Search results for: gas chromatography-mass spectrometry (GC-MS)

Authors: Atchara Sankom, Warapa Mahakarnchanakul, Ronnarit Rittiron, Tanaboon Sajjaanantakul, Thammasak Thongket

Abstract:

Vegetables are frequently contaminated with pesticides residues resulting in the most food safety concern among agricultural products. The objective of this work was to develop a method to detect the organophosphate (OP) pesticides residues in vegetables using Near Infrared (NIR) spectroscopy technique. Low concentration (ppm) of OP pesticides in vegetables were investigated. The experiment was divided into 2 sections. In the first section, Chinese kale spiked with different concentrations of chlorpyrifos pesticide residues (0.5-100 ppm) was chosen as the sample model to demonstrate the appropriate conditions of sample preparation, both for a solution or solid sample. The spiked samples were extracted with acetone. The sample extracts were applied as solution samples, while the solid samples were prepared by the dry-extract system for infrared (DESIR) technique. The DESIR technique was performed by embedding the solution sample on filter paper (GF/A) and then drying. The NIR spectra were measured with the transflectance mode over wavenumber regions of 12,500-4000 cm⁻¹. The QuEChERS method followed by gas chromatography-mass spectrometry (GC-MS) was performed as the standard method. The results from the first section showed that the DESIR technique with NIR spectroscopy demonstrated good accurate calibration result with R² of 0.93 and RMSEP of 8.23 ppm. However, in the case of solution samples, the prediction regarding the NIR-PLSR (partial least squares regression) equation showed poor performance (R² = 0.16 and RMSEP = 23.70 ppm). In the second section, the DESIR technique coupled with NIR spectroscopy was applied to the detection of OP pesticides in vegetables. Vegetables (Chinese kale, cabbage and hot chili) were spiked with OP pesticides (chlorpyrifos ethion and profenofos) at different concentrations ranging from 0.5 to 100 ppm. Solid samples were prepared (based on the DESIR technique), then samples were scanned by NIR spectrophotometer at ambient temperature (25+2°C). The NIR spectra were measured as in the first section. The NIR- PLSR showed the best calibration equation for detecting low concentrations of chlorpyrifos residues in vegetables (Chinese kale, cabbage and hot chili) according to the prediction set of R2 and RMSEP of 0.85-0.93 and 8.23-11.20 ppm, respectively. For ethion residues, the best calibration equation of NIR-PLSR showed good indexes of R² and RMSEP of 0.88-0.94 and 7.68-11.20 ppm, respectively. As well as the results for profenofos pesticide, the NIR-PLSR also showed the best calibration equation for detecting the profenofos residues in vegetables according to the good index of R² and RMSEP of 0.88-0.97 and 5.25-11.00 ppm, respectively. Moreover, the calibration equation developed in this work could rapidly predict the concentrations of OP pesticides residues (0.5-100 ppm) in vegetables, and there was no significant difference between NIR-predicted values and actual values (data from GC-MS) at a confidence interval of 95%. In this work, the proposed method using NIR spectroscopy involving the DESIR technique has proved to be an efficient method for the screening detection of OP pesticides residues at low concentrations, and thus increases the food safety potential of vegetables for domestic and export markets.

Keywords: NIR spectroscopy, organophosphate pesticide, vegetable, food safety

Procedia PDF Downloads 150

Authors: Precious Ehiomogue, Ifechukwude Israel Ahuchaogu, Isiguzo Edwin Ahaneku

Abstract:

Vermicompost is the product of the decomposition process using various species of worms, to create a mixture of decomposing vegetable or food waste, bedding materials, and vemicast. This process is called vermicomposting, while the rearing of worms for this purpose is called vermiculture. Several works have verified the adsorption of toxic metals using vermicompost but the application is still scarce for the retention of organic compounds. This research brings to knowledge the effectiveness of earthworm waste (vermicompost) for the remediation of crude oil contaminated soils. The remediation methods adopted in this study were two soil washing methods namely, batch and column process which represent laboratory and in-situ remediation. Characterization of the vermicompost and crude oil contaminated soil were performed before and after the soil washing using Fourier transform infrared (FTIR), scanning electron microscopy (SEM), X-ray fluorescence (XRF), X-ray diffraction (XRD) and Atomic adsorption spectrometry (AAS). The optimization of washing parameters, using response surface methodology (RSM) based on Box-Behnken Design was performed on the response from the laboratory experimental results. This study also investigated the application of machine learning models [Artificial neural network (ANN), Adaptive neuro fuzzy inference system (ANFIS). ANN and ANFIS were evaluated using the coefficient of determination (R²) and mean square error (MSE)]. Removal efficiency obtained from the Box-Behnken design experiment ranged from 29% to 98.9% for batch process remediation. Optimization of the experimental factors carried out using numerical optimization techniques by applying desirability function method of the response surface methodology (RSM) produce the highest removal efficiency of 98.9% at absorbent dosage of 34.53 grams, adsorbate concentration of 69.11 (g/ml), contact time of 25.96 (min), and pH value of 7.71, respectively. Removal efficiency obtained from the multilevel general factorial design experiment ranged from 56% to 92% for column process remediation. The coefficient of determination (R²) for ANN was (0.9974) and (0.9852) for batch and column process, respectively, showing the agreement between experimental and predicted results. For batch and column precess, respectively, the coefficient of determination (R²) for RSM was (0.9712) and (0.9614), which also demonstrates agreement between experimental and projected findings. For the batch and column processes, the ANFIS coefficient of determination was (0.7115) and (0.9978), respectively. It can be concluded that machine learning models can predict the removal of crude oil from polluted soil using vermicompost. Therefore, it is recommended to use machines learning models to predict the removal of crude oil from contaminated soil using vermicompost.

Keywords: ANFIS, ANN, crude-oil, contaminated soil, remediation and vermicompost

Procedia PDF Downloads 111

Authors: Luana Guerra, Silvio C. Sampaio, Vladimir Pavan Margarido, Ralpho R. Reis

Abstract:

Brazil is the world's largest consumer of pesticides. The excessive use of these compounds has negative impacts on animal and human life, the environment, and food security. Bees, crucial for pollination, are exposed to pesticides during the collection of nectar and pollen, posing risks to their health and the food chain, including honey contamination. Aquatic sediments are also affected, impacting water quality and the microbiota. Therefore, the analysis of aquatic sediments and bee honey is essential to identify environmental contamination and monitor ecosystems. The aim of this study was to use samples of honey from honeybees (Apis mellifera) and aquatic sediment as bioindicators of environmental contamination by pesticides and their relationship with agricultural use in the surrounding areas. The sample collections of sediment and honey were carried out in two stages. The first stage was conducted in the Bituruna municipality region in the second half of the year 2022, and the second stage took place in the regions of Laranjeiras do Sul, Quedas do Iguaçu, and Nova Laranjeiras in the first half of the year 2023. In total, 10 collection points were selected, with 5 points in the first stage and 5 points in the second stage, where one sediment sample and one honey sample were collected for each point, totaling 20 samples. The honey and sediment samples were analyzed at the Laboratory of the Paraná Institute of Technology, with ten samples of honey and ten samples of sediment. The selected extraction method was QuEChERS, and the analysis of the components present in the sample was performed using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). The pesticides Azoxystrobin, Epoxiconazole, Boscalid, Carbendazim, Haloxifope, Fomesafen, Fipronil, Chlorantraniliprole, Imidacloprid, and Bifenthrin were detected in the sediment samples from the study area in Laranjeiras do Sul, Paraná, with Carbendazim being the compound with the highest concentration (0.47 mg/kg). The honey samples obtained from the apiaries showed satisfactory results, as they did not show any detection or quantification of the analyzed pesticides, except for Point 9, which had the fungicide tebuconazole but with a concentration Keywords: contamination, water research, agrochemicals, beekeeping activity

Procedia PDF Downloads 37

Authors: Haley Gershon

Abstract:

The transition from using fossil fuel-based building material to formulating eco-friendly and biobased building materials plays a key role in sustainable building. The growing demand on a global level for biobased materials in the building and construction industries heightens the importance of carbon-14 testing, an analytical method used to determine the percentage of biobased content that comprises a material’s ingredients. This presentation will focus on the use of carbon-14 analysis within the building materials sector. Carbon-14, also known as radiocarbon, is a weakly radioactive isotope present in all living organisms. Any fossil material older than 50,000 years will not contain any carbon-14 content. The radiocarbon method is thus used to determine the amount of carbon-14 content present in a given sample. Carbon-14 testing is performed according to ASTM D6866, a standard test method developed specifically for biobased content determination of material in solid, liquid, or gaseous form, which requires radiocarbon dating. Samples are combusted and converted into a solid graphite form and then pressed onto a metal disc and mounted onto a wheel of an accelerator mass spectrometer (AMS) machine for the analysis. The AMS instrument is used in order to count the amount of carbon-14 present. By submitting samples for carbon-14 analysis, manufacturers of building materials can confirm the biobased content of ingredients used. Biobased testing through carbon-14 analysis reports results as percent biobased content, indicating the percentage of ingredients coming from biomass sourced carbon versus fossil carbon. The analysis is performed according to standardized methods such as ASTM D6866, ISO 16620, and EN 16640. Products 100% sourced from plants, animals, or microbiological material are therefore 100% biobased, while products sourced only from fossil fuel material are 0% biobased. Any result in between 0% and 100% biobased indicates that there is a mixture of both biomass-derived and fossil fuel-derived sources. Furthermore, biobased testing for building materials allows manufacturers to submit eligible material for certification and eco-label programs such as the United States Department of Agriculture (USDA) BioPreferred Program. This program includes a voluntary labeling initiative for biobased products, in which companies may apply to receive and display the USDA Certified Biobased Product label, stating third-party verification and displaying a product’s percentage of biobased content. The USDA program includes a specific category for Building Materials. In order to qualify for the biobased certification under this product category, examples of product criteria that must be met include minimum 62% biobased content for wall coverings, minimum 25% biobased content for lumber, and a minimum 91% biobased content for floor coverings (non-carpet). As a result, consumers can easily identify plant-based products in the marketplace.

Keywords: carbon-14 testing, biobased, biobased content, radiocarbon dating, accelerator mass spectrometry, AMS, materials

Procedia PDF Downloads 158

Authors: Yasinta Ratna Esti Wulandari, Vivitri Dewi Prasasty, Adrianus Rio, Cindy Geniola

Abstract:

Mulberry is a plant that widely cultivated around the world, mostly for silk industry. In recent years, the study showed that the mulberry leaves have an anti-diabetic effect which mostly comes from the compound known as 1-deoxynojirimycin (DNJ). DNJ is a very potent α-glucosidase inhibitor. It will decrease the degradation rate of carbohydrates in digestive tract, leading to slower glucose absorption and reducing the post-prandial glucose level significantly. The mulberry leaves also known as the best source of DNJ. Since then, the DNJ in mulberry leaves had received a considerable attention, because of the increased number of diabetic patients and the raise of people awareness to find a more natural cure for diabetic. The DNJ content in mulberry leaves varied depend on the mulberry species, leaf’s age, and the plant’s growth environment. Few of the mulberry varieties that were cultivated in Indonesiaare Morus alba var. kanva-2, M. alba var. multicaulis, M. bombycis var. lembang, and M. cathayana. The lack of data concerning phytochemicals contained in the Indonesian mulberry leaves are restraining their use in the medicinal field. The aim of this study is to fully utilize the use of mulberry leaves cultivated in Indonesia as a medicinal herb in local, national, or global community, by determining the DNJ and other phytochemical contents in them. This study used eight leaf samples which are the young leaves and mature leaves of both Morus alba var. kanva-2, M. alba var. multicaulis, M. bombycis var. lembang, and M. cathayana. The DNJ content was analyzed using reverse phase high performance liquid chromatography (HPLC). The stationary phase was silica C18 column and the mobile phase was acetonitrile:acetic acid 0.1% 1:1 with elution rate 1 mL/min. Prior to HPLC analysis the samples were derivatized with FMOC to ensure the DNJ detectable by VWD detector at 254 nm. Results showed that the DNJ content in samples are ranging from 2.90-0.07 mg DNJ/ g leaves, with the highest content found in M. cathayana mature leaves (2.90 ± 0.57 mg DNJ/g leaves). All of the mature leaf samples also found to contain higher amount of DNJ from their respective young leaf samples. The phytochemicals in leaf samples was tested using qualitative test. Result showed that all of the eight leaf samples contain alkaloids, phenolics, flavonoids, tannins, and terpenes. The presence of this phytochemicals contribute to the therapeutic effect of mulberry leaves. The pyrolysis-gas chromatography-mass spectrometry (Py-GC-MS) analysis was also performed to the eight samples to quantitatively determine their phytochemicals content. The pyrolysis temperature was set at 400 °C, with capillary column Phase Rtx-5MS 60 × 0.25 mm ID stationary phase and helium gas mobile phase. Few of the terpenes found are known to have anticancer and antimicrobial properties. From all the results, all of four samples of mulberry leaves which are cultivated in Indonesia contain DNJ and various phytochemicals like alkaloids, phenolics, flavonoids, tannins, and terpenes which are beneficial to our health.

Keywords: Morus, 1-deoxynojirimycin, HPLC, Py-GC-MS

Procedia PDF Downloads 330

Authors: Feiyu Yang, Chunfang Ni, Rong Wang, Yun Zou, Wenbin Liu, Chenggong Zhang, Fenjin Sun, Chun Wang

Abstract:

Cocaine is the most frequently used illegal drug globally, with the global annual prevalence of cocaine used ranging from 0.3% to 0.4 % of the adult population aged 15–64 years. Growing consumption trend of abused cocaine and drug crimes are a great concern, therefore urine sample testing has become an important noninvasive sampling whereas cocaine and its metabolites (COCs) are usually present in high concentrations and relatively long detection windows. However, direct analysis of urine samples is not feasible because urine complex medium often causes low sensitivity and selectivity of the determination. On the other hand, presence of low doses of analytes in urine makes an extraction and pretreatment step important before determination. Especially, in gathered taking drug cases, the pretreatment step becomes more tedious and time-consuming. So developing a sensitive, rapid and high-throughput method for detection of COCs in human body is indispensable for law enforcement officers, treatment specialists and health officials. In this work, a new automated magnetic dispersive solid-phase extraction (MDSPE) sampling method followed by high performance liquid chromatography-mass spectrometry (HPLC-MS) was developed for quantitative enrichment of COCs from human urine, using prepared magnetic nanoparticles as absorbants. The nanoparticles were prepared by silanizing magnetic Fe3O4 nanoparticles and modifying them with divinyl benzene and vinyl pyrrolidone, which possesses the ability for specific adsorption of COCs. And this kind of magnetic particle facilitated the pretreatment steps by electromagnetically controlled extraction to achieve full automation. The proposed device significantly improved the sampling preparation efficiency with 32 samples in one batch within 40mins. Optimization of the preparation procedure for the magnetic nanoparticles was explored and the performances of magnetic nanoparticles were characterized by scanning electron microscopy, vibrating sample magnetometer and infrared spectra measurements. Several analytical experimental parameters were studied, including amount of particles, adsorption time, elution solvent, extraction and desorption kinetics, and the verification of the proposed method was accomplished. The limits of detection for the cocaine and cocaine metabolites were 0.09-1.1 ng·mL-1 with recoveries ranging from 75.1 to 105.7%. Compared to traditional sampling method, this method is time-saving and environmentally friendly. It was confirmed that the proposed automated method was a kind of highly effective way for the trace cocaine and cocaine metabolites analyses in human urine.

Keywords: automatic magnetic dispersive solid-phase extraction, cocaine detection, magnetic nanoparticles, urine sample testing

Procedia PDF Downloads 204

Authors: Angel Villabona-Ortíz, Candelaria Tejada-Tovar, Andrea Viera-Devoz

Abstract:

Wastewater treatment is an issue of vital importance in these times where the impacts of human activities are most evident, which have become essential tasks for the normal functioning of society. However, they put entire ecosystems at risk by time destroying the possibility of sustainable development. Various conventional technologies are used to remove pollutants from water. Agroindustrial waste is the product with the potential to be used as a renewable raw material for the production of energy and chemical products, and their use is beneficial since products with added value are generated from materials that were not used before. Considering the benefits that the use of residual biomass brings, this project proposes the use of agro-industrial residues from corn crops for the production of natural adsorbents whose purpose is aimed at the remediation of contaminated water bodies with large loads of nutrients. The adsorption capacity of two biomaterials obtained from the processing of corn stalks was evaluated by batch system tests. Biochar impregnated with sulfuric acid and thermally activated was synthesized. On the other hand, the cellulose was extracted from the corn stalks and chemically modified with cetyltrimethylammonium chloride in order to quaternize the surface of the adsorbent. The adsorbents obtained were characterized by thermogravimetric analysis (TGA), scanning electron microscopy (SEM), infrared spectrometry with Fourier Transform (FTIR), analysis by Brunauer, Emmett and Teller method (BET) and X-ray Diffraction analysis ( XRD), which showed favorable characteristics for the cellulose extraction process. Higher adsorption capacities of the nutrients were obtained with the use of biochar, with phosphate being the anion with the best removal percentages. The effect of the initial adsorbate concentration was evaluated, with which it was shown that the Freundlich isotherm better describes the adsorption process in most systems. The adsorbent-phosphate / nitrate systems fit better to the Pseudo Primer Order kinetic model, while the adsorbent-sulfate systems showed a better fit to the Pseudo second-order model, which indicates that there are both physical and chemical interactions in the process. Multicomponent adsorption tests revealed that phosphate anions have a higher affinity for both adsorbents. On the other hand, the thermodynamic parameters standard enthalpy (ΔH °) and standard entropy (ΔS °) with negative results indicate the exothermic nature of the process, whereas the ascending values of standard Gibbs free energy (ΔG °). The adsorption process of anions with biocarbon and modified cellulose is spontaneous and exothermic. The use of the evaluated biomateriles is recommended for the treatment of industrial effluents contaminated with sulfate, nitrate and phosphate anions.

Keywords: adsorption, biochar, modified cellulose, corn stalks

Procedia PDF Downloads 182

Authors: Aguasanta Miguel Sarmiento, José Miguel Dávila, María Luisa de la Torre

Abstract:

Acid mine drainage (AMD) is an acidic leachate with high levels of metals and sulphates in solution, which seriously affects the durability and strength of metallic materials used in the construction of structural and mechanical components. This paper presents the results of the evolution over time of the reduction in tensile strength and defects in AISI 304 stainless steel in contact with acid mine drainage. For this purpose, a total of 30 bars with a diameter of 8 mm and a length of 14 cm were placed transversely in the course of a stream contaminated by AMD from the sulphide mines of the Iberian Pyritic Belt (SW Spain). This stream has average pH values of 2.6, a potential of 660 mV, and average concentrations of 12 g/L of sulphates, 1.2 g/L of Fe, 191 mg/L of Zn, etc. Every two months of exposure, 6 stainless steel bars were extracted from the acid stream. They were subjected to surface roughness analysis carried out with the help of Mitutoyo Surftest SJ-210 surface roughness tester. The analysis was carried out at three different points on 5 specimens from each series. The average reading of each parameter is calculated in order to ensure the accuracy of the measurements and the surface coverage. Arithmetic mean roughness value (Ra), mean roughness depth (Rz), and root mean square roughness (Rq) were measured. Five specimens from each series were statically tensile tested using universal equipment (Servosis ME 403 of 200kN). The specimens were clamped at their ends with two grips for cylindrical sections, and the tensile force was applied at a constant speed of 0.5 kN/s, according to the requirements of standard UNE-EN ISO 6892-1: 2020. To determine the modulus of elasticity, limits close to 15% and 55% of the maximum load were used, depending on the course of each test. Field Emission Scanning Electron Microscopy (FESEM) was used to observe corrosion products and defects generated by exposure to AMD. Energy dispersive X-ray spectrometry (EDS) was used to analyse the chemical composition of the corrosion products formed. For this purpose, small pieces were cut from the resulting specimens, cleaned, and embedded in epoxy resin. The results show that after only 5 months of exposure of AISI 304 stainless steel to the mining environment, the surface roughness increases significantly, with average depths almost 6 times greater than the initial one. Cracks are observed on the surface of the material, which increases in size with the time of exposure. A large number of grains with a composition of more than 57% Pb and 16% Sn can be observed inside these cracks. Tensile tests show a reduction in the resistance of this material after only two months of exposure. The results show the serious problems that would result from the use of this material for the use of mechanical components in a sulphide mining environment, not only because of the significant reduction in the lifetime of such components, but also because of the implications for human safety.

Keywords: acid mine drainage, corrosion, mechanical properties, stainless steel

Procedia PDF Downloads 16

Authors: Malory Jonata

Abstract:

Chlordecone (CLD) is an organochlorine pesticide used between 1971 and 1993 in both Guadeloupe and Martinique for the control of banana black weevil. The bishomocubane structure which characterizes this chemical compound led to high stability in organic matter and high persistence in the environment. Recently, researchers found that CLD can be degraded by isolated bacteria consortiums and, particularly, by bacteria such as Citrobacter sp 86 and Delsulfovibrio sp 86. Actually, six transformation product families of CLD are known. Moreover, the latest discovery showed that CLD was disappearing faster than first predicted in highly contaminated soil in Guadeloupe. However, the toxicity of transformation products is still unknown, and knowledge has to be deepened on the degradation ways and chemical characteristics of chlordecone and its transformation products. Microbial fuel cells (MFC) are electrochemical systems that can convert organic matter into electricity thanks to electroactive bacteria. These bacteria can exchange electrons through their membranes to solid surfaces or molecules. MFC have proven their efficiency as bioremediation systems in water and soils. They are already used for the bioremediation of several organochlorine compounds such as perchlorate, trichlorophenol or hexachlorobenzene. In this study, a three-electrodes system, inspired by MFC, is used to try to degrade chlordecone using bacteria from a mangrove swamp in Martinique. As we know, some mangrove bacteria are electroactive. Furthermore, the CLD rate seems to decline in mangrove swamp sediments. This study aims to prove that electroactive bacteria from a mangrove swamp in Martinique can degrade CLD thanks to a three-electrodes bioelectrochemical system. To achieve this goal, the tree-electrodes assembly has been connected to a potentiostat. The substrate used is mangrove water and sediments sampled in the mangrove swamp of La Trinité, a coastal city in Martinique, where CLD contamination has already been studied. Electroactive biofilms are formed by imposing a potential relative to Saturated Calomel Electrode using chronoamperometry. Moreover, their comportment has been studied by using cyclic voltametry. Biofilms have been studied under different imposed potentials, several conditions of the substrate and with or without CLD. In order to quantify the evolution of CLD rates in the substrate’s system, gas chromatography coupled with mass spectrometry (GC-MS) was performed on pre-treated samples of water and sediments after short, medium and long-term contact with the electroactive biofilms. Results showed that between -0,8V and -0,2V, the three-electrodes system was able to reduce the chemical in the substrate solution. The first GC-MS analysis result of samples spiked with CLD seems to reveal decreased CLD concentration over time. In conclusion, the designed bioelectrochemical system can provide the necessary conditions for chlordecone degradation. However, it is necessary to improve three-electrodes control settings in order to increase degradation rates. The biological pathways are yet to enlighten by biologicals analysis of electroactive biofilms formed in this system. Moreover, the electrochemical study of mangrove substrate gives new informations on the potential use of this substrate for bioremediation. But further studies are needed to a better understanding of the electrochemical potential of this environment.

Keywords: bioelectrochemistry, bioremediation, chlordecone, mangrove swamp

Procedia PDF Downloads 83

Authors: Prabhat Kashyap, Krishan Kumar

Abstract:

Delhi, the capital of India, is one of the most populated and polluted city among the world. In terms of air quality, Delhi’s air is degrading day by day & becomes worst of any major city in the world. The role of biogenic volatile organic compounds (BVOCs) is not much studied in cities like Delhi as a culprit for degraded air quality. They not only play a critical role in rural areas but also determine the atmospheric chemistry of urban areas as well. Particularly, Isoprene (2-methyl 1,3-butadiene, C5H8) is the single largest emitted compound among other BVOCs globally, that influence the tropospheric ozone chemistry in urban environment as the ozone forming potential of isoprene is very high. It is mainly emitted by vegetation & a small but significant portion is also released by vehicular exhaust of petrol operated vehicles. This study investigates the spatial and temporal variations of quantitative measurements of isoprene emissions along with different traffic tracers in 2 different seasons (post-monsoon & winter) at four different locations of Delhi. For the quantification of anthropogenic and biogenic isoprene, two sites from traffic intersections (Punjabi Bagh & CRRI) and two sites from vegetative locations (JNU & Yamuna Biodiversity Park) were selected in the vicinity of isoprene emitting tree species like Ficus religiosa, Dalbergia sissoo, Eucalyptus species etc. The concentrations of traffic tracers like benzene, toluene were also determined & their robust ratios with isoprene were used to differentiate anthropogenic isoprene with biogenic portion at each site. The ozone forming potential (OFP) of all selected species along with isoprene was also estimated. For collection of intra-day samples (3 times a day) in each season, a pre-conditioned fenceline monitoring (FLM) carbopack X thermal desorption tubes were used and further analysis was done with Gas chromatography attached with mass spectrometry (GC-MS). The results of the study proposed that the ambient air isoprene is always higher in post-monsoon season as compared to winter season at all the sites because of high temperature & intense sunlight. The maximum isoprene emission flux was always observed during afternoon hours in both seasons at all sites. The maximum isoprene concentration was found to be 13.95 ppbv at Biodiversity Park during afternoon time in post monsoon season while the lower concentration was observed as low as 0.07 ppbv at the same location during morning hours in winter season. OFP of isoprene at vegetation sites is very high during post-monsoon because of high concentrations. However, OFP for other traffic tracers were high during winter seasons & at traffic locations. Furthermore, high correlation between isoprene emissions with traffic volume at traffic sites revealed that a noteworthy share of its emission also originates from road traffic.

Keywords: biogenic VOCs, isoprene emission, anthropogenic isoprene, urban vegetation

Procedia PDF Downloads 116

Authors: Aguasanta M. Sarmiento, Jose Miguel Davila, Juan Carlos Fortes, Maria Luisa de la Torre

Abstract:

Acid mine drainage (AMD) is an acidic leachate with high levels of metals and sulphates in solution, which seriously affects the durability and strength of metallic materials used in the construction of structural and mechanical components. This paper presents the results of the evolution over time of the reduction in tensile strength and defects in AISI 304 stainless steel in contact with acid mine drainage. For this purpose, a total of 30 bars with a diameter of 8 mm and a length of 14 cm were placed transversely in the course of a stream contaminated by AMD from the sulphide mines of the Iberian Pyritic Belt (SW Spain). This stream has average pH values of 2.6, a potential of 660 mV and average concentrations of 12 g/L of sulphates, 1.2 g/L of Fe, 191 mg/L of Zn, etc. Every two months of exposure, 6 stainless steel bars were extracted from the acid stream. They were subjected to surface roughness analysis carried out with the help of Mitutoyo Surftest SJ-210 surface roughness tester. The analysis was carried out at three different points on 5 specimens from each series. The average reading of each parameter is calculated in order to ensure the accuracy of the measurements and the surface coverage. Arithmetic mean roughness value (Ra), mean roughness depth (Rz) and root mean square roughness (Rq) were measured. Five specimens from each series were statically tensile tested using universal equipment (Servosis ME 403 of 200kN). The specimens were clamped at their ends with two grips for cylindrical sections, and the tensile force was applied at a constant speed of 0.5 kN/s, according to the requirements of standard UNE-EN ISO 6892-1: 2020. To determine the modulus of elasticity, limits close to 15% and 55% of the maximum load were used, depending on the course of each test. Field Emission Scanning Electron Microscopy (FESEM) was used to observe corrosion products and defects generated by exposure to AMD. Energy dispersive X-ray spectrometry (EDS) was used to analyze the chemical composition of the corrosion products formed. For this purpose, small pieces were cut from the resulting specimens, cleaned and embedded in epoxy resin. The results show that after only 5 months of exposure of AISI 304 stainless steel to the mining environment, the surface roughness increases significantly, with average depths almost 6 times greater than the initial one. Cracks are observed on the surface of the material, which increases in size with the time of exposure. A large number of grains with a composition of more than 57% Pb and 16% Sn can be observed inside these cracks. Tensile tests show a reduction in the resistance of this material after only two months of exposure. The results show the serious problems that would result from the use of this material for the use of mechanical components in a sulphide mining environment, not only because of the significant reduction in the lifetime of such components but also because of the implications for human safety.

Keywords: Acid mine drainage, Corrosion, Mechanical properties, Stainless steel

Procedia PDF Downloads 9

Authors: Tomasz Borowski, Dawid Sołoducha, Agata Markowska-Szczupak, Aneta Wesołowska, Marian Kordas, Rafał Rakoczy

Abstract:

Essential oils (EOs) are fragrant volatile oils obtained from plants. These are used for cooking (for flavor and aroma), cleaning, beauty (e.g., rosemary essential oil is used to promote hair growth), health (e.g. thyme essential oil cures arthritis, normalizes blood pressure, reduces stress on the heart, cures chest infection and cough) and in the food industry as preservatives and antioxidants. Rosemary and thyme essential oils are considered the most eminent herbs based on their history and medicinal properties. They possess a wide range of activity against different types of bacteria and fungi compared with the other oils in both in vitro and in vivo studies. However, traditional uses of EOs are limited due to rosemary and thyme oils in high concentrations can be toxic. In light of the accessible data, the following hypothesis was put forward: Low frequency rotating magnetic field (RMF) increases the antimicrobial potential of EOs. The aim of this work was to investigate the antimicrobial activity of commercial Salvia Rosmarinus L. and Thymus vulgaris L. essential oil from Polish company Avicenna-Oil under Rotating Magnetic Field (RMF) at f = 25 Hz. The self-constructed reactor (MAP) was applied for this study. The chemical composition of oils was determined by gas chromatography coupled with mass spectrometry (GC-MS). Model bacteria Escherichia coli K12 (ATCC 25922) was used. Minimum inhibitory concentrations (MIC) against E. coli were determined for the essential oils. Tested oils in very small concentrations were prepared (from 1 to 3 drops of essential oils per 3 mL working suspensions). From the results of disc diffusion assay and MIC tests, it can be concluded that thyme oil had the highest antibacterial activity against E. coli. Moreover, the study indicates the exposition to the RMF, as compared to the unexposed controls causing an increase in the efficacy of antibacterial properties of tested oils. The extended radiation exposure to RMF at the frequency f= 25 Hz beyond 160 minutes resulted in a significant increase in antibacterial potential against E. coli. Bacteria were killed within 40 minutes in thyme oil in lower tested concentration (1 drop of essential oils per 3 mL working suspension). Rapid decrease (>3 log) of bacteria number was observed with rosemary oil within 100 minutes (in concentration 3 drops of essential oils per 3 mL working suspension). Thus, a method for improving the antimicrobial performance of essential oil in low concentrations was developed. However, it still remains to be investigated how bacteria get killed by the EOs treated by an electromagnetic field. The possible mechanisms relies on alteration in the permeability of ionic channels in ionic channels in the bacterial cell walls that transport in the cells was proposed. For further studies, it is proposed to examine other types of essential oils and other antibiotic-resistant bacteria (ARB), which are causing a serious concern throughout the world.

Keywords: rotating magnetic field, rosemary, thyme, essential oils, Escherichia coli

Procedia PDF Downloads 156

Authors: Simran Baweja, Bhavika Kalal, Surajit Maity

Abstract:

Photoinduced tautomerization reactions have been the centre of attention among the scientific community over the past several decades because of their significance in various biological systems. 7-azaindole (7AI) is considered a model system for DNA base pairing and to understand the role of such tautomerization reactions in mutations. To the best of our knowledge, extensive studies have been carried out on 7-azaindole and its solvent clusters exhibiting proton/ hydrogen transfer in both solution as well as gas phases. Derivatives of the above molecule, like 2,7- and 2,6-diazaindoles are proposed to have even better photophysical properties due to the presence of -aza group on the 2nd position. However, there are studies in the solution phase that suggest the relevance of these molecules, but there are no experimental studies reported in the gas phase yet. In our current investigation, we present the first gas phase spectroscopic data of 2,7-diazaindole (2,7-DAI) and its solvent cluster (2,7-DAI-H2O). In this, we have employed state-of-the-art laser spectroscopic methods such as fluorescence excitation (LIF), dispersed fluorescence (DF), resonant two-photon ionization-time of flight mass spectrometry (2C-R2PI), photoionization efficiency spectroscopy (PIE), IR-UV double resonance spectroscopy, i.e., fluorescence-dip infrared spectroscopy (FDIR) and resonant ion-dip infrared spectroscopy (IDIR) to understand the electronic structure of the molecule. The origin band corresponding to the S1 ← S0 transition of the bare 2,7-DAI is found to be positioned at 33910 cm-1, whereas the origin band corresponding to S1 ← S0 transition of the 2,7-DAI-H2O is positioned at 33074 cm-1. The red-shifted transition in the case of solvent cluster suggests the enhanced feasibility of excited state hydrogen/ proton transfer. The ionization potential for the 2,7-DAI molecule is found to be 8.92 eV which is significantly higher than the previously reported 7AI (8.11 eV) molecule, making it a comparatively complex molecule to study. The ionization potential is reduced by 0.14 eV in the case of 2,7-DAI-H2O (8.78 eV) cluster compared to that of 2,7-DAI. Moreover, on comparison with the available literature values of 7AI, we found the origin band of 2,7-DAI and 2,7-DAI-H2O to be red-shifted by -729 and -280 cm-1 respectively. The ground and excited state N-H stretching frequencies of the 27DAI molecule were determined using fluorescence-dip infrared spectra (FDIR) and resonant ion dip infrared spectroscopy (IDIR), obtained at 3523 and 3467 cm-1, respectively. The lower value of vNH in the electronically excited state of 27DAI implies the higher acidity of the group compared to the ground state. Moreover, we have done extensive computational analysis, which suggests that the energy barrier in the excited state reduces significantly as we increase the number of catalytic solvent molecules (S= H2O, NH3) as well as the polarity of solvent molecules. We found that the ammonia molecule is a better candidate for hydrogen transfer compared to water because of its higher gas-phase basicity. Further studies are underway to understand the excited state dynamics and photochemistry of such N-rich chromophores.

Keywords: excited state hydrogen transfer, supersonic expansion, gas phase spectroscopy, IR-UV double resonance spectroscopy, laser induced fluorescence, photoionization efficiency spectroscopy

Procedia PDF Downloads 75

Authors: Olympia Nisiforou

Abstract:

The cultural heritage of each country represents a unique and irreplaceable witness of the past. Nevertheless, on many occasions, such heritage is extremely vulnerable to natural disasters and reckless behaviors. Even if such exhibits are now located in Museums, they still receive insufficient protection due to improper environmental conditions. These external changes can negatively affect the conditions of the exhibits and contribute to inefficient maintenance in time. Hence, it is imperative to develop an innovative, low-cost system, to monitor indoor air quality systematically, since conventional methods are quite expensive and time-consuming. The present study gives an insight into the indoor air quality of the National Byzantine Museum of Cyprus. In particular, systematic measurements of particulate matter, bio-aerosols, the concentration of targeted chemical pollutants (including Volatile organic compounds (VOCs), temperature, relative humidity, and lighting conditions as well as microbial counts have been performed using conventional techniques. Measurements showed that most of the monitored physiochemical parameters did not vary significantly within the various sampling locations. Seasonal fluctuations of ammonia were observed, showing higher concentrations in the summer and lower in winter. It was found that the outdoor environment does not significantly affect indoor air quality in terms of VOC and Nitrogen oxides (NOX). A cutting-edge portable Gas Chromatography-Mass Spectrometry (GC-MS) system (TORION T-9) was used to identify and measure the concentrations of specific Volatile and Semi-volatile Organic Compounds. A large number of different VOCs and SVOCs found such as Benzene, Toluene, Xylene, Ethanol, Hexadecane, and Acetic acid, as well as some more complex compounds such as 3-ethyl-2,4-dimethyl-Isopropyl alcohol, 4,4'-biphenylene-bis-(3-aminobenzoate) and trifluoro-2,2-dimethylpropyl ester. Apart from the permanent indoor/outdoor sources (i.e., wooden frames, painted exhibits, carpets, ventilation system and outdoor air) of the above organic compounds, the concentration of some of them within the areas of the museum were found to increase when large groups of visitors were simultaneously present at a specific place within the museum. The high presence of Particulate Matter (PM), fungi and bacteria were found in the museum’s areas where carpets were present but low colonial counts were found in rooms where artworks are exhibited. Measurements mentioned above were used to validate an innovative low-cost air-quality monitoring system that has been developed within the present work. The developed system is able to monitor the average concentrations (on a bidaily basis) of several pollutants and presents several innovative features, including the prompt alerting in case of increased average concentrations of monitored pollutants, i.e., exceeding the limit values defined by the user.

Keywords: exibitions, indoor air quality , VOCs, pollution

Procedia PDF Downloads 123

Authors: Malkhaz Jokhadze, Vakhtang Mshvildadze, Levan Makaradze, Ekaterine Mosidze, Salome Barbaqadze, Mariam Murtazashvili, Dali Berashvili, Koba sivsivadze, Lasha Bakuridze, Aliosha Bakuridze

Abstract:

Essential oils are one of the most important groups of biologically active substances present in plants. Due to the chemical diversity of components, essential oils and their preparations have a wide spectrum of pharmacological action. They have bactericidal, antiviral, fungicidal, antiprotozoal, anti-inflammatory, spasmolytic, sedative and other activities. They are expectorant, spasmolytic, sedative, hypotensive, secretion enhancing, antioxidant remedies. Based on preliminary pharmacological studies, we have developed a formulation called “Phytobiotic” containing essential oils, a feed additive for poultry as an alternative to antibiotics. Phytobiotic is a water-soluble powder containing a composition of essential oils of thyme, clary, monarda and auxiliary substances: dry extract of liquorice and inhalation lactose. On this stage of research, the goal was to study the chemical composition of provided phytobiotic, identify the main substances and determine their quantity, investigate the biological activity of phytobiotic through in vitro and in vivo studies. Using gas chromatography-mass spectrometry, 38 components were identified in phytobiotic, representing acyclic-, monocyclic-, bicyclic-, and sesquiterpenes. Together with identification of main active substances, their quantitative content was determined, including acyclic terpene alcohol β-linalool, acyclic terpene ketone linalyl acetate, monocyclic terpenes: D-limonene and γ-terpinene, monocyclic aromatic terpene thymol. Provided phytobiotic has pronounced and at the same time broad spectrum of antibacterial activity. In the cell model, phytobiotic showed weak antioxidant activity, and it was stronger in the ORAC (chemical model) tests. Meanwhile anti-inflammatory activity was also observed. When fowls were supplied feed enriched with phytobiotic, it was observed that gained weight of the chickens in the experimental group exceeded the same data for the control group during the entire period of the experiment. The survival rate of broilers in the experimental group during the growth period was 98% compared to -94% in the control group. As a result of conducted researches probable four different mechanisms which are important for the action of phytobiotics were identified: sensory, metabolic, antioxidant and antibacterial action. General toxic, possible local irritant and allergenic effects of phytobiotic were also investigated. Performed assays proved that formulation is safe.

Keywords: clary, essential oils, monarda, poultry, phytobiotics, thyme

Procedia PDF Downloads 172

Authors: Vadim Allheily, Rudiger Schmitt, Lionel Merlat, Gildas L'Hostis

Abstract:

Inflight devices are nowadays major actors in both military and civilian landscapes. Among others, missiles, mortars, rockets or even drones this last decade are increasingly sophisticated, and it is today of prior manner to develop always more efficient defensive systems from all these potential threats. In this frame, recent High Energy Laser weapon prototypes (HEL) have demonstrated some extremely good operational abilities to shot down within seconds flying targets several kilometers off. Whereas test outcomes are promising from both experimental and cost-related perspectives, the deterioration process still needs to be explored to be able to closely predict the effects of a high-energy laser irradiation on typical structures, heading finally to an effective design of laser sources and protective countermeasures. Laser matter interaction researches have a long history of more than 40 years at the French-German Research Institute (ISL). Those studies were tied with laser sources development in the mid-60s, mainly for specific metrology of fast phenomena. Nowadays, laser matter interaction can be viewed as the terminal ballistics of conventional weapons, with the unique capability of laser beams to carry energy at light velocity over large ranges. In the last years, a strong focus was made at ISL on the interaction process of laser radiation with metal targets such as artillery shells. Due to the absorbed laser radiation and the resulting heating process, an encased explosive charge can be initiated resulting in deflagration or even detonation of the projectile in flight. Drones and Unmanned Air Vehicles (UAVs) are of outmost interests in modern warfare. Those aerial systems are usually made up of polymer-based composite materials, whose complexity involves new scientific challenges. Aside this main laser-matter interaction activity, a lot of experimental and numerical knowledge has been gathered at ISL within domains like spectrometry, thermodynamics or mechanics. Techniques and devices were developed to study separately each aspect concerned by this topic; optical characterization, thermal investigations, chemical reactions analysis or mechanical examinations are beyond carried out to neatly estimate essential key values. Results from these diverse tasks are then incorporated into analytic or FE numerical models that were elaborated, for example, to predict thermal repercussion on explosive charges or mechanical failures of structures. These simulations highlight the influence of each phenomenon during the laser irradiation and forecast experimental observations with good accuracy.

Keywords: composite materials, countermeasure, experimental work, high-energy laser, laser-matter interaction, modeling

Procedia PDF Downloads 263

Authors: K. Verma, v. S. Moholkar

Abstract:

In developed countries, water pollution caused by industrial discharge has emerged as a significant environmental concern over the past decades. However, despite ongoing efforts, a fully effective and sustainable remediation strategy has yet to be identified. This paper describes how enzymatic and sonochemical treatments have demonstrated great promise in degrading bio-refractory pollutants. Mainly, a compelling area of interest lies in the combined technique of sono-enzymatic treatment, which has exhibited a synergistic enhancement effect surpassing that of the individual techniques. This study employed the covalent attachment method to immobilize Laccase from Trametes versicolor onto amino-functionalized magnetic Fe₃O₄ nanoparticles. To comprehensively characterize the synthesized free nanoparticles and the laccase-immobilized nanoparticles, various techniques such as X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), scanning electron microscope (SEM), vibrating sample magnetometer (VSM), and surface area through Brunauer-Emmett-Teller (BET) were employed. The size of immobilized Fe₃O₄@Laccase was found to be 60 nm, and the maximum loading of laccase was found to be 24 mg/g of nanoparticle. An investigation was conducted to study the effect of various process parameters, such as immobilized Fe₃O₄ Laccase dose, temperature, and pH, on the % Chemical oxygen demand (COD) removal as a response. The statistical design pinpointed the optimum conditions (immobilized Fe₃O₄ Laccase dose = 1.46 g/L, pH = 4.5, and temperature = 66 oC), resulting in a remarkable 65.58% COD removal within 60 minutes. An even more significant improvement (90.31% COD removal) was achieved with ultrasound-assisted enzymatic reaction utilizing a 10% duty cycle. The investigation of various kinetic models for free and immobilized laccase, such as the Haldane, Yano, and Koga, and Michaelis-Menten, showed that ultrasound application impacted the kinetic parameters Vmax and Km. Specifically, Vmax values for free and immobilized laccase were found to be 0.021 mg/L min and 0.045 mg/L min, respectively, while Km values were 147.2 mg/L for free laccase and 136.46 mg/L for immobilized laccase. The lower Km and higher Vmax for immobilized laccase indicate its enhanced affinity towards the substrate, likely due to ultrasound-induced alterations in the enzyme's confirmation and increased exposure of active sites, leading to more efficient degradation. Furthermore, the toxicity and Liquid chromatography-mass spectrometry (LC-MS) analysis revealed that after the treatment process, the wastewater exhibited 70% less toxicity than before treatment, with over 25 compounds degrading by more than 75%. At last, the prepared immobilized laccase had excellent recyclability retaining 70% activity up to 6 consecutive cycles. A straightforward manufacturing strategy and outstanding performance make the recyclable magnetic immobilized Laccase (Fe₃O₄ Laccase) an up-and-coming option for various environmental applications, particularly in water pollution control and treatment.

Keywords: kinetic, laccase enzyme, sonoenzymatic, ultrasound irradiation

Procedia PDF Downloads 67

Authors: Soraia V. M. de Sá, Miguel A. Faria, José O. Fernandes, Sara C. Cunha

Abstract:

Cereals play a vital role in fulfilling the nutritional needs of children, supplying essential nutrients crucial for their growth and development. However, concerns arise due to children's heightened vulnerability due to their unique physiology, specific dietary requirements, and relatively higher intake in relation to their body weight. This vulnerability exposes them to harmful food contaminants, particularly mycotoxins, prevalent in cereals. Because of the thermal stability of mycotoxins, conventional industrial food processing often falls short of eliminating them. Children, especially those aged 4 months to 12 years, frequently encounter mycotoxins through the consumption of specialized food products, such as instant foods, breakfast cereals, bars, cookie snacks, fruit puree, and various dairy items. A close monitoring of this demographic group's exposure to mycotoxins is essential, as toxins ingestion may weaken children’s immune systems, reduce their resistance to infectious diseases, and potentially lead to cognitive impairments. The severe toxicity of mycotoxins, some of which are classified as carcinogenic, has spurred the establishment and ongoing revision of legislative limits on mycotoxin levels in food and feed globally. While EU Commission Regulation 1881/2006 addresses well-known mycotoxins in processed cereal-based foods and infant foods, the absence of regulations specifically addressing emerging mycotoxins underscores a glaring gap in the regulatory framework, necessitating immediate attention. Emerging mycotoxins have gained mounting scrutiny in recent years due to their pervasive presence in various foodstuffs, notably cereals and cereal-based products. Alarmingly, exposure to multiple mycotoxins is hypothesized to exhibit higher toxicity than isolated effects, raising particular concerns for products primarily aimed at children. This study scrutinizes the presence of 22 mycotoxins of the diverse range of chemical classes in 148 processed cereal-based foods, including 39 breakfast cereals, 25 infant formulas, 27 snacks, 25 cereal bars, and 32 cookies commercially available in Portugal. The analytical approach employed a modified QuEChERS procedure followed by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) analysis. Given the paucity of information on the risk assessment of children to multiple mycotoxins in cereal and cereal-based products consumed by children of Portugal pioneers the evaluation of this critical aspect. Overall, aflatoxin B1 (AFB1) and aflatoxin G2 (AFG2) emerged as the most prevalent regulated mycotoxins, while enniatin B (ENNB) and sterigmatocystin (STG) were the most frequently detected emerging mycotoxins.

Keywords: cereal-based products, children´s nutrition, food safety, UPLC-MS/MS analysis

Procedia PDF Downloads 71

Authors: Fabio N. Moreno, Letícia B. Marinho, Beatriz D. Ruiz, Maria Helena R. B. Martins

Abstract:

Brazil is a top consumer of pesticides worldwide, and the São Paulo State is one of the highest consumers among the Brazilian federative states. However, representative data about the occurrence of pesticides in surface waters of the São Paulo State is scarce. This paper aims to present the results of pesticides monitoring executed within the Water Quality Monitoring Network of CETESB (The Environmental Agency of the São Paulo State) between the 2018-2022 period. Surface water sampling points (21 to 25) were selected within basins of predominantly agricultural land-use (5 to 85% of cultivated areas). The samples were collected throughout the year, including high-flow and low-flow conditions. The frequency of sampling varied between 6 to 4 times per year. Selection of pesticide molecules for monitoring followed a prioritizing process from EMBRAPA (Brazilian Agricultural Research Corporation) databases of pesticide use. Pesticides extractions in aqueous samples were performed according to USEPA 3510C and 3546 methods following quality assurance and quality control procedures. Determination of pesticides in water (ng L-1) extracts were performed by high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS) and by gas chromatography with nitrogen phosphorus (GC-NPD) and electron capture detectors (GC-ECD). The results showed higher frequencies (20- 65%) in surface water samples for Carbendazim (fungicide), Diuron/Tebuthiuron (herbicides) and Fipronil/Imidaclopride (insecticides). The frequency of observations for these pesticides were generally higher in monitoring points located in sugarcane cultivated areas. The following pesticides were most frequently quantified above the Aquatic life benchmarks for freshwater (USEPA Office of Pesticide Programs, 2023) or Brazilian Federal Regulatory Standards (CONAMA Resolution no. 357/2005): Atrazine, Imidaclopride, Carbendazim, 2,4D, Fipronil, and Chlorpiryfos. Higher median concentrations for Diuron and Tebuthiuron in the rainy months (october to march) indicated pesticide transport through surface runoff. However, measurable concentrations in the dry season (april to september) for Fipronil and Imidaclopride also indicates pathways related to subsurface or base flow discharge after pesticide soil infiltration and leaching or dry deposition following pesticide air spraying. With exception to Diuron, no temporal trends related to median concentrations of the most frequently quantified pesticides were observed. These results are important to assist policymakers in the development of strategies aiming at reducing pesticides migration to surface waters from agricultural areas. Further studies will be carried out in selected points to investigate potential risks as a result of pesticides exposure on aquatic biota.

Keywords: pesticides monitoring, são paulo state, water quality, surface waters

Procedia PDF Downloads 59

Authors: Yana Posmitnaya, Galina Rudnitskaya, Tatyana Lukashenko, Anton Bukatin, Anatoly Evstrapov

Abstract:

An important area of biological and medical research is the study of genetic mutations and polymorphisms that can alter gene function and cause inherited diseases and other diseases. The following methods to analyse DNA fragments are used: capillary electrophoresis and electrophoresis on microfluidic chip (MFC), mass spectrometry with electrophoresis on MFC, hybridization assay on microarray. Electrophoresis on MFC allows to analyse small volumes of samples with high speed and throughput. A soft lithography in polydimethylsiloxane (PDMS) was chosen for operative fabrication of MFCs. A master-form from silicon and photoresist SU-8 2025 (MicroChem Corp.) was created for the formation of micro-sized structures in PDMS. A universal topology which combines T-injector and simple cross was selected for the electrophoretic separation of the sample. Glass K8 and PDMS Sylgard® 184 (Dow Corning Corp.) were used for fabrication of MFCs. Electroosmotic flow (EOF) plays an important role in the electrophoretic separation of the sample. Therefore, the estimate of the quantity of EOF and the ways of its regulation are of interest for the development of the new methods of the electrophoretic separation of biomolecules. The following methods of surface modification were chosen to change EOF: high-frequency (13.56 MHz) plasma treatment in oxygen and argon at low pressure (1 mbar); 1% aqueous solution of polyvinyl alcohol; 3% aqueous solution of Kolliphor® P 188 (Sigma-Aldrich Corp.). The electroosmotic mobility was evaluated by the method of Huang X. et al., wherein the borate buffer was used. The influence of physical and chemical methods of treatment on the wetting properties of the PDMS surface was controlled by the sessile drop method. The most effective way of surface modification of MFCs, from the standpoint of obtaining the smallest value of the contact angle and the smallest value of the EOF, was the processing with aqueous solution of Kolliphor® P 188. This method of modification has been selected for the treatment of channels of MFCs, which are used for the separation of mixture of oligonucleotides fluorescently labeled with the length of chain with 10, 20, 30, 40 and 50 nucleotides. Electrophoresis was performed on the device MFAS-01 (IAI RAS, Russia) at the separation voltage of 1500 V. 6% solution of polydimethylacrylamide with the addition of 7M carbamide was used as the separation medium. The separation time of components of the mixture was determined from electropherograms. The time for untreated MFC was ~275 s, and for the ones treated with solution of Kolliphor® P 188 – ~ 220 s. Research of physical-chemical methods of surface modification of MFCs allowed to choose the most effective way for reducing EOF – the modification with aqueous solution of Kolliphor® P 188. In this case, the separation time of the mixture of oligonucleotides decreased about 20%. The further optimization of method of modification of channels of MFCs will allow decreasing the separation time of sample and increasing the throughput of analysis.

Keywords: electrophoresis, microfluidic chip, modification, nucleic acid, polydimethylsiloxane, soft lithography

Procedia PDF Downloads 413

Authors: Xuechun Kan, Dongdong Li, Fan Li, Peidang Liu

Abstract:

Triple-negative breast cancer (TNBC) is the most malignant subtype of breast cancer with a poor prognosis, and radiotherapy is one of the main treatment methods. However, due to the obvious resistance of tumor cells to radiotherapy, high dose of ionizing radiation is required during radiotherapy, which causes serious damage to normal tissues near the tumor. Therefore, how to improve radiotherapy resistance and enhance the specific killing of tumor cells by radiation is a hot issue that needs to be solved in clinic. Recent studies have shown that silver-based nanoparticles have strong radiosensitization, and silver nanoclusters (AgNCs) also provide a broad prospect for tumor targeted radiosensitization therapy due to their ultra-small size, low toxicity or non-toxicity, self-fluorescence and strong photostability. Aptamer 5TR1 is a 25-base oligonucleotide aptamer that can specifically bind to mucin-1 highly expressed on the membrane surface of TNBC 4T1 cells, and can be used as a highly efficient tumor targeting molecule. In this study, AgNCs were synthesized by DNA template based on 5TR1 aptamer (NC-T5-5TR1), and its role as a targeted radiosensitizer in TNBC radiotherapy was investigated. The optimal DNA template was first screened by fluorescence emission spectroscopy, and NC-T5-5TR1 was prepared. NC-T5-5TR1 was characterized by transmission electron microscopy, ultraviolet-visible spectroscopy and dynamic light scattering. The inhibitory effect of NC-T5-5TR1 on cell activity was evaluated using the MTT method. Laser confocal microscopy was employed to observe NC-T5-5TR1 targeting 4T1 cells and verify its self-fluorescence characteristics. The uptake of NC-T5-5TR1 by 4T1 cells was observed by dark-field imaging, and the uptake peak was evaluated by inductively coupled plasma mass spectrometry. The radiation sensitization effect of NC-T5-5TR1 was evaluated through cell cloning and in vivo anti-tumor experiments. Annexin V-FITC/PI double staining flow cytometry was utilized to detect the impact of nanomaterials combined with radiotherapy on apoptosis. The results demonstrated that the particle size of NC-T5-5TR1 is about 2 nm, and the UV-visible absorption spectrum detection verifies the successful construction of NC-T5-5TR1, and it shows good dispersion. NC-T5-5TR1 significantly inhibited the activity of 4T1 cells and effectively targeted and fluoresced within 4T1 cells. The uptake of NC-T5-5TR1 reached its peak at 3 h in the tumor area. Compared with AgNCs without aptamer modification, NC-T5-5TR1 exhibited superior radiation sensitization, and combined radiotherapy significantly inhibited the activity of 4T1 cells and tumor growth in 4T1-bearing mice. The apoptosis level of NC-T5-5TR1 combined with radiation was significantly increased. These findings provide important theoretical and experimental support for NC-T5-5TR1 as a radiation sensitizer for TNBC.

Keywords: 5TR1 aptamer, silver nanoclusters, radio sensitization, triple-negative breast cancer

Procedia PDF Downloads 60

Authors: Razieh Karimi Aghcheh, Christian Kubicek, Joseph Strauss, Gerhard Braus

Abstract:

Filamentous fungi are of considerable economic and social significance for human health, nutrition and in white biotechnology. These organisms are dominant producers of a range of primary metabolites such as citric acid, microbial lipids (biodiesel) and higher unsaturated fatty acids (HUFAs). In particular, they produce also important but structurally complex secondary metabolites with enormous therapeutic applications in pharmaceutical industry, for example: cephalosporin, penicillin, taxol, zeranol and ergot alkaloids. Several fungal secondary metabolites, which are significantly relevant to human health do not only include antibiotics, but also e.g. lovastatin, a well-known antihypercholesterolemic agent produced by Aspergillus. terreus, or aflatoxin, a carcinogen produced by A. flavus. In addition to their roles for human health and agriculture, some fungi are industrially and commercially important: Species of the ascomycete genus Hypocrea spp. (teleomorph of Trichoderma) have been demonstrated as efficient producer of highly active cellulolytic enzymes. This trait makes them effective in disrupting and depolymerization of lignocellulosic materials and thus applicable tools in number of biotechnological areas as diverse as clothes-washing detergent, animal feed, and pulp and fuel productions. Fungal LaeA/LAE1 (Loss of aflR Expression A) homologs their gene products act at the interphase between secondary metabolisms, cellulase production and development. Lack of the corresponding genes results in significant physiological changes including loss of secondary metabolite and lignocellulose degrading enzymes production. At the molecular level, the encoded proteins are presumably methyltransferases or demethylases which act directly or indirectly at heterochromatin and interact with velvet domain proteins. Velvet proteins bind to DNA and affect expression of secondary metabolites (SMs) genes and cellulases. The dynamic interplay between LaeA/LAE1, velvet proteins and additional interaction partners is the key for an understanding of the coordination of metabolic and morphological functions of fungi and is required for a biotechnological control of the formation of desired bioactive products. Aspergilli and Trichoderma represent different biotechnologically significant species with significant differences in the LaeA/LAE1-Velvet protein machinery and their target proteins. We, therefore, performed a comparative study of the interaction partners of this machinery and the dynamics of the various protein-protein interactions using our robust proteomic and mass spectrometry techniques. This enhances our knowledge about the fungal coordination of secondary metabolism, cellulase production and development and thereby will certainly improve recombinant fungal strain construction for the production of industrial secondary metabolite or lignocellulose hydrolytic enzymes.

Keywords: cellulases, LaeA/1, proteomics, secondary metabolites

Procedia PDF Downloads 270

Authors: Malek Mahmoudi, Jonas Keruckas, Dmytro Volyniuk, Jurate Simokaitiene, Juozas V. Grazulevicius

Abstract:

Host materials have been discovered as one of the most appealing methods for harvesting triplet states in organic materials for application in organic light-emitting diodes (OLEDs). The ideal host-guest system for emission in thermally delayed fluorescence OLEDs with 20% guest concentration for efficient energy transfer has been demonstrated in the present investigation. In this work, 3,3'-bis[9-(4-fluorophenyl) carbazole] (bFPC) has been used as the host, which induces balanced charge carrier transport for high-efficiency OLEDs.For providing a complete characterization of the synthesized compound, photophysical, photoelectrical, charge-transporting, and electrochemical properties of the compound have been examined. Excited-state lifetimes and singlet-triplet energy gaps were measured for characterization of photophysical properties, while thermogravimetric analysis, as well as differential scanning calorimetry measurements, were performed for probing of electrochemical and thermal properties of the compound. The electrochemical properties of this compound were investigated by cyclic voltammetry (CV) method, and ionization potential (IPCV) value of 5.68 eV was observed. UV–Vis absorption and photoluminescence spectrum of a solution of the compound in toluene (10-5 M) showed maxima at 302 and 405 nm, respectively. Photoelectron emission spectrometry was used for the characterization of charge-injection properties of the studied compound in solid. The ionization potential of this material was found to be 5.78 eV, and time-of-flight measurement was used for testing charge-transporting properties and hole mobility estimated using this technique in a vacuum-deposited layer reached 4×10-4 cm2 V-1s-1. Since the compound with high charge mobilities was tested as a host in an organic light-emitting diode. The device was fabricated by successive deposition onto a pre-cleaned indium tin oxide (ITO) coated glass substrate under a vacuum of 10-6 Torr and consisting of an indium-tin-oxide anode, hole injection and transporting layer(MoO3, NPB), emitting layer with bFPC as a host and 4CzIPN (2,4,5,6-tetra(9-carbazolyl)isophthalonitrile) which is a new highly efficient green thermally activated delayed fluorescence (TADF) material as an emitter, an electron transporting layer(TPBi) and lithium fluoride layer topped with aluminum layer as a cathode exhibited the highest maximum current efficiency and power efficiency of 33.9 cd/A and 23.5 lm/W, respectively and the electroluminescence spectrum showed only a peak at 512nm. Furthermore, the new bicarbazole-based compound was tested as a host in thermally activated delayed fluorescence organic light-emitting diodes are reaching luminance of 25300 cd m-2 and external quantum efficiency of 10.1%. Interestingly, the turn-on voltage was low enough (3.8 V), and such a device can be used for highly efficient light sources.

Keywords: thermally-activated delayed fluorescence, host material, ionization energy, charge mobility, electroluminescence

Procedia PDF Downloads 140

Authors: Anupalli Roja Rani, Pavithra Dasari

Abstract:

Background: Among several, breast cancer has emerged as the most common female cancer in developing countries. It is the most common cause of cancer-related deaths worldwide among women. It is a molecularly and clinically heterogeneous disease. Moreover, it is a hormone–dependent tumor in which estrogens can regulate the growth of breast cells by binding with estrogen receptors (ERs). Moreover, the use of natural products in cancer therapeutics is due to their properties of biocompatibility and less toxicity. Plants are the vast reservoirs for various bioactive compounds. Coleus barbatus (Lamiaceae) contains anticancer properties against several cancer cell lines. Method: In the present study, an attempt is being made to enrich the knowledge of the anticancer activity of pure compounds extracted from Coleus barbatus (Andrew). On human breast cancer cell lines MCF-7. Here in, we are assessing the antiproliferative activity of Coleus barbatus (Andrew) plant extracts against MCF 7 and also evaluating their toxicity in normal human mammary cell lines such as Human Mammary Epithelial Cells (HMEC). The active fraction of plant extract was further purified with the help of Flash chromatography, Medium Pressure Liquid Chromatography (MPLC) and preparative High-Performance Liquid Chromatography (HPLC). The structure of pure compounds will be elucidated by using modern spectroscopic methods like Nuclear magnetic resonance (NMR), Electrospray Ionisation Mass Spectrometry (ESI-MS) methods. Later, the growth inhibition morphological assessment of cancer cells and cell cycle analysis of purified compounds were assessed using FACS. The growth and progression of signaling molecules HER2, GRP78 was studied by secretion assay using ELISA and expression analysis by flow cytometry. Result: Cytotoxic effect against MCF-7 with IC50 values were derived from dose response curves, using six concentrations of twofold serially diluted samples, by SOFTMax Pro software (Molecular device) and respectively Ellipticine and 0.5% DMSO were used as a positive and negative control. Conclusion: The present study shows the significance of various bioactive compounds extracted from Coleus barbatus (Andrew) root material. It acts as an anti-proliferative and shows cytotoxic effects on human breast cancer cell lines MCF7. The plant extracts play an important role pharmacologically. The whole plant has been used in traditional medicine for decades and the studies done have authenticated the practice. Earlier, as described, the plant has been used in the ayurveda and homeopathy medicine. However, more clinical and pathological studies must be conducted to investigate the unexploited potential of the plant. These studies will be very useful for drug designing in the future.

Keywords: coleus barbatus, HPLC, MPLC, NMR, MCF7, flash chromatograph, ESI-MS, FACS, ELISA.

Procedia PDF Downloads 114

Authors: Dmitriy Babenko, Sergey Yegorov, Ilya Korshukov, Aidana Sultanbekova, Valentina Barkhanskaya, Tatiana Bashirova, Yerzhan Zhunusov, Yevgeniya Li, Viktoriya Parakhina, Svetlana Kolesnichenko, Yeldar Baiken, Aruzhan Pralieva, Zhibek Zhumadilova, Matthew S. Miller, Gonzalo H. Hortelano, Anar Turmuhambetova, Antonella E. Chesca, Irina Kadyrova

Abstract:

Introduction: The rapidly evolving COVID-19 pandemic, along with the re-emergence of pathogens causing acute respiratory infections (ARI), has necessitated the development of novel diagnostic tools to differentiate various causes of ARI. MALDI-MS, due to its wide usage and affordability, has been proposed as a potential instrument for diagnosing SARS-CoV-2 versus non-SARS-CoV-2 ARI. The aim of this study was to investigate the potential of MALDI-MS in conjunction with a machine learning model to accurately distinguish between symptomatic infections caused by SARS-CoV-2 and non-SARS-CoV-2 during both the early and later phases of the pandemic. Furthermore, this study aimed to analyze mass spectrometry (MS) data obtained from nasal swabs of healthy individuals. Methods: We gathered mass spectra from 252 samples, comprising 108 SARS-CoV-2-positive samples obtained in 2020 (Covid 2020), 7 SARS-CoV- 2-positive samples obtained in 2023 (Covid 2023), 71 samples from symptomatic individuals without SARS-CoV-2 (Control non-Covid ARVI), and 66 samples from healthy individuals (Control healthy). All the samples were subjected to RT-PCR testing. For data analysis, we employed the caret R package to train and test seven machine-learning algorithms: C5.0, KNN, NB, RF, SVM-L, SVM-R, and XGBoost. We conducted a training process using a five-fold (outer) nested repeated (five times) ten-fold (inner) cross-validation with a randomized stratified splitting approach. Results: In this study, we utilized the Covid 2020 dataset as a case group and the non-Covid ARVI dataset as a control group to train and test various machine learning (ML) models. Among these models, XGBoost and SVM-R demonstrated the highest performance, with accuracy values of 0.97 [0.93, 0.97] and 0.95 [0.95; 0.97], specificity values of 0.86 [0.71; 0.93] and 0.86 [0.79; 0.87], and sensitivity values of 0.984 [0.984; 1.000] and 1.000 [0.968; 1.000], respectively. When examining the Covid 2023 dataset, the Naive Bayes model achieved the highest classification accuracy of 43%, while XGBoost and SVM-R achieved accuracies of 14%. For the healthy control dataset, the accuracy of the models ranged from 0.27 [0.24; 0.32] for k-nearest neighbors to 0.44 [0.41; 0.45] for the Support Vector Machine with a radial basis function kernel. Conclusion: Therefore, ML models trained on MALDI MS of nasopharyngeal swabs obtained from patients with Covid during the initial phase of the pandemic, as well as symptomatic non-Covid individuals, showed excellent classification performance, which aligns with the results of previous studies. However, when applied to swabs from healthy individuals and a limited sample of patients with Covid in the late phase of the pandemic, ML models exhibited lower classification accuracy.

Keywords: SARS-CoV-2, MALDI-TOF MS, ML models, nasopharyngeal swabs, classification

Procedia PDF Downloads 108

Authors: Anil Saini, Jatinder Kumar Ratan

Abstract:

Now a day, the key issue for the scientific community is to devise the innovative technologies for sustainable control of urban pollution. In urban cities, a large surface area of the masonry structures, buildings, and pavements is exposed to the open environment, which may be utilized for the control of air pollution, if it is built from the photocatalytically active cement-based constructional materials such as concrete, mortars, paints, and blocks, etc. The photocatalytically active cement is formulated by incorporating a photocatalyst in the cement matrix, and such cement is generally known as self-cleaning cement In the literature, self-cleaning cement has been synthesized by incorporating nanosized-TiO₂ (n-TiO₂) as a photocatalyst in the formulation of the cement. However, the utilization of n-TiO₂ for the formulation of self-cleaning cement has the drawbacks of nano-toxicity, higher cost, and agglomeration as far as the commercial production and applications are concerned. The use of microsized-TiO₂ (m-TiO₂) in place of n-TiO₂ for the commercial manufacture of self-cleaning cement could avoid the above-mentioned problems. However, m-TiO₂ is less photocatalytically active as compared to n- TiO₂ due to smaller surface area, higher band gap, and increased recombination rate. As such, the use of m-TiO₂ in the formulation of self-cleaning cement may lead to a reduction in photocatalytic activity, thus, reducing the self-cleaning, depolluting, and antimicrobial abilities of the resultant cement material. So improvement in the photoactivity of m-TiO₂ based self-cleaning cement is the key issue for its practical applications in the present scenario. The current work proposes the use of surface-fluorinated m-TiO₂ for the formulation of self-cleaning cement to enhance its photocatalytic activity. The calcined dolomite, a constructional material, has also been utilized as co-adsorbent along with the surface-fluorinated m-TiO₂ in the formulation of self-cleaning cement to enhance the photocatalytic performance. The surface-fluorinated m-TiO₂, calcined dolomite, and the formulated self-cleaning cement were characterized using diffuse reflectance spectroscopy (DRS), X-ray diffraction analysis (XRD), field emission-scanning electron microscopy (FE-SEM), energy dispersive x-ray spectroscopy (EDS), X-ray photoelectron spectroscopy (XPS), scanning electron microscopy (SEM), BET (Brunauer–Emmett–Teller) surface area, and energy dispersive X-ray fluorescence spectrometry (EDXRF). The self-cleaning property of the as-prepared self-cleaning cement was evaluated using the methylene blue (MB) test. The depolluting ability of the formulated self-cleaning cement was assessed through a continuous NOX removal test. The antimicrobial activity of the self-cleaning cement was appraised using the method of the zone of inhibition. The as-prepared self-cleaning cement obtained by uniform mixing of 87% clinker, 10% calcined dolomite, and 3% surface-fluorinated m-TiO₂ showed a remarkable self-cleaning property by providing 53.9% degradation of the coated MB dye. The self-cleaning cement also depicted a noteworthy depolluting ability by removing 5.5% of NOx from the air. The inactivation of B. subtiltis bacteria in the presence of light confirmed the significant antimicrobial property of the formulated self-cleaning cement. The self-cleaning, depolluting, and antimicrobial results are attributed to the synergetic effect of surface-fluorinated m-TiO₂ and calcined dolomite in the cement matrix. The present study opens an idea and route for further research for acile and economical formulation of self-cleaning cement.

Keywords: microsized-titanium dioxide (m-TiO₂), self-cleaning cement, photocatalysis, surface-fluorination

Procedia PDF Downloads 170

Authors: Sharmin Sultana, Nicolas Nuns, Pardis Simon, Jean-Marc Giraudon, Jean-Francois Lamonior, Nathalie D. Geyter, Rino Morent

Abstract:

Environmental issues, especially air pollution, have become a huge concern of environmental legislation as a consequence of growing awareness in our global world. In this regard, control of volatile organic compounds (VOCs) emission has become an important issue due to their potential toxicity, carcinogenicity, and mutagenicity. The research of innovative technologies for VOC abatement is stimulated to accommodate the new stringent standards in terms of VOC emission. One emerging strategy is the coupling of 2 existing complementary technologies, namely here non-thermal plasma (NTP) and heterogeneous catalysis, to get a more efficient process for VOC removal in air. The objective of this current work is to investigate the abatement of trichloroethylene (TCE-highly toxic chlorinated VOC) from moist air (RH=15%) as a function of time by combined use of multi-pin-to-plate negative DC corona/glow discharge with Fe-doped cryptomelanes catalyst downstream i.e. post plasma-catalysis (PPC) process. For catalyst alone case, experiments reveal that, initially, Fe doped cryptomelane (regardless the mode of Fe incorporation by co-precipitation (Fe-K-OMS-2)/ impregnation (Fe/K-OMS-2)) exhibits excellent activity to decompose TCE compared to cryptomelane (K-OMS-2) itself. A maximum obtained value of TCE abatement after 6 min is as follows: Fe-KOMS-2 (73.3%) > Fe/KOMS-2 (48.5) > KOMS-2 (22.6%). However, with prolonged operation time, whatever the catalyst under concern, the abatement of TCE decreases. After 111 min time of exposure, the catalysts can be ranked as follows: Fe/KOMS-2 (11%) < K-OMS-2 (12.3%) < Fe-KOMS-2 (14.5%). Clearly, this phenomenon indicates catalyst deactivation either by chlorination or by blocking the active sites. Remarkably, in PPC configuration (energy density = 60 J/L, catalyst temperature = 150°C), experiments reveal an enhanced performance towards TCE removal regardless the type of catalyst. After 6 min time on stream, the TCE removal efficiency amount as follows: K-OMS-2 (60%) < Fe/K-OMS-2 (79%) < Fe-K-OMS-2 (99.3%). The enhanced performances over Fe-K-OMS-2 catalyst are attributed to its high surface oxygen mobility and structural defects leading to high O₃ decomposition efficiency to give active species able to oxidize the plasma processed hazardous\by-products and the possibly remaining VOC into CO₂. Moreover, both undoped and doped catalysts remain strongly capable to abate TCE with time on stream. The TCE removal efficiencies of the PPC processes with Fe/KOMS-2 and KOMS-2 catalysts are not affected by time on stream indicating an excellent catalyst stability. When using the Fe-K-OMS-2 as catalyst, TCE abatement slightly reduces with time on stream. However, it is noteworthy to stress that still a constant abatement of 83% is observed during at least 30 minutes. These results prove that the combination of NTP with catalysts not only increases the catalytic activity but also allows to avoid, to some extent, the poisoning of catalytic sites resulting in an enhanced catalyst stability. In order to better understand the different surface processes occurring in the course of the total TCE oxidation in PPC experiments, a detailed X-ray Photoelectron Spectroscopy (XPS) and Time of Flight-Secondary Ion Mass Spectrometry (ToF-SIMS) study on the fresh and used catalysts is in progress.

Keywords: Fe doped cryptomelane, non-thermal plasma, plasma-catalysis, stability, trichloroethylene

Procedia PDF Downloads 208

Authors: Vladimir Berezin, Andrey Bogoyavlenskiy, Pavel Alexyuk, Madina Alexyuk, Aizhan Turmagambetova, Irina Zaitseva, Nadezhda Sokolova, Elmira Omirtaeva

Abstract:

Introduction: Triterpene saponins of plant origin are one of the most promising candidates for elaboration of novel adjuvants. Due to the combination of immunostimulating activity and the capacity interact with amphipathic molecules with formation of highly immunogenic nanocomplexes, triterpene saponins could serve as a good adjuvant/delivery system for vaccine use. In the research presented adjuvants on the base of nanocomplexes contained triterpene saponins isolated from Glycyrrhiza glabra and Saponaria officinalis plants indigenous to Kazakhstan were elaborated for influenza vaccine use. Methods: Purified triterpene saponins 'Glabilox' and 'SO1' with low toxicity and high immunostimulatory activity were isolated from plants Glycyrrhiza glabra L. and Saponaria officinalis L. by high-performance liquid chromatography (HPLC) and identified using electrospray ionization mass spectrometry (ESI-MS). Influenza virus A/St-Petersburg/5/09 (H1N1) propagated in 9-days old chicken embryos was concentrated and purified by centrifugation in sucrose gradient. Nanocomplexes contained lipids, and triterpene saponins Glabilox or SO1 were prepared by dialysis technique. Immunostimulating activity of experimental vaccine preparations was studied in vaccination/challenge experiments in mice. Results: Humoral and cellular immune responses and protection against influenza virus infection were examined after single subcutaneous and intranasal immunization. Mice were immunized subunit influenza vaccine (HA+NA) or whole virus inactivated influenza vaccine in doses 3.0/5.0/10.0 µg antigen/animal mixed with adjuvant in dose 15.0 µg/animal. Sera were taken 14-21 days following single immunization and mice challenged by A/St-Petersburg/5/09 influenza virus in dose 100 EID₅₀. Study of experimental influenza vaccine preparations in animal immunization experiments has shown that subcutaneous and intranasal immunization with subunit influenza vaccine mixed with nanocomplexes contained Glabilox or SO1 saponins stimulated high levels of humoral immune response (IgM, IgA, IgG1, IgG2a, and IgG2b antibody) and cellular immune response (IL-2, IL-4, IL-10, and IFN-γ cytokines) and resulted 80-90% protection against lethal influenza infection. Also, single intranasal and single subcutaneous immunization with whole virus inactivated influenza vaccine mixed with nanoparticulated adjuvants stimulated high levels of humoral and cellular immune responses and provided 100% protection against lethal influenza infection. Conclusion: The results of study have shown that nanocomplexes contained purified triterpene saponins Glabilox and SO1 isolated from plants indigenous to Kazakhstan can stimulate a broad spectrum of humoral and cellular immune responses and induce protection against lethal influenza infection. Both elaborated adjuvants are promising for incorporation to influenza vaccine intended for subcutaneous and intranasal routes of immunization.

Keywords: influenza vaccine, adjuvants, triterpene saponins, immunostimulating activity

Procedia PDF Downloads 137

Authors: Jelena Filipovic, Marija Bodroza-Solarov, Milenko Kosutic, Nebojsa Novkovic, Vladimir Filipovic, Vesna Vucurovic

Abstract:

Spelt wheat is suitable raw material for extruded products such as pasta, special types of bread and other products of altered nutritional characteristics compared to conventional wheat products. During the process of extrusion, spelt is exposed to high temperature and high pressure, during which raw material is also mechanically treated by shear forces. Spelt wheat is growing without the use of pesticides in harsh ecological conditions and in marginal areas of cultivation. So it can be used for organic and health safe food. Pasta is the most popular foodstuff; its consumption has been observed to rise. Pasta quality depends mainly on the properties of flour raw materials, especially protein content and its quality but starch properties are of a lesser importance. Pasta is characterized by significant amounts of complex carbohydrates, low sodium, total fat fiber, minerals, and essential fatty acids and its nutritional value can be improved with additional functional component. Over the past few decades, wheat pasta has been successfully formulated using different ingredients in pasta to cater health-conscious consumers who prefer having a product rich in protein, healthy lipids and other health benefits. Flaxseed flour is used in the production of bakery and pasta products that have properties of functional foods. However, it should be taken into account that food products retain the technological and sensory quality despite the added flax seed. Flaxseed contains important substances in its composition such as vitamins and minerals elements, and it is also an excellent source of fiber and one of the best sources of ω-3 fatty acids and lignin. In this paper, the quality and identification of spelt extruded product with the addition of flax seed, which is positively contributing to the nutritive and technology changes of the product, is investigated. ω-3 fatty acids are polyunsaturated essential fatty acids, and they must be taken with food to satisfy the recommended daily intake. Flaxseed flour is added in the quantity of 10/100 g of sample and 20/100 g of sample on farina. It is shown that the presence of ω-3 fatty acids in pasta can be clearly distinguished from other fatty acids by gas chromatography with mass spectrometry. Addition of flax seed flour influence chemical content of pasta. The addition of flax seed flour in spelt pasta in the quantities of 20g/100 g significantly increases the share of ω-3 fatty acids, which results in improved ratio of ω-6/ω-3 1:2.4 and completely satisfies minimum daily needs of ω-3 essential fatty acids (3.8 g/100 g) recommended by FDA. Flex flour influenced the pasta quality by increasing of hardness (2377.8 ± 13.3; 2874.5 ± 7.4; 3076.3 ± 5.9) and work of shear (102.6 ± 11.4; 150.8 ± 11.3; 165.0 ± 18.9) and increasing of adhesiveness (11.8 ± 20.6; 9.,98 ± 0.12; 7.1 ± 12.5) of the final product. Presented data point at good indicators of technological quality of spelt pasta with flax seed and that GC-MS analysis can be used in the quality control for flax seed identification. Acknowledgment: The research was financed by the Ministry of Education and Science of the Republic of Serbia (Project No. III 46005).

Keywords: GC-MS analysis, ω-3 fatty acids, flex seed, spelt wheat, daily needs

Procedia PDF Downloads 162

Authors: Bello Gonzalez T. D. J., Setten Van M., Essen Van A., Brouwer M., Veldman K. T.

Abstract:

Resistance to fluoroquinolones (FQ) has evolved increasingly over the years, posing a significant challenge for the treatment of human infections, particularly gastrointestinal tract infections caused by zoonotic bacteria transmitted through the food chain and environment. In broiler chickens, a relatively high proportion of FQ resistance has been observed in Escherichia coli indicator, Salmonella and Campylobacter isolates. We hypothesize that flumequine (Flu), used as a secondary choice for the treatment of poultry infections, could potentially be associated with a high proportion of FQ resistance. To evaluate this hypothesis, we used an in vitro fermentation chicken caeca model. Two continuous single-stage fermenters were used to simulate in real time the physiological conditions of the chicken caeca microbial content (temperature, pH, caecal content mixing, and anoxic environment). A pool of chicken caecal content containing FQ-resistant E. coli obtained from chickens at slaughter age was used as inoculum along with a spiked FQ-susceptible Campylobacter jejuni strain isolated from broilers. Flu was added to one of the fermenters (Flu-fermenter) every 24 hours for two days to evaluate the selection and maintenance of FQ resistance over time, while the other served as a control (C-Fermenter). The experiment duration was 5 days. Samples were collected at three different time points: before, during and after Flu administration. Serial dilutions were plated on Butzler culture media with and without Flu (8mg/L) and enrofloxacin (4mg/L) and on MacConkey culture media with and without Flu (4mg/L) and enrofloxacin (1mg/L) to determine the proportion of resistant strains over time. Positive cultures were identified by mass spectrometry and matrix-assisted laser desorption/ionization (MALDI). A subset of the obtained isolates were used for Whole Genome Sequencing analysis. Over time, E. coli exhibited positive growth in both fermenters, while C. jejuni growth was detected up to day 3. The proportion of Flu-resistant E. coli strains recovered remained consistent over time after antibiotic selective pressure, while in the C-fermenter, a decrease was observed at day 5; a similar pattern was observed in the enrofloxacin-resistant E. coli strains. This suggests that Flu might play a role in the selection and persistence of enrofloxacin resistance, compared to C-fermenter, where enrofloxacin-resistant E. coli strains appear at a later time. Furthermore, positive growth was detected from both fermenters only on Butzler plates without antibiotics. A subset of C. jejuni strains from the Flu-fermenter revealed that those strains were susceptible to ciprofloxacin (MIC < 0.12 μg/mL). A selection of E. coli strains from both fermenters revealed the presence of plasmid-mediated quinolone resistance (PMQR) (qnr-B19) in only one strain from the C-fermenter belonging to sequence type (ST) 48, and in all from Flu-fermenter belonged to ST189. Our results showed that Flu selective impact on PMQR-positive E. coli strains, while no effect was observed in C. jejuni. Maintenance of Flu-resistance was correlated with antibiotic selective pressure. Further studies into antibiotic resistance gene transfer among commensal and zoonotic bacteria in the chicken caeca content may help to elucidate the resistance spread mechanisms.

Keywords: fluoroquinolone-resistance, escherichia coli, campylobacter jejuni, in vitro model

Procedia PDF Downloads 62