Silicon's Influence on Polyphenol and Flavonoid Profiles in Pea (Pisum sativum L.) under Cadmium Exposure in Hydroponics: A Study of Metabolomics, Extraction Efficacy, and Antimicrobial Properties of Extracts.

The current study aimed to investigate the impact of silicon (Si) supplementation in the form of Na2SiO3 on the metabolome of peas under normal conditions and following exposure to cadmium (Cd) stress. Si is known for its ability to enhance stress tolerance in various plant species, including the mitigation of heavy metal toxicity. Cd, a significant contaminant, poses risks to both human health and the environment. The study focused on analyzing the levels of bioactive compounds in different plant parts, including the shoot, root, and pod, to understand the influence of Si supplementation on their biosynthesis. Metabolomic analysis of pea samples was conducted using a targeted HPLC/MS approach, enabling the identification of 15 metabolites comprising 9 flavonoids and 6 phenolic acids. Among the detected compounds, flavonoids, such as flavon and quercetin, along with phenolic acids, including chlorogenic acid and salicylic acid, were found in significant quantities. The study compared Si supplementation at concentrations of 1 and 2 mM, as well as Cd stress conditions, to evaluate their effects on the metabolomic profile. Additionally, the study explored the extraction efficiency of three different methods: accelerated solvent extraction (ASE), supercritical fluid extraction (SFE), and maceration (MAC). The results revealed that SFE was the most efficient method for extracting polyphenolic compounds from the pea samples. Moreover, the study investigated the stability of polyphenolic compounds under different pH conditions, ranging from 4.0 to 6.0, providing insights into the influence of the pH on the extraction and stability of bioactive compounds.

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.

Comparative Studies of Extracts Obtained from Brassica oleracea L. Plants at Different Stages of Growth by Isolation and Determination of Isothiocyanates: An Assessment of Chemopreventive Properties of Broccoli.

The main goal of this work was to develop analytical procedures for the isolation and determination of selected isothiocyanates. As an example, particularly sulforaphane from plants of the Brassicaceae Burnett or Cruciferae Juss family. The applied methodology was mainly based on classical extraction methods and high-performance liquid chromatography coupled with tandem mass spectrometry. Moreover, the effect of temperature on the release of isothiocyanates from plant cells was considered. The cytotoxic activity of the obtained plant extracts against a selected cancer cell line has also been included. The results allow evaluating the usefulness of obtained plant extracts and raw sprouts regarding their content of isothiocyanates-bioactive compounds with chemopreventive properties.

Development and Validation of LC-MS/MS Method for Determination of Cytisine in Human Serum and Saliva.

Cytisine (CYT) is a quinolizidine alkaloid used for nicotine addiction treatment. Recent clinical trial data regarding cytisine confirm its high effectiveness and safety as a smoking cessation treatment. CYT's popularity is growing due to its increased availability and licensing in more countries worldwide. This increased use by smokers has also resulted in an urgent need for continued drug research, including developing appropriate analytical methods for analyzing the drug in biological samples. In this study, a simple, fast, and reliable method combining hydrophilic interaction liquid chromatography and electrospray ionization quadrupole time of flight mass spectrometry (HILIC/ESI-QTOF-MS) for the determination of CYT in human serum and saliva was developed and validated. This was undertaken after the previous pre-treatment of the sample using solid-phase extraction (SPE). A hydrophilic interaction liquid chromatography (HILIC) column with a silica stationary phase was used for chromatographic analysis. In a linear gradient, the mobile phase consisted of acetonitrile (ACN) and formate buffer at pH 4.0. The proposed method was fully validated and demonstrated its sensitivity, selectivity, precision, and accuracy. The method was successfully applied to determine CYT in serum and, for the first time, in saliva. The findings indicate that saliva could be a promising non-invasive alternative to measure the free concentration of CYT.

Implications of Sample Preparation Methods on the MALDI-TOF MS Identification of Spore-Forming Bacillus Species from Food Samples: A Closer Look at Bacillus licheniformis, Peribacillus simplex, Lysinibacillus fusiformis, Bacillus flexus, and Bacillus marisflavi.

This research underscores the criticality of tailored culture conditions and incubation periods for effective and accurate identification of spore-forming bacteria: Bacillus licheniformis, Peribacillus simplex, Lysinibacillus fusiformis, Bacillus flexus, and Bacillus marisflav, isolated from food samples, utilizing the MALDI-TOF MS technique. All isolated strains were confirmed as Gram-positive bacteria from diverse genera through 16S rDNA gene sequencing. To enhance the accuracy of the identification process, the study employed an optimization strategy involving a varied incubation time (ranging from 1 to 48 h) and two distinct sample preparation approaches-direct transfer facilitated by formic acid and protein extraction via ethanol. It was observed that matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) could successfully identify approximately 47% of the samples following a 24 h incubation period. The study emphasizes the critical role of sample preparation methods in enabling precise bacterial identification. Our findings reveal the necessity of tailoring the incubation time for each sample, as the optimum period for accurate identification fluctuated between 1 and 12 h. Further demonstrating the interplay between incubation time and spore quantity, our study used the Schaeffer-Fulton staining method to show that the lowest spore counts were detected between 5 and 8 h of incubation. This provides evidence that spore formation impacts bacterial identification. Our research thus deepens the understanding of spore-forming bacteria identification using MALDI-TOF MS and illuminates the various factors affecting the dependability and accuracy of this technique. Future research may explore additional variables, such as the effect of varying culture media, to further augment identification accuracy and gain a holistic understanding of spore-forming bacterial behavior in food samples. By enhancing our knowledge, these findings can substantially contribute to improving food safety and quality assurance strategies by enabling the more accurate and efficient identification of spore-forming bacteria in the food industry, thereby elevating the standards of food safety.

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.

Accumulation of polystyrene nanoplastics and triclosan by a model tooth-carp fish, Aphaniops hormuzensis (Teleostei: Aphaniidae).

The presence and effects of nanoplastics (NPs; <1 µm) in the aquatic environment are a growing concern. In this study, a model tooth-carp fish, Aphaniops hormuzensis, has been exposed to different concentrations of fluorescent polystyrene nanoplastics (PS-NP) in its diet (up to 5 mg kg-1) over periods of 28 d and the particle accumulation in different tissues determined. Accumulation was observed in both digestive and non-digestive organs, with concentrations greater in the gut, liver and gill (up to 400 µg kg-1 dw) than in the skin and muscle (<180 µg kg-1 dw), but no dependency on exposure time or dose was evident. The presence of the organic contaminant, triclosan (TCS), in the diet and at concentrations up to 0.5 µg kg-1 did not affect PS-NP uptake by A. hormuzensis, while TCS accumulation in the whole body increased with time (up to 10 µg kg-1) and, likewise, appeared to be unaffected by the presence of PS-NPs. These observations suggest that the two contaminants do not interact with each other or that any interactions have no impact on accumulation. The results of this study add to the growing body of evidence that NPs can be translocated by aquatic organisms after ingestion, and reveal that, for the species and conditions employed, nanoplastics are accumulated more readily than a widely used organic chemical.

Comprehensive Study of Si-Based Compounds in Selected Plants (Pisum sativum L., Medicago sativa L., Triticum aestivum L.).

This review describes the role of silicon (Si) in plants. Methods of silicon determination and speciation are also reported. The mechanisms of Si uptake by plants, silicon fractions in the soil, and the participation of flora and fauna in the Si cycle in terrestrial ecosystems have been overviewed. Plants of Fabaceae (especially Pisum sativum L. and Medicago sativa L.) and Poaceae (particularly Triticum aestivum L.) families with different Si accumulation capabilities were taken into consideration to describe the role of Si in the alleviation of the negative effects of biotic and abiotic stresses. The article focuses on sample preparation, which includes extraction methods and analytical techniques. The methods of isolation and the characterization of the Si-based biologically active compounds from plants have been overviewed. The antimicrobial properties and cytotoxic effects of known bioactive compounds obtained from pea, alfalfa, and wheat were also described.

Advanced Mass Spectrometric Techniques for the Comprehensive Study of Synthesized Silicon-Based Silyl Organic Compounds: Identifying Fragmentation Pathways and Characterization.

The primary objective of this study was to synthesize and characterize novel silicon-based silyl organic compounds in order to gain a deeper understanding of their potential applications and interactions with other compounds. Four new artificial silyl organic compounds were successfully synthesized: 1-O-(Trimethylsilyl)-2,3,4,6-tetra-O-acetyl-ß-d-glucopyranose (compound 1), 1-[(1,1-dimethylehtyl)diphenylsilyl]-1H-indole (compound 2), O-tert-butyldiphenylsilyl-(3-hydroxypropyl)oleate (compound 3), and 1-O-tert-Butyldiphenylsilyl-myo-inositol (compound 4). To thoroughly characterize these synthesized compounds, a combination of advanced mass spectrometric techniques was employed, including nanoparticle-assisted laser desorption/ionization mass spectrometry (NALDI-MS), Fourier-transform ion cyclotron resonance mass spectrometry (FT-ICR-MS), and triple quadrupole electrospray tandem mass spectrometry (QqQ ESI-MS/MS). These analytical methods enabled the accurate identification and characterization of the synthesized silyl organic compounds, providing valuable insights into their properties and potential applications. Furthermore, the electrospray ionization-Fourier transform ion cyclotron resonance-tandem mass spectrometry (ESI-FT-ICR-MS/MS) technique facilitated the proposal of fragmentation pathways for the ionized silyl organic compounds, contributing to a more comprehensive understanding of their behavior during mass spectrometric analysis. These findings suggest that mass spectrometric techniques offer a highly effective means of investigating and characterizing naturally occurring silicon-based silyl organic compounds, with potential implications for advancing research in various fields and applications in different industries.

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.

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.

Characterization of the salivary microbiome before and after antibiotic therapy via separation technique.

In the present research, the MALDI-TOF MS technique was applied as a tool to rapidly identify the salivary microbiome. In this fact, it has been monitored the changes occurred in molecular profiles under different antibiotic therapy. Significant changes in the composition of the salivary microbiota were noticed not only in relation to the non antibiotic (non-AT) and antibiotic treatment (AT) groups, but also to the used media, the antibiotic therapy and co-existed microbiota. Each antibiotic generates specific changes in molecular profiles. The highest number of bacterial species was isolated in the universal culture medium (72%) followed by the selective medium (48% and 38%). In the case of non-AT patients, the prevalence of Streptococcus salivarius (25%), Streptococcus vestibularis (19%), Streptococcus oralis (13%), and Staphylococcus aureus (6%) was identified while in the case of AT, Streptococcus salivarius (11%), Streptococcus parasanguinis (11%), Staphylococcus epidermidis (12%), Enterococcus faecalis (9%), Staphylococcus hominis (8%), and Candida albicans (6%) were identified. Notable to specified that the Candida albicans was noticed only in AT samples, indicating a negative impact on the antibiotic therapy. The accuracy of the MALDI-TOF MS technique was performed by the 16S rRNA gene sequencing analysis-as a reference method. Conclusively, such an approach highlighted in the present study can help in developing the methods enabling a faster diagnosis of disease changes at the cellular level before clinical changes occur. Once the MALDI tool allows for the distinguishing of the microbiota of non-AT and AT, it may enable to monitor the diseases treatment and develop a treatment regimen for individual patients in relation to each antibiotic. KEY POINTS: The salivary microbiota of antibiotic-treated patients was more bacteria variety MALDI-TOF MS is a promising tool for recording of reproducible molecular profiles Our data can allow to monitor the treatment of bacterial diseases for patients.

The Association between the Bisphenols Residues in Amniotic Fluid and Fetal Abnormalities in Polish Pregnant Women-Its Potential Clinical Application.

The present study aimed to investigate the relationship between the concentrations of bisphenols residues in the amniotic fluid (AF) samples collected during amniocentesis and fetal chromosomal abnormalities in pregnant women. A total of 33 pregnant Polish women aged between 24 and 44 years, and screened to detect high risk for chromosomal defects in the first trimester, were included in this study. Samples were collected from these patients during routine diagnostic and treatment procedures at mid-gestation. The concentrations of various bisphenols residues in the samples were determined by liquid chromatography coupled with triple quadrupole tandem mass spectrometry (LC-ESI-QqQ-MS/MS). Residues of eight analytes (BPS, BPF, BPA, BPAF, BADGE, BADGE•2H2O, BADGE•H2O•HCl and BADGE•2HCl) were detected in amniotic fluid samples in the range 0.69 ng/mL to 3.38 ng/mL. Fetuses with chromosomal abnormalities showed a slightly higher frequency of occurrence of selected bisphenols residues in the AF samples collected between 15-26 weeks of pregnancies. Finally, the proposed method was applied in the simultaneous determination of several endocrine-disrupting chemicals from bisphenol group in 33 human AF samples. BADGE•H2O•HCl has been identified in the AF samples taken from women older than average in the examined group. The number of detected compounds has been significant for the following analytes: BPS, BPAF, BADGE•H2O•HCl and BADGE. The proposed method may be an attractive alternative for application in large-scale human biomonitoring studies.

"Omic" Approaches to Bacteria and Antibiotic Resistance Identification.

The quick and accurate identification of microorganisms and the study of resistance to antibiotics is crucial in the economic and industrial fields along with medicine. One of the fastest-growing identification methods is the spectrometric approach consisting in the matrix-assisted laser ionization/desorption using a time-of-flight analyzer (MALDI-TOF MS), which has many advantages over conventional methods for the determination of microorganisms presented. Thanks to the use of a multiomic approach in the MALDI-TOF MS analysis, it is possible to obtain a broad spectrum of data allowing the identification of microorganisms, understanding their interactions and the analysis of antibiotic resistance mechanisms. In addition, the literature data indicate the possibility of a significant reduction in the time of the sample preparation and analysis time, which will enable a faster initiation of the treatment of patients. However, it is still necessary to improve the process of identifying and supplementing the existing databases along with creating new ones. This review summarizes the use of "-omics" approaches in the MALDI TOF MS analysis, including in bacterial identification and antibiotic resistance mechanisms analysis.

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.

Determination of 15 human pharmaceutical residues in fish and shrimp tissues by high-performance liquid chromatography-tandem mass spectrometry.

An efficient, reliable, and sensitive multiclass analytical method has been expanded to simultaneously determine 15 human pharmaceutical residues in fish and shrimp tissue samples by ultra-high-performance liquid chromatography-tandem mass spectrometry. The investigated compounds comprise ten classes, namely, analgesic, antibacterial, anticonvulsant, cardiovascular, fluoroquinolones, macrolides, nonsteroidal anti-inflammatory, penicillins, stimulant, and sulfonamide. A simple liquid extraction procedure based on 0.1% formic acid in methanol was developed. Chromatographic conditions were optimized, and mobile phase A was 0.1% ammonium acetate, and mobile phase B was acetonitrile. The mobile phase's gradient program was as follows: 0-2 min, 15% B; 2-5 min, linear to 95% B; 5-10 min, 95% B; and 10-12 min. The limits of detection were from 0.017 to 1.371 µg/kg, while a quantification range was measured from 0.051 to 4.113 µg/kg. Finally, amoxicillin, azithromycin, caffeine, carbamazepine, ciprofloxacin, clarithromycin, diclofenac, erythromycin, furosemide, ibuprofen, ketoprofen, naproxen, sulfamethoxazole, tetracycline, and triclosan were quantifiable in fish and shrimp samples.

Development of the Validated Stability-Indicating Method for the Determination of Vortioxetine in Bulk and Pharmaceutical Formulation by HPLC-DAD, Stress Degradation Kinetics Studies and Detection of Degradation Products by LC-ESI-QTOF-MS.

Vortioxetine (VOR) is a new antidepressant drug used to treat major depressive disorder. In this work, a novel, simple, rapid, accurate, precise, selective, stability-indicating, and fully validated high-performance liquid chromatography method with diode array detection (HPLC-DAD) was developed to determine VOR in bulk and pharmaceutical formulations. A Polar-RP column was used, with a mobile phase consisting of acetonitrile (ACN), methanol (MeOH), acetate buffer pH 3.5, and addition of diethylamine (DEA) in the isocratic elution mode. Assessing the stability of the VOR is fundamental to guarantee the efficacy, safety, and quality of drug products. In this study, the VOR active pharmaceutical ingredient (API) and tablets were subjected to a detailed study of forced degradation, using several degrading agents (acid, alkaline, water, heat, light, and oxidation agents). The developed HPLC-DAD method allows the collection of all the essential data to determine degradation kinetics. It was found that the decomposition of vortioxetine is fragile towards oxidative conditions and photolysis, yielding the first-order and second-order kinetic reaction in the above stress conditions, respectively. The degradation products (DPs) were identified by the high-resolution liquid chromatography coupled with electrospray ionization-quadrupole-time of flight-mass spectrometry (LC-ESI-QTOF-MS) method. The HPLC-DAD method was successfully applied for the quantification of VOR in tablets. Additionally, in silico toxicity prediction of the DPs was performed.

Identification, Structure and Characterization of Bacillus tequilensis Biofilm with the Use of Electrophoresis and Complementary Approaches.

Biofilm is a complex structure formed as a result of the accumulation of bacterial cell clusters on a surface, surrounded by extracellular polysaccharide substances (EPSs). Biofilm-related bacterial infections are a significant challenge for clinical treatment. Therefore, the main goal of our study was to design a complementary approach in biofilm characterization before and after the antibiotic treatment. The 16S rRNA gene sequencing allowed for the identification of Bacillus tequilensis, as a microbial model of the biofilm formation. Capillary electrophoresis demonstrates the capability to characterize and show the differences of the electrophoretic mobility between biofilms untreated and treated with antibiotics: amoxicillin, gentamicin and metronidazole. Electrophoretic results show the clumping phenomenon (amoxicillin and gentamicin) as a result of a significant change on the surface electric charge of the cells. The stability of the dispersion study, the molecular profile analysis, the viability of bacterial cells and the scanning morphology imaging were also investigated. The microscopic and spectrometry study pointed out the degradation/remodeling of the EPSs matrix, the inhibition of the cell wall synthesis and blocking the ribosomal protein synthesis by amoxicillin and gentamicin. However, untreated and treated bacterial cells show a high stability for the biofilm formation system. Moreover, on the basis of the type of the antibiotic treatment, the mechanism of used antibiotics in cell clumping and degradation were proposed.

Investigation of the mechanism of zearalenone metabolization in different systems: Electrochemical and theoretical approaches.

Mycotoxins are toxic metabolites produced by mold fungi, which commonly contaminate cereal crops. These compounds include zearalenone (ZEA), which may disturb the proper functioning of the endocrine system in mammals. The metabolism of ZEA plays a key role in its toxic properties. The type and amount of produced metabolites may contribute to both the reduction and increase in its pathogenic effect. Therefore, it is extremely important to investigate the possible pathways of zearalenone metabolism. The electrochemical approach may prove alternatives for the in vitro and in vivo methods. For this reason, in this study the electrochemical, theoretical and in vitro assays were applied to determine the possible ZEA metabolism mechanism. Electrochemical processes were conducted in the EC/ESI-MS system. The application of HPLC-MS/MS allowed to characterize the obtained electrochemical products specific for phase I and II. Appropriate conditions for redox identification processes were optimized with regard to such parameters as the potential value, mobile phase and working electrode. In addition, the data obtained by instrumental techniques for ZEA metabolism were compared with those obtained by in vitro methods for HepG2 cell lines. Furthermore, results from quantum mechanical calculations allowed to explain the mechanism and the conformational stability of the reduction products.

Determination of the pharmaceuticals-nano/microplastics in aquatic systems by analytical and instrumental methods.

Pharmaceutical residues and nanoplastic and microplastic particles as emerging pollutants in the aquatic environment are a subject of increasing concern in terms of the effect on water sources and marine organisms. There is lack of information about pharmaceutical-nanoplastic and pharmaceutical-microplastic mixtures. The present study aimed to investigate the fate and effect of pharmaceutical residues and nanoplastic and microplastic particles, the results of combinations of pharmaceutical residues with nanoplastic and microplastic particles, and toxic effects of pharmaceutical residues and nanoplastic and microplastic particles. Moreover, the objective was also to introduce analytical methods for pharmaceuticals, along with instrumental techniques for nanoplastic and microplastic particles in aquatic environments and organisms. PhAC alone can affect marine environments and aquatic organisms. When pharmaceutical residues combine with nanoplastic and microplastic particles, the rate of toxicity increases, and the result of this phenomenon constitutes this kind of pollutant in wastewater. Hence, the rate of mortality in organisms enhances. This study aimed to investigate the effect of pharmaceuticals residues and nanoplastic and microplastic particles, and a mixture of pharmaceutical residues and nanoplastic and microplastic particles in aquatic biota. Another object was survey methods for recognizing pharmaceutical residues and nanoplastic and microplastic particles. The findings show that pharmaceutical residues in organisms caused cell structure damage, inflammatory response, and nerve cell apoptosis. This study aimed to investigate the effect of microplastic particles in the human food chain and their impact on human health. Moreover, this review aims to present an innovative methodology based on comprehensive analytical techniques used to determine and identify pharmaceuticals adsorbed on nano- and microplastics in aquatic ecosystems. Finally, this review addresses the knowledge gaps and provides insights into future research strategies to better understand their interactions.