In Vitro and In Silico of Cholinesterases Inhibition and In Vitro and In Vivo Anti-Melanoma Activity Investigations of Extracts Obtained from Selected Berberis Species.

Berberis species have a long history of use in traditional Chinese medicine, Ayurvedic medicine, and Western herbal medicine. The aim of this study was the quantification of the main isoquinoline alkaloids in extracts obtained from various Berberis species by HPLC, in vitro and in silico determination of anti-cholinesterase activity, and in vitro and in vivo investigations of the cytotoxic activity of the investigated plant extracts and alkaloid standards. In particular, Berberis species whose activity had not been previously investigated were selected for the study. In the most investigated Berberis extracts, a high content of berberine and palmatine was determined. Alkaloid standards and most of the investigated plant extracts exhibit significant anti-cholinesterase activity. Molecular docking results confirmed that both alkaloids are more favourable for forming complexes with acetylcholinesterase compared to butyrylcholinesterase. The kinetic results obtained by HPLC-DAD indicated that berberine noncompetitively inhibited acetylcholinesterase, while butyrylcholinesterase was inhibited in a mixed mode. In turn, palmatine exhibited a mixed inhibition of acetylcholinesterase. The cytotoxic activity of berberine and palmatine standards and plant extracts were investigated against the human melanoma cell line (A375). The highest cytotoxicity was determined for extract obtained from Berberis pruinosa cortex. The cytotoxic properties of the extract were also determined in the in vivo investigations using the Danio rerio larvae xenograft model. The obtained results confirmed a significant effect of the Berberis pruinosa cortex extract on the number of cancer cells in a living organism. Our results showed that extracts obtained from Berberis species, especially the Berberis pruinosa cortex extract, can be recommended for further in vivo experiments in order to confirm the possibility of their application in the treatment of neurodegenerative diseases and human melanoma.

Comparison of supercritical CO2 extraction and pressurized fluid extraction for isolation of alkaloids from Anacardium occidentale with the study of its anti-inflammatory activity.

In recent years, there has been a growing interest in the therapeutic potential of natural compounds, particularly of plant origin, owing to their demonstrated anti-inflammatory properties. Among these, Anacardium occidentale, commonly known as cashew, has garnered significant attention due to its reputed health benefits. This study aim to establish a correlation between the bioactive compounds contained in the extracts of Anacardium occidentale and its anti-inflammatory activity. Dried Anacardium occidentale leaves powder was used as the extraction matrix. Extraction techniques are maceration, pressurized fluid extraction (PFE), and supercritical fluid extraction (SFE). The preliminary analysis of extracts was made by LC-MS/MS. The Response Surface Methodology (RSM), Principal Component Analysis (PCA), and heat maps were employed to model the influence of experimental conditions on extraction yield and peak area of specific compounds from the plant. To evaluate anti-inflammatory activity, RAW 264.7 cells were cultured, activated with LPS, and treated with varying concentrations of the plant extracts. Cell proliferation was assessed using the XTT assay. Indeed, Anacardium occidentale extracts contain anacardic acids, cardanols, and cardol, with distinct profiles yielded by SFE and ethanol-based methods. RSM shows that temperature and ethanol, as additives to CO2, significantly affect extraction efficiency in both PFE and SFE. Moreover, this composition with SFE demonstrate higher selectivity for specific group of compounds. The extracts exhibit anti-inflammatory properties without cytotoxicity in macrophages, reducing levels of pro-inflammatory proteins COX-2, COX-1, and TLR4 in activated cells. This suggests their potential as anti-inflammatory agents without adverse effects on cell viability or pro-inflammatory protein levels in non-activated cells. Overall, these findings underscore the promising therapeutic potential of Anacardium occidentale extracts in mitigating inflammation, while also providing crucial insights into optimizing the extraction process for targeted compound isolation. Thus, this makes a good prospect for the development of anti-inflammatory drugs from this plant.

Metal tolerance and Cd phytoremoval ability in Pisum sativum grown in spiked nutrient solution.

In the presented study, the effects of cadmium (Cd) stress and silicon (Si) supplementation on the pea plant (Pisum sativum L.) were investigated. The tendency to accumulate cadmium in the relevant morphological parts of the plant (roots and shoots respectively)-bioaccumulation, the transfer of this element in the plant (translocation) and the physiological parameters of the plant through indicators of oxidative stress were determined. Model studies were carried out at pH values 6.0 and 5.0 plant growth conditions in the hydroponic cultivation. It was shown that Cd accumulates mostly in plant roots at both pH levels. However, the Cd content is higher in the plants grown at lower pH. The Cd translocation factor was below 1.0, which indicates that the pea is an excluder plant. The contamination of the plant growth environment with Cd causes the increased antioxidant stress by the growing parameters of the total phenolic content (TPC), polyphenol oxidase activity (PPO), the malondialdehyde (MDA) and lipid peroxidation (LP). The results obtained showed that the supplementation with Si reduces these parameters, thus lowering the oxidative stress of the plant. Moreover, supplementation with Si leads to a lower content of Cd in the roots and reduces bioaccumulation of Cd in shoots and roots of pea plants.

Determination of Some Isoquinoline Alkaloids in Extracts Obtained from Selected Plants of the Ranunculaceae, Papaveraceae and Fumarioideae Families by Liquid Chromatography and In Vitro and In Vivo Investigations of Their Cytotoxic Activity.

Alkaloids are heterocyclic bases with widespread occurrence in nature. Plants are rich and easily accessible sources of them. Most isoquinoline alkaloids have cytotoxic activity for different types of cancer, including malignant melanoma, the most aggressive type of skin cancer. The morbidity of melanoma has increased worldwide every year. For that reason, developing new candidates for anti-melanoma drugs is highly needed. The aim of this study was to investigate the alkaloid compositions of plant extracts obtained from Macleaya cordata root, stem and leaves, Pseudofumaria lutea root and herb, Lamprocapnos spectabilis root and herb, Fumaria officinalis whole plant, Thalictrum foetidum root and herb, and Meconopsis cambrica root and herb by HPLC-DAD and LC-MS/MS. For determination of cytotoxic properties, human malignant melanoma cell line A375, human Caucasian malignant melanoma cell line G-361, and human malignant melanoma cell line SK-MEL-3 were exposed in vitro to the tested plant extracts. Based on the in vitro experiments, Lamprocapnos spectabilis herb extract was selected for further, in vivo research. The toxicity of the extract obtained from Lamprocapnos spectabilis herb was tested using an animal zebrafish model in the fish embryo toxicity test (FET) for determination of the LC50 value and non-toxic doses. Determination of the influence of the investigated extract on the number of cancer cells in a living organism was performed using a zebrafish xenograft model. Determination of the contents of selected alkaloids in different plant extracts was performed using high performance liquid chromatography (HPLC) in a reverse-phase system (RP) on a Polar RP column with a mobile phase containing acetonitrile, water and ionic liquid. The presence of these alkaloids in plant extracts was confirmed by LC-MS/MS. Preliminary cytotoxic activity of all prepared plant extracts and selected alkaloid standards was examined using human skin cancer cell lines A375, G-361, and SK-MEL-3. The cytotoxicity of the investigated extract was determined in vitro by cell viability assays (MTT). For in vivo determination of investigated extract cytotoxicity, a Danio rerio larvae xenograft model was used. All investigated plant extracts in in vitro experiments exhibited high cytotoxic activity against the tested cancer cell lines. The results obtained using the Danio rerio larvae xenograft model confirmed the anticancer activity of the extract obtained from Lamprocapnos spectabilis herb. The conducted research provides a basis for future investigations of these plant extracts for potential use in the treatment of malignant melanoma.

Determination of Selected Isoquinoline Alkaloids from Chelidonium majus, Mahonia aquifolium and Sanguinaria canadensis Extracts by Liquid Chromatography and Their In Vitro and In Vivo Cytotoxic Activity against Human Cancer Cells.

The search for new substances with cytotoxic activity against various cancer cells, especially cells that are very resistant to currently used chemotherapeutic agents, such as melanoma cells, is a very important scientific aspect. We investigated the cytotoxic effect of Chelidonium majus, Mahonia aquifolium and Sanguinaria canadensis extracts obtained from different parts of these plants collected at various vegetation stages on FaDu, SCC-25, MCF-7, and MDA-MB-231 cancer cells. Almost all the tested extracts showed higher cytotoxicity against these cancer cells than the anticancer drug etoposide. The highest cytotoxicity against the FaDu, SCC-25, MCF-7 and MDA-MB-231 cancer cell lines was obtained for the Sanguinaria candensis extract collected before flowering. The cytotoxicity of extracts obtained from different parts of Chelidonium majus collected at various vegetation stages was also evaluated on melanoma cells (A375, G361 and SK-MEL-3). The highest cytotoxic activity against melanoma A375 cells was observed for the Chelidonium majus root extract, with an IC50 of 12.65 µg/mL. The same extract was the most cytotoxic against SK-MEL-3 cells (IC50 = 1.93 µg/mL), while the highest cytotoxic activity against G361 cells was observed after exposure to the extract obtained from the herb of the plant. The cytotoxic activity of Chelidonium majus extracts against melanoma cells was compared with the cytotoxicity of the following anticancer drugs: etoposide, cisplatin and hydroxyurea. In most cases, the IC50 values obtained for the anticancer drugs were higher than those obtained for the Chelidonium majus extracts. The most cytotoxic extract obtained from the root of Chelidonium majus was selected for in vivo cytotoxic activity investigations using a Danio rerio larvae xenograft model. The model was applied for the first time in the in vivo investigations of the extract's anticancer potential. The application of Danio rerio larvae xenografts in cancer research is advantageous because of the transparency and ease of compound administration, the small size and the short duration and low cost of the experiments. The results obtained in the xenograft model confirmed the great effect of the investigated extract on the number of cancer cells in a living organism. Our investigations show that the investigated plant extracts exhibit very high cytotoxic activity and can be recommended for further experiments in order to additionally confirm their potential use in the treatment of various human cancers.

Synthesis and physicochemical characterization of bovine lactoferrin supersaturated complex with iron (III) ions.

The aim of the study was to investigate the process of Fe3+ binding to bLTF. Moreover, the physicochemical characterization of the respective supersaturated complex was studied. The knowledge should be important for the description of processes that may take place in dairy products fortified with iron. Additionally, the synthesized complex can be utilized as a dietary supplement for the treatment of iron deficiency anemia (IDA). Finally, it was shown that formation of supersaturated iron-protein structures which include LTF often accompanies development of neurodegenerative diseases such as Alzheimer or Parkinson. Thus, the study can reveal some aspects of its pathogenesis process. The methodology of the investigation comprised the utilization of batch sorption study and applying Freundlich and Langmuir models. The complex also was characterized by numerous techniques: spectrometric (ICP-MS), spectroscopic (UV-Vis, ATR-FTIR), electron microscopy (TEM-EDX), SDS-PAGE. Based on obtained results the potential mechanisms of iron interaction with protein were described. Moreover, the molecular docking was applied to visualize possible metal binding sites. The respective complex contains ≈ 33.0 mg/g of iron which is nearly 50 Fe3+ per one protein molecule. The cytotoxicity of the obtained complex was evaluated by MTT reduction and LDH release assays on Caco-2 and nL929 cell lines.

Isoquinoline Alkaloid Contents in Macleaya cordata Extracts and Their Acetylcholinesterase and Butyrylcholinesterase Inhibition.

An important strategy for treating neurodegenerative disorders is to maintain the levels of acetylcholine in the synaptic cleft by blocking the cholinesterases. Searching for new effective compounds with inhibited acetylcholinesterase and butyrylcholinesterase activity is one of the most significant challenges of the modern scientific research. The aim of this study was the optimization of the condition for cholinesterase activity determination by high-performance liquid chromatography coupled with diode array detector (HPLC-DAD) in terms of concentrations of enzymatic reaction mixture components, temperature of incubation, and incubation time. In vitro investigation of acetylcholinesterase and butyrylcholinesterase activity inhibition by some isoquinoline alkaloids and extracts obtained from the aerial part and roots of Macleaya cordata collected in May, July, and September. Acetylcholinesterase and butyrylcholinesterase activity inhibition of the extracts obtained from the plant had not been tested previously. The application of the HPLC method allowed eliminating absorption of interfering components, for example, alkaloids such as sanguinarine and berberine. The HPLC method was successfully applied for the evaluation of the acetylcholinesterase inhibitory activity in samples such as plant extracts, especially those containing colored components adsorbing at the same wavelength as the adsorption wavelength of 5-thio-2-nitro-benzoic acid, which is the product of the reaction between thiocholine (product of the hydrolysis of acetyl/butyrylthiocholine reaction) with Ellman's reagent. Moreover, liquid chromatography coupled with a triple quadrupole mass spectrometer (LC-QqQ-ESI-MS/MS) analysis allowed evaluating the identification of relevant bioactive compounds in the obtained plant extracts. The investigated alkaloids, especially sanguinarine and chelerythrine, and all the Macleaya cordata extracts, especially the extract obtained from the aerial part collected in May, exhibited very high cholinesterase activity inhibition. HPLC-DAD was also applied for the kinetics study of the most active alkaloids sanguinarine and chelerythrine. Our investigations demonstrated that these plant extracts can be recommended for further in vivo experiments to confirm their cholinesterase inhibition activity.

Capillary Zone Electrophoresis in Tandem with Flow Cytometry in Viability Study of Various ATCC Bacterial Strains under Antibiotic Treatment.

The aim of this study was to develop an innovative method of examining bacterial survival using capillary zone electrophoresis (CZE) and flow cytometry (FC) as a reference method. For this purpose, standard strains of bacteria from the ATCC collection were used: Enterococcus faecalis ATCC 14506, Staphylococcus aureus ATCC 11632, Klebsiella pneumoniae ATCC 10031, Pseudomonas aeruginosa ATCC 27853, and Escherichia coli ATCC 25922, as well as seven antibiotics with different antimicrobial mechanisms of action. The ratio of live and dead cells in the tested sample in CZE measurements were calculated using our algorithm that takes into account the detection time. Results showed a high agreement between CZE and FC in the assessment of the percentage of live cells exposed to the stress factor in both antibiotic susceptibility and time-dependent assays. The applied measuring system to assess the effectiveness of antibiotic therapy in in vitro conditions is a method with great potential, and the data obtained with the use of CZE mostly correspond to the expected drug sensitivity according to EUCAST and CLSI guidelines.

Comparison Study of Cytotoxicity of Bare and Functionalized Zinc Oxide Nanoparticles.

In this paper, a study of the cytotoxicity of bare and functionalized zinc oxide nanoparticles (ZnO NPs) is presented. The functionalized ZnO NPs were obtained by various types of biological methods including microbiological (intra- and extracellular with Lactobacillus paracasei strain), phytochemical (Medicago sativa plant extract) and biochemical (ovalbumin from egg white protein) synthesis. As a control, the bare ZnO NPs gained by chemical synthesis (commercially available) were tested. The cytotoxicity was measured through the use of (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) dye as well as lactate dehydrogenase (LDH) assays against murine fibroblast L929 and Caco-2 cell lines. As a complementary method, scanning electron microscopy (SEM) was performed to assess the morphology of the tested cells after treatment with ZnO NPs. The microscopic data confirmed the occurrence of apoptotic blebbing and loss of membrane permeability after the administration of all ZnO NPs. The reactive oxygen species (ROS) concentration during the cell lines' exposure to ZnO NPs was measured fluorometrically. Additionally, the photocatalytic degradation of methylene blue (MB) dye in the different light conditions, as well as the antioxidant activity of bare and functionalized ZnO NPs, is also reported. The addition of all types of tested ZnO NPs to methylene blue resulted in enhanced rates of photo-degradation in the presence of both types of irradiation, but the application of UV light resulted in higher photocatalytic activity of ZnO NPs. Furthermore, bare (chemically synthetized) NPs have been recognized as the strongest photocatalysts. In the context of the obtained results, a mechanism underlying the toxicity of bio-ZnO NPs, including (a) the generation of reactive oxygen species and (b) the induction of apoptosis, is proposed.

Separation and Determination of Chemopreventive Phytochemicals of Flavonoids from Brassicaceae Plants.

The main aim of this study was to develop a method for the isolation and determination of polyphenols-in particular, flavonoids present in various morphological parts of plants belonging to the cabbage family (Brassicaceae). Therefore, a procedure consisting of maceration, acid hydrolysis and measurement of the total antioxidant capacity of plant extracts (using DPPH assay) was conducted. Qualitative analysis was performed employing thin-layer chromatography (TLC), which was presented to be a suitable methodology for the separation and determination of chemopreventive phytochemicals from plants belonging to the cabbage family. The study involved the analysis of 25 vegetal samples, including radish, broccoli, Brussels sprouts, kale, canola, kohlrabi, cabbage, Chinese cabbage, red cabbage, pak choi and cauliflower. In addition, selected flavonoids content in free form and bonded to glycosides was determined by using an RP-UHPLC-ESI-MS/MS method.

Comparative Studies of Selected Criteria Enabling Optimization of the Extraction of Polar Biologically Active Compounds from Alfalfa with Supercritical Carbon Dioxide.

The aim of this research was to provide crucial and useful data about the selection of the optimization criteria of supercritical carbon dioxide extraction of alfalfa at a quarter-technical plant. The correlation between more general output, including total phenolics and flavonoids content, and a more specified composition of polar constituents was extensively studied. In all alfalfa extracts, polar bioactive constituents were analyzed by both spectrometric (general output) and chromatographic (detailed output) analyses. Eight specific phenolic acids and nine flavonoids were determined. The most dominant were salicylic acid (221.41 µg g-1), ferulic acid (119.73 µg g-1), quercetin (2.23 µg g-1), and apigenin (2.60 µg g-1). For all seventeen analyzed compounds, response surface methodology and analysis of variance were used to provide the optimal conditions of supercritical fluid extraction for each individual constituent. The obtained data have shown that eight of those compounds have a similar range of optimal process parameters, being significantly analogous for optimization based on total flavonoid content.

Metabolic Profiling of VOCs Emitted by Bacteria Isolated from Pressure Ulcers and Treated with Different Concentrations of Bio-AgNPs.

Considering the advent of antibiotic resistance, the study of bacterial metabolic behavior stimulated by novel antimicrobial agents becomes a relevant tool to elucidate involved adaptive pathways. Profiling of volatile metabolites was performed to monitor alterations of bacterial metabolism induced by biosynthesized silver nanoparticles (bio-AgNPs). Escherichia coli, Enterococcus faecalis, Klebsiella pneumoniae and Proteus mirabilis were isolated from pressure ulcers, and their cultures were prepared in the presence/absence of bio-AgNPs at 12.5, 25 and 50 µg mL-1. Headspace solid phase microextraction associated to gas chromatography-mass spectrometry was the employed analytical platform. At the lower concentration level, the agent promoted positive modulation of products of fermentation routes and bioactive volatiles, indicating an attempt of bacteria to adapt to an ongoing suppression of cellular respiration. Augmented response of aldehydes and other possible products of lipid oxidative cleavage was noticed for increasing levels of bio-AgNPs. The greatest concentration of agent caused a reduction of 44 to 80% in the variety of compounds found in the control samples. Pathway analysis indicated overall inhibition of amino acids and fatty acids routes. The present assessment may provide a deeper understanding of molecular mechanisms of bio-AgNPs and how the metabolic response of bacteria is untangled.

Determination of Cytotoxic Activity of Sanguinaria canadensis Extracts against Human Melanoma Cells and Comparison of Their Cytotoxicity with Cytotoxicity of Some Anticancer Drugs.

Melanoma is an enormous global health burden, and should be effectively addressed with better therapeutic strategies. Therefore, new therapeutic agents are needed for the management of this disease. The aim of this study was the investigation of cytotoxic activity of some isoquinoline alkaloid standards and extracts obtained from Sanguinaria canadensis-collected before, during, and after flowering-against three different human melanoma cells (A375, G361, SK-MEL-3). The cytotoxicity of these extracts was not previously tested on these melanoma cell lines. Determination of alkaloid contents was performed by HPLC-DAD using Polar RP column and mobile phase containing acetonitrile, water, and 1-butyl-3-methylimidazolium tetrafluoroborate. The cytotoxicity of alkaloid standards was investigated by determination of cell viability and calculation of IC50 values. Significant differences were observed in the alkaloids content and cytotoxic activity of the extracts, depending on the season of collection of the plant material. In the Sanguinaria canadensis extracts high contents of sanguinarine (from 4.8543 to 9.5899 mg/g of dry plant material) and chelerythrine (from 42.7224 to 6.8722 mg/g of dry plant material) were found. For both of these alkaloids, very high cytotoxic activity against the tested cell lines were observed. The IC50 values were in the range of 0.11-0.54 µg/mL for sanguinarine and 0.14 to 0.46 µg/mL for chelerythrine. IC50 values obtained for Sanguinaria canadensis extracts against all tested cell lines were also very low (from 0.88 to 10.96 µg/mL). Cytotoxic activity of alkaloid standards and Sanguinaria canadensis extracts were compared with the cytotoxicity of anticancer drugs-etoposide, cisplatin, and hydroxyurea. In all cases except the one obtained for cisplatin against A375, which was similar to that obtained for Sanguinaria canadensis after flowering against the same cell line, IC50 values obtained for anticancer drugs were higher than the IC50 values obtained for sanguinarine, chelerythrine, and Sanguinaria canadensis extracts. Our results showed that Sanguinaria canadensis extracts and isoquinoline alkaloids, especially sanguinarine and chelerythrine, could be recommended for further in vivo experiments in order to confirm the possibility of their application in the treatment of human melanomas.

Determination of Cytotoxic Activity of Selected Isoquinoline Alkaloids and Plant Extracts Obtained from Various Parts of Mahonia aquifolium Collected in Various Vegetation Seasons.

Melanoma is a serious form of skin cancer that begins in cells known as melanocytes. While it is less common than the other forms of skin cancer, melanoma is more dangerous because of its ability to spread to other organs more rapidly if it is not treated at an early stage. The number of people diagnosed with melanoma has increased over the last few decades. The most widely used treatments include surgery, chemotherapy, and radiation therapy. The search for new drugs to treat various cancers is one of the most important challenges of modern scientific research. Some isoquinoline alkaloids found in different plant species have strong cytotoxic effects on various cancer cells. We tested the effect of isoquinoline alkaloids and extracts obtained from various parts of Mahonia aquifolium collected in various vegetation seasons on human melanoma cancer cells and our data indicated that investigated extract induced significant reduction in cell viability of Human malignant melanoma cells (A375), human Caucasian malignant melanoma cell line (G361), and human malignant melanoma cell line (SKMEL3 cancer cell lines in a dose- and time-dependent manner. Differences in cytotoxic activity were observed for extracts obtained from various parts of Mahonia aquifolium. Significant differences were also obtained in the alkaloids content and cytotoxic activity of the extracts depending on the season of collection of plant material. Our investigations exhibit that these plant extracts can be recommended for further in vivo experiments in order to confirm the possibility of their use in the treatment of human melanomas.

Combination of electrochemical unit and ESI-MS in fragmentation of flavonoids.

INTRODUCTION: Predictive approaches on the activity of natural compounds based on the fragmentation by instrumental techniques are important for consideration of such molecules as drug candidates and defining new structures with promising properties. Since flavonoids are well-known antioxidants, their redox properties can be related to their pharmacological activity. OBJECTIVES: In this work, the potential of electrochemical unit coupled to electrospray ionisation mass spectrometry (ESI-MS) was assessed for fragmentation activity relationships studies of selected flavonoids. METHODOLOGY: Methodology of this research included electrochemical conversion of standards of flavonoids at different pH values and their further analysis with the use of ESI-MS. In addition, signals obtained from the blank samples were also identified and used for interpretation due to electrochemical nature of the ESI source. Half maximal inhibitory concentration (IC50 ) values of flavonoids for 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) antioxidant activity assays were analysed for possible correlation with the structures of flavonoids and products of electrochemical conversion. RESULTS: Fragmentation activity relationships were suggested using the proposed approach and for some of the flavonoids it was not specific enough to determine the input of a particular structural feature to the activity, but for others they were in agreement with those found in the literature. Obtained results showed potential of the proposed approach for application in plant sciences as a fast pre-screening tool for newly isolated bioactive compounds.

Determination of Neonicotinoids in Honey Samples Originated from Poland and Other World Countries.

A method development for determination of neonicotinoid residues in honey samples was developed. The proposed methodology consisted in QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe). That was used for sample preparation and UHPLC/UV (ultra-performance liquid chromatography with ultraviolet detection) utilized for chromatographic analysis. The developed method proved to be sensitive, with LOD (Limit of detection) value in the range of 60.80 to 80.98 ng/g hence LOQ (Limit of quantification) value was in the range of 184.26 to 245.40 ng/g. The method has tested on Polish honey and applied to honey from various countries (Bulgaria, Czech Republic, France, Greece, Italy, Portugal, Romania, Australia, Brazil, Cameroon, Russia, USA and Turkey). Several honey types were tested, while physicochemical properties of all honeys and were investigated. The methodology for general characterization of pollen grains originated from selected plants, to confirm the type of honey was also presented. There was a total lack of the mentioned neonicotinoids in sunflower honey. Except of this, only two samples of rapeseed and two samples of acacia honey (from Poland and Romania) were neonicotinoids free. In 19 samples the targeted pesticides were detected above LOQ. In all other investigated samples, the neonicotinoids were found at least at the LOD or LOQ level.

The Influence of Plant Material Enzymatic Hydrolysis and Extraction Conditions on the Polyphenolic Profiles and Antioxidant Activity of Extracts: A Green and Efficient Approach.

The aim of this study was to develop a new comprehensive extraction protocol based on green technology for the enhanced release of polyphenolic compounds from plant cells. In this work, extracts from yerba mate and yellow lupine seed were obtained by using three different extraction techniques: maceration, supercritical fluid extraction with co-solvent (SFE) and enzyme assisted-supercritical fluid extraction with co-solvent (EA-SFE). Several experimental parameters such as time, type of solvent and co-solvent as well as CO2 flow rate were selected to obtain the highest extraction efficiency. The chemical profiles in the obtained extracts and their biological activity were evaluated. HPLC-MS/MS analysis indicated that the level of phenolic compounds in extracts from yerba mate obtained by EA-SFE was approximately five times higher than for maceration and 3.2 times higher than for SFE. In the case of extracts from yellow lupine seed an approximately 5.6-fold increase was observed in comparison with maceration and SFE with 96% MeOH, and 2.9 times for SFE with 96% EtOH. The developed protocol with a mix of enzymes commonly applied in the agricultural industry significantly raises the efficiency of liberation of secondary metabolites.

Interactions of Whey Proteins with Metal Ions.

Whey proteins tend to interact with metal ions, which have implications in different fields related to human life quality. There are two impacts of such interactions: they can provide opportunities for applications in food and nutraceuticals, but may lead to analytical challenges related to their study and outcomes for food processing, storage, and food interactions. Moreover, interactions of whey proteins with metal ions are complicated, requiring deep understanding, leading to consequences, such as metalloproteins, metallocomplexes, nanoparticles, or aggregates, creating a biologically active system. To understand the phenomena of metal-protein interactions, it is important to develop analytical approaches combined with studies of changes in the biological activity and to analyze the impact of such interactions on different fields. The aim of this review was to discuss chemistry of ß-lactoglobulin, α-lactalbumin, and lactotransferrin, their interactions with different metal ions, analytical techniques used to study them and the implications for food and nutraceuticals.

Distribution of sapogenins in morphological Medicago sativa L. parts: Comparison of various extraction techniques.

Saponins in plant extracts were indirectly determined by estimation of the content of sapogenins. The first step of determination is extraction with high efficiency. One conventional extraction technique (maceration) and two modern ones (accelerated solvent extraction and supercritical fluid extraction) were compared. Methanol and ethanol were used as solvents or co-solvents. The results were supported by statistical analysis. Saponins were extracted from leaves, roots, and sprouts of Medicago sativa. Acid hydrolysis, purification, and determination by high-performance liquid chromatography with evaporative light scattering detector were used. The content of sapogenins was the highest in the roots. Smaller amounts of sapogenins were found in sprouts and the smallest ones in leaves. The main ingredient was medicagenic acid with mean concentration of 621.8 µg/g in roots, 456.7 µg/g in sprouts, and 471.3 µg/g in leaf extract. The highest content of sapogenins in extract was obtained after maceration with methanol; however, this method is nonselective in relation to biologically active compounds. Due to the possibility of using the obtained extracts with sapogenins in the cosmetic or pharmaceutical industry, the selection of extraction techniques and solvents is a very important aspect. Additionally, the chosen technique should be considered eco-friendly and consistent with the assumptions of "green chemistry."

Determination of Selected Isoquinoline Alkaloids from Mahonia Aquifolia; Meconopsis Cambrica; Corydalis Lutea; Dicentra Spectabilis; Fumaria Officinalis; Macleaya Cordata Extracts by HPLC-DAD and Comparison of Their Cytotoxic Activity.

Alkaloids have protective functions for plants and can play an important role in living organisms. Alkaloids may have a wide range of pharmacological activities. Many of them have cytotoxic activity. Nowadays, cancer has become a serious public health problem. Searching for effective drugs with anticancer activity is one of the most significant challenges of modern scientific research. The aim of this study was the investigation of cytotoxic activity of extracts obtained from Corydalis lutea root and herb, Dicentra spectabilis root and herb, Fumaria officinalis, Macleaya cordata leaves and herb, Mahonia aquifolia leaves and cortex, Meconopsis cambrica root and herb on FaDu, SCC-25, MCF-7, and MDA-MB-231 cancer cell lines. The cytotoxic activity of these extracts has not been previously tested for these cell lines. The aim was also to quantify selected alkaloids in the investigated extracts by High Performance Liquid Chromatography (HPLC). The analyses of alkaloid content were performed using HPLC in reversed phase (RP) mode using Polar RP column and mobile phase containing acetonitrile, water and ionic liquid (IL). Cytotoxic effect of the tested plant extracts and respective alkaloid standards were examined using human pharyngeal squamous carcinoma cells (FaDu), human tongue squamous carcinoma cells (SCC-25), human breast adenocarcinoma cell line (MCF-7), human triple-negative breast adenocarcinoma cell line (MDA-MB-231). All investigated plant extracts possess cytotoxic activity against tested cancer cell lines: FaDu, SCC-25, MCF-7, and MDA-MB-231. The highest cytotoxic activity against FaDu, SCC-25, and MCF-7 cell lines was estimated for Macleaya cordata leaf extract, while the highest cytotoxic activity against MDA-MB-231 cell line was obtained for Macleaya cordata herb extract. Differences in cytotoxic activity were observed for extracts obtained from various parts of investigated plants. In almost all cases the cytotoxic activity of investigated plant extracts, especially at the highest concentration against tested cell lines was significantly higher than the activity of anticancer drug etoposide. Our investigations exhibit that these plant extracts can be recommended for further in vivo experiments to confirm their anticancer activity.