In our study, we investigated the chemical composition and cytotoxic activity of essential oils isolated from Dalmatian sage (Salvia officinalis L.) collected along the Adriatic coast of Croatia. Scanning electron microscopy (SEM) was used to examine the morphology of the stem and leaf surfaces. Essential oil excretory glands were detected on both the leaves and stem surfaces. The essential oils were isolated by hydrodistillation, and their chemical composition was determined by gas chromatography and mass spectrometry (GC-MS). Sage essential oils were mixtures of terpene compounds, among which the most common were: α- and β-thujone, camphor, and 1,8-cineol. Cytotoxic activity was tested using MTS assay on multiple cell lines: normal and immortalized fibroblasts (HF77FA and HDF-Tert), immortalized lung line (BEAS-2B), and breast adenocarcinoma (MDA-MB-231). The growth of treated cells was determined relative to control conditions without treatment. The immortalized lung line was the least resistant to the activity of the essential oils, whereas immortalized fibroblasts were the most resistant. Statistical analysis has connected the cytotoxic effect and chemical composition of the studied essential oils. To the best of our knowledge, this work is the first testing of the cytotoxic activity of S. officinalis EO’s on the BEAS-2B, HF77FA, and HDF-Tert cell lines. The presented data on essential oil chemical composition and cytotoxic effect on 4 types of human cells supports pharmacotherapeutic potential this plant is known to have.

Helichrysum italicum (Roth) G. Don (Asteraceae), also known as immortelle, usually grows in the Mediterranean area. The composition of the essential oil (EO) of immortelle is a mixture of various aromatic substances, mainly monoterpenes and sesquiterpenes. Distillation is the most widely used method for extraction of EO immortelle, although the yield is very low (<1%). In this work, we aim to investigate how the use of different distillation methods affects the yield and chemical composition of immortelle EO. For this purpose, we applied two conventional methods: steam distillation (SD) and hydrodistillation (HD), and a modern (environmentally friendly) technique—microwave-assisted distillation (MAD). Wild immortelles from four different locations in Croatia were collected and carefully prepared for extraction. Each sample was then analyzed by gas chromatography–mass spectrometry (GC-MS). GraphPad Prisma statistical software was used to study the statistics between different groups of connections and analyze the data on the number of connections. The results show that HD gives a significantly higher yield (0.31 ± 0.09%) compared to MAD (0.15 ± 0.03%) and SD (0.12 ± 0.04%). On the other hand, the highest number of chemical compounds was identified with MAD (95.75 ± 15.31%), and most of them are subordinate compounds with complex structures. SD isolated EOs are rich in derived acyclic compounds with the highest percentage of ketones. The results show that the application of different distillation methods significantly affects the composition of the obtained immortelle EO, considering the yield of EO, the number of isolated, derived and non-derived compounds, chemotypes and compounds with simple (acyclic) and complex structures.

Helichrysum italicum (Roth) G. Don., immortelle, is a plant species used in ethnomedicine and the food industry as a spice added to food, beverages, and bakery products. It has been shown to possess various biological activities, such as antioxidant and antibacterial activity, making it useful as a natural preservative. We investigated the phytochemical profile and biological activity of H. italicum essential oils from wild-grown plant material collected from natural habitats in the Republic of Croatia and Bosnia and Herzegovina. Using high-resolution scanning electron microscopy (SEM), a visual investigation of plant organs (stem, leaf, and flower) was performed, confirming the presence of essential oil reservoirs on the surface of all examined plant organs. Essential oils were isolated by hydrodistillation in the Clevenger apparatus. The chemical composition of the essential oils was determined using the GC-MS analytical technique. Cytotoxic activity tests were performed in vitro on three cell lines: skin (fibroblast), lung, and breast cancer. Using statistical tools, the synergistic and selective effects of H. italicum essential oil on healthy and tumor cells were correlated to chemical composition and cytotoxic activity. The synergistic and antagonistic effects of H. italicum essential oil’s individual components were simulated by testing pure compounds and their mixture of cytotoxic activity on fibroblasts and breast cancer cells. The results confirm that essential oil’s biological activity is much greater than the sum of the effects of its components. The present data are novel contributions to the body of knowledge on the biological activity of this species used in the food industry.

Microplastics (MPs) are a ubiquitous marine pollutant, and their presence in bivalves is receiving increasing attention due to the associated risks to human health. The steps of pretreatment, detection, and quantification in the analysis of MPs depend on the type of polymer. Research on MPs is challenging because of the varying characteristics of these materials, such as the size, shape, and polymer type. Consequently, there are no standardized methods for their collection, separation, identification, or quantification. This review specifically examines the available bivalve digestion steps, focusing on efficient and time-reducing methods, such as the microwave-assisted (MAW) procedure and its advantages. Recent achievements in the application of pyrolysis gas chromatography–mass spectrometry (Pyr-GC-MS) are presented for the profiling of polymer mass-related microplastics data in marine bivalves. Here, we provide an overview of the abundance, properties, and polymer types of MPs in bivalve species, highlighting the polymer mass fractions. To date, the available mass-based concentrations have revealed nine types of MPs—polyethylene (PE), polypropylene (PP), polyvinyl chloride (PVC), polyethylene terephthalate (PET), polystyrene (PS), polymethyl methacrylate (PMMA), polyamide 66 (PA66), polycarbonate (PC), and polyamide 6 (PA6)—with PE, PP, and PVC being the most common. The total MP levels in bivalves were at ppm levels, ranging from 0.26 µg/g to 36.4 µg/g wet weight. The risk of human ingestion of MPs was assessed through the consumption of bivalves as seafood. The overall potential human health risk value (H) for marine bivalves was classified within the moderate to high hazard category.

The widespread presence of microplastics (MPs) and associated plastic-derived com-pounds, bisphenols (BPs), in the environment – originating from plastic production or adsorption of environmental pollutants – results in unavoidable human exposure. There-fore, assessing their occurrence in human biological samples is essential for understand-ing potential health risks. In this study, pyrolysis gas chromatography-mass spectrometry was used to analyse human intraocular fluids – aqueous humour (AH) and vitreous hu-mour (VH)—to identify 11 microplastic polymer clusters. A method based on dansyl chloride derivatization combined with ultra-performance liquid chromatography-tandem mass spectrometry was employed to screen for five bisphenols (BPs) in blood serum col-lected from the same patients. Potential MPs isolated from ocular samples were qualita-tively assessed by stereomicroscopy. The number of particles ranged from 14 to 69 per gram in AH and from 21 to 34 per gram in VH. Among AH samples, the most frequently detected polymer clusters were C-PMMA, C-PVC, C-PET, and C-PP, with average levels ranging from 0.41 to 0.82 µg/g. The three VH samples contained only C-PMMA, with an average concentration of 0.55±0.12 µg/g. Polymers such as C-PA66 (0.30±0.21 µg/g) and C-PA6 (0.15±0.44 µg/g) were occasionally observed in AH. Analysis of 13 human serum samples demonstrated exposure to bisphenol A (BPA; 3.90–5.63 ng/mL) and bisphenol AF (BPAF; 2.13–4.54 ng/mL), but no clear correlation was observed between serum BP levels and intraocular MPs within the limitations of the present dataset. These findings suggest the presence of MPs and BPs in human biological samples; however, the results should be interpreted with caution owing to potential background contamination and the pilot na-ture of the study.

Praćenjem stanja okoliša putem mjerenja sadržaja različitih onečišćujućih tvari (kontaminanata) u školjkašima ujedno se mogu pratiti biološke promjene i štetni učinci onečišćujućih tvari kojima su ti organizmi izloženi te se može procijeniti utjecaj ljudskih aktivnosti na određeni ekosustav. Akumulacija različitih onečišćujućih tvari u školjkašima namijenjenim za prehranu je stalna opasnost za ljude, kao potrošače na vrhu hranidbenog lanca. Mikroplastika je složeni kontaminant, koji uključuje vrlo širok skup čestica koje u procjeni rizika zbog njihove iznimne raznolikosti (veličina, oblik, topljivost, sastav polimera, adsorbirane kemikalije, biološki materijal, itd.) predstavljaju jedinstveni izazov. Glavni razlog opasnosti konzumacije školjkaša leži u biologiji njihovog organizma budući se oni hrane filtriranjem vode ili mora u kojem se uzgajaju ili prirodno žive, nakupljajući u svom organizmu različite tvari iz okoline. U razumijevanju kemijskog rizika mikroplastike koristi se model profila polimera, odnosno toksičnosti njihovih monomera. Nesumnjivo, analiza mikroplastike uključuje uz kemijsku karakterizaciju i određivanje ukupnog broja čestica kako bi se procijenila izloženost ljudi mikroplastici temeljem podataka o prehrambenim navikama ciljane populacije. U ovom radu dan je pregled literaturnih spoznaja o istraživanjima mikroplastike u dvoljušturnim mekušcima, kemijskoj karakterizaciji mikroplastike i pojavnosti u različitim vrstama školjkaša.