Volatile compounds produced by glandular trichomes (composed of one stalk cell and two elliptically formed head cells according to scanning electron microscope investigation) were isolated from the plants collected in two locations. Additionally, as a part of specialized metabolites, total polyphenols, total tannins, total flavonoids and total phenolic acids were determined spectrophotometrically. In the lipophilic volatile fractions-essential oils, the most abundant compounds identified were hexahydrofarnesyl acetone, caryophyllene oxide and hexadecanoic acid. In total, the class of oxygenated sesquiterpenes and the group of fatty aldehydes, acids and alcoholic compounds dominated in the essential oils. In the hydrophilic volatile fractions-hydrosols, the most abundant compounds identified were trans-p-mentha-1(7),8-dien-2-ol, allo-aromadendrene and (E)-caryophyllene. A group of oxygenated monoterpenes and the sesquiterpene hydrocarbons dominated in the hydrosols. Antioxidant activity of essential oils and hydrosols was tested with two methods: 2,2′-diphenyl-1-picrylhydrazyl (DPPH) and oxygen radical absorbance capacity (ORAC). Essential oils showed higher antioxidant activity than hydrosols and showed similar antioxidant activity to Rosmarinus officinalis essential oil. Obtained results demonstrate that this genus is a potential source of volatiles with antioxidant activity.

Micellar electrokinetic chromatography (MEKC) was used for the separation and determination of two iridoid glycosides, aucubin and catalpol, in several Plantago species growing in Croatia: P. altissima L., P. argentea Chaix, P. coronopus L., P. holosteum Scop. (subsp. depauperata, subsp. holosteum and subsp. scopulorum), P. lagopus L., P. lanceolata L., and P. maritima L. Hot water extraction (HWE) was applied for the isolation of iridoid substances. Significant differences appeared between the iridoid contents in the examined species. The yield of aucubin and catalpol was up to 0.27% and 1.81% of the dry mass of the leaves, respectively. Besides aucubin and catalpol, two related compounds were determined in the plant samples.

A study on the type and distribution of trichomes, morphological traits of the calyx and content of antioxidant compounds was conducted on Teucrium arduini L. populations in Croatia and in Bosnia and Herzegovina. Non-glandular trichomes, peltate trichomes and small capitate trichomes with four secretory head cells were observed on leaves, stems and calyx, whereas long capitate trichomes with a unicellular head cell were observed exclusively on the calyx. Average calyx length was 5.23–6.51 mm, calyx tube length 2.92–3.68 mm, lower lip length 1.25–2.42 mm and upper lip length 1.89–2.99 mm. The concentration of antioxidant compounds in all populations was the highest in leaves: total polyphenols (TP): 8.43–13.55%; tannins (T): 1.79–7.33%; flavonoids (TF): 0.28–0.40%; phenolic acids (TPA): 2.90–5.07%. Inflorescences generally contained the smallest amount of TP (5.89–11.42%), T (0.87–6.50%) and TPA (1.66–2.27%), while the smallest content of TF was obtained for the majority of stem samples (0.06–0.12%). The researched populations did not differ with regard to the type and overall distribution of the trichomes, though the Bosnian populations have a shorter calyx tube and longer upper lip. Some variability was observed in the TP and T content between sampling sites, while quite high variability was noted in the TF and TPA content.