Novel drug therapy of acute hepatic failure induced in rats by a combination of tadalafil and Lepidium sativum.

BACKGROUND: Hepatocyte death and a systemic inflammatory response are the outcome of a complex chain of events mediated by numerous inflammatory cells and chemical mediators. The point of this study was to find out if tadalafil and/or Lepidium sativum (L. sativum) could help people who have been exposed to carbon tetrachloride (CCL4) and are experiencing acute moderate liver failure. This was especially true when the two were used together. METHOD AND MATERIALS: To cause mild liver failure 24 h before sacrifice, a single oral dosage of CCL4 (2.5 mL/kg b.w.) (50% in olive oil) was utilized. Furthermore, immunohistochemical expression of nuclear factor kappa B (NF-κB) as well as histological abnormalities were performed on liver tissue. RESULTS: The results showed that tadalafil and/or L. sativum, especially in combination, performed well to cure acute mild liver failure caused by CCL4. This was demonstrated by a decrease in NF-κB expression in the liver tissue and an improvement in organ damage markers observed in the blood and liver tissues. Furthermore, such therapy reduced interleukin1 beta (IL-1ß) and tumor necrosis factor-alpha (TNF-α) levels in the liver tissue. It's worth noting that the tested combination resulted in greater liver improvement. CONCLUSIONS: According to the findings, tadalafil and L. sativum, particularly in combination, have the ability to protect the liver from the negative effects of CCL4 exposure. Because of its capacity to improve liver function, restore redox equilibrium, and decrease inflammatory mediators, it is a prospective option for mitigating the negative effects of common environmental pollutants such as CCL4.

Garden Cress Seed Oil Abrogates Testicular Oxidative Injury and NF-kB-Mediated Inflammation in Diabetic Mice.

Diabetes mellitus is a metabolic disorder associated with various complications encompassing male reproductive dysfunction. The present study aimed to investigate the therapeutic potential of biologically active Lepidium sativum seed oil (LSO) against the testicular dysfunction associated with streptozotocin (STZ)-induced diabetes. Male adults (n = 24) were divided into four groups: control, LSO-administered, diabetic (D), and LSO-treated diabetic (D+LSO) groups. LSO was extracted from L. sativum seeds, and its chemical composition was determined using GC-MS. Serum testosterone levels, testicular enzymatic antioxidants (catalase (CAT) and superoxide dismutase (SOD)), an oxidative stress (OS) biomarker, malondialdehyde (MDA), pro-inflammatory markers (NF-kB, IL-1, IL-6, and TNF-α), and the expression level of NF-kB were assessed. In addition, histopathological changes were evaluated in testicular tissues. The results obtained showed that the chemical composition of LSO indicated its enrichment mainly with γ-tocopherol (62.1%), followed by 2-methylhexacosane (8.12%), butylated hydroxytoluene (8.04%), 10-Methylnonadecane (4.81%), and δ-tocopherol (3.91%). Moreover, LSO administration in the D+LSO mice significantly increased testosterone levels and ameliorated the observed testicular oxidative damage, inflammatory response, and reduced NF-kB expression compared to the diabetic mice. Biochemical and molecular analyses confirmed the histological results. In conclusion, LSO may prevent the progression of diabetes-induced impairment in the testes through inhibition of the OS- and NF-kB-mediated inflammatory response.

Physiochemical Properties, Lipid Breakdown, ß-Carotenoids, Tocopherols, Vitamins, Amino and Fatty Acid Profiles of Soxhlet Extracted Oil from Different Garden Cress Seed (Lepidium sativum L.) Genotypes in Ethiopia.

Physiochemical properties, lipid breakdown, ß-carotenoids, tocopherols, and vitamins as well as amino and fatty acid profiles of Soxhlet-extracted oil from five different garden cress (Lepidium sativum L.) seed genotypes (namely: CG8, CG7, CG17, CG4, and 207910) across Ethiopia regions were investigated. Results showed that despite the seeds' proximate peak and least values, the extraction yield, viscosity, specific gravity, refractive index, lipid breakdown, and boiling point of garden cress seed oil across the genotypes noticeably varied with promising amino and fatty acid profiles. Further, the genotype CG17 obtained greater quantities of ß-carotenoids, tocopherols and vitamin values compared to the other genotypes.

Bio-Assisted Synthesis and Characterization of Zinc Oxide Nanoparticles from Lepidium sativum and Their Potent Antioxidant, Antibacterial and Anticancer Activities.

Nanotechnology is an emerging area of research that deals with the production, manipulation, and application of nanoscale materials. Bio-assisted synthesis is of particular interest nowadays, to overcome the limitations associated with the physical and chemical means. The aim of this study was to synthesize ZnO nanoparticles (NPs) for the first time, utilizing the seed extract of Lepidium sativum. The synthesized NPs were confirmed through various spectroscopy and imagining techniques, such as XRD, FTIR, HPLC, and SEM. The characterized NPs were then examined for various in vitro biological assays. Crystalline, hexagonal-structured NPs with an average particle size of 25.6 nm were obtained. Biosynthesized ZnO NPs exhibited potent antioxidant activities, effective α-amylase inhibition, moderate urease inhibition (56%), high lipase-inhibition (71%) activities, moderate cytotoxic potential, and significant antibacterial activity. Gene expression of caspase in HepG2 cells was enhanced along with elevated production of ROS/RNS, while membrane integrity was disturbed upon the exposure of NPs. Overall results indicated that bio-assisted ZnO NPs exhibit excellent biological potential and could be exploited for future biomedical applications. particularly in antimicrobial and cancer therapeutics. Moreover, this is the first comprehensive study on Lepidium sativum-mediated synthesis of ZnO nanoparticles and evaluation of their biological activities.

The Phytotoxin Myrigalone A Triggers a Phased Detoxification Programme and Inhibits Lepidium sativum Seed Germination via Multiple Mechanisms including Interference with Auxin Homeostasis.

Molecular responses of plants to natural phytotoxins comprise more general and compound-specific mechanisms. How phytotoxic chalcones and other flavonoids inhibit seedling growth was widely studied, but how they interfere with seed germination is largely unknown. The dihydrochalcone and putative allelochemical myrigalone A (MyA) inhibits seed germination and seedling growth. Transcriptome (RNAseq) and hormone analyses of Lepidium sativum seed responses to MyA were compared to other bioactive and inactive compounds. MyA treatment of imbibed seeds triggered the phased induction of a detoxification programme, altered gibberellin, cis-(+)-12-oxophytodienoic acid and jasmonate metabolism, and affected the expression of hormone transporter genes. The MyA-mediated inhibition involved interference with the antioxidant system, oxidative signalling, aquaporins and water uptake, but not uncoupling of oxidative phosphorylation or p-hydroxyphenylpyruvate dioxygenase expression/activity. MyA specifically affected the expression of auxin-related signalling genes, and various transporter genes, including for auxin transport (PIN7, ABCG37, ABCG4, WAT1). Responses to auxin-specific inhibitors further supported the conclusion that MyA interferes with auxin homeostasis during seed germination. Comparative analysis of MyA and other phytotoxins revealed differences in the specific regulatory mechanisms and auxin transporter genes targeted to interfere with auxin homestasis. We conclude that MyA exerts its phytotoxic activity by multiple auxin-dependent and independent molecular mechanisms.

Sulphoraphane Affinity-Based Chromatography for the Purification of Myrosinase from Lepidium sativum Seeds.

Sulforaphane and other natural isothiocyanates released from the respective plant glucosinolates by the plant enzyme myrosinase (ß-thioglucoside glucohydrolase) show extensive anticancer and antimicrobial effects. In this study, myrosinase from garden cress (Lepidium sativum) seeds was purified to electrophoretic homogeneity by a fast and easy strategy consisting of fractionation by isoelectric precipitation with ammonium sulphate (AS) and affinity chromatography using sulforaphane (SFN) attached to cellulose resin. The overall purification of enzyme with respect to crude extract was 169-fold and recovery of 37%. Under non-reducing conditions, two protein bands exhibiting myrosinase activity with masses of about 114 and 122 kDa, respectively, and a 58 kDa protein band with no activity were detected by SDS-PAGE and zymography on polyacrylamide gel. MALDI-Tof/Tof of tryptic fragments obtained from the respective protein bands detected sequence motifs homologous to the regions responsible for glycoside-substrate binding and similarities to members of the enzyme subfamilies ß-glucosidases and myrosinases GH. The enzyme hydrolyzed both the natural (sinigrin, sinalbin, glucoraphanin) and the synthetic (p-nitrophenol-ß-D-glucopyranoside (pNPG)) substrates. The highest catalytic activity of purified enzyme was achieved against sinigrin. The KM and Vmax values of the enzyme for sinigrin were found to be 0.57 mM, and 1.3 mM/s, respectively. The enzyme was strongly activated by 30 µM ascorbic acid. The optimum temperature and pH for enzyme was 50 °C and pH 6.0, respectively. The purified enzyme could be stored at 4 °C and slightly acidic pH for at least 45 days without a significant decrease in specific activity.

Inuloxin A Inhibits Seedling Growth and Affects Redox System of Lycopersicon esculentum Mill. and Lepidium sativum L.

Allelochemicals are considered an environment-friendly and promising alternative for weed management, although much effort is still needed for understanding their mode of action and then promoting their use in plant allelopathy management practices. Here, we report that Inuloxin A (InA), an allelochemical isolated from Dittrichia viscosa, inhibited root elongation and growth of seedlings of Lycopersicon esculentum and Lepidium sativum at the highest concentrations tested. InA-induced antioxidant responses in the seedlings were investigated by analysing the contents of glutathione (GSH) and ascorbate (ASC), and their oxidized forms, dehydroascorbate (DHA), and glutathione disulphide (GSSG), as well as the redox state of thiol-containing proteins. An increase in ASC, DHA, and GSH levels at high concentrations of InA, after 3 and 6 days, were observed. Moreover, the ASC/DHA + ASC and GSH/GSSG + GSH ratios showed a shift towards the oxidized form. Our study provides the first insight into how the cell redox system responds and adapts to InA phytotoxicity, providing a framework for further molecular studies.

Enhanced Efficiency of the Removal of Cytostatic Anthracycline Drugs Using Immobilized Mycelium of Bjerkandera adusta CCBAS 930.

The aim of this study was to evaluate the bioremoval of anthracycline antibiotics (daunomycin-DNR, doxorubicin-DOX, and mitoxantrone-MTX) by immobilized mycelium of B. adusta CCBAS 930. The activity of oxidoreductases: versatile peroxidases (VP), superoxide dismutase (SOD), catalase (CAT), and glucose oxidase (GOX), and the levels of phenolic compounds (PhC) and free radicals (SOR) were determined during the biotransformation of anthracyclines by B. adusta strain CCBAS 930. Moreover, the phytotoxicity (Lepidium sativum L.), biotoxicity (MARA assay), and genotoxicity of anthracyclines were evaluated after biological treatment. After 120 h, more than 90% of anthracyclines were removed by the immobilized mycelium of B. adusta CCBAS 930. The effective biotransformation of anthracyclines was correlated with detoxification and reduced genotoxicity.

Lepidium sativum Sprouts Grown under Elevated CO2 Hyperaccumulate Glucosinolates and Antioxidants and Exhibit Enhanced Biological and Reduced Antinutritional Properties.

The nutritional and health-promoting properties of plants are largely determined by their tissue chemistry. Tuning growth conditions could affect the accumulation of phytochemicals and, therefore, enhance the biological activities. Herein, the impact of elevated CO2 (eCO2; 620 µmol CO2 mol-1 air) on growth and chemical composition of sprouts of three Lepidium sativum cultivars (Haraz, Khider and Rajab) was investigated. Changes in the sprout actions against some human chronic diseases were evaluated. eCO2 induced biomass accumulation (1.46-, 1.47- and 2-fold in Haraz, Khider and Rajab, respectively) and pigment accumulation and reduced the level of antinutrients in L. sativum cultivars. Compared to the control, eCO2 induced total glucosinolate accumulation (0.40-, 0.90- and 1.29-fold in Khider, Haraz and Rajab, respectively), possibly through increased amino acid production, and their hydrolysis by myrosinase. In line with increased polyphenol production, improved phenylalanine ammonia lyase activity was observed. The antioxidant, anti-inflammatory, hypocholesterolemic, antibacterial and anticancer activities of the produced sprouts were significantly improved by sprouting and eCO2 exposure. PCA indicated that the cultivars showed interspecific responses. Thus, the present study confirms the synergistic effect of sprouting with eCO2 exposure as a promising approach to produce more bioactive L. sativum sprouts.

Short-term physiological and biometrical responses of Lepidium sativum seedlings exposed to PET-made microplastics and acid rain.

Plastics enter in terrestrial natural system primarily by agricultural purposes, while acid rain is the result of anthropogenic activities. The synergistic effects of microplastics and acid rain on plant growth are not known. In this study, different sizes of polyethylene terephthalate (PET) and acid rain are tested on Lepidium sativum, in two separate experimental sets. In the first one we treated plants only with PET, in the second one we used PET and acid rain together. In both experimentations we analyzed: i) plant biometrical parameters (shoot height, leaf number, percentage inhibition of seed germination, fresh biomass), and ii) oxidative stress responses (hydrogen peroxide; ascorbic acid and glutathione). Results carried out from our experiments highlighted that different sizes of polyethylene terephthalate are able to affect plant growth and physiological responses, with or without acid rain supplied during acute toxicity (6 days). SHORT DESCRIPTION: This study showed that different sizes of PET microplastics affect physiological and biometrical responses of Lepidum sativum seedlings, with or without acid rain; roots and leaves responded differently.

Monitoring of hydrolysis products of mustard gas, some sesqui- and oxy-mustards and other chemical warfare agents in a plant material by HPLC-MS/MS.

At present, there is a real threat of chemical warfare agents being used in terrorist acts and military clashes. Sulfur and nitrogen mustards are blister agents with high lethality and rapid disruption of armed forces. These highly poisonous substances are hydrolyzed to the characteristic marker compounds when released into the environment. Analysis of environmental objects allows to establish the fact of alleged use of chemical warfare agents and to reveal their type. However, water and soil samples are not always reliable for retrospective analysis. The resulting chemical warfare agent markers may be washed out from the application site over time by groundwaters or atmospheric condensations. This study shows the potential for using plants as a convenient material for retrospective analysis. Garden cress (Lepidium sativum) was chosen as a model plant for this purpose, since it can be easily and quickly grown hydroponically. The plants were cultivated in the environment of the selected markers to study an accumulation of these compounds by the plants. An effective and fast method of homogenization with subsequent ultrasonic extraction was applied. The extracts were analyzed using a specially developed and validated HPLC-MS/MS approach. Separation of the hydrophilic markers was carried out on a reversed-phase column with a polar endcapping. Sensitive mass spectrometric detection was performed in the multiple reaction monitoring mode. Achieved limits of detection for most markers were in the range of 2-40 ng mL-1. It was discovered from the research that after the removal of markers from the growing medium the plants are able to store and concentrate these markers for at least 5 weeks, ensuring a high retrospectivity of the analysis. The obtained results indicate the perspective of using plants as additional objects of analysis during the investigation of incidents related to the use of chemical warfare agents. However, more complex plants and models should be studied in the future.

Effect of various abiotic stressors on some biochemical indices of Lepidium sativum plants.

In this study, the regulation of ascorbate peroxidase (APX) specific activity, anthocyanin, carotenoid, hydrogen peroxide, lipid peroxidation, and protein levels in cress leaves in response to different abiotic stresses were investigated. The total APX specific activity was significantly elevated after 9 days of drought treatment, short-term (2 h) exposure to 10, 100 and 370 µE of light, long-term exposure (at least 6 days) to 100 mM NaCl versus the specific APX activity in the controls. Furthermore, a significant change in total APX activity was detected in response to treatment with different temperatures; this change was an early response to 4 °C and 30 °C for a maximum of 4 h, while short-term exposure to 35 °C did not change total APX activity. The results of the present study revealed that plants have a wide range of mechanisms to cope with different stresses that possibly involve morphological changes. The results indicated that Lepidium sativum plants launch common protective pathways only under drought, salinity and high light stresses, while other protective mechanisms/strategies could be responsible for increasing the plants tolerance towards temperature and low light. Future studies will investigate changes in the photosynthetic quantum yield and specific target metabolites, proteins, and nonenzymatic antioxidants.

Application of Festuca arundinacea in phytoremediation of soils contaminated with Pb, Ni, Cd and petroleum hydrocarbons.

Phytoremediation is a promising "green technique" used to purify contaminated soils. The performed phytoremediation experiments assisted by the fertilization process involving pots of F.arundinacea grown on soils with diverse concentrations and types of contaminations produced the following decreased percentages after 6 months: Pb (25.4-34.1%), Ni (18.7-23.8%), Cd (26.3-46.7%), TPH (49.4-60.1%). Primarily, TPH biodegradation was occurring as a result of basic bioremediation stimulated by adding optimal volumes of biogenic substances and corrections in the soil reaction, while phytoremediation improved this process by 17.4 - 23.1%. The highest drop in a range of 45.6 - 55.5% was recorded for the group of C12-C18 hydrocarbons, with the lowest one for C25-C36, amounting to 9.1-17.4%. Translocation factor values were: TF<1 and ranged, respectively, for: Pb (0.46-0.53), Ni (0.29-0.33), and Cd (0.21-0.25), which indicate that heavy metals absorbed by Festuca arundinacea they mainly accumulated in the root of the tissue in descending order: Cd

Fungal biodegradation and multi-level toxicity assessment of vinasse from distillation of winemaking by-products.

The wastewaters from distilleries of winemaking by-products, a scarcely studied type of vinasse, were treated by white-rot fungal strains from species Irpex lacteus, Ganoderma resinaceum, Trametes versicolor, Phlebia rufa and Bjerkandera adusta. The main objectives of this study were to evaluate fungal performance during vinasse biodegradation, their enzyme patterns and ecotoxicity evolution throughout treatment. Despite all strains were able to promote strong (>80%) dephenolization and reduction of total organic carbon (TOC), P. rufa was less affected by vinasse toxicity and exhibit better decolorization. In batch cultures at 28 °C and pH 4.0, the first phase of P. rufa biodegradation kinetics was characterized by strong metabolic activity with simultaneous depletion of TOC, phenolics and sugars. The main events of second phase are the increase of peroxidases production after the peak of laccase activity, and strong color removal. At the end of treatment, it was observed highly significant (p < 0.001) abatement of pollution parameters (83-100% removal). Since water reclamation and reuse for e.g. crop irrigation is a priority issue, vinasse ecotoxicity was assessed with bioindicators representing three different phylogenetic and trophic levels: a marine bacterium (Aliivibrio fischeri), a freshwater microcrustacean (Daphnia magna) and a dicotyledonous macrophyte (Lepidium sativum). It was observed significant (p < 0.05) reduction of initial vinasse toxicity, as evaluated by these bioindicators, deserving special mention an almost complete phytotoxicity elimination.

Biodegradation Potential and Ecotoxicity Assessment in Soil Extracts Amended with Phenoxy Acid Herbicide (2,4-D) and a Structurally-Similar Plant Secondary Metabolite (Ferulic Acid).

Phenoxy acid 2,4-D (2,4-dichlorophenoxy acid) is one of the most commonly-used herbicide in agriculture. Biodegradation of 2,4-D can be stimulated by structurally-related plant secondary metabolites such as ferulic acid (FA). The aim of this study is to: (1) assess the potential of indigenous soil bacteria to degrade 2,4-D in the presence of FA by PCR analysis of functional tfdA genes, (2) to determine the influence of 2,4-D and FA on samples ecotoxicity using Phytotoxkit® and Microtox® biotests. The detection of tfdA genes varied depending on the enrichment of samples with FA. FA suppressed detection of the tfdA genes, 100 µM 2,4-D induced higher detection of studied amplicons, while 500 µM 2,4-D delayed their detection. The ecotoxicity response was specific and differed between plants (PE% Lepidium sativum > Sinapis alba > Sorghum saccharatum) and bacteria (PE% up to 99% for Vibrio fischeri). Our findings confirm that 2,4-D and FA had a toxic influence on the used organisms.

Exploring the chelation-based plant strategy for iron oxide nanoparticle uptake in garden cress (Lepidium sativum) using magnetic particle spectrometry.

Although iron is one of Earth's most abundant elements, its availability to plants remains an agricultural challenge, particularly in high pH environments. At high pH, iron forms insoluble ferric oxide-hydroxides that makes it inaccessible to plants. It is estimated that 30% of the world's cropland is too alkaline for optimal plant growth. Staple crops, like rice, are particularly susceptible to iron deficiency, thereby, necessitating the need for continued research in developing iron-based fertilizers. Recent studies have demonstrated the potential of using iron oxide nanoparticles (IONPs) as fertilizers to address iron deficiency in plants, but some studies have generated conflicting results. One of the major challenges associated in investigating IONP plant uptake and translocation is the inability to distinguish between intact IONPs versus leached iron ions. In this study, we utilized a new approach based on magnetic particle spectrometry (MPS) to monitor the uptake and distribution of different sized (10 and 20 nm) chelated IONPs in plants. We exposed garden cress (Lepidium sativum) plants to EDTA-capped IONPs and observed an 8-fold enhancement in total biomass and 1.4 times increase in chlorophyll production compared to plants treated with a commercial chelated iron fertilizer (Fe-EDTA). Moreover, we demonstrated that the uptake and tissue distribution of IONPs can be quantitatively monitored using MPS, and the results of the analysis were validated by atomic absorption spectroscopy, which is the conventional method used to study IONP plant uptake. Our study demonstrates that MPS is a reliable, sensitive, and effective analytical tool for the development of IONP-based fertilizers.

Upgrading the phenolic content, antioxidant and antimicrobial activities of garden cress seeds using solid-state fermentation by Trichoderma reesei.

AIMS: This study aims to evaluate the impact of solid-state fermentation (SSF) by Trichoderma reesei on the phenolic content, antioxidant and antimicrobial activities of garden cress seeds (GCS). METHODS AND RESULTS: The factorial statistical design was employed to optimize the SSF conditions, incubation time, pH, temperature and moisture, for maximum production of the phenolic content and microbial carbohydrate-cleaving enzymes from GCS. The total phenolic content significantly increased from unfermented GCS (401 mg gallic acid equivalent (GAE) 100 g-1 ) to fermented GCS (3600 mg GAE 100 g-1 ) by ninefold. The total antioxidant activity significantly increased in fermented GCS. Fifteen phenolic compounds were detected in fermented GCS with high concentrations compared to 14 in unfermented GCS using high-performance liquid chromatography. A strong correlation between the production of the carbohydrate-cleaving enzymes and the phenolic content of fermented GCS was observed. The phenolic compounds of fermented GCS showed higher antimicrobial activity. CONCLUSIONS: The fermented GCS is a powerful source of phenolic compounds with high antioxidant potentials, which can be used as dietary supplement and antimicrobial agent. SIGNIFICANCE AND IMPACT OF THE STUDY: Solid-state fermentation is a promising technique used for production of added-value bioactive compounds. SSF increased the total phenolic content and antioxidant activity of GCS several folds compared to germination process, which recently studied.

Monochromatic lights-induced trends in antioxidant and antidiabetic polyphenol accumulation in in vitro callus cultures of Lepidium sativum L.

Lepidium sativum L. is an important edible, herbaceous plant with huge medicinal value as cardio-protective, hepatoprotective and antitumor agent. This study was designed and performed to investigate biosynthesis of plant's active ingredients in callus cultures of L. sativum in response to the exposure of multi spectral lights. Optimum biomass accumulation (15.36 g/L DW), total phenolic and flavonoid contents (TPC; 47.43 mg/g; TFC; 9.41 mg/g) were recorded in calli placed under white light (24 h) compared to rest of the treatments. Antioxidant enzymatic activities i.e. superoxide dismutase and peroxidase were found optimum in cultures exposed to green light (SOD; 0.054 nM/min/mg FW, POD; 0.501 nM/min/mg FW). Phytochemical analysis further confirmed the potential influence of white light exposure on enhanced production of plant's metabolites. Significant enhancement level of major metabolic compounds such as chlorogenic acid (7.20 mg/g DW), quercetin (22.08 mg/g DW), kaempferol (7.77 mg/g DW) and minor compounds including ferulic acid, sinapic acid, protocatechuic acid, vanillic acid and caffeic acid were recorded in white light compared to control (photoperiod), whereas blue light increased the p-coumaric acid accumulation. Moreover, callus cultures of this plant under white light (24 h) showed highest in vitro based anti-diabetic and antioxidant activities compared to other conditions. Finding of our current study revealed that multi spectral lights are proved to be an effective strategy for enhancing metabolic quantity of antioxidant and anti-diabetic bioactive compounds in callus cultures of L. sativum L.

Effect of solvent and extraction technique on composition and biological activity of Lepidium sativum extracts.

The aim of this study was to evaluate chemical and biological potential of garden cress (Lepidium sativum L.) to receive valuable plant extracts with potential application in pharmacy or food industry. Four techniques of extraction and three environmentally friendly solvents such as water, supercritical CO2 and ethanol have been tested. Biological activity and chemical profile were evaluated in obtained extracts. GC/MS analysis showed that SFE extract from dried sprouts of L. sativum was especially rich in such glucosinolate derivatives as benzyl cyanide and benzyl thiocyanate. However, the extract obtained from freeze-dried sprouts by SFE with addition of 96% ethanol as co-solvent was especially rich in flavonoids and simultaneously exhibited the best antimicrobial activity. Comparison of MALDI-TOF-MS spectra of all obtained extracts clearly indicates that both SFE and maceration with water are the most selective techniques of extraction due to the lowest level of interfering substances with high molecular masses.

Nerve agent markers screening after accumulation in garden cress (Lepidium sativum) used as a model plant object.

In this research an accumulation of nerve agent markers in garden cress (Lepidium sativum) as a model plant object was studied using LC-QTOF hybrid system. For the determination of methylphosphonic acid and alkyl methylphosphonates, which are specific markers of sarin, soman, VR and VX, simple and sensitive approach was developed. Direct analysis of aqueous extracts on the reversed phase column with polar endcapping allowed to achieve satisfactory retention factor for methylphosphonic acid, which has high polarity and is usually very weakly retained on the ordinary reversed phase columns. Application of the QTOF mass spectrometer with high mass resolution led to the increase in the accuracy of the conducted measurements. The HPLC-HRMS technique developed exclusively for this study has been validated for linearity, limit of detection, limit of quantification, precision, accuracy and matrix effect prior to the analysis of plant extract samples. Hydroponic growth model was employed to examine accumulation of nerve agent markers in garden cress. It was found that after elimination of nerve agent markers from the plant growth medium, garden cress was able to store these substances for at least 5 weeks providing high retrospectivity of the analysis. Moreover, during the cress growth, no metabolization of alkyl methylphosphonates was observed. This allows not only to reveal the fact of nerve agents release into environment, but also to define its type after a long period of time.