Search results for: nickel affinity chromatography
1064 H2 Permeation Properties of a Catalytic Membrane Reactor in Methane Steam Reforming Reaction
Authors: M. Amanipour, J. Towfighi, E. Ganji Babakhani, M. Heidari
Abstract:
Cylindrical alumina microfiltration membrane (GMITM Corporation, inside diameter=9 mm, outside diameter=13 mm, length= 50 mm) with an average pore size of 0.5 micrometer and porosity of about 0.35 was used as the support for membrane reactor. This support was soaked in boehmite sols, and the mean particle size was adjusted in the range of 50 to 500 nm by carefully controlling hydrolysis time, and calcined at 650 °C for two hours. This process was repeated with different boehmite solutions in order to achieve an intermediate layer with an average pore size of about 50 nm. The resulting substrate was then coated with a thin and dense layer of silica by counter current chemical vapour deposition (CVD) method. A boehmite sol with 10 wt.% of nickel which was prepared by a standard procedure was used to make the catalytic layer. BET, SEM, and XRD analysis were used to characterize this layer. The catalytic membrane reactor was placed in an experimental setup to evaluate the permeation and hydrogen separation performance for a steam reforming reaction. The setup consisted of a tubular module in which the membrane was fixed, and the reforming reaction occurred at the inner side of the membrane. Methane stream, diluted with nitrogen, and deionized water with a steam to carbon (S/C) ratio of 3.0 entered the reactor after the reactor was heated up to 500 °C with a specified rate of 2 °C/ min and the catalytic layer was reduced at presence of hydrogen for 2.5 hours. Nitrogen flow was used as sweep gas through the outer side of the reactor. Any liquid produced was trapped and separated at reactor exit by a cold trap, and the produced gases were analyzed by an on-line gas chromatograph (Agilent 7890A) to measure total CH4 conversion and H2 permeation. BET analysis indicated uniform size distribution for catalyst with average pore size of 280 nm and average surface area of 275 m2.g-1. Single-component permeation tests were carried out for hydrogen, methane, and carbon dioxide at temperature range of 500-800 °C, and the results showed almost the same permeance and hydrogen selectivity values for hydrogen as the composite membrane without catalytic layer. Performance of the catalytic membrane was evaluated by applying membranes as a membrane reactor for methane steam reforming reaction at gas hourly space velocity (GHSV) of 10,000 h−1 and 2 bar. CH4 conversion increased from 50% to 85% with increasing reaction temperature from 600 °C to 750 °C, which is sufficiently above equilibrium curve at reaction conditions, but slightly lower than membrane reactor with packed nickel catalytic bed because of its higher surface area compared to the catalytic layer.Keywords: catalytic membrane, hydrogen, methane steam reforming, permeance
Procedia PDF Downloads 2561063 Investigation of NiO/V₂O₅ Powder Composite as Cathode Material for Lithium-Ion Batteries
Authors: Katia Ayouz-Chebout, Fatima Boudeffar, Maha Ayat, Malika Berouaken, Chafiaa Yaddaden, Saloua Merazga, Nouredine Gabouze
Abstract:
Transition metal oxide composites have been widely reported in energy storage and conversion systems. In this regard, an attempt has been made to synthesize NiO@V₂O₅ nanocomposite. The structures and morphology of synthesized powder are investigated by X-ray diffraction, scanning electron microscope (SEM), and Attenuated Total Reflection (ATR). The electrochemical properties and performances as cathode electrodes based on active material NiO@V₂O₅ were studied by cyclic voltammetry (CV), between potential bias [0.01V to 3V], with scanning speed of 0,1mVs⁻¹, the galvanostatic charge/discharge (CDG) for 100 cycles was also measured.Keywords: composite nanobelts, vanadium pentoxide, nickel oxide, Li-ion batteries
Procedia PDF Downloads 231062 Synthesis and Characterization of Molecularly Imprinted Polymer as a New Adsorbent for the Removal of Pyridine from Organic Medium
Authors: Opeyemi Elujulo, Aderonke Okoya, Kehinde Awokoya
Abstract:
Molecularly imprinted polymers (MIP) for the adsorption of pyridine (PYD) was obtained from PYD (the template), styrene (the functional monomer), divinyl benzene (the crosslinker), benzoyl peroxide (the initiator), and water (the porogen). When the template was removed by solvent extraction, imprinted binding sites were left in the polymer material that are capable of selectively rebinding the target molecule. The material was characterized by Fourier transform infrared spectroscopy and differential scanning calorimetry. Batch adsorption experiments were performed to study the adsorption of the material in terms of adsorption kinetics, isotherms, and thermodynamic parameters. The results showed that the imprinted polymer exhibited higher affinity for PYD compared to non-imprinted polymer (NIP).Keywords: molecularly imprinted polymer, bulk polymerization, environmental pollutant, adsorption
Procedia PDF Downloads 1421061 Comparative Analysis of Benzene, Toluene, Ethylbenzene, and Xylene Concentrations at Roadside and Urban Background Sites in Leicester and Lagos Using Thermal Desorption-Gas Chromatography-Mass Spectrometry
Authors: Emmanuel Bernard, Rebecca L. Cordell, Akeem A. Abayomi, Rose Alani, Paul S. Monks
Abstract:
This study investigates the prevalence and extent of BTEX (Benzene, Toluene, Ethylbenzene, and Xylene) contamination in Leicester, United Kingdom, and Lagos, Nigeria, through field measurements at roadside (RS) and urban background (UB) sites. Using thermal desorption gas chromatography mass spectrometry (TD-GC-MS), BTEX concentrations were quantified. In Leicester, the average RS concentration was 24.9 ± 8.9 μg/m³, and the UB concentration was 12.7 ± 5.7 μg/m³. In Lagos, the RS concentration was significantly higher at 106 ± 39.3 μg/m³, and the UB concentration was 20.1 ± 8.9 μg/m³. The RS concentration in Lagos was approximately 4.3 times higher than in Leicester, while the UB concentration was about 1.6 times higher. These disparities are attributed to differences in road infrastructure, traffic regulation compliance, fuel and oil quality, and local activities. In Leicester, the highest UB concentration (20.5 ± 1.7 μg/m³) was at Knighton Village, near the heavily polluted RS Wigston roundabout. In Lagos, the highest concentration (172.1 ± 12.2 μg/m³) was at Ojuelegba, a major transportation hub. Correlation analysis revealed strong positive relationships between the concentrations of BTEX compounds in both cities, suggesting common sources such as vehicular emissions and industrial activities. The ratios of toluene to benzene (T:B) and m/p xylene to ethylbenzene (m/p X:E) were analysed to infer source contributions and the photochemical age of air masses. The T:B ratio in Leicester ranged from 0.44 to 0.71, while in Lagos, it ranged from 1.36 to 2.17. The m/p X:E ratio in Leicester ranged from 2.11 to 2.19, like other UK cities, while in Lagos, it ranged from 1.65 to 2.32, indicating relatively fresh emissions. This study highlights significant differences in BTEX concentrations between Leicester and Lagos, emphasizing the need for tailored pollution control strategies to address the specific sources and conditions in different urban environments.Keywords: BTEX contamination, urban air quality, thermal desorption GC-MS, roadside emissions, urban background sites, vehicular emissions, pollution control strategies
Procedia PDF Downloads 481060 Assessment of the Quality of a Mixture of Vegetable Oils from Kazakhstan Origin
Authors: Almas Mukhametov, Dina Dautkanova, Moldir Yerbulekova, Gulim Tuyakova, Raziya Zhakudaeva, Makpal Seisenaly, Asemay Kazhymurat
Abstract:
The composition of samples of mixtures of vegetable oils of Kazakhstan origin, consisting of sunflower, safflower and linseed oils, has been experimentally substantiated. With an approximate optimal ratio of w-6:w-3 fatty acids in 80:15:05 triacylglycerols, providing its therapeutic and prophylactic properties. The resulting mixture can be used in the development of functional products. The result was also identified and evaluated by physical and chemical quality indicators, the content of vitamin E, and the concentration of ions of copper (Cu), iron (Fe), cadmium (Cd), lead (Pb), arsenic (As), nickel (Ni), as well as mercury (Hg).Keywords: vegetable oil, sunflower, safflower, linseed, mixture, fatty acid composition, heavy metals
Procedia PDF Downloads 1891059 Molecular Topology and TLC Retention Behaviour of s-Triazines: QSRR Study
Authors: Lidija R. Jevrić, Sanja O. Podunavac-Kuzmanović, Strahinja Z. Kovačević
Abstract:
Quantitative structure-retention relationship (QSRR) analysis was used to predict the chromatographic behavior of s-triazine derivatives by using theoretical descriptors computed from the chemical structure. Fundamental basis of the reported investigation is to relate molecular topological descriptors with chromatographic behavior of s-triazine derivatives obtained by reversed-phase (RP) thin layer chromatography (TLC) on silica gel impregnated with paraffin oil and applied ethanol-water (φ = 0.5-0.8; v/v). Retention parameter (RM0) of 14 investigated s-triazine derivatives was used as dependent variable while simple connectivity index different orders were used as independent variables. The best QSRR model for predicting RM0 value was obtained with simple third order connectivity index (3χ) in the second-degree polynomial equation. Numerical values of the correlation coefficient (r=0.915), Fisher's value (F=28.34) and root mean square error (RMSE = 0.36) indicate that model is statistically significant. In order to test the predictive power of the QSRR model leave-one-out cross-validation technique has been applied. The parameters of the internal cross-validation analysis (r2CV=0.79, r2adj=0.81, PRESS=1.89) reflect the high predictive ability of the generated model and it confirms that can be used to predict RM0 value. Multivariate classification technique, hierarchical cluster analysis (HCA), has been applied in order to group molecules according to their molecular connectivity indices. HCA is a descriptive statistical method and it is the most frequently used for important area of data processing such is classification. The HCA performed on simple molecular connectivity indices obtained from the 2D structure of investigated s-triazine compounds resulted in two main clusters in which compounds molecules were grouped according to the number of atoms in the molecule. This is in agreement with the fact that these descriptors were calculated on the basis of the number of atoms in the molecule of the investigated s-triazine derivatives.Keywords: s-triazines, QSRR, chemometrics, chromatography, molecular descriptors
Procedia PDF Downloads 3931058 Laboratory Scale Production of Bio-Based Chemicals from Industrial Waste Feedstock in South Africa
Authors: P. Mandree, S. O. Ramchuran, F. O'Brien, L. Sethunya, S. Khumalo
Abstract:
South Africa is identified as one of the five emerging waste management markets, globally. The waste sector in South Africa influences the areas of energy, water and food at an economic and social level. Recently, South African industries have focused on waste valorization and diversification of the current product offerings in an attempt to reduce industrial waste, target a zero waste-to-landfill initiative and recover energy. South Africa has a number of waste streams including industrial and agricultural biomass, municipal waste and marine waste. Large volumes of agricultural and forestry residues, in particular, are generated which provides significant opportunity for production of bio-based fuels and chemicals. This could directly impact development of a rural economy. One of the largest agricultural industries is the sugar industry, which contributes significantly to the country’s economy and job creation. However, the sugar industry is facing challenges due to fluctuations in sugar prices, increasing competition with low-cost global sugar producers, increasing energy and agricultural input costs, lower consumption and aging facilities. This study is aimed at technology development for the production of various bio-based chemicals using feedstock from the sugar refining process. Various indigenous bacteria and yeast species were assessed for the potential to produce platform chemicals in flask studies and at 30 L fermentation scale. Quantitative analysis of targeted bio-based chemicals was performed using either gas chromatography or high pressure liquid chromatography to assess production yields and techno-economics in order to compare performance to current commercial benchmark processes. The study also creates a decision platform for the research direction that is required for strain development using Industrial Synthetic Biology.Keywords: bio-based chemicals, biorefinery, industrial synthetic biology, waste valorization
Procedia PDF Downloads 1191057 Isolation and Structural Elucidation of 20 Hydroxyecdystone from Vitex doniana Sweet Stem Bark
Authors: Mustapha A. Tijjani, Fanna I. Abdulrahman, Irfan Z. Khan, Umar K. Sandabe, Cong Li
Abstract:
Air dried sample V. doniana after collection and identification was extracted with ethanol and further partition with chloroform, ethyl acetate and n-butanol. Ethanolic extract (11.9g) was fractionated on a silica gel accelerated column chromatography using solvents such as n-hexane, ethyl acetate and methanol. Each eluent fractions (150ml aliquots) were collected and monitored with thin layer chromatography. Fractions with similar Rf values from same solvents system were pooled together. Phytochemical test of all the fractions were performed using standard procedure. Complete elution yielded 48 fractions (150ml/fraction) which were pooled to 24 fractions base on the Rf values. It was further recombined and 12 fractions were obtained on the basis on Rf values and coded Vd1 to Vd12 fractions. Vd8 was further eluted with ethylacetate and methanol and gave fourteen sub fractions Vd8-a, -Vd8-m. Fraction Vd8-a (56mg) gave a white crystal compound coded V1. It was further checked on TLC and observed under ultraviolet lamp and was found to give a single spot. The Rf values were calculated to be 0.433. The melting point was determined using Gallenkamp capillary melting point apparatus and found to be 241-243°C uncorrected. Characterization of the isolated compound coded V1 was done using FT-infra-red spectroscopy, HNMR, 13CNMR(1and 2D) and HRESI-MS. The IR spectrum of compound V1 shows prominent peaks that corresponds to OHstr (3365cm-1) and C=0 (1652cm-1) etc. This spectrum suggests that among the functional moiety in compound V1 are the carbonyl and hydroxyl group. The 1H NMR (400 MHz) spectrum of compound V1 in DMSO-d6 displayed five singlet signals at δ 0.72 (3H, s, H-18), 0.79 (3H, s, H-19), 1.03 (3H, s, H-21), 1.04 (3H, s, H-26), 1.06 (3H, s, H-27) each integrating for three protons indicating the five methyl functional groups present in the compound. It further showed a broad singlet at δ 5.58 integrated for 1 H due to an olefinic H-atom adjacent to the carbonyl carbon atom. Three signals at δ 3.10 (d, J = 9.0 Hz, H-22), 3.59 (m, 1H, 2H-a) and 3.72 (m, 1H, 3H-e), each integrating for one proton is due to oxymethine protons indicating that three oxymethine H-atoms are present in the compound. These all signals are characteristic to the ecdysteroid skeletons. The 13C-NMR spectrum showed the presence of 27 carbon atoms, suggesting that may be steroid skeleton. The DEPT-135 experiment showed the presence of five CH3, eight CH2, and seven CH groups, and seven quaternary C-atoms. The molecular formula was established as C27H44O7 by high resolution electron spray ionization-mass spectroscopy (HRESI-MS) positive ion mode m/z 481.3179. The signals in mass spectrum are 463, 445, and 427 peaks corresponding to losses of one, two, three, or four water molecules characteristic for ecdysterone skeleton reported in the literature. Based on the spectral analysis (HNMR, 13CNMR, DEPT, HMQC, IR, HRESI-MS) the compound V1 is thus concluded to have ecdysteriod skeleton and conclusively conforms with 2β, 3β 14α, 20R, 22R, 25-hexahydroxy-5 β cholest-7-ene-6- one, or 2, 3, 14, 20, 22, 25 hexahydroxy cholest-7-ene-6-one commonly known as 20-hydroxyecdysone.Keywords: vitex, phytochemical, purification, isolation, chromatography, spectroscopy
Procedia PDF Downloads 3561056 Cytotoxicological Evaluation of a Folate Receptor Targeting Drug Delivery System Based on Cyclodextrins
Authors: Caroline Mendes, Mary McNamara, Orla Howe
Abstract:
For chemotherapy, a drug delivery system should be able to specifically target cancer cells and deliver the therapeutic dose without affecting normal cells. Folate receptors (FR) can be considered key targets since they are commonly over-expressed in cancer cells and they are the molecular marker used in this study. Here, cyclodextrin (CD) has being studied as a vehicle for delivering the chemotherapeutic drug, methotrexate (MTX). CDs have the ability to form inclusion complexes, in which molecules of suitable dimensions are included within the CD cavity. In this study, β-CD has been modified using folic acid so as to specifically target the FR molecular marker. Thus, the system studied here for drug delivery consists of β-CD, folic acid and MTX (CDEnFA:MTX). Cellular uptake of folic acid is mediated with high affinity by folate receptors while the cellular uptake of antifolates, such as MTX, is mediated with high affinity by the reduced folate carriers (RFCs). This study addresses the gene (mRNA) and protein expression levels of FRs and RFCs in the cancer cell lines CaCo-2, SKOV-3, HeLa, MCF-7, A549 and the normal cell line BEAS-2B, quantified by real-time polymerase chain reaction (real-time PCR) and flow cytometry, respectively. From that, four cell lines with different levels of FRs, were chosen for cytotoxicity assays of MTX and CDEnFA:MTX using the MTT assay. Real-time PCR and flow cytometry data demonstrated that all cell lines ubiquitously express moderate levels of RFC. These experiments have also shown that levels of FR protein in CaCo-2 cells are high, while levels in SKOV-3, HeLa and MCF-7 cells are moderate. A549 and BEAS-2B cells express low levels of FR protein. FRs are highly expressed in all the cancer cell lines analysed when compared to the normal cell line BEAS-2B. The cell lines CaCo-2, MCF-7, A549 and BEAS-2B were used in the cell viability assays. 48 hours treatment with the free drug and the complex resulted in IC50 values of 93.9 µM ± 9.2 and 56.0 µM ± 4.0 for CaCo-2 for free MTX and CDEnFA:MTX respectively, 118.2 µM ± 10.8 and 97.8 µM ± 12.3 for MCF-7, 36.4 µM ± 6.9 and 75.0 µM ± 8.5 for A549 and 132.6 µM ± 12.1 and 288.1 µM ± 16.3 for BEAS-2B. These results demonstrate that MTX is more toxic towards cell lines expressing low levels of FR, such as the BEAS-2B. More importantly, these results demonstrate that the inclusion complex CDEnFA:MTX showed greater cytotoxicity than the free drug towards the high FR expressing CaCo-2 cells, indicating that it has potential to target this receptor, enhancing the specificity and the efficiency of the drug.Keywords: cyclodextrins, cancer treatment, drug delivery, folate receptors, reduced folate carriers
Procedia PDF Downloads 3011055 Screening of Plant Growth Promoting Rhizobacteria in the Rhizo- and Endosphere of Sunflower (Helianthus anus) and Their Role in Enhancing Growth and Yield Attriburing Trairs and Colonization Studies
Authors: A. Majeed, M.K. Abbasi, S. Hameed, A. Imran, T. Naqqash, M. K. Hanif
Abstract:
Plant growth-promoting rhizobacteria (PGPR) are free-living soil bacteria that aggressively colonize the rhizosphere/plant roots, and enhance the growth and yield of plants when applied to seed or crops. Root associated (endophytic and rhizospheric) PGPR were isolated from Sunflower (Helianthus anus) grown in soils collected from 16 different sites of sub division Dhirkot, Poonch, Azad Jammu & Kashmir, Pakistan. A total of 150 bacterial isolates were isolated, purified, screened in vitro for their plant growth promoting (PGP) characteristics. 11 most effective isolates were selected on the basis of biochemical assays (nitrogen fixation, phosphate solubilization, growth hormone production, biocontrol assay, and carbon substrates utilization assay through gas chromatography (GCMS), spectrophotometry, high performance liquid chromatography HPLC, fungal and bacterial dual plate assay and BIOLOG GN2/GP2 microplate assay respectively) and were tested on the crop under controlled and field conditions. From the inoculation assay, the most promising 4 strains (on the basis of increased root/shoot weight, root/shoot length, seed oil content, and seed yield) were than selected for colonization studies through confocal laser scanning and transmission electron microscope. 16Sr RNA gene analysis showed that these bacterial isolates belong to Pseudononas, Enterobacter, Azospirrilum, and Citobacter genera. This study is the clear evident that such isolates have the potential for application as inoculants adapted to poor soils and local crops to minimize the chemical fertilizers harmful for soil and environmentKeywords: PGPR, nitrogen fixation, phosphate solubilization, colonization
Procedia PDF Downloads 3401054 An Investigation of Machinability of Inconel 718 in EDM Using Different Cryogenic Treated Tools
Authors: Pradeep Joshi, Prashant Dhiman, Shiv Dayal Dhakad
Abstract:
Inconel 718 is a family if Nickel-Chromium based Superalloy; it has very high oxidation and corrosion resistance. Inconel 718 is widely being used in aerospace, engine, turbine etc. due to its high mechanical strength and creep resistance. Being widely used, its machining should be easy but in real its machining is very difficult, especially by using traditional machining methods. It becomes easy to machine only by using non Traditional machining such as EDM. During EDM machining there is wear of both tool and workpiece, the tool wear is undesired because it changes tool shape, geometry. To reduce the tool wear rate (TWR) cryogenic treatment is performed on tool before the machining operation. The machining performances of the process are to be evaluated in terms of MRR, TWR which are functions of Discharge current, Pulse on-time, Pulse Off-time.Keywords: EDM, cyrogenic, TWR, MRR
Procedia PDF Downloads 4571053 Effect of Phthalates on Male Infertility: Myth or Truth?
Authors: Rashmi Tomar, A. Srinivasan, Nayan K. Mohanty, Arun K. Jain
Abstract:
Phthalates have been used as additives in industrial products since the 1930s, and are universally considered to be ubiquitous environmental contaminants. The general population is exposed to phthalates through consumer products, as well as diet and medical treatments. Animal studies showing the existence of an association between some phthalates and testicular toxicity have generated public and scientific concern about the potential adverse effects of environmental changes on male reproductive health. Unprecedented declines in fertility rates and semen quality have been reported during the last half of the 20th century in developed countries and increasing interest exists on the potential relationship between exposure to environmental contaminants, including phthalates, and human male reproductive health Studies. Phthalates may be associated with altered endocrine function and adverse effects on male reproductive development and function, but human studies are limited. The aim of the present study was detection of phthalate compounds, estimation of their metabolites in infertile & fertile male. Blood and urine samples were collected from 150 infertile patients & 75 fertile volunteers recruited through Department of Urology, Safdarjung Hospital, New Delhi. Blood have been collected in separate glass tubes from the antecubital vein of the patients, serum have been separate and estimate the phthalate level in serum samples by Gas Chromatography / Mass Spectrometry using NIOSH / OSHA detailed protocol. Urine of Infertile & Fertile Subjects was collected & extracted using solid phase extraction method, analysis by HPLC. In conclusion, to the best of our knowledge the present study based on human is first to show the presence of phthalate in human serum samples and their metabolites in urine samples. Significant differences were observed between several phthalates in infertile and fertile healthy individuals.Keywords: Gas Chromatography, HPLC, male infertility, phthalates, serum, toxicity, urine
Procedia PDF Downloads 3631052 Anti-proliferative Activity and HER2 Receptor Expression Analysis of MCF-7 (Breast Cancer Cell) Cells by Plant Extract Coleus Barbatus (Andrew)
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 1141051 Revealing the Genome Based Biosynthetic Potential of a Streptomyces sp. Isolate BR123 Presenting Broad Spectrum Antimicrobial Activities
Authors: Neelma Ashraf
Abstract:
Actinomycetes, particularly genus Streptomyces is of great importance due to their role in the discovery of new natural products, particularly antimicrobial secondary metabolites in the medicinal science and biotechnology industry. Different Streptomyces strains were isolated from Helianthus annuus plants and tested for antibacterial and antifungal activities. The most promising five strains were chosen for further investigation, and growth conditions for antibiotic synthesis were optimised. The supernatants were extracted in different solvents, and the extracted products were analyzed using liquid chromatography-mass spectrometry (LC-MS) and biological testing. From one of the potent strains Streptomyces globusus sp. BR123, a compound lavendamycin was identified using these analytical techniques. In addition, this potent strain also produces a strong antifungal polyene compound with a quasimolecular ion of 2072. Streptomyces sp. BR123 was genome sequenced because of its promising antimicrobial potential in order to identify the gene cluster responsible for analyzed compound “lavendamycin”. The genome analysis yielded candidate genes responsible for the production of this potent compound. The genome sequence of 8.15 Mb of Streptomyces sp. isolate BR123 with a GC content of 72.63% and 8103 protein coding genes was attained. Many antimicrobial, antiparasitic, and anticancerous compounds were detected through multiple biosynthetic gene clusters predicted by in-Silico analysis. Though, the novelty of metabolites was determined through the insignificant resemblance with known biosynthetic gene clusters. The current study gives insight into the bioactive potential of Streptomyces sp. isolate BR123 with respect to the synthesis of bioactive secondary metabolites through genomic and spectrometric analysis. Moreover, the comparative genome study revealed the connection of isolate BR123 with other Streptomyces strains, which could expand the knowledge of this genus and the mechanism involved in the discovery of new antimicrobial metabolites.Keywords: streptomyces, secondary metabolites, genome, biosynthetic gene clusters, high performance liquid chromatography, mass spectrometry
Procedia PDF Downloads 701050 Phenolic Composition and Antioxidant Activity of Sorbus L. Fruits and Leaves
Authors: Raudone Lina, Raudonis Raimondas, Gaivelyte Kristina, Pukalskas Audrius, Janulis Valdimaras, Viskelis Pranas
Abstract:
Sorbus L. species are widely distributed in the Northern hemisphere and have been used for medicinal purposes in various traditional medicine systems and as food ingredients. Various Sorbus L. raw materials, fruits, leaves, inflorescences, barks, possess diuretic, anti-inflammatory, hypoglycemic, anti-diarrheal and vasoprotective activities. Phenolics, to whom main pharmacological activities are attributed, are compounds of interest due to their notable antioxidant activity. The aim of this study was to determine the antioxidant profiles of fruits and leaves of selected Sorbus L. species (S. anglica, S. aria f. latifolia, S. arranensis, S. aucuparia, S. austriaca, S. caucasica, S. commixta, S. discolor, S. gracilis, S. hostii, S. semi-incisa, S. tianschanica) and to identify the phenolic compounds with potent contribution to antioxidant activity. Twenty two constituents were identified in Sorbus L. species using ultra high performance liquid chromatography coupled to quadruple and time-of-flight mass spectrometers (UPLC–QTOF–MS). Reducing activity of individual constituents was determined using high performance liquid chromatography (HPLC) coupled to post-column FRAP assay. Signicantly greatest trolox equivalent values corresponding up to 45% of contribution to antioxidant activity were assessed for neochlorogenic and chlorogenic acids, which were determined as markers of antioxidant activity in samples of leaves and fruits. Characteristic patterns of antioxidant profiles obtained using HPLC post-column FRAP assay significantly depend on specific Sorbus L. species and raw materials and are suitable for equivalency research of Sorbus L. fruits and leaves. Selecting species and target plant organs with richest phenolic composition and strongly expressed antioxidant power is the first step in further research of standardized extracts.Keywords: FRAP, antioxidant, phenolic, Sorbus L., chlorogenic acid, neochlorogenic acid
Procedia PDF Downloads 4581049 Ethylene Sensitivity in Orchids and Its Control Using 1-MCP: A Review
Authors: Parviz Almasi
Abstract:
Ethylene is produced as a gaseous growth regulator in all plants and their constructive parts such as roots, stems, leaves, flowers and fruits. It is considered a multifunctional phytohormone that regulates both growths including flowering, fruit ripening, inhibition of root growth, and senescence such as senescence of leaves and flowers and etc. In addition, exposure to external ethylene is caused some changes that are often undesirable and harmful. Some flowers are more sensitive to others and when exposed to ethylene; their aging process is hastened. 1-MCP is an exogenous and endogenous ethylene action inhibitor, which binds to the ethylene receptors in the plants and prevents ethylene-dependent reactions. The binding affinity of 1- MCP for the receptors is about 10 times more than ethylene. Hence, 1-MCP can be a potential candidate for controlling of ethylene injury in horticultural crops. This review integrates knowledge of ethylene biosynthesis in the plants and also a mode of action of 1-MCP in preventing of ethylene injury.Keywords: ethylene injury, biosynthesis, ethylene sensitivity, 1-MCP
Procedia PDF Downloads 1011048 Determination of Rare Earth Element Patterns in Uranium Matrix for Nuclear Forensics Application: Method Development for Inductively Coupled Plasma Mass Spectrometry (ICP-MS) Measurements
Authors: Bernadett Henn, Katalin Tálos, Éva Kováss Széles
Abstract:
During the last 50 years, the worldwide permeation of the nuclear techniques induces several new problems in the environmental and in the human life. Nowadays, due to the increasing of the risk of terrorism worldwide, the potential occurrence of terrorist attacks using also weapon of mass destruction containing radioactive or nuclear materials as e.g. dirty bombs, is a real threat. For instance, the uranium pellets are one of the potential nuclear materials which are suitable for making special weapons. The nuclear forensics mainly focuses on the determination of the origin of the confiscated or found nuclear and other radioactive materials, which could be used for making any radioactive dispersive device. One of the most important signatures in nuclear forensics to find the origin of the material is the determination of the rare earth element patterns (REE) in the seized or found radioactive or nuclear samples. The concentration and the normalized pattern of the REE can be used as an evidence of uranium origin. The REE are the fourteen Lanthanides in addition scandium and yttrium what are mostly found together and really low concentration in uranium pellets. The problems of the REE determination using ICP-MS technique are the uranium matrix (high concentration of uranium) and the interferences among Lanthanides. In this work, our aim was to develop an effective chemical sample preparation process using extraction chromatography for separation the uranium matrix and the rare earth elements from each other following some publications can be found in the literature and modified them. Secondly, our purpose was the optimization of the ICP-MS measuring process for REE concentration. During method development, in the first step, a REE model solution was used in two different types of extraction chromatographic resins (LN® and TRU®) and different acidic media for environmental testing the Lanthanides separation. Uranium matrix was added to the model solution and was proved in the same conditions. Methods were tested and validated using REE UOC (uranium ore concentrate) reference materials. Samples were analyzed by sector field mass spectrometer (ICP-SFMS).Keywords: extraction chromatography, nuclear forensics, rare earth elements, uranium
Procedia PDF Downloads 3091047 Spatial Distribution of Heavy Metals in Khark Island-Iran Using Geographic Information System
Authors: Abbas Hani, Maryam Jassasizadeh
Abstract:
The concentrations of Cd, Pb, and Ni were determined from 40 soil samples collected in surface soils of Khark Island. Geostatistic methods and GIS were used to identify heavy metal sources and their spatial pattern. Principal component analysis coupled with correlation between heavy metals showed that level of mentioned heavy metal was lower than the standard level. Then the data obtained from the soil analyzing were studied for the purposes of normal distribution. The best way of interior finding for cadmium and nickel was ordinary kriging and the best way of interpolation of lead was inverse distance weighted. The result of this study help us to understand heavy metals distribution and make decision for remediation of soil pollution.Keywords: geostatistics, ordinary kriging, heavy metals, GIS, Khark
Procedia PDF Downloads 1681046 Development of Long and Short Range Ordered Domains in a High Specific Strength Steel
Authors: Nikhil Kumar, Aparna Singh
Abstract:
Microstructural development when annealed at different temperatures in a high aluminum and manganese light weight steel has been examined. The FCC matrix of the manganese (Mn)-rich and nickel (Ni)-rich areas in the studied Fe-Mn-Al-Ni-C-light weight steel have been found to contain anti phase domains. In the Mn-rich region short order range of domains manifested by the diffuse scattering in the electron diffraction patterns was observed. Domains in the Ni-rich region were found to be arranged periodically validated through lattice imaging. The nature of these domains can be tuned with annealing temperature resulting in profound influence in the mechanical properties.Keywords: Anti-phase domain boundaries, BCC, FCC, Light Weight Steel
Procedia PDF Downloads 1411045 Inhibition of Echis ocellatus Venom Metalloprotease by Flavonoid-Rich Ethyl Acetate Sub-fraction of Moringa oleifera Leaves (Lam.): in vitro and in silico Approaches
Authors: Adeyi Akindele Oluwatosin, Mustapha Kaosarat Keji, Ajisebiola Babafemi Siji, Adeyi Olubisi Esther, Damilohun Samuel Metibemu, Raphael Emuebie Okonji
Abstract:
Envenoming by Echis ocellatus is potentially life-threatening due to severe hemorrhage, renal failure, and capillary leakage. These effects are attributed to snake venom metalloproteinases (SVMPs). Due to drawbacks in the use of antivenom, natural inhibitors from plants are of interest in studies of new antivenom treatment. Antagonizing effects of bioactive compounds of Moringa oleifera, a known antisnake plant, are yet to be tested against SVMPs of E. ocellatus (SVMP-EO). Ethanol crude extract of M. oleifera was partitioned using n-hexane and ethyl acetate. Each partition was fractionated using column chromatography and tested against SVMP-EO purified through ion-exchange chromatography with EchiTab-PLUS polyvalent anti-venom as control. Phytoconstituents of ethyl acetate fraction were screened against the catalytic site of crystal of BaP1-SVMP, while drug-likeness and ADMET toxicity of compound were equally determined. The molecular weight of isolated SVMP-EO was 43.28 kDa, with a specific activity of 245 U/ml, a percentage yield of 62.83 %, and a purification fold of 0.920. The Vmax and Km values are 2 mg/ml and 38.095 μmol/ml/min, respectively, while the optimal pH and temperature are 6.0 and 40°C, respectively. Polyvalent anti-venom, crude extract, and ethyl acetate fraction of M. oleifera exhibited a complete inhibitory effect against SVMP-EO activity. The inhibitions of the P-1 and P-II metalloprotease’s enzymes by the ethyl acetate fraction are largely due to methanol, 6, 8, 9-trimethyl-4-(2-phenylethyl)-3-oxabicyclo[3.3.1]non-6-en-1-yl)- and paroxypropione, respectively. Both compounds are potential drug candidates with little or no concern of toxicity, as revealed from the in-silico predictions. The inhibitory effects suggest that this compound might be a therapeutic candidate for further exploration for treatment of Ocellatus’ envenoming.Keywords: Echis ocellatus, Moringa oleifera, anti-venom, metalloproteases, snakebite, molecular docking
Procedia PDF Downloads 1491044 Quality Assessment of the Essential Oil from Eucalyptus globulus Labill of Blida (Algeria) Origin
Authors: M. A. Ferhat, M. N. Boukhatem, F. Chemat
Abstract:
Eucalyptus essential oil is extracted from Eucalyptus globulus of the Myrtaceae family and is also known as Tasmanian blue gum or blue gum. Despite the reputation earned by aromatic and medicinal plants of Algeria. The objectives of this study were: (i) the extraction of the essential oil from the leaves of Eucalyptus globulus Labill., Myrtaceae grown in Algeria, and the quantification of the yield thereof, (ii) the identification and quantification of the compounds in the essential oil obtained, and (iii) the determination of physical and chemical properties of EGEO. The chemical constituents of Eucalyptus globulus essential oil (EGEO) of Blida origin has not previously been investigated. Thus, the present study has been conducted for the determination of chemical constituents and different physico-chemical properties of the EGEO. Chemical composition of the EGEO, grown in Algeria, was analysed by Gas Chromatography-Mass Spectrometry. The chemical components were identified on the basis of Retention Time and comparing with mass spectral database of standard compounds. Relative amounts of detected compounds were calculated on the basis of GC peak areas. Fresh leaves of E. globulus on steam distillation yielded 0.96% (v/w) of essential oil whereas the analysis resulted in the identification of a total of 11 constituents, 1.8 cineole (85.8%), α-pinene (7.2%), and β-myrcene (1.5%) being the main components. Other notable compounds identified in the oil were β-pinene, limonene, α-phellandrene, γ-terpinene, linalool, pinocarveol, terpinen-4-ol, and α-terpineol. The physical properties such as specific gravity, refractive index and optical rotation and the chemical properties such as saponification value, acid number and iodine number of the EGEO were examined. The oil extracted has been analyzed to have 1.4602-1.4623 refractive index value, 0.918-0.919 specific gravity (sp.gr.), +9 - +10 optical rotation that satisfy the standards stipulated by European Pharmacopeia. All the physical and chemical parameters were in the range indicated by the ISO standards. Our findings will help to access the quality of the Eucalyptus oil which is important in the production of high value essential oils that will help to improve the economic condition of the community as well as the nation.Keywords: chemical composition, essential oil, eucalyptol, gas chromatography
Procedia PDF Downloads 3281043 Recrystallization Behavior and Microstructural Evolution of Nickel Base Superalloy AD730 Billet during Hot Forging at Subsolvus Temperatures
Authors: Marcos Perez, Christian Dumont, Olivier Nodin, Sebastien Nouveau
Abstract:
Nickel superalloys are used to manufacture high-temperature rotary engine parts such as high-pressure disks in gas turbine engines. High strength at high operating temperatures is required due to the levels of stress and heat the disk must withstand. Therefore it is necessary parts made from materials that can maintain mechanical strength at high temperatures whilst remain comparatively low in cost. A manufacturing process referred to as the triple melt process has made the production of cast and wrought (C&W) nickel superalloys possible. This means that the balance of cost and performance at high temperature may be optimized. AD730TM is a newly developed Ni-based superalloy for turbine disk applications, with reported superior service properties around 700°C when compared to Inconel 718 and several other alloys. The cast ingot is converted into billet during either cogging process or open die forging. The semi-finished billet is then further processed into its final geometry by forging, heat treating, and machining. Conventional ingot-to-billet conversion is an expensive and complex operation, requiring a significant amount of steps to break up the coarse as-cast structure and interdendritic regions. Due to the size of conventional ingots, it is difficult to achieve a uniformly high level of strain for recrystallization, resulting in non-recrystallized regions that retain large unrecrystallized grains. Non-uniform grain distributions will also affect the ultrasonic inspectability response, which is used to find defects in the final component. The main aim is to analyze the recrystallization behavior and microstructural evolution of AD730 at subsolvus temperatures from a semi-finished product (billet) under conditions representative of both cogging and hot forging operations. Special attention to the presence of large unrecrystallized grains was paid. Double truncated cones (DTCs) were hot forged at subsolvus temperatures in hydraulic press, followed by air cooling. SEM and EBSD analysis were conducted in the as-received (billet) and the as-forged conditions. AD730 from billet alloy presents a complex microstructure characterized by a mixture of several constituents. Large unrecrystallized grains present a substructure characterized by large misorientation gradients with the formation of medium to high angle boundaries in their interior, especially close to the grain boundaries, denoting inhomogeneous strain distribution. A fine distribution of intragranular precipitates was found in their interior, playing a key role on strain distribution and subsequent recrystallization behaviour during hot forging. Continuous dynamic recrystallization (CDRX) mechanism was found to be operating in the large unrecrystallized grains, promoting the formation intragranular DRX grains and the gradual recrystallization of these grains. Evidences that hetero-epitaxial recrystallization mechanism is operating in AD730 billet material were found. Coherent γ-shells around primary γ’ precipitates were found. However, no significant contribution to the overall recrystallization during hot forging was found. By contrast, strain presents the strongest effect on the microstructural evolution of AD730, increasing the recrystallization fraction and refining the structure. Regions with low level of deformation (ε ≤ 0.6) were translated into large fractions of unrecrystallized structures (strain accumulation). The presence of undissolved secondary γ’ precipitates (pinning effect), prior to hot forging operations, could explain these results.Keywords: AD730 alloy, continuous dynamic recrystallization, hot forging, γ’ precipitates
Procedia PDF Downloads 2001042 LIFirr with an Indicator of Microbial Activity in Paraffinic Oil
Authors: M. P. Casiraghi, C. M. Quintella, P. Almeida
Abstract:
Paraffinic oils were submitted to microbial action. The microorganisms consisted of bacteria of the genera Pseudomonas sp and Bacillus lincheniforms. The alterations in interfacial tension were determined using a tensometer and applying the hanging drop technique at room temperature (299 K ±275 K). The alteration in the constitution of the paraffins was evaluated by means of gas chromatography. The microbial activity was observed to reduce interfacial tension by 54 to 78%, as well as consuming the paraffins C19 to C29 and producing paraffins C36 to C44. The LIFirr technique made it possible to determine the microbial action quickly.Keywords: paraffins, biosurfactants, LIFirr, microbial activity
Procedia PDF Downloads 5261041 Predicting the Adsorptive Capacities of Biosolid as a Barrier in Soil to Remove Industrial Contaminants
Authors: H. Aguedal, H. Hentit, A. Aziz, D. R. Merouani, A. Iddou
Abstract:
The major environmental risk of soil pollution is the contamination of groundwater by infiltration of organic and inorganic pollutants that can cause a serious pollution. To protect the groundwater, in this study, we proceeded to test the reliability of a bio solid as barrier to prevent the migration of a very dangerous pollutant ‘Cadmium’ through the different soil layers. The follow-up the influence of several parameters, such as: turbidity, pluviometry, initial concentration of cadmium and the nature of soil, allow us to find the most effective manner to integrate this barrier in the soil. From the results obtained, we noted the effective intervention of the barrier. Indeed, the recorded passing quantities are lowest for the highest rainfall; we noted that the barrier has a better affinity towards higher concentrations; the most retained amounts of cadmium has been in the top layer of the two types of soil, while the lowest amounts of cadmium are recorded in the inner layers of soils.Keywords: adsorption of cadmium, barrier, groundwater pollution, protection
Procedia PDF Downloads 3641040 Gas Chromatography-Analysis, Antioxidant, Anti-Inflammatory, and Anticancer Activities of Some Extracts and Fractions of Linum usitatissimum
Authors: Eman Abdullah Morsi, Hend Okasha, Heba Abdel Hady, Mortada El-Sayed, Mohamed Abbas Shemis
Abstract:
Context: Linum usitatissimum (Linn), known as Flaxseed, is one of the most important medicinal plants traditionally used for various health as nutritional purposes. Objective: Estimation of total phenolic and flavonoid contents as well as evaluate the antioxidant using α, α-diphenyl-β-picrylhydrazyl (DPPH), 2-2'azinobis (3-ethylbenzthiazoline-6-sulphonic acid (ABTS) and total antioxidant capacity (TAC) assay and investigation of anti-inflammatory by Bovine serum albumin (BSA) and anticancer activities of hepatocellular carcinoma cell line (HepG2) and breast cancer cell line (MCF7) have been applied on hexane, ethyl acetate, n-butanol and methanol extracts and also, fractions of methonal extract (hexane, ethyl acetate and n-butanol). Materials and Methods: Phenolic and flavonoid contents were detected using spectrophotometric and colorimetric assays. Antioxidant and anti-inflammatory activities were estimated in-vitro. Anticancer activity of extracts and fractions of methanolic extract were tested on (HepG2) and (MCF7). Results: Methanolic extract and its ethyl acetate fraction contain higher contents of total phenols and flavonoids. In addition, methanolic extract had higher antioxidant activity. Butanolic and ethyl acetate fractions yielded higher percent of inhibition of protein denaturation. Meanwhile, ethyl acetate fraction and methanolic extract had anticancer activity against HepG2 and MCF7 (IC50=60 ± 0.24 and 29.4 ± 0.12µg.mL⁻¹) and (IC50=94.7 ± 0.21 and 227 ± 0.48µg.mL⁻¹), respectively. In Gas chromatography-mass spectrometry (GC-MS) analysis, methanolic extract has 32 compounds, whereas; ethyl acetate and butanol fractions contain 40 and 36 compounds, respectively. Conclusion: Flaxseed contains totally different biologically active compounds that have been found to possess good variable activities, which can protect human body against several diseases.Keywords: phenolic content, flavonoid content, HepG2, MCF7, hemolysis-assay, flaxseed
Procedia PDF Downloads 1261039 Assessing the Mass Concentration of Microplastics and Nanoplastics in Wastewater Treatment Plants by Pyrolysis Gas Chromatography−Mass Spectrometry
Authors: Yanghui Xu, Qin Ou, Xintu Wang, Feng Hou, Peng Li, Jan Peter van der Hoek, Gang Liu
Abstract:
The level and removal of microplastics (MPs) in wastewater treatment plants (WWTPs) has been well evaluated by the particle number, while the mass concentration of MPs and especially nanoplastics (NPs) remains unclear. In this study, microfiltration, ultrafiltration and hydrogen peroxide digestion were used to extract MPs and NPs with different size ranges (0.01−1, 1−50, and 50−1000 μm) across the whole treatment schemes in two WWTPs. By identifying specific pyrolysis products, pyrolysis gas chromatography−mass spectrometry were used to quantify their mass concentrations of selected six types of polymers (i.e., polymethyl methacrylate (PMMA), polypropylene (PP), polystyrene (PS), polyethylene (PE), polyethylene terephthalate (PET), and polyamide (PA)). The mass concentrations of total MPs and NPs decreased from 26.23 and 11.28 μg/L in the influent to 1.75 and 0.71 μg/L in the effluent, with removal rates of 93.3 and 93.7% in plants A and B, respectively. Among them, PP, PET and PE were the dominant polymer types in wastewater, while PMMA, PS and PA only accounted for a small part. The mass concentrations of NPs (0.01−1 μm) were much lower than those of MPs (>1 μm), accounting for 12.0−17.9 and 5.6− 19.5% of the total MPs and NPs, respectively. Notably, the removal efficiency differed with the polymer type and size range. The low-density MPs (e.g., PP and PE) had lower removal efficiency than high-density PET in both plants. Since particles with smaller size could pass the tertiary sand filter or membrane filter more easily, the removal efficiency of NPs was lower than that of MPs with larger particle size. Based on annual wastewater effluent discharge, it is estimated that about 0.321 and 0.052 tons of MPs and NPs were released into the river each year. Overall, this study investigated the mass concentration of MPs and NPs with a wide size range of 0.01−1000 μm in wastewater, which provided valuable information regarding the pollution level and distribution characteristics of MPs, especially NPs, in WWTPs. However, there are limitations and uncertainties in the current study, especially regarding the sample collection and MP/NP detection. The used plastic items (e.g., sampling buckets, ultrafiltration membranes, centrifugal tubes, and pipette tips) may introduce potential contamination. Additionally, the proposed method caused loss of MPs, especially NPs, which can lead to underestimation of MPs/NPs. Further studies are recommended to address these challenges about MPs/NPs in wastewater.Keywords: microplastics, nanoplastics, mass concentration, WWTPs, Py-GC/MS
Procedia PDF Downloads 2811038 A High-Throughput Enzyme Screening Method Using Broadband Coherent Anti-stokes Raman Spectroscopy
Authors: Ruolan Zhang, Ryo Imai, Naoko Senda, Tomoyuki Sakai
Abstract:
Enzymes have attracted increasing attentions in industrial manufacturing for their applicability in catalyzing complex chemical reactions under mild conditions. Directed evolution has become a powerful approach to optimize enzymes and exploit their full potentials under the circumstance of insufficient structure-function knowledge. With the incorporation of cell-free synthetic biotechnology, rapid enzyme synthesis can be realized because no cloning procedure such as transfection is needed. Its open environment also enables direct enzyme measurement. These properties of cell-free biotechnology lead to excellent throughput of enzymes generation. However, the capabilities of current screening methods have limitations. Fluorescence-based assay needs applicable fluorescent label, and the reliability of acquired enzymatic activity is influenced by fluorescent label’s binding affinity and photostability. To acquire the natural activity of an enzyme, another method is to combine pre-screening step and high-performance liquid chromatography (HPLC) measurement. But its throughput is limited by necessary time investment. Hundreds of variants are selected from libraries, and their enzymatic activities are then identified one by one by HPLC. The turn-around-time is 30 minutes for one sample by HPLC, which limits the acquirable enzyme improvement within reasonable time. To achieve the real high-throughput enzyme screening, i.e., obtain reliable enzyme improvement within reasonable time, a widely applicable high-throughput measurement of enzymatic reactions is highly demanded. Here, a high-throughput screening method using broadband coherent anti-Stokes Raman spectroscopy (CARS) was proposed. CARS is one of coherent Raman spectroscopy, which can identify label-free chemical components specifically from their inherent molecular vibration. These characteristic vibrational signals are generated from different vibrational modes of chemical bonds. With the broadband CARS, chemicals in one sample can be identified from their signals in one broadband CARS spectrum. Moreover, it can magnify the signal levels to several orders of magnitude greater than spontaneous Raman systems, and therefore has the potential to evaluate chemical's concentration rapidly. As a demonstration of screening with CARS, alcohol dehydrogenase, which converts ethanol and nicotinamide adenine dinucleotide oxidized form (NAD+) to acetaldehyde and nicotinamide adenine dinucleotide reduced form (NADH), was used. The signal of NADH at 1660 cm⁻¹, which is generated from nicotinamide in NADH, was utilized to measure the concentration of it. The evaluation time for CARS signal of NADH was determined to be as short as 0.33 seconds while having a system sensitivity of 2.5 mM. The time course of alcohol dehydrogenase reaction was successfully measured from increasing signal intensity of NADH. This measurement result of CARS was consistent with the result of a conventional method, UV-Vis. CARS is expected to have application in high-throughput enzyme screening and realize more reliable enzyme improvement within reasonable time.Keywords: Coherent Anti-Stokes Raman Spectroscopy, CARS, directed evolution, enzyme screening, Raman spectroscopy
Procedia PDF Downloads 1411037 Novel Aminoglycosides to Target Resistant Pathogens
Authors: Nihar Ranjan, Derrick Watkins, Dev P. Arya
Abstract:
Current methods in the study of antibiotic activity of ribosome targeted antibiotics are dependent on cell based bacterial inhibition assays or various forms of ribosomal binding assays. These assays are typically independent of each other and little direct correlation between the ribosomal binding and bacterial inhibition is established with the complementary assay. We have developed novel high-throughput capable assays for ribosome targeted drug discovery. One such assay examines the compounds ability to bind to a model ribosomal RNA A-site. We have also coupled this assay to other functional orthogonal assays. Such analysis can provide valuable understanding of the relationships between two complementary drug screening methods and could be used as standard analysis to correlate the affinity of a compound for its target and the effect the compound has on a cell.Keywords: bacterial resistance, aminoglycosides, screening, drugs
Procedia PDF Downloads 3711036 Functional Surfaces and Edges for Cutting and Forming Tools Created Using Directed Energy Deposition
Authors: Michal Brazda, Miroslav Urbanek, Martina Koukolikova
Abstract:
This work focuses on the development of functional surfaces and edges for cutting and forming tools created through the Directed Energy Deposition (DED) technology. In the context of growing challenges in modern engineering, additive technologies, especially DED, present an innovative approach to manufacturing tools for forming and cutting. One of the key features of DED is its ability to precisely and efficiently deposit Fully dense metals from powder feedstock, enabling the creation of complex geometries and optimized designs. Gradually, it becomes an increasingly attractive choice for tool production due to its ability to achieve high precision while simultaneously minimizing waste and material costs. Tools created using DED technology gain significant durability through the utilization of high-performance materials such as nickel alloys and tool steels. For high-temperature applications, Nimonic 80A alloy is applied, while for cold applications, M2 tool steel is used. The addition of ceramic materials, such as tungsten carbide, can significantly increase the tool's resistance. The introduction of functionally graded materials is a significant contribution, opening up new possibilities for gradual changes in the mechanical properties of the tool and optimizing its performance in different sections according to specific requirements. In this work, you will find an overview of individual applications and their utilization in the industry. Microstructural analyses have been conducted, providing detailed insights into the structure of individual components alongside examinations of the mechanical properties and tool life. These analyses offer a deeper understanding of the efficiency and reliability of the created tools, which is a key element for successful development in the field of cutting and forming tools. The production of functional surfaces and edges using DED technology can result in financial savings, as the entire tool doesn't have to be manufactured from expensive special alloys. The tool can be made from common steel, onto which a functional surface from special materials can be applied. Additionally, it allows for tool repairs after wear and tear, eliminating the need for producing a new part and contributing to an overall cost while reducing the environmental footprint. Overall, the combination of DED technology, functionally graded materials, and verified technologies collectively set a new standard for innovative and efficient development of cutting and forming tools in the modern industrial environment.Keywords: additive manufacturing, directed energy deposition, DED, laser, cutting tools, forming tools, steel, nickel alloy
Procedia PDF Downloads 501035 Hydrochemistry and Stable Isotopes (ẟ18O and ẟ2H) Tools Applied to the Study of Karst Aquifers in Wonderfonteinspruit Valley: North West, South Africa
Authors: Naziha Mokadem, Rainier Dennis, Ingrid Dennis
Abstract:
In South Africa, Karst aquifers are receiving greater attention since they provide large supplies of water which is used for domestic and agricultural purposes as well as for industry. Accordingly, a better insight into the origin of water mineralization and the geochemical processes controlling the recharge of the aquifer is crucial. Analyses of geochemical and environmental isotopes could lead to relevant information regarding karstification and infiltration processes, groundwater chemistry and isotopy. A study was conducted in a typical karst landscape of Wonderfonteinspruit catchment, also known as Wonderfonteinspruit Valley in North-western -South Africa. Furthermore, fifty-two samples were collected from (35 boreholes, 5 surface waters, 4 Dams, 4 springs, 1 canal, 2 pipelines, 1 cave) within the study area for hydrochemistry and 2H and 18O analysis. The determination of the anions (Cl-, SO42-, NO2, NO3-) were performed using Metrohm ion chromatography, model: 761 compact IC, with a precision of ± 0.001 mg/l. While, the cations (Na+, Mg2+, K+, Ca2+) were determined using Metrohm ion chromatography, Model: ICP-MS 7500 series. The alkalinity (Alk) was determined by pH meter with volumetric titration using HCL to pH 4.5; 4.2; and 8.2. In addition, 18O and 2H relative to the Vienna-Standard Mean Ocean Water (RVSMOW), were determined by picarro L2130-I Isotopic H2O (Cavity Ringdown laser spectrometer, Picarro Ltd). The hydrochemical analysis of Wonderfonteinspruit groundwater showed a dominance of the cations Ca-Mg and the anion HCO3. Piper diagram shows that the groundwater sample of study area is characterized by four hydrochemical facies: Two main groups: (1) Ca–Mg–Cl–SO4; (2) Ca–Mg–HCO3 and two minor groups: (3) Ca–Mg–Cl; (4) Na–K–HCO3. The majority of boreholes of Malmani (Transvaal Supergroup) aquifer are plotted in Ca–Mg–HCO3.Oxygen-18 (18O‰SMOW) and deuterium (D‰SMOW) isotopic data indicate that the aquifer’s recharge is influenced by two phenomena; precipitation rates for most of the samples and river flow (Wonderfonteinspruit, Middelvieinspruit, Renfonteinspruit) for some samples.Keywords: South Africa, Wonderfonteinspruit Valley, isotopic, hydrochemical, carbonate aquifers
Procedia PDF Downloads 154