The alphabet of sea fennel: Comprehensive phytochemical characterisation of Croatian populations of Crithmum maritimum L.

Extreme environmental conditions affect the synthesis and accumulation of bioactive metabolites in halophytic plants. The aim of this study was to investigate the presence and quantity of key health-promoting phytochemicals in Croatian sea fennel, one of the most popular Mediterranean halophytes with a wide range of uses. The EOs were characterised by a high content of limonene (up to 93%), while the fatty acid profile shows a low content of oleic acid and the presence of valuable linoleic acid (ω-6) and linolenic acid (ω-3) in high percentages. The dominances of lutein and α-tocopherol were also confirmed in all samples. The results confirm the great variability in the chemistry of sea fennel populations in the Mediterranean region, with significant differences in the composition of the Croatian samples compared to the others, as well as the presence and high concentrations of the analysed bioactive compounds that contribute to the plant's health-promoting attributes.

Volatiles from French and Croatian Sea Fennel Ecotypes: Chemical Profiles and the Antioxidant, Antimicrobial and Antiageing Activity of Essential Oils and Hydrolates.

Sea fennel is a halophytic plant rich in valuable nutritional components and is characterized by pleasant organoleptic properties. While its essential oils (EOs) are well investigated, there are no reports on the volatiles from their corresponding hydrolates, which are the main by-products of EO isolation, as well as on their biological activity. Therefore, the composition and biological activities of EOs and corresponding hydrolates of sea fennel from Atlantic (French, FRA) and Mediterranean (Croatian, CRO) ecotypes were investigated and compared. The EO from the CRO sample was characterized by an abundance of sabinene and limonene, while that from the FRA ecotype was rich in dillapiol and carvacryl methyl ether. The CRO hydrolate was rich in terpinen-4-ol and 10-(acetylmethyl)-3-carene, while dillapiol, thymyl methyl ether and γ-terpinene were the main compounds in the FRA sea fennel hydrolate. The biological activities of the EOs and hydrolates were evaluated for their antioxidant (with DPPH, NO, FRAP and ORAC bioassays), antimicrobial (against some Gram+ and Gram- spoilage bacteria) and antiageing (tyrosinase, elastase and collagenase inhibition) activities. Both EOs showed low reducing powers and antiradical activities while the ability of both hydrolates to quench NO was slightly higher (35-39% if inhibition). The FRA EO showed low activity against Staphylococcus aureus (8 mm), while CRO moderately inhibited the growth of P. aeruginosa (8 mm), but strongly inhibited the other two bacterial strains. While the French EO showed no antityrosinase and anticollagenase activity, the Croatian oil significantly inhibited both enzymes (IC50 of 650 µg/mL and IC50 of 2570 µg/mL, respectively) probably due to the dominance of limonene and sabinene. Neither EO exhibited antielastase properties, while the hydrolates from both ecotypes showed no antiageing activity, regardless of the enzyme tested. The EOs from the aerial parts of sea fennel from FRA and CRO differed greatly in composition, resulting in different activities. The Croatian samples appeared to have better biological properties and are therefore good candidates for applications as preservatives or antiageing agents.

Conventional vs. Microwave-Assisted Hydrodistillation: Influence on the Chemistry of Sea Fennel Essential Oil and Its By-Products.

The main objectives of this study were to investigate the effects of the applied essential oil (EO) isolation method, conventional hydro-distillation (HD), and microwave-assisted hydro-distillation (MHD) on the chemical profile of sea fennel (Crithmum maritimum L.) essential oil and to investigate the main constituents present in the liquid by-products of EOs isolation (hydrolate and residual wastewater). Headspace-solid phase microextraction (HS-SPME) was used to isolate hydrolate components, while gas chromatography coupled with mass spectrometry (GC-MS) was used to detect and analyse the chemical constituents of the essential oils and hydrolates. The phenolic composition of the wastewater extracts was analysed by high performance liquid chromatography (HPLC). The EO obtained by MHD had a higher yield of limonene and sabinene. The chemical composition of the hydrolates differed from the EO compositions. The content of terpinen-4-ol in the MHD hydrolate was higher, while several compounds were detected in relatively high proportions only in the HD hydrolate. MHD also resulted in a higher phenolic content of the wastewater, where an increase in the concentration of chlorogenic acid was also observed. It can be concluded that the isolation method had a great influence on the profile of sea fennel EOs, especially on their corresponding hydrolates and residual wastewater extracts. Due to their valuable chemical composition, these by-products can be a cost-effective source of bioactive compounds that have great potential for use in various industries.

Phytochemical and Cytogenetic Study of Two Centaurea Species from Croatia: the Particular Case of Diploid and Tetraploid C. salonitana.

Natural wild populations of C. rupestris and C. salonitana were studied to determine possible relationships between the volatile oil (VO) composition and ploidy level. The chemical composition of the volatile oil was investigated using the GC/MS technique. The predominant components of the VO of diploid and tetraploid C. salonitana were hexadecanoic acid and α-linoleic acids, while in C. rupestris they were germacrene D and ß-caryophyllene in one population and heptacosane and germacrene D, in another. The nuclear DNA amounts (2 C DNA), determined by flow cytometry, were 3.54 pg for C. rupestris, 3.39 pg for the diploid and 6.79 pg for the tetraploid population of C. salonitana. Evidence that the degree of ploidy solely influences the chemical composition of the essential oil of C. salonitana was not found. The results presented are the first data to be reported on the DNA content of the studied Centaurea populations from Croatia, as well as on the chemical composition of C. salonitana volatile oil.

Chemical Profiling of Sea Fennel (Crithmum maritimum L., Apiaceae) Essential Oils and Their Isolation Residual Waste-Waters.

Sea fennel (Crithmum maritimum L.) is a wild-growing halophyte used in cuisine, traditional medicine or cosmetic products for its beneficial nutritive value and pleasant sensory characteristics. This study aimed to investigate sea fennel essential oils (EOs) from different parts of the plant (flowers, leaves and stems) and the corresponding hydrodistillation by-products (residual water) to validate their potential use and application in different industries. EOs were analyzed by gas chromatography coupled with mass spectrometry (GC-MS), while the phenolic profile of the residual water was analyzed by high-performance liquid chromatography (HPLC) and spectrophotometric methods. The EO analysis confirmed the presence of 14 compounds, dominated by sabinene (from 42.55 to 51.47%) and limonene (from 36.28 to 43.58%), while among the 12 detected phenolics, chlorogenic acid and its isomers (cryptochlorogenic and neochlorogenic acid) were found in the highest concentrations. Total phenolic, flavonoid and tannin contents were concentrated in the order flowers > leaves > stems. Although the sea fennel samples showed differences in chemical profiles, overall they were rich in bioactive compounds with relatively high amounts of key compounds with already proved good biological properties, especially in waste-water, indicating great potential for re-use in accordance with green processing technology trends.

Volatile Oil Chemical Composition of Wild, Edible Centaurea scabiosa L. and Its Cytotoxic Activity.

Centaurea species are well known as a source of phytopharmaceuticals having both beneficial and harmful influences on human health. Centaurea scabiosa L. is a wild edible plant used in Mediterranean cuisine in the Dalmatian region of Croatia. We have assessed the volatile oil's chemical composition using GC/MS chromatography and its cytotoxic activity on human fibroblasts using the MTT test. Data on chromosome number, obtained by classical karyological methods, and genome size, assessed by flow cytometry, of the same plant material of C. scabiosa, were also given. The major chemical compounds found in C. scabiosa volatile oil were heptacosane, caryophyllene oxide, alloaromadendrene epoxide, α-cyperone, and α-bisabolol. This volatile oil showed no cytotoxicity on human fibroblasts in a dose range of 0.01-1 g/L. The chromosome number of a C. scabiosa sample from Croatia showed 2n = 20 + 2B chromosomes. The total genome DNA amount of 2C = 3.3 ± 0.01 pg or 1 Cx = 1628 Mbp presents the first report on the genome size of this species from Croatia. The presented results support the idea of using this plant in the human diet. To our knowledge, this is the first report on edible C. scabiosa species in general and in particular from Croatia.

Polyphenol Composition, Anticholinesterase and Antioxidant Potential of the Extracts of Clinopodium vulgare L.

Clinopidium vulgare L. (wild basil, Lamiaceae) is a well-known medicinal plant used in the traditional medicine in many countries. Medicinal plants present potential sources of bioactive compounds. Many of them are rich in polyphenol compounds that show biological potential in terms of protecting biological molecules from oxidation and in inhibition of cholinesterase enzymes, which may be significant in the treatment of diseases related to oxidative stress. In this work, we examined the chemical composition of Clinopodium vulgare L. hot water and methanol extract using spectroscopic and HPLC/DAD techniques. Using DPPH and FRAP methods the antioxidant activity was analyzed. The ability to protect proteins and lipids from oxidation was also determined as well as the ability of extracts to inhibit cholinesterase enzymes using Ellman's method. Analyzed extracts were rich in polyphenol compounds. Among 16 identified and quantified phenolic compounds dominant were: rosmarinic (26.63 and 34.21 mg/g) and ellagic acid (23.11 and 29.31 mg/g) of hot water and methanol extract, respectively. They show good antioxidant activity and good potential in protecting lipids from oxidation. The ability of extracts to inhibit enzyme acetylcholinesterase was weak, while inhibition of the butyrylcholinesterase was missing. Extracts show prooxidant activity in terms of protecting proteins from oxidation.

Quantitatively Unraveling Hierarchy of Factors Impacting Virgin Olive Oil Phenolic Profile and Oxidative Stability.

A single phenolic group and even a compound play different roles in the sensory properties and stability of virgin olive oil (VOO), which in turn are strongly influenced by several factors. Understanding the causes of differences in phenolic compound composition and oxidative stability (OS) in VOOs is essential for targeted and timely harvest and processing while maintaining desired oil quality. The phenolic profile and OS of two monocultivar VOOs (Oblica and Leccino) grown in two geographical sites of different altitudes (coastal plain and hilly hinterland) were analyzed throughout the ripening period over two years. Concentration of secoiridoids was 30% higher in the Oblica than in the Leccino VOOs, which in turn had significantly higher values of OS. Both cultivars had more than twice as high concentrations of the two most abundant phenolic compounds, the dialdehyde form of decarboxymethyl oleuropein aglycone and the dialdehyde form of decarboxymethyl ligstroside aglycone, and OS values in a colder growing site of higher altitude. Among the studied monocultivar VOOs, the secoiridoid group did not behave equally during ripening. The hierarchy of different influencing factors was investigated using multivariate statistics and revealed: cultivar > geographical site > harvest period > growing season. In addition, the possibility of traceability of VOO using molecular markers was investigated by establishing SSR profiles of oils of the studied cultivars and comparing them with SSR profiles of leaves.

Application of Phenotyping Methods in Detection of Drought and Salinity Stress in Basil (Ocimum basilicum L.).

Basil is one of the most widespread aromatic and medicinal plants, which is often grown in drought- and salinity-prone regions. Often co-occurrence of drought and salinity stresses in agroecosystems and similarities of symptoms which they cause on plants complicates the differentiation among them. Development of automated phenotyping techniques with integrative and simultaneous quantification of multiple morphological and physiological traits enables early detection and quantification of different stresses on a whole plant basis. In this study, we have used different phenotyping techniques including chlorophyll fluorescence imaging, multispectral imaging, and 3D multispectral scanning, aiming to quantify changes in basil phenotypic traits under early and prolonged drought and salinity stress and to determine traits which could differentiate among drought and salinity stressed basil plants. Ocimum basilicum "Genovese" was grown in a growth chamber under well-watered control [45-50% volumetric water content (VWC)], moderate salinity stress (100 mM NaCl), severe salinity stress (200 mM NaCl), moderate drought stress (25-30% VWC), and severe drought stress (15-20% VWC). Phenotypic traits were measured for 3 weeks in 7-day intervals. Automated phenotyping techniques were able to detect basil responses to early and prolonged salinity and drought stress. In addition, several phenotypic traits were able to differentiate among salinity and drought. At early stages, low anthocyanin index (ARI), chlorophyll index (CHI), and hue (HUE2 D ), and higher reflectance in red (R Red ), reflectance in green (R Green ), and leaf inclination (LINC) indicated drought stress. At later stress stages, maximum fluorescence (F m ), HUE2 D , normalized difference vegetation index (NDVI), and LINC contribute the most to the differentiation among drought and non-stressed as well as among drought and salinity stressed plants. ARI and electron transport rate (ETR) were best for differentiation of salinity stressed plants from non-stressed plants both at early and prolonged stress.

Comparative Study of Chemical Composition, Cholinesterase Inhibition and Antioxidant Potential of Mentha pulegium L. Essential Oil.

Perennial plant Mentha pulegium L. (pennyroyal, Lamiaceae) can be found in Europe and Mediterranean. In areas where it thrives, M. pulegium is used in nutrition and as medicinal plant. Essential oil of M. pulegium is also a frequent constituent of foods and fragrances, because of mint-like odor. Regarding the use of M. pulegium in traditional medicine and nutrition, as well as fact that essential oils are potential sources of bioactive components, this study was conducted to examine the chemical composition of essential oil of M. pulegium wild growing in Bosnia and Herzegovina and its biological activity. The chemical profile testing was made using GC/MS and GC/FID technique. Potential of cholinesterase inhibition was tested by Ellman's assay. The antioxidant activity was tested by DPPH and FRAP assay. The dominant components in analyzed oil were pulegone 54.4 %, p-menthone 14.0 % and piperitenone 12.8 %. Good antioxidant activity and moderate cholinesterase inhibition potential of tested essential oil indicates to possibility of its use in treatment of diseases related to free radicals, Alzheimer disease and as lipid protecting antioxidant.

UPLC-MS/MS Phytochemical Analysis of Two Croatian Cistus Species and Their Biological Activity.

Aqueous extracts of two Cistus species wild growing in Croatia-Cistus creticus (CC) and Cistus salviifolius (CS)-have been assessed with UPLC-MS/MS, showing 43 different phytochemicals, with flavonol glycosides: myricetin-3-hexoside and myricetin-rhamnoside, predominate ones in CC and myricetin-3-hexoside in CS. Antioxidant potential tested with the FRAP method showed no difference between CS and CC aqueous extracts, while higher phenolic content of CC comparing to CS, determined with a Folin-Cicolateu reagent correlated to its higher antioxidant capacity observed by the DPPH method. Both extracts were assessed for antimicrobial activity, using disc-diffusion and broth microdilution assays, targeting the opportunistic pathogens, associated with food poisoning, urinary, respiratory tract, blood stream and wound infections in humans. Antimicrobial assays revealed that fungi were in general more sensitive to both Cistus aqueous extracts, comparing to the bacteria where two extracts showed very similar activity. The most potent activity was observed against A. baumannii for both extracts. The extracts were tested on human lung cancer (A549) cell line using the MTT assay, showing very similar antiproliferative activity. After 72 h treatment with CC and CS aqueous extracts in concentration of 0.5 g/L, the viability of the cells were 37% and 50% respectively, compared to non-treated cells.

Terpenes, Phenylpropanoids, Sulfur and Other Essential Oil Constituents as Inhibitors of Cholinesterases.

Essential oils constituents are a diverse family of low molecular weight organic compounds with comprehensive biological activity. According to their chemical structure, these active compounds can be divided into four major groups: terpenes, terpenoids, phenylpropenes, and "others". In addition, they may contain diverse functional groups according to which they can be classified as hydrocarbons (monoterpenes, sesquiterpenes, and aliphatic hydrocarbons); oxygenated compounds (monoterpene and sesquiterpene alcohols, aldehydes, ketones, esters, and other oxygenated compounds); and sulfur and/or nitrogen containing compounds (thioesters, sulfides, isothiocyanates, nitriles, and others). Compounds that act as cholinesterase inhibitors still represent the only pharmacological treatment of Alzheimer´s disease. Numerous in vitro studies showed that some compounds, found in essential oils, have a promising cholinesterase inhibitory activity, such as α-pinene, δ-3-carene, 1,8-cineole, carvacrol, thymohydroquinone, α- and ß-asarone, anethole, etc. This review summarizes the most relevant research published to date on essential oil constituents and their acetylcholinesterase/butyrylcholinesterase inhibitory potential as well as their structure related activity, synergistic and antagonistic effects.

Differentiation Between Unfiltered and Filtered Oblica and Leccino cv. Virgin Olive Oils.

The quality parameters, a variety of microcomponents, and the sensory characteristics of Oblica and Leccino cv. virgin olive oils (VOOs) were evaluated before and after filtration process adopted in order to estimate the individual varietal compositional changes. The dynamics of the formation of hydrolytic and oxidative changes in unfiltered (UF) and filtered (F) oils was asses by comparing level of free fatty acids (FFA), peroxide value (PV), and spectrophotometric indices periodically during 1 year of oil storage. An analysis of phenolics, tocopherols, and fatty acids was determined by chromatographic (HPLC and GC) and spectrometric methods, oxidative stability by Rancimat method while sensory analyses of obtained olive oils were performed by a trained professional panel. Single monovarietal VOO loses phenols in different rate with the applied filtration. Total secoiridoids decreases significantly in "Oblica" VOOs while no changes in their concentrations were found between unfiltered and filtered "Leccino" oils. Intensity of desired sensory properties decreases with filtration. In "Leccino" VOOs decrease of oxidative stability was more pronounced. After 12 months of storage, filtered "Leccino" VOOs had significantly lower FFA values than observed for the unfiltered counterparts. Further, there were no significant changes in PV and K270 values between unfiltered and filtered oils of both studied varietal oils. Storage time influenced more studied quality parameters than filtration, during which PV of unfiltered oils faster deteriorate. The highest changes between stored and corresponding fresh samples were exhibited in unfiltered "Oblica" VOOs. PRACTICAL APPLICATION: Quality enhancement of olive oil is constantly being done professionally and scientifically. The information provided in this study can be used in the industry of olive oil for improve the phenolic content, oxidative stability, and the sensory quality of virgin olive oils. The findings of stability test could be guidelines for mindful leading of the oil finishing up to bottling.

Phytochemical Composition, Antioxidant Potential and Cholinesterase Inhibition Potential of Extracts from Mentha pulegium L.

The aim of this work was to perform the phytochemical analysis as well as antioxidant and anticholinesterase potential of hot water and methanolic extracts from Mentha pulegium L., Lamiaceae, wild growing in Bosnia and Herzegovina. It is a plant that is traditionally used in Bosnia and Herzegovina medicine to treat gastrointestinal and neurological disorders. By HPLC-DAD analysis of tested extracts, 15 phenol compounds were identified and quantified. Rosmarinic acid was the most abundant compound in both extracts, followed by ellagic acid, eriodictyol, naringenin and chlorogenic acid. Both extracts were demonstrated good radical scavenging potential, reducing power and ability to inhibit lipid oxidation. The tested extracts also showed low ability to inhibit protein oxidation and low or no acetylcholinesterase and butyrylcholinesterase inhibition potential. The promising radical scavenging potential, reducing power and lipid protection potential of tested extracts indicates that the extracts of M. pulegium could be useful in preventing free radical linked diseases, as well as food protecting antioxidants.

Phytochemical Composition and Antioxidant Activities of the Essential Oil and Extracts of Satureja subspicata Vis. Growing in Bosnia and Herzegovina.

The phytochemical composition and the antioxidant activities of the essential oil, as well as methanol and hot water extracts of endemic Satureja subspicata Vis. growing in Bosnia and Herzegovina (BiH), were described. ß-Caryophyllene, cis-ß-ocimene, and α-pinene, identified by GC/MS and GC-FID, were the dominant oil components. The major compound of both of extracts, identified by HPLC-DAD, was rosmarinic acid. The analyzed essential oil showed moderate antioxidant activity. In this first report on the extracts of S. subspicata growing in BiH, the obtained results showed a high content of rosmarinic acid, as well as considerable amount of total phenols and flavonoids. Compared to the hot water extract, the methanol extract exhibits higher antioxidant potential, measured by DPPH and FRAP assay (IC50 = 0.45 g/l and 1879.43 equiv. Fe2+ µm), while the hot water extract showed higher potential in inhibition of linoleic acid oxidation (51.7% and 61.5% for 1 and 10 g/l). A good antioxidant potential of the tested extracts indicates their potential use as antioxidants, particularly for lipid protection, and partly explains the justification of the use of this plant in traditional medicine of BiH.

Qualitative HPLC-DAD/ESI-TOF-MS Analysis, Cytotoxic, and Apoptotic Effects of Croatian Endemic Centaurea ragusina L. Aqueous Extracts.

Centaurea ragusina L., an endemic Croatian plant species, revealed a good cytotoxic activity of aqueous extracts (AE) on human bladder (T24) and human glioblastoma (A1235) cancer cell lines. The chemical constituents were tentatively identified using high performance liquid chromatography HPLC-DAD/ESI-TOF-MS in negative ionization mode. The main compounds of herba extract were sesquiterpene lactones: solstitialin A 3,13-diacetate and epoxyrepdiolide; organic acid: quinic acid. The main compounds of flower extract were organic acids: quinic acid, citric acid, and malic acid; sesquiterpene lactone: cynaropicrin; phenolic compounds: chlorogenic acid and phenylpropanoid: syringin. The AE of C. ragusina were investigated for correlation of their effects on human bladder (T24) and human glioblastoma (A1235) cancer cell lines using the MTT assay. Although both extracts showed significant dose- and time-dependent cytotoxic activity against both cancer cell lines, the flower extract exhibited slightly higher activity. In order to determine type of cell death induced by treatment, cell lines were exposed subsequently to a treatment with both flower and herba AE. The majority of the cells died by induced apoptosis treatment. Flower AE (26.25%), compared to a leaf AE (22.15%) showed slightly higher percentage of an apoptosis in T24 cells, when compared to a non-treated cells (0.04%).

Phytochemical and Cytogenetic Characterization of Centaurea solstitialis L. (Asteraceae) from Croatia.

The cytogenetic characterization of Centaurea solstitialis L. (Asteraceae) showed a chromosome number of 2n = 16. Karyotype is composed by four pairs of metacentric, two pairs of submetacentric and two pairs of subtelocentric chromosomes. Physical mapping of two rDNA probes revealed two loci of 35S and one locus of 5S rRNA genes. Chromomycin fluorochrome banding revealed that all rDNA loci were GC rich. The genome size (2C-value) of 1.95 pg classes this species in the group of very small genomes. Chemical composition of C. solstitialis volatile oil (VO) from Croatia, studied with gas chromatography-mass spectrometry showed dominant components as it follows: hexadecanoic acid, α-linolenic acid, germacrene D and heptacosane. Antioxidant capacity, measured by ferric reducing power assay and 2,2-diphenyl-1-picrylhydrazyl methods, as well as inhibition of acetyl- and butyrylcholinesterase of VO was lower comparing to a standard solutions. Volatile oil tested with disc diffusion method showed good inhibitory potential against Pseudomonas aeruginosa, Escherichia coli and all tested fungi: Candida albicans, Penicillium funiculosum and Aspergillus fumigatus. The microdilution method showed best activity against Chronobacter sakazakii and A. fumigatus.

Campanula portenschlagiana ROEM. et SCHULT.: chemical and antimicrobial activities.

The phytochemical profile and the antimicrobial effects of the volatile oil and the aqueous extract of Campanula portenschlagiana, a wild growing plant endemic to Croatia, were described. In the volatile oil, 53 compounds were identified by GC-FID and GC/MS analyses. Diterpene alcohols constituted the major compound class with labda-13(16),14-dien-8-ol as the main compound. The aqueous extract was characterized by the total phenolic content. The antimicrobial potential of the volatile oil and the aqueous extract was evaluated against a diverse range of microorganisms comprising food-spoilage and food-borne pathogens. The volatile oil exhibited interesting and promising antimicrobial effects against the tested species, which were generally more pronounced against Gram-negative bacteria. In addition, the inhibitory effect of this volatile oil was also evaluated against eleven extended-spectrum ß-lactamase (ESBL)-producing isolates. The results suggest that the C. portenschlagiana volatile oil might be used as antimicrobial agent against ESBL-producing isolates and Gram-negative bacteria.

Phytochemical profiles of volatile constituents from Centaurea ragusina leaves and flowers and their antimicrobial effects.

The volatile oils obtained from the leaves and flowers of the Croatian endemic plant Centaurea ragusina L. were evaluated for antimicrobial activity. The chemical compositions, determined by GC and GC-MS, were complex, with sesquiterpenes the most abundant class of compound. Nonterpene hydrocarbons were also identified in high quantities, while acids and ketones were found in high quantities only in the leaf volatile oil. Aldehydes, alcohols, esters and other compounds were identified in lower quantities. Antimicrobial effects of the volatile samples were investigated by using the disc diffusion and microdilution assays. They were found to inhibit a wide range of bacteria and fungi, causing in vitro growth inhibition at a concentration of 250 microg/mL. The volatile oils displayed great antibacterial potential with MIC values ranging from 16 to 128 microg/mL against the Gram-positive bacteria Bacillus cereus, Clostridium perfringens, Enterococcus faecalis and Staphylococcus aureus, and lesser activity against the Gram-negative bacteria Cronobacter sakazakii, Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa, as well as fungal strains (MIC = 32-128 microg/mL).

Screening for acetylcholinesterase inhibition and antioxidant activity of selected plants from Croatia.

The methanol, ethyl acetate and chloroform extracts of selected Croatian plants were tested for their acetylcholinesterase (AChE) inhibition and antioxidant activity. Assessment of AChE inhibition was carried out using microplate reader at 1 mg mL⁻¹. Antioxidant capacities were determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging test and ferric reducing/antioxidant power assay (FRAP). Total phenol content (TPC) of extracts were determined using Folin-Ciocalteu colorimetric method. Out of 48 extracts, only methanolic extract of the Salix alba L. cortex exerted modest activity towards AChE, reaching 50.80% inhibition at concentration of 1 mg mL⁻¹. All the other samples tested had activity below 20%. The same extract performed the best antioxidative activity using DPPH and FRAP method, too. In essence, among all extracts used in the screening, methanolic extracts showed the best antioxidative activity as well as highest TPC.