Search results for: planar chromatography

Authors: Ifeanyichukwu Edeh, Tim Overton, Steve Bowra

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Currently biodiesel is produced from plant oils or animal’s fats by a liquid-phase catalysed transesterification process at low temperature. Although biodiesel is renewable and to a large extent sustainable, inherent properties such as poor cold flow, low oxidation stability, low cetane value restrict application to blends with fossil fuels. An alternative to biodiesel is renewable diesel produced by catalytic hydrotreating of oils and fats and is considered a drop in fuel because its properties are similar to petroleum diesel. In addition to developing alternative productions routes there is continued interest in reducing the cost of the feed stock, waste cooking oils and fats are increasingly used as the feedstocks due to low cost. However, use of oils and fat are highly adulterated resulting in high free fatty acid content which turn impacts on the efficiency of FAME production. Therefore, in light of the need to develop, alternative lipid feed stocks and related efficient catalysis the present study investigates the potential of producing renewable diesel from the lipids-extracted from activated sludge, a waste water treatment by-product, through catalytic hydrothermal decarboxylation. The microbial lipids were first extracted from the activated sludge using the Folch et al method before hydrothermal decarboxylation reactions were carried out using palladium (Pd/C) and platinum (Pt/C) on activated carbon as the catalysts in a batch reactor. The impact of three temperatures 290, 300, 330 °C and residence time between 30 min and 4hrs was assessed. At the end of the reaction, the products were recovered using organic solvents and characterized using gas chromatography (GC). The principle products of the reaction were pentadecane and heptadecane. The highest yields of pentadecane and heptadecane from lipid-extract were 23.23% and 15.21%, respectively. These yields were obtained at 290 °C and residence time 1h using Pt/C. To the best of our knowledge, the current work is the first investigation on the hydrothermal decarboxylation of lipid-extract from activated sludge.

Keywords: activated sludge, lipid, hydrothermal decarboxylation, renewable diesel

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Authors: Gargi Prasad, Ashiho A. Mao, Deepu Vijayan, S. Mandal

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The main objective of the present study was to optimize and develop an efficient protocol for in vitro propagation of a medicinally important orchid Cephalantheropsis obcordata (Lindl.) Ormerod along with genetic stability analysis of regenerated plants. This plant has been traditionally used in Chinese folk medicine and the decoction of whole plant is known to possess anticancer activity. Nodal segments used as explants were inoculated on Murashige and Skoog (MS) medium supplemented with various concentrations of isopentenyl adenine (2iP). The rooted plants were successfully acclimatized in the greenhouse with 100% survival rate. Inter-simple sequence repeats (ISSR) markers were used to assess the genetic fidelity of in vitro raised plants and the mother plant. It was revealed that monomorphic bands showing the absence of polymorphism in all in vitro raised plantlets analyzed, confirming the genetic uniformity among the regenerants. Phytochemical analysis was done to compare the antioxidant activities and HPLC fingerprinting assay of 80% aqueous ethanol extract of the leaves and stem of in vitro and in vivo grown C. obcordata. The extracts of the plants were examined for their antioxidant activities by using free radical 1, 1-diphenyl-2-picryl hydrazyl (DPPH) scavenging method, 2,2’-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging ability, reducing power capacity, estimation of total phenolic content, flavonoid content and flavonol content. A simplified method for the detection of ascorbic acid, phenolic acids and flavonoids content was also developed by using reversed phase high-performance liquid chromatography (HPLC). This is the first report on the micropropagation, genetic integrity study and quantitative phytochemical analysis of in vitro regenerated plants of C. obcordata.

Keywords: Cephalantheropsis obcordata, genetic fidelity, ISSR markers, HPLC

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Authors: Thamer Alshuwaili, Yonglin Ren, Bob Du, Manjree Agarwal

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The warehouse beetle, Trogoderma variabile Ballion (Coleoptera: Dermestidae), is a major pest of packaged and processed stored products. Warehouse beetle is the common name which was given by Okumura (1972). This pest has been reported to infest 119 different commodities, and it is distributed throughout the tropical and subtropical parts of the world. Also, it is difficult to control because of the insect's ability to stay without food for long times, and it can survive for years under dry conditions and low-moisture food, and it has also developed resistance to many insecticides. The young larvae of these insects can cause damage to seeds, but older larvae prefer to feed on whole grains. The percentage of damage caused by these insects range between 30-70% in the storage. T. variabile is the species most responsible for causing significant damage in grain stores worldwide. Trogoderma spp. is a huge problem for cereal grains, and there are many countries, such as the USA, Australia, China, Kenya, Uganda and Tanzania who have specific quarantine regulations against possible importation. Also, grain stocks can be almost completely destroyed because of the massive populations the insect may develop. However, the purpose of the current research was to optimize conditions to collect volatile organic compound from Trogoderma variabile at different life stages by using headspace solid phase microextraction (SPME) coupled with gas chromatography-mass spectrometry (GC-MS) and flame ionization detection (FID). Using SPME technique to extract volatile from insects is an efficient, straightforward and nondestructive method. Result of the study shows that 15 insects were optimal number for larvae and adults. Selection of the number of insects depend on the height of the peak area and the number of peaks. Sixteen hours were optimized as the best extraction time for larvae and 8 hours was the optimal number of adults.

Keywords: Trogoderma variabile, warehouse beetle , GC-MS, Solid phase microextraction

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Authors: W. Jahouach-Rabai, J. Aribi, Z. Azzouz-Berriche, R. Lahsni, F. Hosni

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Gamma irradiation applied in removing pharmaceutical contaminants from wastewater is an effective advanced oxidation process (AOP), considered as an alternative to conventional water treatment technologies. In this purpose, the degradation efficiency of several detected contaminants under gamma irradiation was evaluated. In fact, radiolysis of organic pollutants in aqueous solutions produces powerful reactive species, essentially hydroxyl radical ( ·OH), able to destroy recalcitrant pollutants in water. Pharmaceuticals considered in this study are aqueous solutions of paracetamol, ibuprofen, and diclofenac at different concentrations 0.1-1 mmol/L, which were treated with irradiation doses from 3 to 15 kGy. The catalytic oxidation of these compounds by gamma irradiation was investigated using hydrogen peroxide (H₂O₂) as a convenient oxidant. Optimization of the main parameters influencing irradiation process, namely irradiation doses, initial concentration and oxidant volume (H₂O₂) were investigated, in the aim to release high degradation efficiency of considered pharmaceuticals. Significant modifications attributed to these parameters appeared in the variation of degradation efficiency, chemical oxygen demand removal (COD) and concentration of radio-induced radicals, confirming them synergistic effect to attempt total mineralization. Pseudo-first-order reaction kinetics could be used to depict the degradation process of these compounds. A sophisticated analytical study was released to quantify the detected radio-induced radicals (electron paramagnetic resonance spectroscopy (EPR) and high performance liquid chromatography (HPLC)). All results showed that this process is effective for the degradation of many pharmaceutical products in aqueous solutions due to strong oxidative properties of generated radicals mainly hydroxyl radical. Furthermore, the addition of an optimal amount of H₂O₂ was efficient to improve the oxidative degradation and contribute to the high performance of this process at very low doses (0.5 and 1 kGy).

Keywords: AOP, COD, hydroxyl radical, EPR, gamma irradiation, HPLC, pharmaceuticals

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Authors: Hanan Khojah, RuAngelie Edrada-Ebel

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Neurodegenerative disease including Alzheimer’s disease is a major cause of long-term disability. Oxidative stress is frequently implicated as one of the key contributing factors to neurodegenerative diseases. Protection against neuronal damage remains a great challenge for researchers. Ficus carica (commonly known as fig) is a species of great antioxidant nutritional value comprising a protective mechanism against innumerable health disorders related to oxidative stress as well as Alzheimer’s disease. The purpose of this work was to characterize the non-polar active metabolites in Ficus carica endocarp, mesocarp, and exocarp. Crude extracts were prepared using several extraction solvents, which included 1:1 water: ethylacetate, acetone and methanol. The dried extracts were then solvent partitioned between equivalent amounts of water and ethylacetate. Purification and fractionation were accomplished by high-throughput chromatography. The isolated metabolites were tested on their effect on human neuroblastoma cell line by cell viability test and cell cytotoxicity assay with acrolein. Molecular weights of the active metabolites were determined via LC–HRESIMS and GC-EIMS. Metabolomic profiling was performed to identify the active metabolites by using differential expression analysis software (Mzmine) and SIMCA for multivariate analysis. Structural elucidation and identification of the interested active metabolites were studied by 1-D and 2-D NMR. Significant differences in bioactivity against a concentration-dependent assay on acrolein radicals were observed between the three fruit parts. However, metabolites obtained from mesocarp and the endocarp demonstrated bioactivity to scavenge ROS radical. NMR profiling demonstrated that aliphatic compounds such as γ-sitosterol tend to induce neuronal bioactivity and exhibited bioactivity on the cell viability assay. γ-Sitosterol was found in higher concentrations in the mesocarp and was considered as one of the major phytosterol in Ficus carica.

Keywords: alzheimer, Ficus carica, γ-Sitosterol, metabolomics

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Authors: George Madalin Danila, Mihaiella Cretu, Cristian Puscasu

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The recycling of used antifreeze/coolant is a widely discussed and intricate issue. Complying with government regulations for the proper disposal of hazardous waste poses a significant challenge for today's automotive and industrial industries. In recent years, global focus has shifted toward Earth's fragile ecology, emphasizing the need to restore and preserve the natural environment. The business and industrial sectors have undergone substantial changes to adapt and offer products tailored to these evolving markets. The global antifreeze market size was evaluated at US 5.4 billion in 2020 to reach USD 5,9 billion by 2025 due to the increased number of vehicles worldwide, but also to the growth of HVAC systems. This study presents the evaluation of an ion exchange resin-based installation designed for the recycling of engine coolants, specifically ethylene glycol (EG) and propylene glycol (PG). The recycling process aims to restore the coolant to meet the stringent ASTM standards for both new and recycled coolants. A combination of physical-chemical methods, gas chromatography-mass spectrometry (GC-MS), and inductively coupled plasma mass spectrometry (ICP-MS) was employed to analyze and validate the purity and performance of the recycled product. The experimental setup included performance tests, namely corrosion to glassware and the tendency to foaming of coolant, to assess the efficacy of the recycled coolants in comparison to new coolant standards. The results demonstrate that the recycled EG coolants exhibit comparable quality to new coolants, with all critical parameters falling within the acceptable ASTM limits. This indicates that the ion exchange resin method is a viable and efficient solution for the recycling of engine coolants, offering an environmentally friendly alternative to the disposal of used coolants while ensuring compliance with industry standards.

Keywords: engine coolant, glycols, recycling, ion exchange resin, circular economy

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Authors: Martin Arguelles Perez, Woosoon Yim, Alexander Barzilov

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High-fidelity radiation monitoring is an essential component in the enhancement of the situational awareness capabilities of the Department of Energy’s Office of Environmental Management (DOE-EM) personnel. In this paper, multiple units of unmanned aerial vehicles (UAVs) each equipped with a cadmium zinc telluride (CZT) gamma-ray sensor are used for radiation source localization, which can provide vital real-time data for the EM tasks. To achieve this goal, a fully autonomous system of multicopter-based UAV swarm in 3D tetrahedron formation is used for surveying the area of interest and performing radiation source localization. The CZT sensor used in this study is suitable for small-size multicopter UAVs due to its small size and ease of interfacing with the UAV’s onboard electronics for high-resolution gamma spectroscopy enabling the characterization of radiation hazards. The multicopter platform with a fully autonomous flight feature is suitable for low-altitude applications such as radiation contamination sites. The conventional approach uses a single UAV mapping in a predefined waypoint path to predict the relative location and strength of the source, which can be time-consuming for radiation localization tasks. The proposed UAV swarm-based approach can significantly improve its ability to search for and track radiation sources. In this paper, two approaches are developed using (a) 2D planar circular (3 UAVs) and (b) 3D tetrahedron formation (4 UAVs). In both approaches, accurate estimation of the gradient vector is crucial for heading angle calculation. Each UAV carries the CZT sensor; the real-time radiation data are used for the calculation of a bulk heading vector for the swarm to achieve a UAV swarm’s source-seeking behavior. Also, a spinning formation is studied for both cases to improve gradient estimation near a radiation source. In the 3D tetrahedron formation, a UAV located closest to the source is designated as a lead unit to maintain the tetrahedron formation in space. Such a formation demonstrated a collective and coordinated movement for estimating a gradient vector for the radiation source and determining an optimal heading direction of the swarm. The proposed radiation localization technique is studied by computer simulation and validated experimentally in the indoor flight testbed using gamma sources. The technology presented in this paper provides the capability to readily add/replace radiation sensors to the UAV platforms in the field conditions enabling extensive condition measurement and greatly improving situational awareness and event management. Furthermore, the proposed radiation localization approach allows long-term measurements to be efficiently performed at wide areas of interest to prevent disasters and reduce dose risks to people and infrastructure.

Keywords: radiation, unmanned aerial system(UAV), source localization, UAV swarm, tetrahedron formation

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Authors: Efstratios Karavasilis, Foteini Christidi, Georgios Velonakis, Agapi Plousi, Kalliopi Platoni, Nikolaos Kelekis, Ioannis Evdokimidis, Efstathios Efstathopoulos

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Introduction: Advanced structural and functional neuroimaging techniques contribute to the study of anatomical and functional brain connectivity and its role in the pathophysiology and symptoms’ heterogeneity in several neurodegenerative disorders, including multiple sclerosis (MS). Aim: In the present study, we applied multiparametric neuroimaging techniques to investigate the structural and functional cortico-cerebellar changes in MS patients. Material: We included 51 MS patients (28 with clinically isolated syndrome [CIS], 31 with relapsing-remitting MS [RRMS]) and 51 age- and gender-matched healthy controls (HC) who underwent MRI in a 3.0T MRI scanner. Methodology: The acquisition protocol included high-resolution 3D T1 weighted, diffusion-weighted imaging and echo planar imaging sequences for the analysis of volumetric, tractography and functional resting state data, respectively. We performed between-group comparisons (CIS, RRMS, HC) using CAT12 and CONN16 MATLAB toolboxes for the analysis of volumetric (cerebellar gray matter density) and functional (cortico-cerebellar resting-state functional connectivity) data, respectively. Brainance suite was used for the analysis of tractography data (cortico-cerebellar white matter integrity; fractional anisotropy [FA]; axial and radial diffusivity [AD; RD]) to reconstruct the cerebellum tracts. Results: Patients with CIS did not show significant gray matter (GM) density differences compared with HC. However, they showed decreased FA and increased diffusivity measures in cortico-cerebellar tracts, and increased cortico-cerebellar functional connectivity. Patients with RRMS showed decreased GM density in cerebellar regions, decreased FA and increased diffusivity measures in cortico-cerebellar WM tracts, as well as a pattern of increased and mostly decreased functional cortico-cerebellar connectivity compared to HC. The comparison between CIS and RRMS patients revealed significant GM density difference, reduced FA and increased diffusivity measures in WM cortico-cerebellar tracts and increased/decreased functional connectivity. The identification of decreased WM integrity and increased functional cortico-cerebellar connectivity without GM changes in CIS and the pattern of decreased GM density decreased WM integrity and mostly decreased functional connectivity in RRMS patients emphasizes the role of compensatory mechanisms in early disease stages and the disintegration of structural and functional networks with disease progression. Conclusions: In conclusion, our study highlights the added value of multimodal neuroimaging techniques for the in vivo investigation of cortico-cerebellar brain changes in neurodegenerative disorders. An extension and future opportunity to leverage multimodal neuroimaging data inevitably remain the integration of such data in the recently-applied mathematical approaches of machine learning algorithms to more accurately classify and predict patients’ disease course.

Keywords: advanced neuroimaging techniques, cerebellum, MRI, multiple sclerosis

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Authors: Y. A. Prada, Fanny Guzman, Rafael Cabanzo, John J. Castillo, Enrique Mejia-Ospino

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In this work, we show the important results about of synthesis of photoluminiscents gold nanoclusters using a small peptide as template for biosensing applications. Interestingly, we design one peptide (NBC2854) homologue to conservative domain from 215 250 residue of a galactolectin protein which can recognize the proteophosphoglycans (PPG) from Leishmania. Peptide was synthetized by multiple solid phase synthesis using FMoc group methodology in acid medium. Finally, the peptide was purified by High-Performance Liquid Chromatography using a Vydac C-18 preparative column and the detection was at 215 nm using a Photo Diode Array detector. Molecular mass of this peptide was confirmed by MALDI-TOF and to verify the α-helix structure we use Circular Dichroism. By means of the methodology used we obtained a novel fluorescents gold nanoclusters (AuNC) using NBC2854 as a template. In this work, we described an easy and fast microsonic method for the synthesis of AuNC with ≈ 3.0 nm of hydrodynamic size and photoemission at 630 nm. The presence of cysteine residue in the C-terminal of the peptide allows the formation of Au-S bond which confers stability to Peptide-based gold nanoclusters. Interactions between the peptide and gold nanoclusters were confirmed by X-ray Photoemission and Raman Spectroscopy. Notably, from the ultrafine spectra shown in the MALDI-TOF analysis which containing only 3-7 KDa species was assigned to Au₈-₁₈[NBC2854]₂ clusters. Finally, we evaluated the Peptide-gold nanocluster as an optical biosensor based on fluorescence spectroscopy and the fluorescence signal of PPG (0.1 µg-mL⁻¹ to 1000 µg-mL⁻¹) was amplified at the same wavelength emission (≈ 630 nm). This can suggest that there is a strong interaction between PPG and Pep@AuNC, therefore, the increase of the fluorescence intensity can be related to the association mechanism that take place when the target molecule is sensing by the Pep@AuNC conjugate. Further spectroscopic studies are necessary to evaluate the fluorescence mechanism involve in the sensing of the PPG by the Pep@AuNC. To our best knowledge the fabrication of an optical biosensor based on Pep@AuNC for sensing biomolecules such as Proteophosphoglycans which are secreted in abundance by parasites Leishmania.

Keywords: biosensing, fluorescence, Leishmania, peptide-gold nanoclusters, proteophosphoglycans

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Authors: Samiyah Tasleem, Muhammad Abdul Haq, Syed Baqir Shyum Naqvi, Muhammad Abid Husnain, Sajjad Haider Naqvi

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The objective of the present study was: a) to investigate the antimicrobial activity of honey samples of different floral sources of Pakistan, b) to recover the phenolic acids in them as a possible contributing factor of antimicrobial activity. Six honey samples from different floral sources, namely: Trachysperm copticum, Acacia species, Helianthus annuus, Carissa opaca, Zizyphus and Magnifera indica were used. The antimicrobial activity was investigated by the disc diffusion method against eight freshly isolated clinical isolates (Staphylococci aureus, Staphylococci epidermidis, Streptococcus faecalis, Pseudomonas aeruginosa, Klebsiella pneumonia, Escherichia coli, Proteus vulgaris and Candida albicans). Antimicrobial activity of honey was compared with five commercial antibiotics, namely: doxycycline (DO-30ug/mL), oxytetracycline (OT-30ug/mL), clarithromycin (CLR–15ug/mL), moxifloxacin (MXF-5ug/mL) and nystatin (NT – 100 UT). The fractions responsible for antimicrobial activity were extracted using ethyl acetate. Solid phase extraction (SPE) was used to recover the phenolic acids of honey samples. Identification was carried out via High-Performance Liquid Chromatography (HPLC). The results indicated that antimicrobial activity was present in all honey samples and found comparable to the antibiotics used in the study. In the microbiological assay, the ethyl acetate honey extract was found to exhibit a very promising antimicrobial activity against all the microorganisms tested, indicating the existence of phenolic compounds. Six phenolic acids, namely: gallic, caffeic, ferulic, vanillic, benzoic and cinnamic acids were identified besides some unknown substance by HPLC. In conclusion, Pakistani honey samples showed a broad spectrum antibacterial and promising antifungal activity. Identification of six different phenolic acids showed that Pakistani honey samples are rich sources of phenolic compounds that could be the contributing factor of antimicrobial activity.

Keywords: Pakistani honey, antimicrobial activity, Phenolic acids eg.gallic, caffeic, ferulic, vanillic, benzoic and cinnamic acids

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Authors: Jelka Pleadin, Greta Krešić, Tina Lešić, Ana Vulić, Renata Barić, Tanja Bogdanović, Dražen Oraić, Ana Legac, Snježana Zrnčić

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Fish has been acknowledged as an integral component of a well-balanced diet, providing a healthy source of energy, high-quality proteins, vitamins, essential minerals and, especially, n-3 long-chain polyunsaturated fatty acids (n-3 LC PUFA), mainly eicosapentaenoic acid (20:5 n-3 EPA), and docosahexaenoicacid, (22:6 n-3 DHA), whose pleiotropic effects in terms of health promotion and disease prevention have been increasingly recognised. In this study, the fatty acids composition of three commercially important farmed fish species: sea bream (Sparus aurata), sea bass (Dicentrarchus labrax) and dentex (Dentex dentex) was investigated. In total, 60 fish samples were retrieved during 2015 (n = 30) and 2016 (n = 30) from different locations in the Adriatic Sea. Methyl esters of fatty acids were analysed using gas chromatography (GC) with flame ionization detection (FID). The results show that the most represented fatty acid in all three analysed species is oleic acid (C18:1n-9, OA), followed by linoleic acid (C18:2n-6, LA) and palmitic acid (C16:0, PA). Dentex was shown to have two to four times higher eicosapentaenoic (EPA) and docosahexaenoic (DHA) acid content as compared to sea bream and sea bass. The recommended n-6/n-3 ratio was determined in all fish species but obtained results pointed to statistically significant differences (p < 0.05) in fatty acid composition among the analysed fish species and their potential as a dietary source of valuable fatty acids. Sea bass and sea bream had a significantly higher proportion of n-6 fatty acids, while dentex had a significantly higher proportion of n-3 (C18:4n-3, C20:4n-3, EPA, DHA) fatty acids. A higher hypocholesterolaemic and hypercholesterolaemic fatty acids (HH) ratio was determined for sea bass and sea bream, which comes as the consequence of a lower share of SFA determined in these two species in comparison to dentex. Since the analysed fish species vary in their fatty acids composition consumption of diverse fish species would be advisable. Based on the established lipid quality indicators, dentex, a fish species underutilised by the aquaculture, seems to be a highly recommendable and important source of fatty acids recommended to be included into the human diet.

Keywords: dentex, fatty acids, farmed fish, sea bass, sea bream

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Authors: Siyabonga Aubrey Mhlongo, Linda Lunga Sibali, Phumlane Selby Mdluli, Peter Papoh Ndibewu, Kholofelo Clifford Malematja

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This paper reports on the successful extraction and quantification of an antibacterial and antifungal agent present in some consumer products (Triclosan: C₁₂H₇Cl₃O₂)generally found in wastewater or effluents using molecularly imprinted membrane adsorbent (MIMs) followed by quantification and removal on a high-performance liquid chromatography (HPLC). Triclosan is an antibacterial and antifungal agent present in some consumer products like toothpaste, soaps, detergents, toys, and surgical cleaning treatments. The MIMs was fabricated usingpolyvinylidene fluoride (PVDF) polymer with selective micro composite particles known as molecularly imprinted polymers (MIPs)via a phase inversion by immersion precipitation technique. This resulted in an improved hydrophilicity and mechanical behaviour of the membranes. Wastewater samples were collected from the Umbogintwini Industrial Complex (UIC) (south coast of Durban, KwaZulu-Natal in South Africa). central UIC effluent treatment plant and pre-treated before analysis. Experimental parameters such as sample size, contact time, stirring speed were optimised. The resultant MIMs had an adsorption efficiency of 97% of TCS with reference to NIMs and bare membrane, which had 92%, 88%, respectively. The analytical method utilized in this review had limits of detection (LoD) and limits of quantification (LoQ) of 0.22, 0.71µgL-1 in wastewater effluent, respectively. The percentage recovery for the effluent samples was 68%. The detection of TCS was monitored for 10 consecutive days, where optimum TCS traces detected in the treated wastewater was 55.0μg/L inday 9 of the monitored days, while the lowest detected was 6.0μg/L. As the concentrations of analytefound in effluent water samples were not so diverse, this study suggested that MIMs could be the best potential adsorbent for the development and continuous progress in membrane technologyand environmental sciences, lending its capability to desalination.

Keywords: molecularly imprinted membrane, triclosan, phase inversion, wastewater

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Authors: Sofiqul Islam, V. Murugan, Prema Kumari, Hari

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Gentamicin is a broad spectrum water-soluble aminoglycoside antibiotics produced by the fermentation process of microorganism known as Micromonospora purpurea. It is widely used for the treatment of infection caused by both gram positive and gram negative bacteria. Gentamicin consists of a mixture of aminoglycoside components like C1, C1a, C2a, and C2. The molecular structure of Gentamicin and its related substances showed that it has lack of presence of chromophore group in the molecule due to which the detection of such components were quite critical and challenging. In this study, a simple Reversed Phase-High Performance Liquid Chromatographic (RP-HPLC) method using ultraviolet (UV) detector was developed and validated for quantification of the related substances present in Gentamicin drug substances. The method was achieved by using Thermo Scientific Hypersil Gold analytical column (150 x 4.6 mm, 5 µm particle size) with isocratic elution composed of methanol: water: glacial acetic acid: sodium hexane sulfonate in the ratio 70:25:5:3 % v/v/v/w as a mobile phase at a flow rate of 0.5 mL/min, column temperature was maintained at 30 °C and detection wavelength of 330 nm. The four components of Gentamicin namely Gentamicin C1, C1a, C2a, and C2 were well separated along with the related substance present in Gentamicin. The Limit of Quantification (LOQ) values were found to be at 0.0075 mg/mL. The accuracy of the method was quite satisfactory in which the % recovery was resulted between 95-105% for the related substances. The correlation coefficient (≥ 0.995) shows the linearity response against concentration over the range of Limit of Quantification (LOQ). Precision studies showed the % Relative Standard Deviation (RSD) values less than 5% for its related substance. The method was validated in accordance with the International Conference of Harmonization (ICH) guideline with various parameters like system suitability, specificity, precision, linearity, accuracy, limit of quantification, and robustness. This proposed method was easy and suitable for use for the quantification of related substances in routine analysis of Gentamicin formulations.

Keywords: reversed phase-high performance liquid chromatographic (RP-HPLC), high performance liquid chromatography, gentamicin, isocratic, ultraviolet

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Authors: Chanchal Biswas, Anrin Bhattacharyya, Gopes Chandra Das, Mahua Ghosh Chaudhuri, Rajib Dey

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Boiler coals with low fixed carbon and higher ash content have always challenged the metallurgists to develop a suitable method for their utilization. In the present study, an attempt is made to establish an energy effective method for the reduction of iron ore fines in the form of nuggets by using ‘Syngas’. By devolatisation (expulsion of volatile matter by applying heat) of boiler coal, gaseous product (enriched with reducing agents like CO, CO2, H2, and CH4 gases) is generated. Iron ore nuggets are reduced by this syngas. For that reason, there is no direct contact between iron ore nuggets and coal ash. It helps to control the minimization of the sulphur intake of the reduced nuggets. A laboratory scale devolatisation furnace designed with reduction facility is evaluated after in-depth studies and exhaustive experimentations including thermo-gravimetric (TG-DTA) analysis to find out the volatile fraction present in boiler grade coal, gas chromatography (GC) to find out syngas composition in different temperature and furnace temperature gradient measurements to minimize the furnace cost by applying one heating coil. The nuggets are reduced in the devolatisation furnace at three different temperatures and three different times. The pre-reduced nuggets are subjected to analytical weight loss calculations to evaluate the extent of reduction. The phase and surface morphology analysis of pre-reduced samples are characterized using X-ray diffractometry (XRD), energy dispersive x-ray spectrometry (EDX), scanning electron microscopy (SEM), carbon sulphur analyzer and chemical analysis method. Degree of metallization of the reduced nuggets is 78.9% by using boiler grade coal. The pre-reduced nuggets with lesser sulphur content could be used in the blast furnace as raw materials or coolant which would reduce the high quality of coke rate of the furnace due to its pre-reduced character. These can be used in Basic Oxygen Furnace (BOF) as coolant also.

Keywords: alternative ironmaking, coal gasification, extent of reduction, nugget making, syngas based DRI, solid state reduction

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Authors: Reema Tiwari, U. C. Kulshrestha

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As a key regulator of atmospheric oxidative capacity and secondary aerosol formations, the signatures of reactive nitrogen (Nr) emissions are becoming increasingly evident in the cascade of air pollution, acidification, and eutrophication of the ecosystem. However, their accurate estimates in N budget remains limited by the photochemical conversion processes where occurrence of differential atmospheric residence time of gaseous (NOₓ, HNO₃, NH₃) and particulate (NO₃⁻, NH₄⁺) Nr species becomes imperative to their spatio temporal evolution on a synoptic scale. The present study attempts to quantify such interactions under tropical conditions when low anticyclonic winds become favorable to the advections from west during winters. For this purpose, a diurnal sampling was conducted using low volume sampler assembly where ambient concentrations of Nr trace gases along with their ionic fractions in the aerosol samples were determined with UV-spectrophotometer and ion chromatography respectively. The results showed a spatial gradient of the gaseous precursors with a much pronounced inter site variability (p < 0.05) than their particulate fractions. Such observations were confirmed for their limited photochemical conversions where less than 1 ratios of day and night measurements (D/N) for the different Nr fractions suggested an influence of boundary layer dynamics at the background site. These phase conversion processes were further corroborated with the molar ratios of NOₓ/NOᵧ and NH₃/NHₓ where incomplete titrations of NOₓ and NH₃ emissions were observed irrespective of their diurnal phases along the sampling transect. Their calculations with equilibrium based approaches for an NH₃-HNO₃-NH₄NO₃ system, on the other hand, were characterized by delays in equilibrium attainment where plots of their below deliquescence Kₘ and Kₚ values with 1000/T confirmed the role of lower temperature ranges in NH₄NO₃ aerosol formation. These results would help us in not only resolving the changing atmospheric inputs of reduced (NH₃, NH₄⁺) and oxidized (NOₓ, HNO₃, NO₃⁻) Nr estimates but also in understanding the dependence of Nr mixing ratios on their local meteorological conditions.

Keywords: diurnal ratios, gas-aerosol interactions, spatial gradient, thermodynamic equilibrium

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Authors: Amanina Iymia Jeffree, Reena Thriumani, Mohammad Iqbal Omar, Ammar Zakaria, Yumi Zuhanis Has-Yun Hashim, Ali Yeon Md Shakaff

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Metabolite profiling is a strategy to be approached in the pattern recognition method focused on three types of cancer cell line that driving the most to death specifically lung, breast, and colon cancer. The purpose of this study was to discriminate the VOCs pattern among cancerous and control group based on metabolite profiling. The sampling was executed utilizing the cell culture technique. All culture flasks were incubated till 72 hours and data collection started after 24 hours. Every running sample took 24 minutes to be completed accordingly. The comparative metabolite patterns were identified by the implementation of headspace-solid phase micro-extraction (HS-SPME) sampling coupled with gas chromatography-mass spectrometry (GCMS). The optimizations of the main experimental variables such as oven temperature and time were evaluated by response surface methodology (RSM) to get the optimal condition. Volatiles were acknowledged through the National Institute of Standards and Technology (NIST) mass spectral database and retention time libraries. To improve the reliability of significance, it is of crucial importance to eliminate background noise which data from 3rd minutes to 17th minutes were selected for statistical analysis. Targeted metabolites, of which were annotated as known compounds with the peak area greater than 0.5 percent were highlighted and subsequently treated statistically. Volatiles produced contain hundreds to thousands of compounds; therefore, it will be optimized by chemometric analysis, such as principal component analysis (PCA) as a preliminary analysis before subjected to a pattern classifier for identification of VOC samples. The volatile organic compound profiling has shown to be significantly distinguished among cancerous and control group based on metabolite profiling.

Keywords: in vitro cancer cell line, metabolite profiling, pattern recognition, volatile organic compounds

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Authors: Mengsha Yin

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Extractable organic matter (EOM) from thirty-six biogenic gas source rocks from the Sanhu Depression in Qaidam Basin in northwestern China were obtained via Soxhlet extraction. Twenty-nine of them were conducted SARA (Saturates, Aromatics, Resins and Asphaltenes) separation for bulk composition analysis. Saturated and aromatic fractions of all the extractions were analyzed by Gas Chromatography-Mass Spectrometry (GC-MS) to investigate the compound compositions. More abundant n-alkanes, naphthalene, phenanthrene, dibenzothiophene and their alkylated products occur in samples in shallower depths. From 2000m downward, concentrations of these compounds increase sharply, and concentration ratios of more-over-less biodegradation susceptible compounds coincidently decrease dramatically. ∑iC15-16, 18-20/∑nC15-16, 18-20 and hopanoids/∑n-alkanes concentration ratios and mono- and tri-aromatic sterane concentrations and concentration ratios frequently fluctuate with depth rather than trend with it, reflecting effects from organic input and paleoenvironments other than biodegradation. Saturated and aromatic compound distributions on the saturates and aromatics total ion chromatogram (TIC) traces of samples display different degrees of biodegradation. Dramatic and simultaneous variations in compound concentrations and their ratios at 2000m and their changes with depth underneath cooperatively justified the crucial control of burial depth on organic matter biodegradation scales in source rocks and prompted the proposition that 2000m is the bottom depth boundary for active microbial activities in this study. The study helps to better curb the conditions where effective source rocks occur in terms of depth in the Sanhu biogenic gas fields and calls for additional attention to source rock pore size estimation during biogenic gas source rock appraisals.

Keywords: pore space, Sanhu depression, saturated and aromatic hydrocarbon compound concentration, source rock organic matter biodegradation, total ion chromatogram

Procedia PDF Downloads 153

Authors: Madalin Enache, Simona Neagu, Roxana Cojoc, Ioana Gomoiu, Delia Ionela Dobre, Ancuta Roxana Trifoi

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Various types of halophilic and halotolerant microorganisms able to be cultivated in laboratory on culture media with a wide range of sodium chloride content are isolated from several salted environments. The extracellular enzymes of these microorganisms showed the enzymatic activity in these spectrums of salinity thus being attractive for several biotechnological processes developed at high ionic strength. In present work, a number of amylase, protease, esterase, lipase, cellulase, pectinase, xilanases and innulinase were identified for more than 50th bacterial strains isolated from water samples and sapropelic mud from four saline and hypersaline lakes located in Romanian plain. On the other hand, the cellulase and pectinase activity were also detected in some halotolerant microorganisms isolated from secondary agricultural flow of grapes processing. The preliminary data revealed that from totally tested strains seven harbor proteases activity, eight amylase activity, four for esterase and another four for lipase, three for pectinase and for one strain were identified either cellulase or pectinase activity. There were no identified enzymes able to hydrolase innulin added to culture media. Several strains isolated from sapropelic mud showed multiple extracellular enzymatic activities, namely three strains harbor three activities and another seven harbor two activities. The data revealed that amylase and protease activities were frequently detected if compare with other tested enzymes. In the case of pectinase were investigated, their ability to be used for increasing resveratrol recovery from material resulted after grapes processing. In this way, the resulted material from grapes processing was treated with microbial supernatant for several times (two, four and 24 hours) and the content of resveratrol was detected by High Performance Liquid Chromatography method (HPLC). The preliminary data revealed some positive results of this treatment.

Keywords: halophilic microorganisms, enzymes, pectinase, salinity

Procedia PDF Downloads 187

Authors: Omolola R. Ayepola, Nicole L. Brooks, Oluwafemi O. Oguntibeju

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The liver plays an important role in the regulation of blood glucose and is a target organ of hyperglycaemia. Hyperglycemia plays a crucial role in the onset of various liver diseases and may culminate into hepatopathy if untreated. Alteration in antioxidant defense and increase in oxidative stress that results in tissue injury is characteristic of diabetes. We evaluated the protective effects of kolaviron-a biflavonoid complex, on hepatic antioxidants, lipid peroxidation and apoptosis in the liver of diabetic rats. To induce type I diabetes, rats were injected with streptozotocin intraperitoneally at a single dose of 50 mg/kg. Oral treatment of diabetic rats with kolaviron (100 mg/kg) started on the 6th day after diabetes induction and continued for 6 weeks (5 times weekly). Diabetic rats exhibited a significant increase in the peroxidation of hepatic lipids as observed from the elevated level of malondialdehyde (MDA) estimated by High-Performance Liquid Chromatography. In addition, Oxygen Radical Absorbance Capacity (ORAC), ratio of reduced to oxidized glutathione (GSH/GSSG) and catalase (CAT) activity was decreased in the liver of diabetic rats. TUNEL assay revealed increased apoptotic cell death in the liver of diabetic rats. Examination of Pancreatic beta-cells by immunohistochemical methods revealed beta cell degeneration and reduction in beta cell/ islet area in the diabetic controls. Kolaviron-treatment increased the area of insulin immunoreactive beta-cells significantly. Kolaviron attenuated lipid peroxidation and apoptosis in the liver of diabetic rats, increased CAT activity GSH levels and the resultant GSH: GSSG. The ORAC of kolaviron-treated diabetic liver was restored to near-normal values. Kolaviron protects the liver against oxidative and apoptotic damage induced by hyperglycemia. The antidiabetic effect of kolaviron may also be related to its beneficial effects on beta-cell function.

Keywords: diabetes mellitus, kolaviron, oxidative stress, liver, apoptosis

Procedia PDF Downloads 381

Authors: L. Kolatorova, J. Vitku, K. Adamcova, M. Simkova, M. Hill, A. Parizek, M. Duskova

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Endocrine disruptors (EDs) are substances leaching from various industrial products, which are able to interfere with the endocrine system. Their harmful effects on human health are generally well-known, and exposure during fetal development may have lasting effects. Fetal exposure and transplacental transport of bisphenol A (BPA) have been recently studied; however, less is known about alternatives such as bisphenol S (BPS), bisphenol F (BPF) and bisphenol AF (BPAF), which have started to appear in consumer products. The human organism is usually exposed to the mixture of EDs, out of which parabens are otherwise known to transfer placenta. The usage of many cosmetic, pharmaceutical and consumer products during the pregnancy that may contain parabens and bisphenols has led to the need for investigation. The aim of the study was to investigate the transplacental transport of BPA, its alternatives, and parabens, and to study their relation to fetal steroidogenesis. BPA, BPS, BPF, BPAF, methylparaben, ethylparaben, propylparaben, butylparaben, benzylparaben and 15 steroids including estrogens, corticoids, androgens and immunomodulatory ones were determined in 27 maternal (37th week of gestation) and cord plasma samples using liquid chromatography - tandem mass spectrometry methods. The statistical evaluation of the results showed significantly higher levels of BPA (p=0.0455) in cord plasma compared to maternal plasma. The results from multiple regression models investigated that in cord plasma, methylparaben, propylparaben and the sum of all measured parabens were inversely associated with testosterone levels. To our best knowledge, this study is the first attempt to determine the levels of alternative bisphenols in the maternal and cord blood, and also the first study reporting the simultaneous detection of bisphenols, parabens, and steroids in these biological fluids. Our study confirmed the transplacental transport of BPA, with likely accumulation in the fetal compartment. The negative association of cord blood parabens and testosterone levels highlights their possible risks, especially for the development of male fetuses. Acknowledgements: This work was supported by the project MH CR 17-30528 A from the Czech Health Research Council, MH CZ - DRO (Institute of Endocrinology - EÚ, 00023761) and by the MEYS CR (OP RDE, Excellent research - ENDO.CZ).

Keywords: bisphenol, endocrine disruptor, paraben, pregnancy, steroid

Procedia PDF Downloads 177

Authors: Suhaila Qari, Samia Moharram, Safaa Alowaidi

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Objectives: To determine the fatty acids profiles in female fish, R. sarba from the Red Sea during the spawning season. Methods: Monthly individual Rhabdosargus sarba were obtained from Bangalah market in Jeddah, Red Sea and transported to the laboratory in ice aquarium. The total length, standard length and weight were measured, fishes were dissected. Ovaries were removed, weighed and 10 ml of concentrated hydrochloric acid were added to 10g of the ovary in a conical flask and immersed in boiling water until the sample was dissolved and the fat was seen to collect on the surface. The conical was cooled and the fat was extracted by shaking with 30 ml of diethyl ether. The extract was bowled after allowing the layers to separate into a weighed flask. The extraction was repeated three times more and distilled off the solvent then the fat dried at 100oC, cooled and weighed. Then 50 mg of lipid was put in a tube, 5 ml of methanolic sulphuric acid was added and 2 ml of benzene, the tube well closed and placed in water bath at 90oC for an hour and half. After cooling, 8 ml water and 5 ml petroleum was added shacked strongly and the ethereal layer was separated in a dry tube, evaporated to dryness. The fatty acid methyl esters were analyzed using a Hewlett Packard (HP 6890) chromatography, asplit /splitless injector and flame ionization detector (FID). Results: In female Rhabdosargus sarba, a total of 29 fatty acids detected in ovaries throughout the spawning season. The main fatty acid group in total lipid was saturated fatty acid (SFA, 28.9%), followed by 23.5% of polyunsaturated fatty acids (PUFA) and 12.9% of monounsaturated fatty acids (MUFA). The dominant SFA were palmitic and stearic, the major MUFA were palmitoleic and oleic, and the major PUFA were C18:2 and C22:2. During spawning stages no significant differences in total SFA, MUFA and PUFA, the highest value of SFA was in late spawning (36.78%). However, the highest value of MUFA and PUFA was in spawning (16.70% and 24.96% respectively). During spawning season there were a significant differences in total SFA between March (late spawning stage) and December (nearly ripe stage), (P < 0.05).

Keywords: sparidae, Rhabdosargus sarba, fish, fatty acids, spawning, gonads, red sea

Procedia PDF Downloads 796

Authors: Marisol Rodriguez-Duarte, Aide Saenz-Galindo, Carolina Flores-Gallegos, Raul Rodriguez-Herrera, Juan Ascacio-Valdes

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The plant known as tarbush (Fluorensia cernua) is a plant originating in northern Mexico, mainly in the states of Coahuila, Durango, San Luis Potosí, Zacatecas and Chihuahua. It is a branched shrub that belongs to the family Asteraceae, has oval leaves of 6 to 11 cm in length and also has small yellow flowers. In Mexico, the tarbush is a very appreciated plant because it has been used as a traditional medicinal agent, for the treatment of gastrointestinal diseases, skin infections and as a healing agent. This plant has been used mainly as an infusion. Due to its traditional use, the content and type of phytochemicals present in the plant are currently unknown and are responsible for its biological properties, so its recovery and identification is very important because the compounds that it contains have relevant applications in the field of food, pharmaceuticals and medicine. The objective of this work was to determine the best extraction condition of phytochemical compounds (mainly polyphenolic compounds) from the leaf using ultrasound/microwave assisted extraction (U/M-AE). To reach the objective, U/M-AE extractions were performed evaluating three mass/volume ratios (1:8, 1:12, 1:16), three ethanol/water solvent concentrations (0%, 30% and 70%), ultrasound extraction time of 20 min and 5 min at 70°C of microwave treatment. All experiments were performed using a fractional factorial experimental design. Once the best extraction condition was defined, the compounds were recovered by liquid column chromatography using Amberlite XAD-16, the polyphenolic fraction was recovered with ethanol and then evaporated. The recovered polyphenolic compounds were quantified by spectrophotometric techniques and identified by HPLC/ESI/MS. The results obtained showed that the best extraction condition of the compounds was using a mass/volume ratio of 1:8 and solvent ethanol/water concentration of 70%. The concentration obtained from polyphenolic compounds using this condition was 22.74 mg/g and finally, 16 compounds of polyphenolic origin were identified. The results obtained in this work allow us to postulate the Mexican plant known as tarbush as a relevant source of bioactive polyphenolic compounds of food, pharmaceutical and medicinal interest.

Keywords: U/M-AE, tarbush, polyphenols, identification

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Authors: R. Raudonis, L. Raudonė, V. Janulis, P. Viškelis

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Pharmacological and clinical studies demonstrated that phenolic compounds particularly flavonoids and phenolic acids are responsible for a wide spectrum of therapeutic activities. Flavonoids and phenolic acids are regarded as natural antioxidants that play an important role in protecting cells from oxidative stress. Qualitatively prepared dry extracts possess high stability and concentration of bio active compounds, facility of standardization and quality control. The aim of this work was to determine the phenolic and antioxidant profiles of Hippophaë rhamnoides L., Betula pendula Roth., Tilia cordata Mill., Sorbus aucuparia L. leaves dry extracts and to identify markers of antioxidant activity. Extracts were analyzed using high-performance liquid chromatography (HPLC) with FRAP post-column assay. Dry extracts are versatile forms possessing wide area of applications, final product ensure consistent phytochemical and functional properties. Seven flavonoids: rutin, hyperoside, isorhamnetin 3-O-rutinoside, isorhamnetin 3-O-glucoside, quercetin, kaempferol, isorhamnetin were identified in dry extract of Hippophaë rhamnoides L. leaves. Predominant compounds were flavonol glycosides which were chosen as markers for quantitative control of dry extracts. Chlorogenic acid, hyperoside, rutin, quercetin, isorhamnetin were prevailing compounds in Betula pendula Roth. leaves extract, whereas strongest ferric reducing activity was determined for chlorogenic acid and hyperoside. Notable amounts of protocatechuic acid and flavonol glycosides, rutin, hyperoside, quercitrin, isoquercitrin were identified in the chromatographic profile of Tilia cordata Mill. Neochlorogenic and chlorogenic acids were significantly dominant compounds in antioxidant profile in dry extract of Sorbus aucuparia L. leaves. Predominant compounds of antioxidant profiles could be proposed as functional markers of quality of phenolic rich raw materials. Dry extracts could be further used for manufacturing of pharmaceutical and nutraceuticals.

Keywords: dry extract, FRAP, antioxidant activity, phenolic

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Authors: S. Junaid S. Qazi, Denice C. Bay, Raymond Chew, Raymond J. Turner

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EmrE, a member of the small multidrug resistance protein family in bacteria is considered to be the archetypical member of its family. It confers host resistance to a wide variety of quaternary cation compounds (QCCs) driven by proton motive force. Generally, purification yield is a challenge in all membrane proteins because of the difficulties in their expression, isolation and solubilization. EmrE is extremely hydrophobic which make the purification yield challenging. We have purified EmrE protein using two different approaches: organic solvent membrane extraction and hexahistidine (his6) tagged Ni-affinity chromatographic methods. We have characterized changes present between ligand affinity of untagged and his6-tagged EmrE proteins in similar membrane mimetic environments using biophysical experimental techniques. Purified proteins were solubilized in a buffer containing n-dodecyl-β-D-maltopyranoside (DDM) and the conformations in the proteins were explored in the presence of four QCCs, methyl viologen (MV), ethidium bromide (EB), cetylpyridinium chloride (CTP) and tetraphenyl phosphonium (TPP). SDS-Tricine PAGE and dynamic light scattering (DLS) analysis revealed that the addition of QCCs did not induce higher multimeric forms of either proteins at all QCC:EmrE molar ratios examined under the solubilization conditions applied. QCC binding curves obtained from the Trp fluorescence quenching spectra, gave the values of dissociation constant (Kd) and maximum specific one-site binding (Bmax). Lower Bmax values to QCCs for his6-tagged EmrE shows that the binding sites remained unoccupied. This lower saturation suggests that the his6-tagged versions provide a conformation that prevents saturated binding. Our data demonstrate that tagging an integral membrane protein can significantly influence the protein.

Keywords: small multidrug resistance (SMR) protein, EmrE, integral membrane protein folding, quaternary ammonium compounds (QAC), quaternary cation compounds (QCC), nickel affinity chromatography, hexahistidine (His6) tag

Procedia PDF Downloads 373

Authors: B. González-Yebra, B. Flores-Nieto, P. Aguilar-Salinas, M. Preciado Puga, A. L. González Yebra

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The monitoring of populations occupationally exposed to organic solvents has been an important issue for several shoe factories for years since the International Agency for Research on Cancer (IARC) has advised on the potential carcinogenic risk of chemicals related to occupations. In order to detect if exposition to organic solvents used in some Mexican shoe factories contributes to oral carcinogenesis, we performed monitoring in three factories. Occupational exposure was determined by using monitors 3M. Organic solvents were assessed by gas chromatography. Then, we recruited 30 shoe workers (30.2 ± 8.4 years) and 10 unexposed subjects (43.3 ± 11.2 years) for the micronuclei (MN) test and immunodetection of some cancer biomarkers (ki-67, p16, caspase-3) in scraped oral epithelial cells. Monitored solvents detected were acetone, benzene, hexane, methyl ethyl ketone, and toluene in acceptable levels according to Official Mexican Norm. We found by MN test higher incidence of nuclear abnormalities (karyorrhexis, pycnosis, karyolysis, condensed chromatin, and macronuclei) in the exposed group than the non-exposed group. On the other hand, we found, a negative expression for Ki-67 and p16 in exfoliated epithelial cells from exposed and non-exposed to organic solvents subjects. Only caspase-3 shown positive patter of expression in 9/30 (30%) exposed subjects, and we detected high karyolysis incidence in caspase-3 subjects (p = 0.021). The absence of expression of proliferation markers p16 and ki-67 and presence of apoptosis marker caspase-3 are indicating the absence of malignancy in oral epithelial cells and low risk for oral cancer. It is a fact that the MN test is a very effective method to detect nuclear abnormalities in exfoliated buccal cells from subjects that have been exposed to organic solvents in the shoe industry. However, in order to improve this tool and predict cancer risk is it is mandatory to implement complementary tests as other biomarkers that can help to detect malignancy in individuals occupationally exposed.

Keywords: biomarkers, oral cancer, organic solvents, shoe industries

Procedia PDF Downloads 133

Authors: Ozan Kahraman, Hao Feng

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Ultrasonic atomization (UA) is a useful technique for producing a liquid spray for various processes, such as spray drying. Ultrasound generates small droplets (a few microns in diameter) by disintegration of the liquid via cavitation and/or capillary waves, with low range velocity and narrow droplet size distribution. In recent years, UA has been investigated as an alternative for enabling or enhancing ultrasound-mediated unit operations, such as evaporation, separation, and purification. The previous studies on the UA separation of a solvent from a bulk solution were limited to ethanol-water systems. More investigations into ultrasound-mediated separation for other liquid systems are needed to elucidate the separation mechanism. This study was undertaken to investigate the effects of the operational parameters on the ultrasound-mediated separation of three miscible liquid pairs: ethanol-, methanol-, and butanol-water. A 2.4 MHz ultrasonic mister with a diameter of 18 mm and rating power of 24 W was installed on the bottom of a custom-designed cylindrical separation unit. Air was supplied to the unit (3 to 4 L/min.) as a carrier gas to collect the mist. The effects of the initial alcohol concentration, viscosity, and temperature (10, 30 and 50°C) on the atomization rates were evaluated. The alcohol concentration in the collected mist was measured with high performance liquid chromatography and a refractometer. The viscosity of the solutions was determined using a Brookfield digital viscometer. The alcohol concentration of the atomized mist was dependent on the feed concentration, feed rate, viscosity, and temperature. Increasing the temperature of the alcohol-water mixtures from 10 to 50°C increased the vapor pressure of both the alcohols and water, resulting in an increase in the atomization rates but a decrease in the separation efficiency. The alcohol concentration in the mist was higher than that of the alcohol-water equilibrium at all three temperatures. More importantly, for ethanol, the ethanol concentration in the mist went beyond the azeotropic point, which cannot be achieved by conventional distillation. Ultrasound-mediated separation is a promising non-equilibrium method for separating and purifying alcohols, which may result in significant energy reductions and process intensification.

Keywords: azeotropic mixtures, distillation, evaporation, purification, seperation, ultrasonic atomization

Procedia PDF Downloads 176

Authors: Laribi-Habchi Hasiba, Bouanane-Darenfed Amel, Drouiche Nadjib, Pausse André, Mameri Nabil

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Chitin, a linear 1, 4-linked N-acetyl-d-glucosamine (GlcNAc) polysaccharide is the major structural component of fungal cell walls, insect exoskeletons and shells of crustaceans. It is one of the most abundant naturally occurring polysaccharides and has attracted tremendous attention in the fields of agriculture, pharmacology and biotechnology. Each year, a vast amount of chitin waste is released from the aquatic food industry, where crustaceans (prawn, crab, Shrimp and lobster) constitute one of the main agricultural products. This creates a serious environmental problem. This linear polymer can be hydrolyzed by bases, acids or enzymes such as chitinase. In this context an extracellular chitinase (ChiA-65) was produced and purified from a newly isolated LHH100. Pure protein was obtained after heat treatment and ammonium sulphate precipitation followed by Sephacryl S-200 chromatography. Based on matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF/MS) analysis, the purified enzyme is a monomer with a molecular mass of 65,195.13 Da. The sequence of the 27 N-terminal residues of the mature ChiA-65 showed high homology with family-18 chitinases. Optimal activity was achieved at pH 4 and 75◦C. Among the inhibitors and metals tested p-chloromercuribenzoic acid, N-ethylmaleimide, Hg2+ and Hg + completelyinhibited enzyme activity. Chitinase activity was high on colloidal chitin, glycol chitin, glycol chitosane, chitotriose and chitooligosaccharide. Chitinase activity towards synthetic substrates in the order of p-NP-(GlcNAc) n (n = 2–4) was p-NP-(GlcNAc)2> p-NP-(GlcNAc)4> p-NP-(GlcNAc)3. Our results suggest that ChiA-65 preferentially hydrolyzed the second glycosidic link from the non-reducing end of (GlcNAc) n. ChiA-65 obeyed Michaelis Menten kinetics the Km and kcat values being 0.385 mg, colloidal chitin/ml and5000 s−1, respectively. ChiA-65 exhibited remarkable biochemical properties suggesting that this enzyme is suitable for bioconversion of chitin waste.

Keywords: Bacillus licheniformis LHH100, characterization, extracellular chitinase, purification

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Authors: Palaniyandi Karuppaiya, Hsin Sheng Tsay, Fang Chen

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Medicinal herbs do represent a huge and noteworthy reservoir for novel anticancer drugs discovery. Dysosma pleiantha (Hance) Woodson (Berberidaceae), one of the oldest traditional Chinese medicinal herb, highly prized by the mountain tribes of Taiwan and China for its medicinal properties contained pharmaceutically important antitumor compounds podophyllotoxin, quercetin and kaempferol. Among lignans, podophyllotoxin is an active antitumor compound and has now been modified to produce clinically useful drugs etoposide and teniposide. In recent years, natural populations of D. peliantha have declined considerably due to anthropogenic activities such as habitat destruction and commercial exploitation for medicinal applications. As to its overall conservation status, D. pleiantha has been ranked as threatened on the China Species Red List. In the present study, an efficient in vitro callus culture system of D. pleiantha was established on Gamborg’s medium with various combinations and concentrations of different auxins and cytokinins under dark condition. Best callus induction was recorded in 2 mg/L 2, 4 - Dichlorophenoxyacetic acid (2,4-D) along with 0.2 mg/L kinetin and the maximum callus proliferation was achieved at 1 mg/L 2,4-D. Among the explants tested, maximum callus induction (86 %) was achieved from tender leaves. Hence, in subsequent experiments, leaf callus was further investigated for suitable callus biomass and production level of anticancer compounds under the influence of different additives. A maximum fresh callus biomass (8.765 g) was recorded in callus proliferation medium contained 500 mg/L casein hydrolysate. High performance liquid chromatography results revealed that the addition of different concentrations of peptone (1, 2 and 4 g/L) in callus proliferation medium enhanced podophyllotoxin (16 fold), quercetin (12 fold) and kaempferol (5 fold) accumulation than control. Thus, the established in vitro callus culture under the influence of different additives may offer an alternative source of enhanced production of podophyllotoxin, kaempferol and quecertin without harming natural plant population.

Keywords: dysosma pleiantha, kaempferol, podophyllotoxin, quercetin

Procedia PDF Downloads 276

Authors: Bruna Lima Gomes, Vanessa Caroline De Barros Bonato, Luciano Freschi, Eduardo Purgatto

Abstract:

Plant hormones are long known to be tightly associated with fruit development and are involved in controlling various aspects of fruit ripening. For fleshy fruits, ripening is characterized for changes in texture, color, aroma and other parameters that markedly contribute to its quality. Ethylene is one of the major players regulating the ripening-related processes, but emerging evidences suggest that auxin is also part of this dynamic control. Thus, the aim of this study was providing new insights into the auxin role during ripening and the hormonal interplay between auxin and ethylene. For that, tomato fruits (Micro-Tom) were collected at mature green stage and separated in four groups: one for indole-3-acetic acid (IAA) treatment, one for ethylene, one for a combination of IAA and ethylene, and one for control. Hormone solution was injected through the stylar apex, while mock samples were injected with buffer only. For ethylene treatments, fruits were exposed to gaseous hormone. Then, fruits were left to ripen under standard conditions and to assess ripening development, hue angle was reported as color indicator and ethylene production was measured by gas chromatography. The transcript levels of three ripening-related ethylene receptors (LeETR3, LeETR4 and LeETR6) were evaluated by RT-qPCR. Results showed that ethylene treatment induced ripening, stimulated ethylene production, accelerated color changes and induced receptor expression, as expected. Nonetheless, auxin treatment showed the opposite effect once fruits remained green for longer time than control group and ethylene perception has changed, taking account the reduced levels of receptor transcripts. Further, treatment with both hormones revealed that auxin effect in delaying ripening was predominant, even with higher levels of ethylene. Altogether, the data suggest that auxin modulates several aspects of the tomato fruit ripening modifying the ethylene perception. The knowledge about hormonal control of fruit development will help design new strategies for effective manipulation of ripening regarding fruit quality and brings a new level of complexity on fruit ripening regulation.

Keywords: ethylene, auxin, fruit ripening, hormonal crosstalk

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Authors: Yohann R. J. Thomas, Sébastien Solan

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Conventional battery technology includes the assembly of electrode/separator/electrode by standard techniques such as stacking or winding, depending on the format size. In that type of batteries, coating or pasting techniques are only used for the electrode process. The processes are suited for large scale production of batteries and perfectly adapted to plenty of application requirements. Nevertheless, as the demand for both easier and cost-efficient production modes, flexible, custom-shaped and efficient small sized batteries is rising. Thin-film, printable batteries are one of the key areas for printed electronics. In the frame of European BASMATI project, we are investigating the feasibility of a new design of lithium-ion battery: interdigitated planar core design. Polymer substrate is used to produce bendable and flexible rechargeable accumulators. Direct fully printed batteries lead to interconnect the accumulator with other electronic functions for example organic solar cells (harvesting function), printed sensors (autonomous sensors) or RFID (communication function) on a common substrate to produce fully integrated, thin and flexible new devices. To fulfill those specifications, a high resolution printing process have been selected: Aerosol jet printing. In order to fit with this process parameters, we worked on nanomaterials formulation for current collectors and electrodes. In addition, an advanced printed polymer-electrolyte is developed to be implemented directly in the printing process in order to avoid the liquid electrolyte filling step and to improve safety and flexibility. Results: Three different current collectors has been studied and printed successfully. An ink of commercial copper nanoparticles has been formulated and printed, then a flash sintering was applied to the interdigitated design. A gold ink was also printed, the resulting material was partially self-sintered and did not require any high temperature post treatment. Finally, carbon nanotubes were also printed with a high resolution and well defined patterns. Different electrode materials were formulated and printed according to the interdigitated design. For cathodes, NMC and LFP were efficaciously printed. For anodes, LTO and graphite have shown to be good candidates for the fully printed battery. The electrochemical performances of those materials have been evaluated in a standard coin cell with lithium-metal counter electrode and the results are similar with those of a traditional ink formulation and process. A jellified plastic crystal solid state electrolyte has been developed and showed comparable performances to classical liquid carbonate electrolytes with two different materials. In our future developments, focus will be put on several tasks. In a first place, we will synthesize and formulate new specific nano-materials based on metal-oxyde. Then a fully printed device will be produced and its electrochemical performance will be evaluated.

Keywords: high resolution digital printing, lithium-ion battery, nanomaterials, solid-state electrolytes

Procedia PDF Downloads 246