Search results for: column chromatography

Authors: Asma Bashir, Neha Farid, Kashif Ali, Kiran Fatima

Abstract:

Background: Bacteriocins are a subclass of antimicrobial peptides that are becoming extremely important in treatments. It is possible to utilise bacteriocins in place of or in addition to traditional antibiotics. It is possible to treat a variety of infections, including Vancomycin-Resistant Staphylococcus aureus (VRSA) and Methicillin-Resistant Staphylococcus aureus (MRSA), using the targeted spectrum of activity of these microorganisms. Method: This study aimed to examine the efficiency of antibiotics and bacteriocin against VRSA and MRSA. The effects of bacteriocins, such as enterocin KAE01, enterocin KAE03, enterocin KAE05, and enterocin KAE06 isolated from Enterococcus faecium strains, alone and in combination with vancomycin and methicillin antibiotics were examined. The selection technique utilized the minimum inhibitory concentrations (MICs) against Gram-positive indicator strain ATCC 6538 Methicillin-Resistant Staphylococcus aureus (MRSA) and indicator strain KSA 02 Vancomycin-Resistant Staphylococcus aureus (VRSA). Results: We report the isolation and identification of enterocins KAE01, KAE03, KAE05, and KAE06 from food isolates of Enterococcus faecium (KAE01, KAE03, KAE05, and KAE06). After isolating the protein, it was partially purified with ammonium sulphate precipitation and purified with fast protein liquid chromatography (FPLC) procedures. Combinations of enterocin KAE01, 1 citric acid, 1 lactic acid, and microcin J25, 1 reuterin, 1 citric acid, and microcin J25, 1 reuterin, 1 lactic acid shown synergistic benefits (FIC index = 0.5) against Vancomycin-Resistant Staphylococcus aureus (VRSA). In addition, a moderately synergistic (FIC index = 0.75) interaction was seen between pediocin PA-1, 1 citric acid, 1 lactic acid, and reuterin 1 citric acid, 1 lactic acid against L. ivanovii HPB28. In the presence of acids, nisin Z exhibited a modestly synergistic effect (FIC index = 0.625-0.75); however, it exhibited additive effects (FIC index = 1) when combined with reuterin or pediocin PA-1 against L. ivanovii HPB28. The efficacy of synergistic consortiums against Gram-positive bacteria was examined. Conclusion: Combining antimicrobials with various modes of action boosted efficacy and expanded the spectrum of inhibition, particularly against multidrug-resistant pathogens, according to our research.

Keywords: Enterococcus faecium, bacteriocin, antimicrobial resistance, antagonistic activity, vancomycin-resistant Staphylococcus aureus, methicillin-resistant Staphylococcus aureus

Procedia PDF Downloads 144

Authors: Sujata Sharma, Avinash Singh, Lovely Gautam, Pradeep Sharma, Mau Sinha, Asha Bhushan, Punit Kaur, Tej P. Singh

Abstract:

Peptidyl-tRNA hydrolase (Pth) Pth is an essential bacterial enzyme that catalyzes the release of free tRNA and peptide moeities from peptidyl tRNAs during stalling of protein synthesis. In order to design inhibitors of Pth from Pseudomonas aeruginosa (PaPth), we have determined the structures of PaPth in its native state and in the bound states with two compounds, amino acylate-tRNA analogue (AAtA) and 5-azacytidine (AZAC). The peptidyl-tRNA hydrolase gene from Pseudomonas aeruginosa was amplified by Phusion High-Fidelity DNA Polymerase using forward and reverse primers, respectively. The E. coliBL21 (λDE3) strain was used for expression of the recombinant peptidyl-tRNA hydrolase from Pseudomonas aeruginosa. The protein was purified using a Ni-NTA superflow column. The crystallization experiments were carried out using hanging drop vapour diffusion method. The crystals diffracted to 1.50 Å resolution. The data were processed using HKL-2000. The polypeptide chain of PaPth consists of 194 amino acid residues from Met1 to Ala194. The centrally located β-structure is surrounded by α-helices from all sides except the side that has entrance to the substrate binding site. The structures of the complexes of PaPth with AAtA and AZAC showed the ligands bound to PaPth in the substrate binding cleft and interacted with protein atoms extensively. The residues that formed intermolecular hydrogen bonds with the atoms of AAtA included Asn12, His22, Asn70, Gly113, Asn116, Ser148, and Glu161 of the symmetry related molecule. The amino acids that were involved in hydrogen bonded interactions in case of AZAC included, His22, Gly113, Asn116, and Ser148. As indicated by fittings of two ligands and the number of interactions made by them with protein atoms, AAtA appears to be a more compatible with the structure of the substrate binding cleft. However, there is a further scope to achieve a better stacking than that of O-tyrosyl moiety because it is not still ideally stacked. These observations about the interactions between the protein and ligands have provided the information about the mode of binding of ligands, nature and number of interactions. This information may be useful for the design of tight inhibitors of Pth enzymes.

Keywords: peptidyl tRNA hydrolase, Acinetobacter baumannii, Pth enzymes, O-tyrosyl

Procedia PDF Downloads 427

Authors: Manohar Salla, Mark S. Butler, Ruby Pelingon, Geraldine Kaeslin, Daniel E. Croker, Janet C. Reid, Jong Min Baek, Paul V. Bernhardt, Elizabeth M. J. Gillam, Matthew A. Cooper, Avril A. B. Robertson

Abstract:

MCC950 is a potent and selective inhibitor of the NOD-like receptor pyrin domain-containing protein 3 (NLRP3) inflammasome that shows early promise for treatment of inflammatory diseases. The identification of major metabolites of lead molecule is an important step during drug development process. It provides an information about the metabolically labile sites in the molecule and thereby helping medicinal chemists to design metabolically stable molecules. To identify major metabolites of MCC950, the compound was incubated with human liver microsomes and subsequent analysis by (+)- and (−)-QTOF-ESI-MS/MS revealed a major metabolite formed due to hydroxylation on 1,2,3,5,6,7-hexahydro-s-indacene moiety of MCC950. This major metabolite can lose two water molecules and three possible regioisomers were synthesized. Co-elution of major metabolite with each of the synthesized compounds using HPLC-ESI-SRM-MS/MS revealed the structure of the metabolite (±) N-((1-hydroxy-1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-4-(2-hydroxypropan-2-yl)furan-2-sulfonamide. Subsequent synthesis of individual enantiomers and coelution in HPLC-ESI-SRM-MS/MS using a chiral column revealed the metabolite was R-(+)- N-((1-hydroxy-1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-4-(2-hydroxypropan-2-yl)furan-2-sulfonamide. To study the possible cytochrome P450 enzyme(s) responsible for the formation of major metabolite, MCC950 was incubated with a panel of cytochrome P450 enzymes. The result indicated that CYP1A2, CYP2A6, CYP2B6, CYP2C9, CYP2C18, CYP2C19, CYP2J2 and CYP3A4 are most likely responsible for the formation of the major metabolite. The biological activity of the major metabolite and the other synthesized regioisomers was also investigated by screening for for NLRP3 inflammasome inhibitory activity and cytotoxicity. The major metabolite had 170-fold less inhibitory activity (IC50-1238 nM) than MCC950 (IC50-7.5 nM). Interestingly, one regioisomer had shown nanomolar inhibitory activity (IC50-232 nM). However, no evidence of cytotoxicity was observed with any of these synthesized compounds when tested in human embryonic kidney 293 cells (HEK293) and human liver hepatocellular carcinoma G2 cells (HepG2). These key findings give an insight into the SAR of the hexahydroindacene moiety of MCC950 and reveal a metabolic soft spot which could be blocked by chemical modification.

Keywords: Cytochrome P450, inflammasome, MCC950, metabolite, microsome, NLRP3

Procedia PDF Downloads 247

Authors: Anthony Credoz, Nathalie Nief, Remy Hedacq, Salvador Jordana, Laurent Cazes

Abstract:

Groundwater monitoring is classically performed in a network of piezometers in industrial sites. Groundwater flow parameters, such as direction, sense and velocity, are deduced from indirect measurements between two or more piezometers. Groundwater sampling is generally done on the whole column of water inside each borehole to provide concentration values for each piezometer location. These flow and concentration values give a global ‘static’ image of potential plume of contaminants evolution in the shallow aquifer with huge uncertainties in time and space scales and mass discharge dynamic. TOTAL R&D Subsurface Environmental team is challenging this classical approach with an innovative dynamic way of characterization of shallow aquifer groundwater. The current study aims at optimizing the tools and methodologies for (i) a direct and multilevel measurement of groundwater velocities in each piezometer and, (ii) a calculation of potential flux of dissolved contaminant in the shallow aquifer. Lab-scale experiments have been designed to test commercial and R&D tools in a controlled sandbox. Multiphysics modeling were performed and took into account Darcy equation in porous media and Navier-Stockes equation in the borehole. The first step of the current study focused on groundwater flow at porous media/piezometer interface. Huge uncertainties from direct flow rate measurements in the borehole versus Darcy flow rate in the porous media were characterized during experiments and modeling. The structure and location of the tools in the borehole also impacted the results and uncertainties of velocity measurement. In parallel, direct-push tool was tested and presented more accurate results. The second step of the study focused on mass flux of dissolved contaminant in groundwater. Several active and passive commercial and R&D tools have been tested in sandbox and reactive transport modeling has been performed to validate the experiments at the lab-scale. Some tools will be selected and deployed in field assays to better assess the mass discharge of dissolved contaminants in an industrial site. The long-term subsurface environmental strategy is targeting an in-situ, real-time, remote and cost-effective monitoring of groundwater.

Keywords: dynamic characterization, groundwater flow, lab-scale, mass flux

Procedia PDF Downloads 163

Authors: Fereydoon Bondarian, Samira Shaygan

Abstract:

Today, many nutritionists recommend the consumption of plants, fruits, and vegetables to provide the antioxidants needed by the body because the use of plant antioxidants usually causes fewer side effects and better treatment. Natural antioxidants increase the power of plasma antioxidants and reduce the incidence of some diseases, such as cancer. Bad lifestyles and environmental factors play an important role in increasing the incidence of cancer. In this study, different extracts of white teas taken from two types of tea available in Iran (clone 100 and Chinese hybrid) due to the presence of a hydroxyl functional group in their structure to inhibit free radicals and anticancer properties, using 3 aqueous, methanolic and aqueous-methanolic methods were used. The total polyphenolic content was calculated using the Folin-Ciocalcu method, and the percentage of inhibition and trapping of free radicals in each of the extracts was calculated using the DPPH method. With the help of high-performance liquid chromatography, a small amount of each catechin in the tea samples was obtained. Clone 100 white tea was found to be the best sample of tea in terms of all the examined attributes (total polyphenol content, antioxidant properties, and individual amount of each catechin). The results showed that aqueous and aqueous-methanolic extracts of Clone 100 white tea have the highest total polyphenol content with 27.59±0.08 and 36.67±0.54 (equivalent gallic acid per gram dry weight of leaves), respectively. Due to having the highest level of different groups of catechin compounds, these extracts have the highest property of inhibiting and trapping free radicals with 66.61±0.27 and 71.74±0.27% (mg/l) of the extracted sample against ascorbic acid). Using the MTT test, the inhibitory effect of clone 100 white tea extract in inhibiting the growth of HCT-116 colon cancer cells was investigated and the best time and concentration treatments were 500, 150 and 1000 micrograms in 8, 16 and 24 hours, respectively. To investigate gene expression changes, selected genes, including tumorigenic genes, proto-oncogenes, tumor suppressors, and genes involved in apoptosis, were selected and analyzed using the real-time PCR method and in the presence of concentrations obtained for white tea. White tea extract at a concentration of 1000 μg/ml 3 times 16, 8, and 24 hours showed the highest growth inhibition in cancer cells with 53.27, 55.8, and 86.06%. The concentration of 1000 μg/ml aqueous extract of white tea under 24-hour treatment increased the expression of tumor suppressor genes compared to the normal sample.

Keywords: catechin, gene expression, suppressor genes, colon cell line

Procedia PDF Downloads 54

Authors: Ali Zaker, Zhi Chen

Abstract:

Due to the concerns about the depletion of fossil fuel sources and the deteriorating environment, the attempt to investigate the production of renewable energy will play a crucial role as a potential to alleviate the dependency on mineral fuels. One particular area of interest is the generation of bio-oil through sewage sludge (SS) pyrolysis. SS can be a potential candidate in contrast to other types of biomasses due to its availability and low cost. However, the presence of high molecular weight hydrocarbons and oxygenated compounds in the SS bio-oil hinders some of its fuel applications. In this context, catalytic pyrolysis is another attainable route to upgrade bio-oil quality. Among different catalysts (i.e., zeolites) studied for SS pyrolysis, activated chars (AC) are eco-friendly alternatives. The beneficial features of AC derived from SS comprise the comparatively large surface area, porosity, enriched surface functional groups, and presence of a high amount of metal species that can improve the catalytic activity. Hence, a sludge-based AC catalyst was fabricated in a single-step pyrolysis reaction with NaOH as the activation agent and was compared with HZSM5 zeolite in this study. The thermal decomposition and kinetics were invested via thermogravimetric analysis (TGA) for guidance and control of pyrolysis and catalytic pyrolysis and the design of the pyrolysis setup. The results indicated that the pyrolysis and catalytic pyrolysis contains four obvious stages, and the main decomposition reaction occurred in the range of 200-600°C. The Coats-Redfern method was applied in the 2nd and 3rd devolatilization stages to estimate the reaction order and activation energy (E) from the mass loss data. The average activation energy (Em) values for the reaction orders n = 1, 2, and 3 were in the range of 6.67-20.37 kJ for SS; 1.51-6.87 kJ for HZSM5; and 2.29-9.17 kJ for AC, respectively. According to the results, AC and HZSM5 both were able to improve the reaction rate of SS pyrolysis by abridging the Em value. Moreover, to generate and examine the effect of the catalysts on the quality of bio-oil, a fixed-bed pyrolysis system was designed and implemented. The composition analysis of the produced bio-oil was carried out via gas chromatography/mass spectrometry (GC/MS). The selected SS to catalyst ratios were 1:1, 2:1, and 4:1. The optimum ratio in terms of cracking the long-chain hydrocarbons and removing oxygen-containing compounds was 1:1 for both catalysts. The upgraded bio-oils with AC and HZSM5 were in the total range of C4-C17, with around 72% in the range of C4-C9. The bio-oil from pyrolysis of SS contained 49.27% oxygenated compounds, while with the presence of AC and HZSM5 dropped to 13.02% and 7.3%, respectively. Meanwhile, the generation of benzene, toluene, and xylene (BTX) compounds was significantly improved in the catalytic process. Furthermore, the fabricated AC catalyst was characterized by BET, SEM-EDX, FT-IR, and TGA techniques. Overall, this research demonstrated AC is an efficient catalyst in the pyrolysis of SS and can be used as a cost-competitive catalyst in contrast to HZSM5.

Keywords: catalytic pyrolysis, sewage sludge, activated char, HZSM5, bio-oil

Procedia PDF Downloads 175

Authors: Jason Gica, Yi-Hsieng Samuel Wu, Deng-Jye Yang, Yi-Chen Chen

Abstract:

Mycotoxins, harmful secondary metabolites produced by certain fungi species, pose significant risks to animals and humans worldwide. Their stable properties lead to contamination during grain harvesting, transportation, and storage, as well as in processed food products. The prevalence of mycotoxin contamination has attracted significant attention due to its adverse impact on food safety and global trade. The secondary contamination pathway from animal products has been identified as an important route of exposure, posing health risks for livestock and humans consuming contaminated products. Pork, one of the highly consumed meat products in Taiwan according to the National Food Consumption Database, plays a critical role in the nation's diet and economy. Given its substantial consumption, pork processing products are a significant component of the food supply chain and a potential source of mycotoxin contamination. This study is paramount for formulating effective regulations and strategies to mitigate mycotoxin-related risks in the food supply chain. By establishing a reliable analytical method, this research contributes to safeguarding public health and enhancing the quality of pork processing products. The findings will serve as valuable guidance for policymakers, food industries, and consumers to ensure a safer food supply chain in the face of emerging mycotoxin challenges. An innovative and efficient analytical approach is proposed using Ultra-High Performance Liquid Chromatography coupled with Temperature Control Fluorescence Detector Light (UHPLC-TCFLD) to determine multiple mycotoxins in pork meat samples due to its exceptional capacity to detect multiple mycotoxins at the lowest levels of concentration, making it highly sensitive and reliable for comprehensive mycotoxin analysis. Additionally, its ability to simultaneously detect multiple mycotoxins in a single run significantly reduces the time and resources required for analysis, making it a cost-effective solution for monitoring mycotoxin contamination in pork processing products. The research aims to optimize the efficient mycotoxin QuEChERs extraction method and rigorously validate its accuracy and precision. The results will provide crucial insights into mycotoxin levels in pork processing products.

Keywords: multiple-mycotoxin analysis, pork processing products, QuEChERs, UHPLC-TCFLD, validation

Procedia PDF Downloads 62

Authors: Anuranjani Kumar, Madhu Bala

Abstract:

Ionising radiation exposure induces generation of free radicals and the oxidative DNA damage. SBL-1, a radioprotective leaf extract prepared from leaves Hippophae rhamnoides L. (Common name; Seabuckthorn), showed > 90% survival in mice population that was treated with lethal dose (10 Gy) of ⁶⁰Co gamma irradiation. In this study, early effects of pre-treatment with or without SBL-1 in blood peripheral blood lymphocytes (PBMCs) were investigated by cell viability assays (trypan blue and MTT). The quantitative in vitro study of Hoescht/PI staining was performed to check the apoptosis/necrosis in PBMCs irradiated at 2 Gy with or without pretreatment of SBL-1 (at different concentrations) up to 24 and 48h. Comet assay was performed in vivo, to detect the DNA strands breaks and its repair mechanism on peripheral blood lymphocytes at lethal dose (10 Gy). For this study, male mice (wt. 28 ± 2g) were administered radioprotective dose (30mg/kg body weight) of SBL-1, 30 min prior to irradiation. Animals were sacrificed at 24h and 48h. Blood was drawn through cardiac puncture, and blood lymphocytes were separated using histopaque column. Both neutral and alkaline comet assay were performed using standardized technique. In irradiated animals, alkaline comet assay revealed single strand breaks (SSBs) that showed significant (p < 0.05) increase in percent DNA in tail and Olive tail moment (OTM) at 24 h while at 48h the percent DNA in tail further increased significantly (p < 0.02). The double strands breaks (DSBs) increased significantly (p < 0.01) at 48 h in neutral assay, in comparison to untreated control. The animals pre-treated with SBL-1 before irradiation showed significantly (p < 0.05) less DSBs at 48 h treatment in comparison to irradiated group of animals. The SBL-1 alone treated group itself showed no toxicity. The antioxidant potential of SBL-1 were also investigated by in vitro biochemical assays such as DPPH (p < 0.05), ABTS, reducing ability (p < 0.09), hydroxyl radical scavenging (p < 0.05), ferric reducing antioxidant power (FRAP), superoxide radical scavenging activity (p < 0.05), hydrogen peroxide scavenging activity (p < 0.05) etc. SBL-1 showed strong free radical scavenging power that plays important role in the studies of radiation-induced injuries. The SBL-1 treated PBMCs showed significant (p < 0.02) viability in trypan blue assay at 24-hour incubation.

Keywords: radiation, SBL-1, SSBs, DSBs, FRAP, PBMCs

Procedia PDF Downloads 151

Authors: Meri Khakhutaishvili, Indira Djaparidze, Maia Vanidze, Aleko Kalandia

Abstract:

Cultivation of blueberry in Georgia started in 21st century. There are more than 20 species of blueberry cultivated in this region from all other the world. The species are mostly planted on acidic soil, previously occupied by tea plantations. Many of the plantations have pretty good yield. It is known that changing the location of a plant to a new soil or climate effects chemical compositions of the plant. However, even though these plants are brought from other countries, no research has been conducted to fully examine the blueberry fruit cultivated in Georgia. Shota Rustaveli National Science Foundation Grant FR/335/10-160/14, gave us an opportunity to continue our previous works and conduct research on several berries, among them of course the chemical composition of stored Blueberry. We were able to conduct the first study that included examining qualitative and quantitative features of bioactive compounds in Georgian Blueberry. This experiments were held in the ‘West Georgia Regional Chromatography center’ (Grant AP/96/13) of our university, that is equipped with modern equipment like HPLC UV-Vis, RI-detector, HPLC-conductivity detector, UPLC-MS-detector. Biochemical analysis was conducted using different physico-chemical and instrumental methods. Separation-identification and quantitative analysis were conducted using UPLC-MS (Waters Acquity QDa detector), HPLC (Waters Brceze 1525, UV-Vis 2489 detectors), pH-meters (Mettler Toledo). Refractrometer -Misco , Spectrometer –Cuvette Changer (Mettler Toledo UV5A), C18 Cartridge Solid Phase Extraction (SPE) Waters Sep-Pak C18 (500 mg), Chemicals – stability radical- 2,2-Diphenil-1-picrilhydrazyl (Aldrich-germany), Acetonitrile, Methanol, Acetic Acid (Merck-Germany), AlCl3, Folin Ciocalteu reagent (preparation), Standarts –Callic acid, Quercetin. Carbohydrate HPLC-RI analysis used systems acetonitrile-water (80-20). UPLC-MS analysis used systems- solvent A- Water +1 % acetic acid და solvent -B Methanol +1% acetic acid). It was concluded that the amount of sugars was in range of 5-9 %, mostly glucose and fructose. Also, the amount of organic acids was 0.2-1.2% most of which was malic and citric acid. Anthocians were also present in the sample 200-550mg/100g. We were able to identify up to 15 different compounds, most of which were products of delphinidine and cyanide. All species have high antioxidant level(DPPH). By rapidly freezing the sample and then keeping it in specific conditions allowed us to keep the sample for 12 months.

Keywords: antioxidants, bioactive, blueberry, storage

Procedia PDF Downloads 208

Authors: Lara Marie Pangan Lo, Soo Im Chung, Yao Cheng Zhang, Xingyue Jin, Mi Young Kang

Abstract:

Being the world’s most consumed grain crop, rice (Oryza sativa L.) demands’ have increase and this prompted the development of new rice cultivars with high bio-functional properties than the commonly used white rice. Ordinary rice variety is already known to be a potential source for a number of nutritional as well as bioactive compounds. To further enhance the rice’s nutritive values, germination is done in addition to making it more tasty and palatable when cooked. Pigmented rice, on the other hand, has become increasingly popular in the recent years for their greater antioxidant potential and other nutraceutical properties which can help alleviate the occurrence of the increasing incidence of metabolic diseases. Combining these two (2) parameters, this research study is sought to quantitatively determine the pre-germinated and germinated quantities of the major bioactive compounds of South Korea’s newly developed purplish pigmented rice grain cultivar Superjami (SJ) and red pigmented rice grain Superhongmi (SH) and compare them against the non-pigmented Normal Brown (NB) rice variety. Powdered rice grain cultivars were subjected to 72-hour germination period and the quantities of GABA, γ-oryzanol, ferulic acid, tocopherol and tocotrienol homologues were compared against their pre-germinated condition using γ- amino butyric acid (GABA) analysis and High Performance Liquid Chromatography (HPLC). Results revealed the effectiveness of germination in enhancing the bioactive components in all rice samples. GABA contents in germinated rice cultivars increased by more than 10-fold following the order: SJ >SH >NB. In addition, purple rice variety (SJ) has higher total γ-oryzanol and ferulic acid contents which increased by > 2-fold after germination followed by the red cultivar SH then the control, NB. Germinated varieties also possess higher total tocotrienol content than their pre-germinated state. As for the total tocopherol content, SJ has higher quantity, but the red-pigmented SH (0.16 mg/kg) is shown to have lower total tocopherol content than the control rice NB (0.86 mg/kg). However, all tocopherol and tocotrienol homologues were present only in small amounts ( < 3.0 mg/kg) in all pre-germinated and germinated samples. In general, all of the analyzed pigmented rice cultivars were found to possess higher bioactive compounds than the control NB rice variety. Also, regardless of their strain, germinated rice samples have higher bioactive compounds than their pre-germinated counterparts. This only shows the effectiveness of germinating rice in enhancing bioactive constituents. Overall, these results suggest the potential of the pigmented rice varieties as natural source of nutraceuticals in bio-functional food development.

Keywords: bioactive compounds, germinated rice, superhongmi, superjami

Procedia PDF Downloads 395

Authors: Momita Das, Sofia Banu, Jibon Kotoky

Abstract:

Environmental contamination of natural resources with persistent organic pollutants is of great world-wide apprehension. Polycyclic aromatic hydrocarbons (PAHs) are among the organic pollutants, released due to various anthropogenic activities. Due to their toxic, carcinogenic and mutagenic properties, PAHs are of environmental and human concern. Presently, bioremediation has evolved as the most promising biotechnology for cleanup of such contaminants because of its economical and less cost effectiveness. In the present study, distribution of 16 USEPA priority PAHs was determined in the soil samples collected from fifteen different sites of Guwahati City, the Gateway of the North East Region of India. The total concentrations of 16 PAHs (Σ16 PAHs) ranged from 42.7-742.3 µg/g. Higher concentration of total PAHs was found more in the Industrial areas compared to all the sites (742.3 µg/g and 628 µg/g). It is noted that among all the PAHs, Naphthalene, Acenaphthylene, Anthracene, Fluoranthene, Chrysene and Benzo(a)Pyrene were the most available and contain the higher concentration of all the PAHs. Since microbial activity has been deemed the most influential and significant cause of PAH removal; further, twenty-three bacteria were isolated from the most contaminated sites using the enrichment process. These strains were acclimatized to utilize naphthalene and anthracene, each at 100 µg/g concentration as sole carbon source. Among them, one Gram-positive strain (JMG-01) was selected, and biodegradation ability and initial catabolic genes of PAHs degradation were investigated. Based on 16S rDNA analysis, the isolate was identified as Bacillus cereus strain JMG-01. Topographic images obtained using Scanning Electron Microscope (SEM) and Atomic Force Microscope (AFM) at scheduled time intervals of 7, 14 and 21 days, determined the variation in cell morphology during the period of degradation. AFM and SEM micrograph of biomass showed high filamentous growth leading to aggregation of cells in the form of biofilm with reference to the incubation period. The percentage degradation analysis using gas chromatography and mass analyses (GC-MS) suggested that more than 95% of the PAHs degraded when the concentration was at 500 µg/g. Naphthalene, naphthalene-2-methy, benzaldehyde-4-propyl, 1, 2, benzene di-carboxylic acid and benzene acetic acid were the major metabolites produced after degradation. Moreover, PCR experiments with specific primers for catabolic genes, ndo B and Cat A suggested that JMG-01 possess genes for PAHs degradation. Thus, the study concludes that Bacillus cereus strain JMG-01 has efficient biodegrading ability and can trigger the clean-up of PAHs contaminated soil.

Keywords: AFM, Bacillus cereus strain JMG-01, degradation, polycyclic aromatic hydrocarbon, SEM

Procedia PDF Downloads 268

Authors: A. Sekhar, N. Ramakrishna Raju

Abstract:

This paper is a result of ground improvement using deep vibro techniques with combination of sand and stone columns performed on a highly liquefaction susceptible site (70 to 80% sand strata and balance silt) with low bearing capacities due to high settlements located (earth quake zone V as per IS code) at Madhepura, Bihar state in northern part of India. Initially, it was envisaged with bored cast in-situ/precast piles, stone/sand columns. However, after detail analysis to address both liquefaction and improve bearing capacities simultaneously, it was analyzed the deep vibro techniques with combination of sand and stone columns is excellent solution for given site condition which may be first time in India. First after detail soil investigation, pre eCPT test was conducted to evaluate the potential depth of liquefaction to densify silty sandy soils to improve factor of safety against liquefaction. Then trail test were being carried out at site by deep vibro compaction technique with sand and stone columns combination with different spacings of columns in triangular shape with different timings during each lift of vibro up to ground level. Different spacings and timing was done to obtain the most effective spacing and timing with vibro compaction technique to achieve maximum densification of saturated loose silty sandy soils uniformly for complete treated area. Then again, post eCPT test and plate load tests were conducted at all trail locations of different spacings and timing of sand and stone columns to evaluate the best results for obtaining the required factor of safety against liquefaction and the desired bearing capacities with reduced settlements for construction of industrial structures. After reviewing these results, it was noticed that the ground layers are densified more than the expected with improved factor of safety against liquefaction and achieved good bearing capacities for a given settlements as per IS codal provisions. It was also worked out for cost-effectiveness of lightly loaded single storied structures by using deep vibro technique with sand column avoiding stone. The results were observed satisfactory for resting the lightly loaded foundations. In this technique, the most important is to mitigating liquefaction with improved bearing capacities and reduced settlements to acceptable limits as per IS: 1904-1986 simultaneously up to a depth of 19M. To our best knowledge it was executed first time in India.

Keywords: ground improvement, deep vibro techniques, liquefaction, bearing capacity, settlement

Procedia PDF Downloads 191

Authors: Neha Sahu, Awantika Singh, Brijesh Kumar, K. R. Arya

Abstract:

Taraxacum officinale Weber or dandelion (Asteraceae) is an important Indian traditional herb used to treat liver detoxification, digestive problems, spleen, hepatic and kidney disorders, etc. The plant is well known to possess important phenolic and flavonoids to serve as a potential source of antioxidative and chemoprotective agents. Biosynthesis of bioactive compounds through in vitro cultures is a requisite for natural resource conservation and to provide an alternative source for pharmaceutical applications. Thus an efficient and reproducible protocol was developed for in vitro biosynthesis of bioactive antioxidative compounds from leaf derived callus and in vitro regenerated cultures of Taraxacum officinale using MS media fortified with various combinations of auxins and cytokinins. MS media containing 0.25 mg/l 2, 4-D (2, 4-Dichloro phenoxyacetic acid) with 0.05 mg/l 2-iP [N6-(2-Isopentenyl adenine)] was found as an effective combination for the establishment of callus with 92 % callus induction frequency. Moreover, 2.5 mg/l NAA (α-Naphthalene acetic acid) with 0.5 mg/l BAP (6-Benzyl aminopurine) and 1.5 mg/l NAA showed the optimal response for in vitro plant regeneration with 80 % regeneration frequency and rooting respectively. In vitro regenerated plantlets were further transferred to soil and acclimatized. Quantitative variability of accumulated bioactive compounds in cultures (in vitro callus, plantlets and acclimatized) were determined through UPLC-MS/MS (ultra-performance liquid chromatography-triple quadrupole-linear ion trap mass spectrometry) and compared with wild plants. The phytochemical determination of in vitro and wild grown samples showed the accumulation of 6 compounds. In in vitro callus cultures and regenerated plantlets, two major antioxidative compounds i.e. chlorogenic acid (14950.0 µg/g and 4086.67 µg/g) and umbelliferone (10400.00 µg/g and 2541.67 µg/g) were found respectively. Scopoletin was found to be highest in vitro regenerated plants (83.11 µg/g) as compared to wild plants (52.75 µg/g). Notably, scopoletin is not detected in callus and acclimatized plants, but quinic acid (6433.33 µg/g) and protocatechuic acid (92.33 µg/g) were accumulated at the highest level in acclimatized plants as compared to other samples. Wild grown plants contained highest content (948.33 µg/g) of flavonoid glycoside i.e. luteolin-7-O-glucoside. Our data suggests that in vitro callus and regenerated plants biosynthesized higher content of antioxidative compounds in controlled conditions when compared to wild grown plants. These standardized cultural conditions may be explored as a sustainable source of plant materials for enhanced production and adequate supply of oxidative polyphenols.

Keywords: anti-oxidative compounds, in vitro cultures, Taraxacum officinale, UPLC-MS/MS

Procedia PDF Downloads 200

Authors: Zoltan Sebestyen, Eszter Barta-Rajnai, Emma Jakab, Zsuzsanna Czegeny

Abstract:

Extraction of the energy content of lignocellulosic biomass is one of the possible pathways to reduce the greenhouse gas emission derived from the burning of the fossil fuels. The application of the bioenergy can mitigate the energy dependency of a country from the foreign natural gas and the petroleum. The diversity of the plant materials makes difficult the utilization of the raw biomass in power plants. This problem can be overcome by the application of thermochemical techniques. Pyrolysis is the thermal decomposition of the raw materials under inert atmosphere at high temperatures, which produces pyrolysis gas, biooil and charcoal. The energy content of these products can be exploited by further utilization. The differences in the chemical and physical properties of the raw biomass materials can be reduced by the use of torrefaction. Torrefaction is a promising mild thermal pretreatment method performed at temperatures between 200 and 300 °C in an inert atmosphere. The goal of the pretreatment from a chemical point of view is the removal of water and the acidic groups of hemicelluloses or the whole hemicellulose fraction with minor degradation of cellulose and lignin in the biomass. Thus, the stability of biomass against biodegradation increases, while its energy density increases. The volume of the raw materials decreases so the expenses of the transportation and the storage are reduced as well. Biooil is the major product during pyrolysis and an important by-product during torrefaction of biomass. The composition of biooil mostly depends on the quality of the raw materials and the applied temperature. In this work, thermoanalytical techniques have been used to study the qualitative and quantitative composition of the pyrolysis and torrefaction oils of a woody (black locust) and two herbaceous samples (rape straw and wheat straw). The biooil contains C5 and C6 anhydrosugar molecules, as well as aromatic compounds originating from hemicellulose, cellulose, and lignin, respectively. In this study, special emphasis was placed on the formation of the lignin monomeric products. The structure of the lignin fraction is different in the wood and in the herbaceous plants. According to the thermoanalytical studies the decomposition of lignin starts above 200 °C and ends at about 500 °C. The lignin monomers are present among the components of the torrefaction oil even at relatively low temperatures. We established that the concentration and the composition of the lignin products vary significantly with the applied temperature indicating that different decomposition mechanisms dominate at low and high temperatures. The evolutions of decomposition products as well as the thermal stability of the samples were measured by thermogravimetry/mass spectrometry (TG/MS). The differences in the structure of the lignin products of woody and herbaceous samples were characterized by the method of pyrolysis-gas chromatography/mass spectrometry (Py-GC/MS). As a statistical method, principal component analysis (PCA) has been used to find correlation between the composition of lignin products of the biooil and the applied temperatures.

Keywords: pyrolysis, torrefaction, biooil, lignin

Procedia PDF Downloads 325

Authors: Soad Abubakr Abdelgalil, Gaber Attia Abo-Zaid, Mohamed Mohamed Yousri Kaddah

Abstract:

Background: The Environmental Protection Agency has listed eggshell waste as the 15th most significant food industry pollution hazard. The utilization of eggshell waste as a source of renewable energy has been a hot topic in recent years. Therefore, finding a sustainable solution for the recycling and valorization of eggshell waste by investigating its potential to produce acid phosphatase (ACP) and organic acids by the newly-discovered B. sonorensis was the target of the current investigation. Results: The most potent ACP-producing B. sonorensis strain ACP2 was identified as a local bacterial strain obtained from the effluent of paper and pulp industries on basis of molecular and morphological characterization. The use of consecutive statistical experimental approaches of Plackett-Burman Design (PBD), and Orthogonal Central Composite Design (OCCD), followed by pH-uncontrolled cultivation conditions in a 7 L bench-top bioreactor, revealed an innovative medium formulation that substantially improved ACP production, reaching 216 U L⁻¹ with ACP yield coefficient Yp/x of 18.2 and a specific growth rate (µ) of 0.1 h⁻¹. The metals Ag+, Sn+, and Cr+ were the most efficiently released from eggshells during the solubilization process by B. sonorensis. The uncontrolled pH culture condition is the most suited and favored setting for improving the ACP and organic acids production simultaneously. Quantitative and qualitative analyses of produced organic acids were carried out using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Lactic acid, citric acid, and hydroxybenzoic acid isomer were the most common organic acids produced throughout the cultivation process. The findings of thermogravimetric analysis (TGA), differential scan calorimeter (DSC), scanning electron microscope (SEM), energy-dispersive spectroscopy (EDS), Fourier-Transform Infrared Spectroscopy (FTIR), and X-Ray Diffraction (XRD) analysis emphasize the significant influence of organic acids and ACP activity on the solubilization of eggshells particles. Conclusions: This study emphasized robust microbial engineering approaches for the large-scale production of a newly discovered acid phosphatase accompanied by organic acids production from B. sonorensis. The biovalorization of the eggshell waste and the production of cost-effective ACP and organic acids were integrated into the current study, and this was done through the implementation of a unique and innovative medium formulation design for eggshell waste management, as well as scaling up ACP production on a bench-top scale.

Keywords: chicken eggshells waste, bioremediation, statistical experimental design, batch fermentation

Procedia PDF Downloads 370

Authors: W. Mohammed Saeed, A. J. Mcbain, S. M. Cruickshank, C. A. O’Neill

Abstract:

In the gut, probiotics have been shown to protect epithelial cells from pathogenic bacteria through a number of mechanisms: 1-Increasing epithelial barrier function, 2-Modulation of the immune response especially innate immune response, 3-Inhibition of pathogen adherence and down regulation of virulence factors. Since probiotics have positive impacts on the gut, their potential effects on other body tissues, such as skin have begun to be investigated. The purpose of this project is to characterize the potential of probiotic bacteria lysate as therapeutic agent for preventing or reducing the S. aureus infection. Normal human primary keratinocytes (KCs) were exposed to S. aureus (106/ml) in the presence or absence of L. rhamnosus GG lysate (extracted from 108cfu/ml). The viability of the KCs was measured after 24 hours using a trypan blue exclusion assay. When KCs were treated with S aureus alone, only 25% of the KCs remained viable at 24 hours post infection. However, in the presence of L. rhamnosus GG lysate the viability of pathogen infected KCs increased to 58% (p=0.008, n=3). Furthermore, when KCs co-exposed, pre- exposed or post-exposed to L. rhamnosus GG lysate, the viability of the KCs increased to ≈60%, the L. rhamnosus GG lysate was afforded equal protection in different conditions. These data suggests that two possible separate mechanisms are involved in the protective effects of L. rhamnosus GG such as reducing S. aureus growth, or inhibiting of pathogenic adhesion. Interestingly, a lysate of L rhamnosus GG provided significant reduction in S. aureus growth and adhesion of S. aureus that being viable following 24 hours incubation with S aureus. Therefore, a series of Liquid Chromatography (RP-LC) methods were adopted to partially purify the lysate in combination with functional assays to elucidate in which fractions the efficacious molecules were contained. In addition, the Mass Spectrometry-based protein sequencing was used to identify putative proteins in the fractions. The data presented from purification process demonstrated that L. rhamnosus GG lysate has the potential to protect keratinocytes from the toxic effects of the skin pathogen, S. aureus. Three potential mechanisms were identified: inhibition of pathogen growth; competitive exclusion; and displacement of the pathogen from keratinocyte binding sites. In this study, ‘moonlight’ proteins were identified in the current study’s MS/MS data for L. rhamnosus GG lysate, which could elucidate the ability of lysate in the competitive exclusion and displacement of S. aureus from keratinocyte binding sites. Taken together, it can be speculated that L. rhamnosus GG lysate utilizes different mechanisms to protect keratinocytes from S. aureus toxicity. The present study indicates that the proteinaceous substances are involved in anti-adhesion activity. This is achieved by displacing the pathogen and preventing the severity of pathogen infection and the moonlight proteins might be involved in inhibiting the adhesion of pathogens.

Keywords: lysate, fractions, adhesion, L. rhamnosus GG, S. aureus toxicity

Procedia PDF Downloads 289

Authors: Maged El-Sayed Mohamed

Abstract:

Eucalyptus camaldulensis is one on the most well-known species of the genus Eucalyptus in the Middle east, its importance relay on the high production of its unique volatile constituents which exhibits many medicinal and pharmacological activities. The gall-forming wasp (Leptocybe invasa) has recently come into sight as the main pest attacking E. camaldulensis and causing severe injury. The wasp lays its eggs in the petiole and midrib of leaves and stems of young shoots of E. camaldulensis, which leads to gall formation. Gall formation by L. invasa damages growing shoot and leaves of Eucalyptus, resulting in abscission of leaves and drying. AIM: This study is an attempt to investigate the effect of the gall wasp (Leptocybe invasa) attack on the volatile constitutes of E. camaldulensis. This could help in the control of this wasp through stimulating plant defenses or production of a new allelochemicals or insecticide. The study of volatile constitutes of Eucalyptus before and after attack by the wasp can help the re-use and recycle of the infected Eucalyptus trees for new pharmacological and medicinal activities. Methodology: The fresh gall wasp-attacked and healthy leaves (100 g each) were cut and immediately subjected to hydrodistillation using Clevenger-type apparatus for 3 hours. The volatile fractions isolated were analyzed using Gas chromatography/mass spectrometry (GC/MS). Kovat’s retention indices (RI) were calculated with respect to a set of co-injected standard hydrocarbons (C10-C28). Compounds were identified by comparing their spectral data and retention indices with Wiley Registry of Mass Spectral Data 10th edition (April 2013), NIST 11 Mass Spectral Library (NIST11/2011/EPA/NIH) and literature data. Results: Fifty-nine components representing 89.13 and 88.60% of the total volatile fraction content respectively were quantitatively analyzed. Twenty-six major compounds at an average concentration greater than 0.1 ± 0.02% have been used for the statistical comparison. From those major components, twenty-one were found in both the attacked and healthy Eucalyptus leaves’ fractions in different concentration and five components, mono terpene p-Mentha-2-4(8) diene and the sesquiterpenes δ-elemene, β-elemene, E-caryophyllene and Bicyclogermacrene, were unique and only produced in the attacked-leaves’ fraction. CONCLUSION: Newly produced components or those commonly found in the volatile fraction and changed in concentration could represent a part of the plant defense mechanisms or might be an element of the plant allelopathic and communication mechanisms. Identification of the components of the gall wasp-damaged leaves can help in their recycling for different physiological, pharmacological and medicinal uses.

Keywords: Eucalyptus camaldulensis, eucalyptus recycling, gall wasp, Leptocybe invasa, plant defense mechanisms, Terpene fraction

Procedia PDF Downloads 354

Authors: A. M. Isakhanyan, F. N. Tkhruni, N. N. Yakimovich, Z. I. Kuvaeva, T. V. Khachatryan

Abstract:

Introduction: At present, the simbiotics based on different genera and species of lactic acid bacteria (LAB) and yeasts are used. One of the basic properties of probiotics is presence of antimicrobial activity and therefore selection of LAB and yeast strains for their co-cultivation with the aim of increasing of the activity is topical. Since probiotic yeast and bacteria have different mechanisms of action, natural synergies between species, higher viability and increasing of antimicrobial activity might be expected from mixing both types of probiotics. Endemic strains of LAB Enterococcus faecium БТK-64, Lactobaccilus plantarum БТK-66, Pediococcus pentosus БТK-28, Lactobacillus rhamnosus БТK-109 and Kluyveromyces lactis БТX-412, Saccharomycopsis sp. БТX- 151 strains of yeast, with probiotic properties and hight antimicrobial activity, were selected. Strains are deposited in "Microbial Depository Center" (MDC) SPC "Armbiotechnology". Methods: LAB and yeast strains were isolated from different dairy products from rural households of Armenia. The genotyping by 16S rRNA sequencing for LAB and 26S RNA sequencing for yeast were used. Combined cultivation of LAB and yeast strains was carried out in the nutrient media on the basis of milk whey, in anaerobic conditions (without shaker, in a thermostat at 37oC, 48 hours). The complex preparations were obtained by purification of cell free culture broth (CFC) broth by the combination of ion-exchange chromatography and gel filtration methods. The spot-on-lawn method was applied for determination of antimicrobial activity and expressed in arbitrary units (AU/ml). Results. The obtained data showed that at the combined growth of bacteria and yeasts, the cultivation conditions (medium composition, time of growth, genera of LAB and yeasts) affected the display of antimicrobial activity. Purification of CFC broth allowed obtaining partially purified antimicrobial complex preparation which contains metabiotics from both bacteria and yeast. The complex preparation inhibited the growth of pathogenic and conditionally pathogenic bacteria, isolated from various internal organs from diseased animals and poultry with greater efficiency than the preparations derived individually alone from yeast and LAB strains. Discussion. Thus, our data shown perspectives of creation of a new class of antimicrobial preparations on the basis of combined cultivation of endemic strains of LAB and yeast. Obtained results suggest the prospect of use of the partially purified complex preparations instead antibiotics in the agriculture and for food safety. Acknowledgments: This work was supported by the RA MES State Committee of Science and Belarus National Foundation for Basic Research in the frames of the joint Armenian - Belarusian joint research project 13РБ-064.

Keywords: co-cultivation, antimicrobial activity, biosafety, metabiotics, lactic acid bacteria, yeast

Procedia PDF Downloads 334

Authors: Mustafa M. Donma, Orkide Donma

Abstract:

The impact of metabolic syndrome (MetS) on cognition and functions of the brain is being investigated. Iron deficiency and deficiencies of B9 (folate) as well as B12 (cobalamin) vitamins are best-known nutritional anemias. They are associated with cognitive disorders and learning difficulties. The antidepressant effects of vitamin D are known and the deficiency state affects mental functions negatively. The aim of this study is to investigate possible correlations of MetS with serum brain-derived neurotrophic factor (BDNF), iron, folate, cobalamin and vitamin D in pediatric patients. 30 children, whose age- and sex-dependent body mass index (BMI) percentiles vary between 85 and 15, 60 morbid obese children with above 99^th percentiles constituted the study population. Anthropometric measurements were taken. BMI values were calculated. Age- and sex-dependent BMI percentile values were obtained using the appropriate tables prepared by the World Health Organization (WHO). Obesity classification was performed according to WHO criteria. Those with MetS were evaluated according to MetS criteria. Serum BDNF was determined by enzyme-linked immunosorbent assay. Serum folate was analyzed by an immunoassay analyzer. Serum cobalamin concentrations were measured using electrochemiluminescence immunoassay. Vitamin D status was determined by the measurement of 25-hydroxycholecalciferol [25-hydroxy vitamin D3, 25(OH)D] using high performance liquid chromatography. Statistical evaluations were performed using SPSS for Windows, version 16. The p values less than 0.05 were accepted as statistically significant. Although statistically insignificant, lower folate and cobalamin values were found in MO children compared to those observed for children with normal BMI. For iron and BDNF values, no alterations were detected among the groups. Significantly decreased vitamin D concentrations were noted in MO children with MetS in comparison with those in children with normal BMI (p ≤ 0.05). The positive correlation observed between iron and BDNF in normal-BMI group was not found in two MO groups. In THE MetS group, the partial correlation among iron, BDNF, folate, cobalamin, vitamin D controlling for waist circumference and BMI was r = -0.501; p ≤ 0.05. None was calculated in MO and normal BMI groups. In conclusion, vitamin D should also be considered during the assessment of pediatric MetS. Waist circumference and BMI should collectively be evaluated during the evaluation of MetS in children. Within this context, BDNF appears to be a key biochemical parameter during the examination of obesity degree in terms of mental functions, cognition and learning capacity. The association observed between iron and BDNF in children with normal BMI was not detected in MO groups possibly due to development of inflammation and other obesity-related pathologies. It was suggested that this finding may contribute to mental function impairments commonly observed among obese children.

Keywords: brain-derived neurotrophic factor, iron, vitamin B9, vitamin B12, vitamin D

Procedia PDF Downloads 118

Authors: Barrera C. Monserrat, Sierra-Alvarez Reyes, Pat-Espadas Aurora, Moreno Andrade Ivan

Abstract:

Antimony is a toxic and carcinogenic metalloid considered a pollutant of priority interest by the United States Environmental Protection Agency. It is present in the environment in two oxidation states: antimonite (Sb (III)) and antimony (Sb (V)). Sb (III) is toxic to several aquatic organisms, but the potential inhibitory effect of Sb species for microorganisms has not been extensively evaluated. The fate and possible toxic impact of antimony on aerobic and anaerobic wastewater treatment systems are unknown. For this reason, the objective of this study was to evaluate the microbial toxicity of Sb (V) and Sb (III) in aerobic and anaerobic environments. Sb(V) and Sb(III) were used as potassium hexahydroxoantimonate (V) and potassium antimony tartrate, respectively (Sigma-Aldrich). The toxic effect of both Sb species in anaerobic environments was evaluated on methanogenic activity and the inhibition of hydrogen production of microorganisms from a wastewater treatment bioreactor. For the methanogenic activity, batch experiments were carried out in 160 mL serological bottles; each bottle contained basal mineral medium (100 mL), inoculum (1.5 g of VSS/L), acetate (2.56 g/L) as substrate, and variable concentrations of Sb (V) or Sb (III). Duplicate bioassays were incubated at 30 ± 2°C on an orbital shaker (105 rpm) in the dark. Methane production was monitored by gas chromatography. The hydrogen production inhibition tests were carried out in glass bottles with a working volume of 0.36 L. Glucose (50 g/L) was used as a substrate, pretreated inoculum (5 g VSS/L), mineral medium and varying concentrations of the two species of antimony. The bottles were kept under stirring and at a temperature of 35°C in an AMPTSII device that recorded hydrogen production. The toxicity of Sb on aerobic microorganisms (from a wastewater activated sludge treatment plant) was tested with a Microtox standardized toxicity test and respirometry. Results showed that Sb (III) is more toxic than Sb (V) for methanogenic microorganisms. Sb (V) caused a 50% decrease in methanogenic activity at 250 mg/L. In contrast, exposure to Sb (III) resulted in a 50% inhibition at a concentration of only 11 mg/L, and an almost complete inhibition (95%) at 25 mg/L. For hydrogen-producing microorganisms, Sb (III) and Sb (V) inhibited 50% of this production with 12.6 mg/L and 87.7 mg/L, respectively. The results for aerobic environments showed that 500 mg/L of Sb (V) do not inhibit the Allivibrio fischeri (Microtox) activity or specific oxygen uptake rate of activated sludge. In the case of Sb (III), this caused a loss of 50% of the respiration of the microorganisms at concentrations below 40 mg/L. The results obtained indicate that the toxicity of the antimony will depend on the speciation of this metalloid and that Sb (III) has a significantly higher inhibitory potential compared to Sb (V). It was shown that anaerobic microorganisms can reduce Sb (V) to Sb (III). Acknowledgments: This work was funded in part by grants from the UA-CONACYT Binational Consortium for the Regional Scientific Development and Innovation (CAZMEX), the National Institute of Health (NIH ES- 04940), and PAPIIT-DGAPA-UNAM (IN105220).

Keywords: aerobic inhibition, antimony reduction, hydrogen inhibition, methanogenic toxicity

Procedia PDF Downloads 161

Authors: Pantea Salehizadeh, Martin P. Bucknall, Robert Driscoll, Jayashree Arcot, George Srzednicki

Abstract:

Banana is one of the most important crops produced in large quantities in tropical and sub-tropical countries. Of the total plant material grown, approximately 40% is considered waste and left in the field to decay. This practice allows fungal diseases such as Sigatoka Leaf Spot to develop, limiting plant growth and spreading spores in the air that can cause respiratory problems in the surrounding population. The pseudostem is considered a waste residue of production (60 to 80 tonnes/ha/year), although it is a good source of dietary fiber and volatile organic compounds (VOC’s). Strategies to process banana pseudostem into palatable, nutritious and marketable food materials could provide significant social and economic benefits. Extraction of VOC’s with desirable odor from dried and fresh pseudostem could improve the smell of products from the confectionary and bakery industries. Incorporation of banana pseudostem flour into bakery products could provide cost savings and improve nutritional value. The aim of this study was to determine the effects of drying methods and different banana species on the profile of volatile aroma compounds in dried banana pseudostem. The banana species analyzed were Musa acuminata and Musa balbisiana. Fresh banana pseudostem samples were processed by either freeze-drying (FD) or heat pump drying (HPD). The extraction of VOC’s was performed at ambient temperature using vacuum distillation and the resulting, mostly aqueous, distillates were analyzed using headspace solid phase microextraction (SPME) gas chromatography – mass spectrometry (GC-MS). Optimal SPME adsorption conditions were 50 °C for 60 min using a Supelco 65 μm PDMS/DVB Stableflex fiber1. Compounds were identified by comparison of their electron impact mass spectra with those from the Wiley 9 / NIST 2011 combined mass spectral library. The results showed that the two species have notably different VOC profiles. Both species contained VOC’s that have been established in literature to have pleasant appetizing aromas. These included l-Menthone, D-Limonene, trans-linlool oxide, 1-Nonanol, CIS 6 Nonen-1ol, 2,6 Nonadien-1-ol, Benzenemethanol, 4-methyl, 1-Butanol, 3-methyl, hexanal, 1-Propanol, 2-methyl- acid، 2-Methyl-2-butanol. Results show banana pseudostem VOC’s are better preserved by FD than by HPD. This study is still in progress and should lead to the optimization of processing techniques that would promote the utilization of banana pseudostem in the food industry.

Keywords: heat pump drying, freeze drying, SPME, vacuum distillation, VOC analysis

Procedia PDF Downloads 326

Authors: Benoit Atge, Alice Dhersin, Oscar Da Silva Cacao, Beatrice Martinez, Dominique Ducint, Catherine Verdun-Esquer, Isabelle Baldi, Mathieu Molimard, Antoine Villa, Mireille Canal-Raffin

Abstract:

Introduction: Healthcare workers' exposure to antineoplastic drugs (AD) is a burning issue for occupational medicine practitioners. Biological monitoring of occupational exposure (BMOE) is an essential tool for assessing AD contamination of healthcare workers. In addition to BMOE, surface sampling is a useful tool in order to understand how workers get contaminated, to identify sources of environmental contamination, to verify the effectiveness of surface decontamination way and to ensure monitoring of these surfaces. The objective of this work was to develop a complete tool including a kit for surface sampling and a quantification analytical method for AD traces detection. The development was realized with the three following criteria: the kit capacity to sample in every professional environment (healthcare services, veterinaries, etc.), the detection of very low AD traces with a validated analytical method and the easiness of the sampling kit use regardless of the person in charge of sampling. Material and method: AD mostly used in term of quantity and frequency have been identified by an analysis of the literature and consumptions of different hospitals, veterinary services, and home care settings. The kind of adsorbent device, surface moistening solution and mix of solvents for the extraction of AD from the adsorbent device have been tested for a maximal yield. The AD quantification was achieved by an ultra high-performance liquid chromatography method coupled with tandem mass spectrometry (UHPLC-MS/MS). Results: With their high frequencies of use and their good reflect of the diverse activities through healthcare, 15 AD (cyclophosphamide, ifosfamide, doxorubicin, daunorubicin, epirubicin, 5-FU, dacarbazin, etoposide, pemetrexed, vincristine, cytarabine, methothrexate, paclitaxel, gemcitabine, mitomycin C) were selected. The analytical method was optimized and adapted to obtain high sensitivity with very low limits of quantification (25 to 5000ng/mL), equivalent or lowest that those previously published (for 13/15 AD). The sampling kit is easy to use, provided with a didactic support (online video and protocol paper). It showed its effectiveness without inter-individual variation (n=5/person; n= 5 persons; p=0,85; ANOVA) regardless of the person in charge of sampling. Conclusion: This validated tool (sampling kit + analytical method) is very sensitive, easy to use and very didactic in order to control the chemical risk brought by AD. Moreover, BMOE permits a focal prevention. Used in routine, this tool is available for every intervention of occupational health.

Keywords: surface contamination, sampling kit, analytical method, sensitivity

Procedia PDF Downloads 127

Authors: Auteleyeva Laura, Maikanov Balgabai, Smagulova Ayana

Abstract:

In order to find and select detoxification methods, patent and information research was conducted, as a result of which 68 patents for inventions were found, among them from the near abroad - 14 (Russia), from far abroad: China – 27, USA - 6, South Korea–1, Germany - 2, Mexico – 4, Yugoslavia – 7, Austria, Taiwan, Belarus, Denmark, Italy, Japan, Canada for 1 security document. Aflatoxin B₁ in various nuts was determined by two methods: enzyme immunoassay "RIDASCREEN ® FAST Aflatoxin" with determination of optical density on a microplate spectrophotometer RIDA®ABSORPTION 96 with RIDASOFT® software Win.NET (Germany) and the method of high-performance liquid chromatography (HPLC Corporation Water, USA) according to GOST 307112001. For experimental contamination of nuts, the cultivation of strain A was carried out. flavus KWIK-STIK on the medium of Chapek (France) with subsequent infection of various nuts (peanuts, peanuts with shells, badam, walnuts with and without shells, pistachios).Based on our research, we have selected 2 detoxification methods: method 1 – combined (5% citric acid solution + microwave for 640 W for 3 min + UV for 20 min) and a chemical method with various leaves of plants: Artemisia terra-albae, Thymus vulgaris, Callogonum affilium, collected in the territory of Akmola region (Artemisia terra-albae, Thymus vulgaris) and Western Kazakhstan (Callogonum affilium). The first stage was the production of ethanol extracts of Artemisia terraea-albae, Thymus vulgaris, Callogonum affilium. To obtain them, 100 g of vegetable raw materials were taken, which was dissolved in 70% ethyl alcohol. Extraction was carried out for 2 hours at the boiling point of the solvent with a reverse refrigerator using an ultrasonic bath "Sapphire". The obtained extracts were evaporated on a rotary evaporator IKA RV 10. At the second stage, the three samples obtained were tested for antimicrobial and antifungal activity. Extracts of Thymus vulgaris and Callogonum affilium showed high antimicrobial and antifungal activity. Artemisia terraea-albae extract showed high antimicrobial activity and low antifungal activity. When testing method 1, it was found that in the first and third experimental groups there was a decrease in the concentration of aflatoxin B1 in walnut samples by 63 and 65%, respectively, but these values also exceeded the maximum permissible concentrations, while the nuts in the second and third experimental groups had a tart lemon flavor; When testing method 2, a decrease in the concentration of aflatoxin B1 to a safe level was observed by 91% (0.0038 mg/kg) in nuts of the 1st and 2nd experimental groups (Artemisia terra-albae, Thymus vulgaris), while in samples of the 2nd and 3rd experimental groups, a decrease in the amount of aflatoxin in 1 to a safe level was observed.

Keywords: nuts, aflatoxin B1, my, mycotoxins

Procedia PDF Downloads 84

Authors: Sevde Altuntas, Fatih Buyukserin

Abstract:

Alzheimer’s disease is responsible for irreversible neural damage of brain parts. One of the disease markers is Amyloid-β 1-42 protein that accumulates in the brain in the form plaques. The basic problem for detection of the protein is the low amount of protein that cannot be detected properly in body liquids such as blood, saliva or urine. To solve this problem, tests like ELISA or PCR are proposed which are expensive, require specialized personnel and can contain complex protocols. Therefore, Surface-enhanced Raman Spectroscopy (SERS) a good candidate for detection of Amyloid-β 1-42 protein. Because the spectroscopic technique can potentially allow even single molecule detection from liquid and solid surfaces. Besides SERS signal can be improved by using nanopattern surface and also is specific to molecules. In this context, our study proposes to fabricate diagnostic test models that utilize Au-coated nanopatterned polycarbonate (PC) surfaces modified with Thioflavin - T to detect low concentrations of Amyloid-β 1-42 protein in water and artificial saliva medium by the enhancement of protein SERS signal. The nanopatterned PC surface that was used to enhance SERS signal was fabricated by using Anodic Alumina Membranes (AAM) as a template. It is possible to produce AAMs with different column structures and varying thicknesses depending on voltage and anodization time. After fabrication process, the pore diameter of AAMs can be arranged with dilute acid solution treatment. In this study, two different columns structures were prepared. After a surface modification to decrease their surface energy, AAMs were treated with PC solution. Following the solvent evaporation, nanopatterned PC films with tunable pillared structures were peeled off from the membrane surface. The PC film was then modified with Au and Thioflavin-T for the detection of Amyloid-β 1-42 protein. The protein detection studies were conducted first in water via this biosensor platform. Same measurements were conducted in artificial saliva to detect the presence of Amyloid Amyloid-β 1-42 protein. SEM, SERS and contact angle measurements were carried out for the characterization of different surfaces and further demonstration of the protein attachment. SERS enhancement factor calculations were also completed via experimental results. As a result, our research group fabricated diagnostic test models that utilize Au-coated nanopatterned polycarbonate (PC) surfaces modified with Thioflavin-T to detect low concentrations of Alzheimer’s Amiloid – β protein in water and artificial saliva medium. This work was supported by The Scientific and Technological Research Council of Turkey (TUBITAK) Grant No: 214Z167.

Keywords: alzheimer, anodic aluminum oxide, nanotopography, surface enhanced Raman spectroscopy

Procedia PDF Downloads 286

Authors: Gómez S. Jennifer, Méndez V. Camila, Moncayo M. Diana, Vega M. Lizeth

Abstract:

The cape gooseberry is a climacteric fruit; Colombia is one of the principal exporters in the world. The environmental condition of temperature and relative moisture decreases the titratable acidity and pH. These conditions and fruit maturation result in the fungal proliferation of Botrytis cinerea disease. Plastic packaging for fresh cape gooseberries was used for mechanical damage protection but created a suitable atmosphere for fungal growth. Beta-cyclodextrins are currently implemented as coatings for the encapsulation of hydrophobic compounds, for example, with bioactive compounds from essential oils such as cinnamaldehyde, which has a high antimicrobial capacity. However, it is a volatile substance. In this article, the casting method was used to obtain a polylactic acid (PLA) polymer film containing the beta-cyclodextrin-cinnamaldehyde inclusion complex, generating an insert that allowed the controlled release of the antifungal substance in packed cape gooseberries to decrease contamination by Botrytis cinerea in a latent state during storage. For the encapsulation technique, three ratios for the cinnamaldehyde: beta-cyclodextrin inclusion complex were proposed: (25:75), (40:60), and (50:50). Spectrophotometry, colorimetry in L*a*b* coordinate space and scanning electron microscopy (SEM) were made for the complex characterization. Subsequently, two ratios of tween and water (40:60) and (50:50) were used to obtain the polylactic acid (PLA) film. To determine mechanical and physical parameters of colourimetry in L*a*b* coordinate space, atomic force microscopy and stereoscopy were done to determine the transparency and flexibility of the film; for both cases, Statgraphics software was used to determine the best ratio in each of the proposed phases, where for encapsulation it was (50:50) with an encapsulation efficiency of 65,92%, and for casting the ratio (40:60) obtained greater transparency and flexibility that permitted its incorporation into the polymeric packaging. A liberation assay was also developed under ambient temperature conditions to evaluate the concentration of cinnamaldehyde inside the packaging through gas chromatography for three weeks. It was found that the insert had a controlled release. Nevertheless, a higher cinnamaldehyde concentration is needed to obtain the minimum inhibitory concentration for the fungus Botrytis cinerea (0.2g/L). The homogeneity of the cinnamaldehyde gas phase inside the packaging can be improved by considering other insert configurations. This development aims to impact emerging food preservation technologies with the controlled release of antifungals to reduce the affectation of the physico-chemical and sensory properties of the fruit as a result of contamination by microorganisms in the postharvest stage.

Keywords: antifungal, casting, encapsulation, postharvest

Procedia PDF Downloads 71

Authors: L. C. Sánchez, L. C. Corrales, A. G. Lancheros, E. Castañeda, Y. Ariza, L. S. Fuentes, L. Sierra, J. L. Cuervo

Abstract:

The importance of environment friendly alternatives in agricultural processes is the reason why the research group Ceparium, the Colegio Mayor de Cundinamarca University, Colombia, investigated the genus Bacillus and its applicability for improving crops of economic importance in Colombia. In this investigation, we presented a study in which the genus Bacillus plays a leading role as beneficial microorganism. The objective was to identify the biochemical potential of three indigenous species of Bacillus, which were able to carry out actions for biological control against pathogens and pests or promoted growth to improve productivity of crops in Colombia. The procedures were performed in three phases: first, the production of biomass of an indigenous strain and a reference strain starting from culture media for production of spores and toxins were made. Spore count was done in a Neubauer chamber, concentrations of spores of Bacillus sphaericus were prepared and a bioassay was done at the Laboratory of Entomology at the University Jorge Tadeo Lozano of Plutella xylostella larvae, insect pest of crucifers in several Colombian regions. The second phase included the extraction in the liquid state fermentation, a secondary metabolite that has antibiosis action against fungi, call iturin B, and was obtained from strains of Bacillus subtilis. The molecule was identified using High Resolution Chromatography (HPLC) and its biocontrol effect on Fusarium sp fungus causes vascular wilt in economically important plant varieties, was confirmed using testing of antagonism in Petri dish. In the third phase, an initial procedure in that let recover and identify microorganisms of the genus Bacillus from the rhizosphere in two aromatic herbs, Rosmarinus officinalis and Thymus vulgaris L. was used. Subsequently, testing of antagonism against Fusarium sp were made and an assay was done under greenhouse conditions to observe biocontrol and growth promoting action by comparing growth in length and dry weight. In the first experiment, native Bacillus sphaericus was lethal to 92% Plutella xylostella larvae in 10 DDA. In the second experiment, iturin B was identified and biological control of Fusarium sp was demonstrated. In the third study, all strains demonstrated biological control and the B14 strain identified as Bacillus megaterium increased root length and productivity of the two plants in terms of weight. It was concluded that the native microorganisms of the genus Bacillus has a great biochemical potential that provides a beneficial interactions with plants, improve their growth and development and therefore a greater impact on production.

Keywords: genus bacillus, biological control, PGPRs, biochemical potential

Procedia PDF Downloads 433

Authors: Ana M. Antolín, Sandra Contreras, Francesc Medina, Didier Tichit

Abstract:

Introduction: High levels of nitrates (> 50 ppm NO3-) in drinking water are potentially risky to human health. In the recent years, the trend of nitrate concentration in groundwater is rising in the EU and other countries. Conventional catalytic nitrate reduction processes into N2 and H2O lead to some toxic intermediates and by-products, such as NO2-, NH4+, and NOx gases. Alternatively, photocatalytic nitrate removal using solar irradiation and heterogeneous catalysts is a very promising and ecofriendly technique. It has been scarcely performed and more research on highly efficient catalysts is still needed. In this work, different nanocatalysts supported on Aeroxide Titania P25 (P25) have been prepared varying: 0.5-4 % wt. Ag); Pt (2, 4 % wt.); Pt precursor (H2PtCl6/K2PtCl6); and impregnation order of both metals. Pt was chosen in order to increase the selectivity to N2 and decrease that to NO2-. Catalysts were characterized by nitrogen physisorption, X-Ray diffraction, UV-visible spectroscopy, TEM and X Ray-Photoelectron Spectroscopy. The aim was to determine the influence of the composition and the preparation method of the catalysts on the conversion and selectivity in the nitrate reduction, as well as going through an overall and better understanding of the process. Nanocatalysts synthesis: For the mono and bimetallic catalysts preparation, wise-drop wetness impregnation of the precursors (AgNO3, H2PtCl6, K2PtCl6) followed by a reduction step (NaBH4) was used to obtain the metal colloids. Results and conclusions: Denitration experiments were performed in a 350 mL PTFE batch reactor under inert standard operational conditions, ultraviolet irradiations (λ=254 nm (UV-C); λ=365 nm (UV-A)), and presence/absence of hydrogen gas as a reducing agent, contrary to most studies using oxalic or formic acid. Samples were analyzed by Ionic Chromatography. Blank experiments using respectively P25 (dark conditions), hydrogen only and UV irradiations without hydrogen demonstrated a clear influence of the presence of hydrogen on nitrate reduction. Also, they demonstrated that UV irradiation increased the selectivity to N2. Interestingly, the best activity was obtained under ultraviolet lamps, especially at a closer wavelength to visible light irradiation (λ = 365 nm) and H2. 2% Ag/P25 leaded to the highest NO3- conversion among the monometallic catalysts. However, nitrite quantities have to be diminished. On the other hand, practically no nitrate conversion was observed with the monometallics based on Pt/P25. Therefore, the amount of 2% Ag was chosen for the bimetallic catalysts. Regarding the bimetallic catalysts, it is observed that the metal impregnation order, amount and Pt precursor highly affects the results. Higher selectivity to the desirable N2 gas is obtained when Pt was firstly added, especially with K2PtCl6 as Pt precursor. This suggests that when Pt is secondly added, it covers the Ag particles, which are the most active in this reaction. It could be concluded that Ag allows the nitrate reduction step to nitrite, and Pt the nitrite reduction step toward the desirable N2 gas.

Keywords: heterogeneous catalysis, hydrogenation, nanocatalyst, nitrate removal, photocatalysis

Procedia PDF Downloads 268

Authors: Vlad Andrei Raducanu, Mariana Lucia Angelescu, Ion Cinca, Vasile Danut Cojocaru, Doina Raducanu

Abstract:

By using smart digital tools, such as generative design (GD) and digital fabrication (DF), problems of high actuality concerning resources optimization (materials, energy, time) can be solved and applications or products of free-form type can be created. In the new digital technology materials are active, designed in response to a set of performance requirements, which impose a total rethinking of old material practices. The article presents the design procedure key steps of a free-form architectural object - a column type one with connections to get an adaptive 3D surface, by using the parametric design methodology and by exploiting the properties of conventional metallic materials. In parametric design the form of the created object or space is shaped by varying the parameters values and relationships between the forms are described by mathematical equations. Digital parametric design is based on specific procedures, as shape grammars, Lindenmayer - systems, cellular automata, genetic algorithms or swarm intelligence, each of these procedures having limitations which make them applicable only in certain cases. In the paper the design process stages and the shape grammar type algorithm are presented. The generative design process relies on two basic principles: the modeling principle and the generative principle. The generative method is based on a form finding process, by creating many 3D spatial forms, using an algorithm conceived in order to apply its generating logic onto different input geometry. Once the algorithm is realized, it can be applied repeatedly to generate the geometry for a number of different input surfaces. The generated configurations are then analyzed through a technical or aesthetic selection criterion and finally the optimal solution is selected. Endless range of generative capacity of codes and algorithms used in digital design offers various conceptual possibilities and optimal solutions for both technical and environmental increasing demands of building industry and architecture. Constructions or spaces generated by parametric design can be specifically tuned, in order to meet certain technical or aesthetical requirements. The proposed approach has direct applicability in sustainable architecture, offering important potential economic advantages, a flexible design (which can be changed until the end of the design process) and unique geometric models of high performance.

Keywords: parametric design, algorithmic procedures, free-form architectural object, sustainable architecture

Procedia PDF Downloads 367

Authors: Callistus I. Iheme, Kenneth E. Asika, Emmanuel I. Ugwor, Chukwuka U. Ogbonna, Ugonna H. Uzoka, Nneamaka A. Chiegboka, Chinwe S. Alisi, Obinna S. Nwabueze, Amanda U. Ezirim, Judeanthony N. Ogbulie

Abstract:

Antimicrobial resistance (AMR) is a major threat to the global health sector. Zinc oxide nanocomposites (ZnONCs), composed of zinc oxide nanoparticles and phytochemicals from Azadirachta indica aqueous leaf extract, were assessed for their physico-chemicals, in silico and in vitro antimicrobial properties on multidrug-resistant Escherichia coli enzymes. Gas chromatography coupled with mass spectroscope (GC-MS) analysis on the ZnONCs revealed the presence of twenty volatile phytochemical compounds, among which is scoparone. Characterization of the ZnONCs was done using ultraviolet-visible spectroscopy (UV-vis), energy dispersive spectroscopy (EDX), transmission electron microscopy (TEM), scanning electron microscopy (SEM), and x-ray diffractometer (XRD). Dehydrogenase enzyme converts colorless 2,3,5-triphenyltetrazolium chloride to the red triphenyl formazan (TPF). The rate of formazan formation in the presence of ZnONCs is proportional to the enzyme activities. The color formation is extracted and determined at 500 nm, and the percentage of enzyme activity is calculated. To determine the bioactive components of the ZnONCs, characterize their binding to enzymes, and evaluate the enzyme-ligand complex stability, respectively Discrete Fourier Transform (DFT) analysis, docking, and molecular dynamics simulations will be employed. The results showed arrays of ZnONCs nanorods with maximal absorption wavelengths of 320 nm and 350 nm and thermally stable at the temperature range of 423.77 to 889.69 ℃. In vitro study assessed the dehydrogenase inhibitory properties of the ZnONCs, conjugate of ZnONCs and ampicillin (ZnONCs-amp), the aqueous leaf extract of A. indica, and ampicillin (standard drug). The findings revealed that at the concentration of 500 μm/mL, 57.89 % of the enzyme activities were inhibited by ZnONCs compared to 33.33% and 21.05% of the standard drug (Ampicillin), and the aqueous leaf extract of the A. indica respectively. The inhibition of the enzyme activities by the ZnONCs at 500 μm/mL was further enhanced to 89.74 % by conjugating with Ampicillin. In silico study on the ZnONCs revealed scoparone as the most viable competitor of nicotinamide adenine dinucleotide (NAD⁺) for the coenzyme binding pocket on E. coli malate and histidinol dehydrogenase. From the findings, it can be concluded that the scoparone components of the nanocomposites in synergy with the zinc oxide nanoparticles inhibited E. coli malate and histidinol dehydrogenase by competitively binding to the NAD⁺ pocket and that the conjugation of the ZnONCs with ampicillin further enhanced the antimicrobial efficiency of the nanocomposite against multidrug resistant E. coli.

Keywords: antimicrobial resistance, dehydrogenase activities, E. coli, zinc oxide nanocomposites

Procedia PDF Downloads 38

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 146