Search results for: two-dimensional HPLC

Authors: Samuel K. Degife, Kamal K. Pant, Sapna Jain

Abstract:

The world´s population and industrialization of countries continued to grow in an alarming rate irrespective of the security for food, energy supply, and pure water availability. As a result, the global energy consumption is observed to increase significantly. Fossil energy resources that mainly comprised of crude oil, coal, and natural gas have been used by mankind as the main energy source for almost two centuries. However, sufficient evidences are revealing that the consumption of fossil resource as transportation fuel emits environmental pollutants such as CO2, NOx, and SOx. These resources are dwindling rapidly besides enormous amount of problems associated such as fluctuation of oil price and instability of oil-rich regions. Biomass is a promising renewable energy candidate to replace fossil-based transportation fuel and chemical production. The present study aims at valorization of hexose sugars (glucose and fructose) using zeolite based catalysts in imidazolium based ionic liquid (1-butyl-3-methylimidazolium chloride, [BMIM] Cl) reaction media. The catalytic effect chromium impregnated H-ZSM-5 (Cr/H-ZSM-5) was studied for dehydration of hexose sugars. The wet impregnation method was used to prepare Cr/H-ZSM-5 catalyst. The characterization of the prepared catalyst was performed using techniques such as Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction analysis (XRD), Temperature-programmed desorption of ammonia (NH3-TPD) and BET-surface area analysis. The dehydration product, 5-hydroxymethylfurfural (5-HMF), was analyzed using high-performance liquid chromatography (HPLC). Cr/H-ZSM-5 was effective in dehydrating fructose with 87% conversion and 55% yield 5-HMF at 180 oC for 30 min of reaction time compared with H-ZSM-5 catalyst which yielded only 31% of 5-HMF at identical reaction condition.

Keywords: chromium, hexose, ionic liquid, , zeolite

Procedia PDF Downloads 145

Authors: Nishtha, Richa, Anju Rao

Abstract:

Plants are the source of herbal medicine and medicinal value of the plants lies in the bioactive phytochemical constituents that produce definite physiological effects on human body. Angiospermic families are known to produce such phytochemical constituents which are termed as secondary plant metabolites. These metabolites include alkaloids, saponins, phenolic compounds, flavonoids, tannins, terpenoids and so on. Solanaceae is one of the important families of Angiosperms known for medicinally important alkaloids such as hyoscyamine, scopolamine, solanine, nicotine, capsaicin etc. Medicinally important species of this family mostly belong to the genera of Datura,Atropa,Solanum,Withania and Nicotiana.Six species such as Datura metel, Solanum torvum, Physalis minima, Cestrum nocturnum, Cestrum diurnum and Nicotiana plumbaginifolia have been collected from different localities of Chandigarh and adjoining areas.Field and anatomical studies helped to identify the plants and their parts used for the study of secondary plant metabolites. Preliminary phytochemcial studies have been done on various parts of plants such as roots, stem and leaves by making aqueous and alcoholic extracts from their powdered forms which showed the presence of alkaloids in almost all the species followed by steroids, flavonoids, terpenoids, tannins etc. HPLC profiles of leaves of Datura metel showed the presence of active compounds such as scopalamine and hyoscyamine and Solanum torvum showed the presence of solanine and solasodine. These alkaloids are important source of drug based medicine used in pharmacognosy. The respective compounds help in treating vomiting, nausea, respiratory disorders, dizziness, asthma and many heart problems.

Keywords: alkaloids, flavanoids, phytochemical constituents, pharmacognosy, secondary metabolites

Procedia PDF Downloads 416

Authors: Zebiri Saliha

Abstract:

Ochratoxin A (OTA) is a mycotoxin produced by certain species of the genera Aspergillus and Penicillium, primarily found in cereals, coffee, and grapevine products. Its accumulation in the body can lead to nephrotoxic, teratogenic, immunosuppressive, and carcinogenic effects. The objective of this study is to investigate the contamination of consumed wheat and its derivatives by toxic fungi in Algeria. For this purpose, an analysis of 200 samples was conducted, including 90 samples of durum wheat and common wheat and 110 samples of wheat derivatives collected from mills (semolina and flour manufacturers). The results revealed an average fungal contamination rate ranging from 60% to 100%. The identified fungal isolates primarily belonged to the genera Aspergillus (70%), Penicillium (27.5%), Alternaria (40%), and Mucor (19.4%). The density of the fungal flora was higher in products intended for animal consumption, such as durum wheat flour (2525 CFU/g), wheat scraps (3175 CFU/g), and wheat bran (2950 CFU/g). Conversely, low fungal density was observed in fine semolina (900 CFU/g) and flour (800 CFU/g) intended for human consumption. The genus Penicillium was isolated in 46% of the analyzed samples of durum wheat derivatives and in 62.7% of the analyzed samples of common wheat derivatives. The Aspergillus genus dominated the majority of the analyzed samples. Molecular identification of Aspergillus and Penicillium isolates by sequencing ITS1-5.8S-ITS2 regions of DNAr and a part of the calmodulin (CaM) gene indicated that the species involved in the production of OTA in wheat and its derivatives were mainly Aspergillus ochraceus, A. westerdijkia, A. alliaceus, A. carbonarius, and Penicillium islandicus. The amounts of OTA produced by these species were determined by HPLC-FLD and ranged between 0,8.9 and 3033μg/g. Given that food safety and quality are major concerns today, understanding the microbial biodiversity of wheat is crucial because it is a staple food in Algeria.

Keywords: wheat derivatives, Aspergillus, microbial biodiversity, OTA

Procedia PDF Downloads 12

Authors: Basanti Ekka, Talis Juhna

Abstract:

Dairy industrial effluents originated by the typical processing activities are composed of various organic and inorganic constituents, and these include proteins, fats, inorganic salts, antibiotics, detergents, sanitizers, pathogenic viruses, bacteria, etc. These contaminants are harmful to not only human beings but also aquatic flora and fauna. Because consisting of large classes of contaminants, the specific targeted removal methods available in the literature are not viable solutions on the industrial scale. Therefore, in this on-going research, a series of coagulation, electrochemical, and catalytic methods will be employed. The bulk coagulation and electrochemical methods can wash off most of the contaminants, but some of the harmful chemicals may slip in; therefore, specific catalysts designed and synthesized will be employed for the removal of targeted chemicals. In the context of Latvian dairy industries, presently, work is under progress on the characterization of dairy effluents by total organic carbon (TOC), Inductively Coupled Plasma Mass Spectrometry (ICP-MS)/ Inductively Coupled Plasma Optical Emission Spectrometry (ICP-OES), High-Performance Liquid Chromatography (HPLC), Gas Chromatography-Mass Spectrometry (GC-MS), and Mass Spectrometry. After careful evaluation of the dairy effluents, a cost-effective natural coagulant will be employed prior to advanced electrochemical technology such as electrocoagulation and electro-oxidation as a secondary treatment process. Finally, graphene oxide (GO) based hybrid materials will be used for post-treatment of dairy wastewater as graphene oxide has been widely applied in various fields such as environmental remediation and energy production due to the presence of various oxygen-containing groups. Modified GO will be used as a catalyst for the removal of remaining contaminants after the electrochemical process.

Keywords: catalysis, dairy wastewater, electrochemical method, graphene oxide

Procedia PDF Downloads 110

Authors: Ana M. T. Mata, Alexiane Ligneul

Abstract:

Textile effluents are usually heavily loaded with organic carbon and color compounds, the latter being azo dyes in an estimated 70% of the case effluent posing a major challenge in environmental protection. In this study, the ecotoxicity of simulated textile effluent after biological treatment with anaerobic and aerobic phase (aerobic granular sludge, AGS) and after advanced oxidation processes (AOP) namely ozonation and UV irradiation as post-treatment, were tested to evaluate the fitness of this treatments for ecotoxicity abatement. AGS treatment achieved an 80% removal in both COD and color. AOP was applied with the intention to mineralize the metabolites resulting from biodecolorization of the azo dye Acid Red 14, especially the stable aromatic amine (4-amino-1-naphthalenesulfonic acid, 4A1NS). The ecotoxicity evaluation was based on growth inhibition of the algae Pseudokirchneriella subcapitata following OECD TG 201 except regarding the medium, MBL medium was used instead. Five replicate control cultures and samples were performed with an average STD of 2.7% regarding specific algae growth rate determination. It was found that untreated textile effluent holds an inhibition of specific growth rate of 82%. AGS treatment by itself is able to lower ecotoxicity to 53%. This is probably due to the high color removal of the treatment. AOP post-treatment with Ozone and UV irradiation improves the ecotoxicity abatment to 49 and 43% inhibition respectively, less significantly than previously thought. Since over 85% of 4A1NS was removed by either of the AOP (followed by HPLC), an individual ecotoxicity test of 4A1NS was performed showing that 4A1NS does not inhibit algae growth (0% inhibition). It was concluded that AGS treatment is able by itself to achieve a significant ecotoxicity abatement of textile effluent. The cost-benefit of AOP as a post-treatment have to be better accessed since their application resulted in an improvement of only 10% regarding ecotoxicity effluent removal. It was also found that the 4A1NS amine had no apparent effect on ecotoxicity. Further studies will be conducted to study where ecotoxicity is coming from after AGS biological treatment and how to eliminate it.

Keywords: textile wastewate, ecotoxicity, aerobic granular sludge, AOP

Procedia PDF Downloads 139

Authors: Gaurav Bhanjana, Ganga Ram Chaudhary, Sandeep Kumar, Neeraj Dilbaghi

Abstract:

Main sources of endocrine disrupting compounds in the ecosystem are hormones, pesticides, phthalates, flame retardants, dioxins, personal-care products, coplanar polychlorinated biphenyls (PCBs), bisphenol A, and parabens. These endocrine disrupting compounds are responsible for learning disabilities, brain development problems, deformations of the body, cancer, reproductive abnormalities in females and decreased sperm count in human males. Although discharge of these chemical compounds into the environment cannot be stopped, yet their amount can be retarded through proper evaluation and detection techniques. The available techniques for determination of these endocrine disrupting compounds mainly include high performance liquid chromatography (HPLC), mass spectroscopy (MS) and gas chromatography-mass spectrometry (GC–MS). These techniques are accurate and reliable but have certain limitations like need of skilled personnel, time consuming, interference and requirement of pretreatment steps. Moreover, these techniques are laboratory bound and sample is required in large amount for analysis. In view of above facts, new methods for detection of endocrine disrupting compounds should be devised that promise high specificity, ultra sensitivity, cost effective, efficient and easy-to-operate procedure. Nowadays, electrochemical sensors/biosensors modified with nanomaterials are gaining high attention among researchers. Bioelement present in this system makes the developed sensors selective towards analyte of interest. Nanomaterials provide large surface area, high electron communication feature, enhanced catalytic activity and possibilities of chemical modifications. In most of the cases, nanomaterials also serve as an electron mediator or electrocatalyst for some analytes.

Keywords: electrochemical, endocrine disruptors, microscopy, nanoparticles, sensors

Procedia PDF Downloads 245

Authors: Hanen Bouaziz, Moez Rafrafi, Ghada Ben Salah, Kamel Jamoussi, Tahia Boudawara, Najiba Zeghal

Abstract:

The present study investigated the protective role of Hyparrhenia hirta against sodium nitrate (NaNO3)-induced nephrotoxicity. A high-performance liquid chromatography coupled with a mass spectrometer (HPLC-MS) method was developed to separate and identify flavonoids in Hyparrhenia hirta. Seven flavonoids were identified as 3-O-methylquercetin, luteolin-7-O-glucoside, luteolin, apigenin-7-O-glucoside, apigenin-8-C-glucoside, luteolin-8-C-glucoside and luteolin-6-C-glucoside. Wistar rats were randomly divided into three groups: a control group and two treated groups during 50 days with NaNO3 administered either alone in drinking water or co-administered with Hyparrhenia hirta. NaNO3 treatment induced a significant increase in plasma levels of creatinine, urea and uric while urinary level decreased significantly. Nephrotoxicity induced by NaNO3 was characterized by significant increase in creatinine clearance. In parallel, a significant increase in malondialdehyde level along with a concomitant decrease in total glutathione content and superoxide dismutase, catalase and glutathione peroxidase activities were observed in the kidney after NaNO3 treatment. The histopathological changes in kidney after NaNO3 administration were shrunken. There were renal tubule cell degeneration and infiltration of mononuclear cells. Most glomeruli revealed shrinkage, a wide capsular space and a peri-glomerular mononuclear cells infiltration. Hyparrhenia hirta supplementation showed a remarkable amelioration of the abnormalities cited above. The results concluded that the treatment with Hyparrhenia hirta had a significant role in protecting the animals from nitrate-induced kidney dysfunction.

Keywords: flavonoids, hyparrhenia hirta, kidney, nitrate toxicity, oxidative stress, rat

Procedia PDF Downloads 416

Authors: Valiollah Babaeipour, Mahdi Rahaie

Abstract:

food supplements are rich in specific nutrients and bioactive compounds that eliminate free radicals and improve cellular metabolism. The major bioactive compounds are found in bran and cereal sprouts. Secondary metabolites of these microorganisms have antioxidant properties that can be used alone or in combination with chemotherapy and radiation therapy to treat cancer. Biologically active compounds such as benzoquinone derivatives extracted from fermented wheat germ extract (FWGE) have several positive effects on the overall state of human health and strengthen the immune system. The present work describes the discontinuous fermentation of raw wheat germ for FWGE production through the simultaneous culture process using the probiotic strains of Saccharomyces cerevisiae, Lactobacillus plantarum, and the possibility of using solid waste. To increase production efficiency, first to select important factors in the optimization of each fermentation process, using a factorial statistical scheme of stirring fraction (120 to 200 rpm), dilution of solids to solvent (1 to 8-12), fermentation time (16 to 24 hours) and strain to wheat germ ratio (20% to 50%) were studied and then simultaneous culture was performed to increase the yields of 2 and 6 dimethoxybenzoquinone (2,6-DMBQ). Since 2 and 6 dimethoxy benzoquinone were fermented as the main biologically active compound in wheat germ extract, UV-Vis analysis was performed to confirm the presence of 2 and 6 dimethoxy benzoquinone in the final product. In addition, 2,6-DMBQ of some products was isolated in a non-polar C-18 column and quantified using high performance liquid chromatography (HPLC). Based on our findings, it can be concluded that the increase of 2 and 6 dimethoxybenzoquinone in the simultaneous culture of Saccharomyces cerevisiae - Lactobacillus plantarum compared to pure culture of Saccharomyces cerevisiae (from 1.89 mg / g) to 28.9% (2.66 mg / g) Increased.

Keywords: wheat germ, FWGE, saccharomyces cerevisiae, lactobacillus plantarum, co-culture, 2, 6-DMBQ

Procedia PDF Downloads 93

Authors: X. Han, S. Duan, C. Liu, C. Zhou, W. Zhu, L. Kong

Abstract:

The ionic imprinting technique refers to the three-dimensional rigid structure with the fixed pore sizes, which was formed by the binding interactions of ions and functional monomers and used ions as the template, it has a high level of recognition to the ionic template. The preparation of monolithic column by the in-situ polymerization need to put the compound of template, functional monomers, cross-linking agent and initiating agent into the solution, dissolve it and inject to the column tube, and then the compound will have a polymerization reaction at a certain temperature, after the synthetic reaction, we washed out the unread template and solution. The monolithic columns are easy to prepare, low consumption and cost-effective with fast mass transfer, besides, they have many chemical functions. But the monolithic columns have some problems in the practical application, such as low-efficiency, quantitative analysis cannot be performed accurately because of the peak shape is wide and has tailing phenomena; the choice of polymerization systems is limited and the lack of theoretical foundations. Thus the optimization of components and preparation methods is an important research direction. During the preparation of ionic imprinted monolithic columns, pore-forming agent can make the polymer generate the porous structure, which can influence the physical properties of polymer, what’ s more, it can directly decide the stability and selectivity of polymerization reaction. The compounds generated in the pre-polymerization reaction could directly decide the identification and screening capabilities of imprinted polymer; thus the choice of pore-forming agent is quite critical in the preparation of imprinted monolithic columns. This article mainly focuses on the research that when using different pore-forming agents, the impact of zinc ion imprinted monolithic column on the enrichment performance of zinc ion.

Keywords: high performance liquid chromatography (HPLC), ionic imprinting, monolithic column, pore-forming agent

Procedia PDF Downloads 188

Authors: Maira C. M. Fonseca, Maria Aparecida N. Sediyama, Rosana Goncalves R. das Dores, Sanzio Mollica Vidigal, Alberto C. P. Dias

Abstract:

Among Brazilian plant diversity, many species are used as food and considered minor crops (non-conventional plant foods) (NCPF). Arrowroot (Maranta arundinacea) is a NCPF from which starch is extracted from rhizome do not have gluten. Thus, arrowroot flower starch can be consumed by celiac people. Additional, some medicinal and functional proprieties are assigned to arrowroot leaves which currently are underutilized. In Brazil, it’s cultivated mainly by small scale farmers and there is no specific recommendation for fertilization. This work aimed to determinate the best fertilization for rhizome production and to verify its influence in phenolic composition and antioxidant activity of leaf extracts. Two arrowroot varieties, “Common” and “Seta”, were cultivated in organic system at state of Minas Gerais, Brazil, using cattle manure with three levels of nitrogen (N) (0, 300 and 900 kg N ha-1). The experiment design was in randomized block with four replicates. The highest production of rhizomes in both varieties, “Common” (38198.24 kg ha-1) and “Seta” (43567.71 kg ha-1), were obtained with the use of 300 kg N ha-1. With this fertilization, the total aerial part, petiole and leaf production in the varieties were respectively: “Common” (190.312 kg ha-1; 159.312 kg ha-1; 31.100 kg ha-1) and “Seta” (207.656 kg ha-1; 180.539 kg ha-1; 27.062 kg ha-1). Methanolic leaf extracts were analysed by HPLC-DAD. The major phenolic compounds found were caffeioylquinic acids, p-coumaric derivatives and flavonoids. In general, the production of these compounds significantly decreases with the increase levels of nitrogen (900 kg N ha-1). With 300 kg N ha-1 the phenolic production was similar to control. The antioxidant activity was evaluated using DPPH method and was detected around 60% of radical scavenging when 0.1 mg/mL of plant extracts were used. We concluded that fertilization with 300 kg N ha-1 increased arrowroot rhizome production, maintaining phenolic compounds yield at leaves.

Keywords: antioxidant activity, non-conventional plants, organic fertilization, phenolic compounds

Procedia PDF Downloads 172

Authors: Raymond Dominic Uzoh

Abstract:

Starch fillers were extracted from three plant sources namely amora tuber (a wild variety of Irish potato), sweet potato and yam starch and their particle size, pH, amylose, and amylopectin percentage decomposition determined accordingly by high performance liquid chromatography (HPLC). The starch was introduced into natural rubber in liquid phase (through gelatinization) by the latex compounding method and compounded according to standard method. The prepared starch/natural rubber composites was characterized by Instron Universal testing machine (UTM) for tensile mechanical properties. The composites was further characterized by x-ray diffraction and crosslink density analysis. The particle size determination showed that amora starch granules have the highest particle size (156 × 47 μm) followed by yam starch (155× 40 μm) and then the sweet potato starch (153 × 46 μm). The pH test also revealed that amora starch has a near neutral pH of 6.9, yam 6.8, and sweet potato 5.2 respectively. Amylose and amylopectin determination showed that yam starch has a higher percentage of amylose (29.68), followed by potato (22.34) and then amora starch with the lowest value (14.86) respectively. The tensile mechanical properties testing revealed that yam starch produced the best tensile mechanical properties followed by amora starch and then sweet potato starch. The structure, crystallinity/amorphous nature of the product composite was confirmed by x-ray diffraction, while the nature of crosslinking was confirmed by swelling test in toluene solvent using the Flory-Rehner approach. This research study has rendered a workable strategy for enhancing interfacial interaction between a hydrophilic filler (starch) and hydrophobic polymeric matrix (natural rubber) yielding moderately good tensile mechanical properties for further exploitation development and application in the rubber processing industry.

Keywords: natural rubber, fillers, starch, amylose, amylopectin, crosslink density

Procedia PDF Downloads 143

Authors: Doostishoar Farzad, Forootanfar Hamid, Hasan-Bikdashti Morvarid, Faramarzi Mohammad Ali, Ameri Atefe

Abstract:

Introduction: Chili peppers and related plants in the family of capsaicum produce a mixture of capsaicins represent anticarcinogenic, antimutagenic, and chemopreventive properties. Vanillylamine, the main product of capsaicin hydrolysis is applied as a precursor for manufacturing of natural vanillin (a famous flavor). It is also used in the production of synthetic capsaicins harboring a wide variety of physiological and biological activities such as antibacterial and anti-inflammatory effects as well as enhancing of adrenal catecholamine secretion, analgesic, and antioxidative activities. The ability of some lipases, such as Novozym 677 BG and Novozym 435 and also some proteases e.g. trypsine and penicillin acylase, in capsaicin hydrolysis and green synthesis of vanillylamine has been investigated. In the present study the optimized culture broth of a newly isolated lipase-producing bacterial strain (Stenotrophomonas maltophilia) applied for the hydrolysis of capsaicin. Materials and methods: In order to compare hydrolytic activity of optimized and basal culture broth through capsaicin 2 mL of each culture broth (as sources of lipase) was introduced to capsaicin solution (500 mg/L) and then the reaction mixture (total volume of 3 mL) was incubated at 40 °C and 120 rpm. Samples were taken every 2 h and analyzed for vanillylamine formation using HPLC. Same reaction mixture containing boiled supernatant (to inactivate lipase) designed as blank and each experiment was done in triplicate. Results: 215 mg/L of vanillylamine was produced after the treatment of capsaicin using the optimized medium for 18 h, while only 61 mg/L of vanillylamine was detected in presence of the basal medium under the same conditions. No capsaicin conversion was observed in the blank sample, in which lipase activity was suppressed by boiling of the sample for 10 min. Conclusion: The application of optimized broth culture for the hydrolysis of capsaicin led to a 43% conversion of that pungent compound to vanillylamine.

Keywords: Capsaicin, green synthesis, lipase, stenotrophomonas maltophilia

Procedia PDF Downloads 457

Authors: Efri Mardawati, Ronny Purwadi, Made Tri Ari Penia Kresnowati, Tjandra Setiadi

Abstract:

EFB are mainly composed of cellulose (≈ 43%), hemicellulose (≈ 23%) and lignin (≈20%). The palm oil empty fruit bunches (EFB) is the lignosellulosic waste from crude palm oil industries mainly compose of (≈ 43%), hemicellulose (≈ 23%) and lignin (≈20%). Xylan, a polymer made of pentose sugar xylose and the most abundant component of hemicellulose in plant cell wall. Further xylose can be used as a raw material for production of a wide variety of chemicals such as xylitol, which is extensively used in food, pharmaceutical and thin coating applications. Currently, xylose is mostly produced from xylan via chemical hydrolysis processes. However, these processes are normally conducted at a high temperature and pressure, which is costly, and the required downstream processes are relatively complex. As an alternative method, enzymatic hydrolysis of xylan to xylose offers an environmentally friendly biotechnological process, which is performed at ambient temperature and pressure with high specificity and at low cost. This process is catalysed by xylanolytic enzymes that can be produced by some fungal species such as Aspergillus niger, Penicillium crysogenum, Tricoderma reseei, etc. Fungal that will be used to produce crude xylanase enzyme in this study is T. Viride ITB CC L.67. It is the purposes of this research to study the influence of pretreatment of EFB for the enzymatic hydrolysis process, optimation of temperature and pH of the hydrolysis process, the influence of substrate and enzyme concentration to the enzymatic hydrolysis process, the dynamics of hydrolysis process and followingly to study the kinetics of this process. Xylose as the product of enzymatic hydrolysis process analyzed by HPLC. The results show that the thermal pretreatment of EFB enhance the enzymatic hydrolysis process. The enzymatic hydrolysis can be well approached by the Michaelis Menten kinetic model, and kinetic parameters are obtained from experimental data.

Keywords: oil palm empty fruit bunches (EFB), xylose, enzymatic hydrolysis, kinetic modelling

Procedia PDF Downloads 363

Authors: Lola Brasseur, Guillaume Caulier, Marie Demeyer, Pascal Gerbaux, Igor Eeckhaut

Abstract:

Seawater being an ideal dispersing agent, chemical communication stays predominant in marine ecosystems. However, if many molecules acting in chemical heterospecific communication have already been well described in terrestrial ecosystems, only three of these molecules were identified in marine ecosystems. Echinoderms and their symbiotic organisms constitute very good models to study heterospecific chemical communication because each class synthesizes a specific type of molecules and symbioses with echinoderms as hosts are very usual. In this study, the chemical communication that allows the commensal shrimps Tuleariocaris holthuisi Hipeau-Jacquotte, 1965 to live with their host Echinometra mathaei (Blainville, 1825) was investigated. The chemoreception of the shrimp was characterized using olfactometers and it was demonstrated that hosts and synthetic hydroxynaphthoquinones are attractive to the symbiotic shrimps. Hydroxynaphthoquinonic pigments also known as spinochromes are by the way synthesized by sea urchin and involved in all probability in a lot of mechanisms. To our knowledge, this study is the first highlighting the ecological function of naphthoquinones as kairomones. Chemical extractions were also performed on sea urchins in order to analyze and identify their specific hydroxynaphthoquinones using HPLC-ESI-MS. Accurate mass identification and elemental composition have been performed on various organs (gonads, coelomic liquid, digestive system and test) in different morphotypes of Echinometra mathaei for a better understanding of the molecular diversity of these semiochemicals. Moreover, some experiments were performed to investigate the dependence of T. holthuisi for their host. First, the analyses showed that the molecules involved in shrimp pigmentation are the same that the ones involved in E. mathaei, suggesting a potential feeding on the host. Secondly, a substantial shrimp depigmentation and an increase of the mortality rate were demonstrated after the symbionts-host separation which could mean a potential implication of spinochromes in the shrimp metabolism.

Keywords: crustacean, sea urchin, spinochrome, symbiosis

Procedia PDF Downloads 162

Authors: Avinash Vellore Sunder, Shikha Shah, Pramod P. Wangikar

Abstract:

The enzymatic conversion of nitriles to carboxylic acids by nitrilases has gained significance in the green synthesis of several pharmaceutical precursors and fine chemicals. While nitrilases have been characterized from different sources, the industrial application requires the identification of nitrilases that possess higher substrate tolerance, wider specificity and better thermostability, along with the development of an efficient bioprocess for producing large amounts of nitrilase. To produce large amounts of nitrilase, we developed a fed-batch fermentation process on defined media for the high cell density cultivation of E. coli cells expressing the well-studied nitrilase from Alcaligenes fecalis. A DO-stat feeding approach was employed combined with an optimized post-induction strategy to achieve nitrilase titer of 2.5*105 U/l and 78 g/l dry cell weight. We also identified 16 novel nitrilase sequences from genome mining and analysis of substrate binding residues. The nitrilases were expressed in E. coli and their biocatalytic potential was evaluated on a panel of 22 industrially relevant nitrile substrates using high-throughput screening and HPLC analysis. Nine nitrilases were identified to exhibit high activity on structurally diverse nitriles including aliphatic and aromatic dinitriles, heterocyclic, -hydroxy and -keto nitriles. With fed-batch biotransformation, whole-cell Zobelia galactanivorans nitrilase achieved yields of 2.4 M nicotinic acid and 1.8 M isonicotinic acid from 3-cyanopyridine and 4-cyanopyridine respectively within 5 h, while Cupravidus necator nitrilase enantioselectively converted 740 mM mandelonitrile to (R)–mandelic acid. The nitrilase from Achromobacter insolitus could hydrolyze 542 mM iminodiacetonitrile in 1 h. The availability of highly active nitrilases along with bioprocesses for enzyme production expands the toolbox for industrial biocatalysis.

Keywords: biocatalysis, isonicotinic acid, iminodiacetic acid, mandelic acid, nitrilase

Procedia PDF Downloads 198

Authors: Faisal A. Salih

Abstract:

Vinpocetine (Vin) is an ethyl ester of apovincamic acid and is a semisynthetic derivative of vincamine, an alkaloid from plants of the genus Periwinkle (plant) vinca minor. It was found that this compound stimulates cerebral metabolism: it increases the uptake of glucose and oxygen, as well as the consumption of these substances by the brain tissue. Vinpocetine enhances the flow of blood in the brain and has a vasodilating, antihypertensive, and antiplatelet effect. Vinpocetine seems to improve the human ability to acquire new memories and restore memories that have been lost. This drug has been clinically used for the treatment of cerebrovascular disorders such as stroke and dementia memory disorders, as well as in ophthalmology and otorhinolaryngology. It has no side effects, and no toxicity has been reported when using vinpocetine for a long time. For the quantitative determination of Vin in dosage forms, the HPLC methods are generally used. A promising alternative is potentiometry with Vin- selective electrode, which does not require expensive equipment and materials. Another advantage of the potentiometric method is that the pills and solutions for injections can be used directly without separation from matrix components, which reduces both analysis time and cost. In this study, it was found that the choice of a good plasticizer an electrode with the following membrane composition: PVC (32.8 wt.%), ortho-nitrophenyl octyl ether (66.6 wt.%), tetrakis-4-chlorophenyl borate (0.6 wt.%) exhibits excellent analytical performance: lower detection limit (LDL) 1.2•10⁻⁷ M, linear response range (LRR) 1∙10⁻³–3.9∙10⁻⁶ M, the slope of the electrode function 56.2±0.2 mV/decade). Vin masses per average tablet weight determined by direct potentiometry (DP) and potentiometric titration (PT) methods for the two different sets of 10 tablets were (100.35±0.2–100.36±0.1) mg for two sets of blister packs. The mass fraction of Vin in individual tablets, determined using DP, was (9.87 ± 0.02–10.16 ±0.02) mg, while the RSD was (0.13–0.35%). The procedure has very good reproducibility, and excellent compliance with the declared amounts was observed.

Keywords: vinpocetine, potentiometry, ion selective electrode, pharmaceutical analysis

Procedia PDF Downloads 41

Authors: Hatem I. Mokhtar, Randa A. Abd-El-Salam, Ghada M. Hadad

Abstract:

An improved nano-composite of self-doped polyaniline nanofibers and silica-coated magnetite nanoparticles were prepared and evaluated for suitability to magnetic dispersive micro solid-phase extraction. The work focused on optimization of the composite capacity to extract four fluoroquinolones (FQs) antibiotics, ciprofloxacin, enrofloxacin, danofloxacin, and difloxacin from water and improvement of composite stability towards acid and atmospheric degradation. Self-doped polyaniline nanofibers were prepared by oxidative co-polymerization of aniline with anthranilic acid. Magnetite nanopariticles were prepared by alkaline co-precipitation and coated with silica by silicate hydrolysis on magnetite nanoparticles surface at pH 6.5. The composite was formed by self-assembly by mixing self-doped polyaniline nanofibers with silica-coated magnetite nanoparticles dispersions in ethanol. The composite structure was confirmed by transmission electron microscopy (TEM). Self-doped polyaniline nanofibers and magnetite chemical structures were confirmed by FT-IR while silica coating of the magnetite was confirmed by Energy Dispersion X-ray Spectroscopy (EDS). Improved stability of the composite magnetic component was evidenced by resistance to degrade in 2N HCl solution. The adsorption capacity of self-doped polyaniline nanofibers based composite was higher than previously reported corresponding composite prepared from polyaniline nanofibers instead of self-doped polyaniline nanofibers. Adsorption-pH profile for the studied FQs on the prepared composite revealed that the best pH for adsorption was in range of 6.5 to 7. Best extraction recovery values were obtained at pH 7 using phosphate buffer. The best solvent for FQs desorption was found to be 0.1N HCl in methanol:water (8:2; v/v) mixture. 20 mL of Spiked water sample with studied FQs were preconcentrated using 4.8 mg of composite and resulting extracts were analysed by HPLC-UV method. The prepared composite represented a suitable adsorbent phase for magnetic dispersive micro-solid phase application.

Keywords: fluoroquinolones, magnetic dispersive micro extraction, nano-composite, self-doped polyaniline nanofibers

Procedia PDF Downloads 98

Authors: Mindaugas Liaudanskas, Monika Tallat-Kelpsaite, Darius Kviklys, Jonas Viskelis, Pranas Viskelis, Norbertas Uselis, Juozas Lanauskas, Valdimaras Janulis

Abstract:

Apples are an important source of various biologically active compounds used for human health. Phenolic compounds detected in apples are natural antioxidants and have antimicrobial, anti-inflammatory, anticarcinogenic, and cardiovascular protective activity. The quantitative composition of phenolic compounds in apples may be affected by various factors. It is important to investigate it in order to provide the consumer with high-quality well-known composition apples and products made out of it. The objective of this study was to evaluate phenolic compounds quantitative composition in apple fruits grown in a different geographical region. In this study, biological replicates of apple cv. 'Ligol', grown in Lithuania, Latvia, Poland, and Estonia, were investigated. Three biological replicates were analyzed; one of each contained 10 apples. Samples of lyophilized apple fruits were extracted with 70% ethanol (v/v) for 20 min at 40∘C temperature using the ultrasonic bath. The ethanol extracts of apple fruits were analyzed by the high-performance liquid chromatography method. The study found that the geographical location of apple-trees had an impact on the composition of phenolic compounds in apples. The number of quercetin glycosides varied from 314.78±9.47 µg/g (Poland) to 648.17±5.61 µg/g (Estonia). The same trend was also observed with flavan-3-ols (from 829.56±47.17 µg/g to 2300.85±35.49 µg/g), phloridzin (from 55.29±1.7 µg/g to 208.78±0.35 µg/g), and chlorogenic acid (from 501.39±28.84 µg/g to 1704.35±22.65 µg/g). It was observed that the amount of investigated phenolic compounds tended to increase from apples grown in the southern location (Poland) (1701.02±75.38 µg/g) to apples grown northern location (Estonia) (4862.15±56.37 µg/g). Apples (cv. 'Ligol') grown in Estonia accumulated approx. 2.86 times higher amount of phenolic compounds than apples grown in Poland. Acknowledgment: This work was supported by a grant from the Research Council of Lithuania, project No. S-MIP-17-8.

Keywords: apples, cultivar 'Ligol', geographical regions, HPLC, phenolic compounds

Procedia PDF Downloads 153

Authors: Djebbar Atmani, Dina Kilani, Tristan Richard

Abstract:

Medicinal plants are an important source for the discovery of potential new substances for use in medicine and food. Pistacia lentiscus, Fraxinus angustifolia and Clematis flammula, plants growing in the Mediterranean basin, are widely used in traditional medicine. Therefore, the present study was designed to investigate their antioxidant, anti-inflammatory, antidiabetic, anti-mutagenic/genotoxic and neuroprotective potential and identification of active compounds using appropriate methodology. Plant extracts and fractions exhibited high scavenging capacity against known radicals, enhanced superoxide dismutase and catalase activitiesand restored blood glucose levels, in vivo, to normal values, in agreement with the in vitro enzymatic inhibition data, through inhibition of amylase and glucosidase activities. Administration of Pistacia lentiscus extracts significantly decreased carrageenan-induced mice paw oedema and reduced effectively IL-1β levels in cell culture, whereas Fraxinus angustifolia extracts showed good healing capacity against wounds when applied topically on rabbits. Pistacia lentiscus and Fraxinus angustifolia extracts showed good neuro-protection and restored cognitive functions in mice, while Clematis flammula extracts showed potent anti-ulcerogenic activity associated to a promising anti-mutagenic/genotoxic activity. HPLC-MS and NMR analyses allowed the identification and structural elucidation of several known and new anthocyanins, flavonols and flavanols. Therefore, Pistacia lentiscus, Fraxinus angustifolia and Clematis flammulacould be used in palliative treatments against inflammatory conditions and diabetes complications, as well as against deterioration of cognitive functions.

Keywords: pistacia lentiscus, clematis flammula, fraxinus angustifolia, phenolic compounds, biological activity

Procedia PDF Downloads 44

Authors: Cristian Moisa, Andreea Lupitu, Adriana Csakvari, Dana G. Radu, Leonard Marian Olariu, Georgeta Pop, Dorina Chambre, Lucian Copolovici, Dana Copolovici

Abstract:

Green synthesis of nanoparticles (NPs) represents a promising, accessible, eco-friendly, and safe process with significant applications in biotechnology, pharmaceutical sciences, and farming. The aim of our study was to obtain silver nanoparticles, using plant wastes extracts resulted in the essential oils extraction process: Thymus vulgaris L., Origanum vulgare L., Lavandula angustifolia L., and in hemp processing for seed and fibre, Cannabis sativa. Firstly, we obtained aqueous extracts of thyme, oregano, lavender, and hemp (two monoicous and one dioicous varieties), all harvested in western part of Romania. Then, we determined the chemical composition of the extracts by liquid-chromatography coupled with diode array and mass spectrometer detectors. The compounds identified in the extracts were in agreement with earlier published data, and the determination of the antioxidant activity of the obtained extracts by DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) assays confirmed their antioxidant activity due to their total polyphenolic content evaluated by Folin-Ciocalteu assay. Then, the silver nanoparticles (AgNPs) were successfully biosynthesised, as was demonstrated by UV-VIS, FT-IR spectroscopies, and SEM, by reacting AgNO₃ solution and plant extracts. AgNPs were spherical in shape, with less than 30 nm in diameter, and had a good bactericidal activity against Gram-positive (Staphylococcus aureus) and Gram-negative bacteria (Escherichia coli, Klebsiella pneumoniae, Pseudomonas fluorescens).

Keywords: plant wastes extracts, chemical composition, high performance liquid chromatography mass spectrometer, HPLC-MS, scanning electron microscopy, SEM, silver nanoparticles

Procedia PDF Downloads 152

Authors: Garzon V., Bustos R., Salvador J. P., Marco M. P., Pinacho D. G.

Abstract:

Bacterial resistance to antimicrobial treatment has increased significantly in recent years, making it a public health problem. Large numbers of bacteria are resistant to all or nearly all known antimicrobials, creating the need for the development of new types of antimicrobials or the use of “last line” antimicrobial drug therapies for the treatment of multi-resistant bacteria. Some of the chemical groups of antimicrobials most used for the treatment of infections caused by multiresistant bacteria in the clinic are Glycopeptide (Vancomycin), Polymyxin (Colistin), Lipopeptide (Daptomycin) and Carbapenem (Meropenem). Molecules that require therapeutic drug monitoring (TDM). Due to the above, a methodology based on nanobiotechnology based on an optical and electrochemical biosensor is being developed, which allows the evaluation of the plasmatic levels of some antimicrobials such as glycopeptide, polymyxin, lipopeptide and carbapenem quickly, at a low cost, with a high specificity and sensitivity and that can be implemented in the future in public and private health hospitals. For this, the project was divided into five steps i) Design of specific anti-drug antibodies, produced in rabbits for each of the types of antimicrobials, evaluating the results by means of an immunoassay analysis (ELISA); ii) quantification by means of an electrochemical biosensor that allows quantification with high sensitivity and selectivity of the reference antimicrobials; iii) Comparison of antimicrobial quantification with an optical type biosensor; iv) Validation of the methodologies used with biosensor by means of an immunoassay. Finding as a result that it is possible to quantify antibiotics by means of the optical and electrochemical biosensor at concentrations on average of 1,000ng/mL, the antibodies being sensitive and specific for each of the antibiotic molecules, results that were compared with immunoassays and HPLC chromatography. Thus, contributing to the safe use of these drugs commonly used in clinical practice and new antimicrobial drugs.

Keywords: antibiotics, electrochemical biosensor, optical biosensor, therapeutic drug monitoring

Procedia PDF Downloads 49

Authors: Younus Mohammad, Anita B. Fallah, Ben J. Boyd, Shakila B. Rizwan

Abstract:

N-acylethanolamines (NAEs) are endogenous lipids, which have neuromodulatory properties. NAEs have shown neuroprotective properties in various neurodegenerative diseases including Alzheimer's disease, Parkinson's disease and ischemic stroke. However, NAEs are eliminated rapidly in vivo by enzymatic hydrolysis. We propose to encapsulate NAEs in liquid crystalline nanoparticles (cubosomes) to increase their biological half-life and explore their therapeutic potential. Recently, we have reported the co-formulation and nanostructural characterization of cubosomes containing the NAE, oleoylethanolamide and a synthetic cubosome forming lipid phytantriol. Here, we report on the formulation of cubosomes with the NAE, linoleoylethanolamide (LEA) as the core cubosome forming lipid. LEA-cubosomes were formulated in the presence of three different steric stabilisers: two brain targeting ligands, Tween 80 and Pluronic P188 and a control, Pluronic F127. Size, morphology and internal structure of formulations were characterized by dynamic light scattering (DLS), cryogenic transmission electron microscopy (Cryo–TEM) and small angle X–ray scattering (SAXS), respectively. Chemical stability of LEA in formulations was investigated using high-performance liquid chromatography (HPLC). Cytotoxicity of formulations towards human cerebral microvascular endothelial cell line (hCMEC/D3) was also investigated using an MTT (3-[4, 5- dimethylthiazol-2-yl]-2, 5-diphenyl tetrazolium bromide) assay. All cubosome formulations had mean particle size of less than 250 nm and were uniformly distributed with polydispersity indices less than 0.2. Cubosomes produced had a bicontinuous cubic internal structure with an Im3m space group but different lattice parameters, indicating the different modes of interaction between the stabilisers and LEA. LEA in formulations was found to be chemically stable. At concentrations of up to 20 µg/mL LEA in the presence of all the stabilisers, greater than 80% cell viability was observed.

Keywords: blood-brain barrier, cubosomes, linoleoyl ethanolamide, N-acylethanolamines (NAEs)

Procedia PDF Downloads 178

Authors: Khayra Zerrouki, Noureddine Djebli, Esra Eroglu, Afife Mat, Ozhan Gul

Abstract:

Neurodegenerative diseases of the human brain comprise a variety of disorders that affect an increasing percentage of the population. Alzheimer’s disease (AD) is a complex, multifactorial, heterogeneous mental illness, which is characterized by an age-dependent loss of memory and an impairment of multiple cognitive functions, but this 10 last years it concerns the population most and most young. Hypericum perforatum has traditionally been used as an external anti-inflammatory and healing remedy for the treatment of swellings, wounds and burns, diseases of the alimentary tract and psychological disorders. It is currently of great interest due to new and important therapeutic applications. In this study, the chemical composition of methanolic extract of Hypericum perforatum (HPM) was analysed by using high performance liquid chromatography – diode array detector (HPLC-DAD). The in vitro antioxidant activity of HPM was evaluated by using several antioxidant tests. HSM exhibits inhibitory capacity against posphatidylcholine liposome peroxidation, induced with iron and ascorbic acid, scavenge DPPH and superoxide radicals and act as reductants. The cytotoxic activity of HSM was also determined by using MTT cell viability assay on HeLa and NRK-52E cell lines. The in vivo activity studies in Swiss mice were determined by using behavioral, memory tests and histological study. According to tests results HPM that may be relevant to the treatment of cognitive disorders. The results of chemical analysis showed a hight level of hyperforin and quercitin that had an important antioxidant activity proved in vitro with the DPPH, anti LPO and SOD; this antioxidant activity was confirmed in vivo after the non-toxic results by means of improvement in behavioral and memory than the reducing shrunken in pyramidal cells of mice brains.

Keywords: AlCl3, alzheimer, mice, neuroprotective, neurotoxicity, phytotherapy

Procedia PDF Downloads 470

Authors: Wasim Sajjad, Manzoor Ahmad, Sundas Qadir, Muhammad Rafiq, Fariha Hasan, Richard Tehan, Kerry L. McPhail, Aamer Ali Shah

Abstract:

Aim: This study aims to investigate the possible radiation protective role of a compatible solute in the tolerance of radio-halophilic bacterium against stresses, like desiccation and exposure to ionizing radiation. Methods and Results: Nine different radio-resistant bacteria were isolated from desert soil, where strain WMA-LM19 was chosen for detailed studies on the basis of its high tolerance for ultraviolet radiation among all these isolates. 16S rRNA gene sequencing indicated that the bacterium was closely related to Stenotrophomonas sp. (KT008383). A bacterial milking strategy was applied for extraction of intracellular compatible solutes in 70% (v/v) ethanol, which were purified by high-performance liquid chromatography (HPLC). The compound was characterized as ectoine by 1H and 13C nuclear magnetic resonance (NMR), and mass spectrometry (MS). Ectoine demonstrated more efficient preventive activity (54.80%) to erythrocyte membranes and also inhibited oxidative damage to proteins and lipids in comparison to the standard ascorbic acid. Furthermore, a high level of ectoine-mediated protection of bovine serum albumin against ionizing radiation (1500-2000 Jm-2) was observed, as indicated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis. Conclusion: The results indicated that ectoine can be used as a potential mitigator and radio-protective agent to overcome radiation- and salinity-mediated oxidative damage in extreme environments. Significance and Impact of the Study: This study shows that ectoine from radio-halophiles can be used as a potential source in topical creams as sunscreen. The investigation of ectoine as UV protectant also changes the prospective that radiation resistance is specific only to molecular adaptation.

Keywords: ectoine, anti-oxidant, stenotrophomonas sp., ultraviolet radiation

Procedia PDF Downloads 185

Authors: Kasipandi Muniyandi, Parimelazhagan Thangaraj

Abstract:

Plants are important prospective wealth of a country, combination of local health care information about a specific plant together with data published by several groups of scientists, can help in deciding whether it should be considered acceptable for medicinal use. In the developed countries, too, plant-derived drugs may be of importance. The wide variety of ailments that are being treated with Plectranthus is an indication of the medicinal value of the genus. A number of species are not toxic and so may be taken orally, whilst others are used topically on the skin or as enemas. This study was designed to evaluate the different properties of Plectranthus stocksii and the aerial parts were collected and extracted with petroleum ether, chloroform, ethyl acetate, acetone and methanol by Soxhlet apparatus and finally macerated with hot water. The quantification assays revealed that, leaf methanol extract showed higher total phenolic (415.41 mg GAE/ g extract) and tannin (177.53 mg GAE/ g extract) contents whereas leaf ethyl acetate exhibited higher flavonoids (777.11 mg RE/ g extract) content. The antioxidant efficiency of the extracts was analyzed by various radical scavenging assays. Among the different antioxidant assays, leaf ethyl acetate extract showed higher free radical scavenging activities against DPPH (IC50 = 3.46 µg/mL), ABTS (27417.65 µM TE/ g extract), FRAP (152.17 mM Fe(II)E/ mg extract) NO• radical (21.46%) and Superoxide radical (IC50 = 24.16 µg/mL) assays. All the parts P. stocksii extracts showed significant protection against OH• induced DNA damage at 50 µg concentration. The HPLC analysis of leaf ethyl acetate extract revealed the presence of Quercetin (30.29 µg/mg of extract) was the major compound. Anticancer activity of leaf ethyl acetate extract showed better IC50 values were 48.87 and 36.08 µg/ mL against MCF-7 and Caco-2 respectively. From this study, P. stocksii can act as a potent antioxidant and cytotoxic antimicrobial agent. The scope for drug development from this plant is endless and there is undoubtedly a call for further research in pharmaceutical industries.

Keywords: antioxidant, cytotoxicity, phenolics, plectranthus stocksii

Procedia PDF Downloads 352

Authors: A. Cervantes-Diaz, B. Sevilla-Moran, Manuel Alcami, Al Mokhtar Lamsabhi, J. L. Alonso-Prados, P. Sandin-España

Abstract:

Profoxydim is a post-emergence herbicide belonging to the cyclohexanedione oxime family, used to control weeds in rice crops. The use of soil organic amendments has increased significantly in the last decades, and their effects on the behavior of many herbicides are still unknown. Additionally, it is known that photolysis is an important degradation process to be considered when evaluating the persistence of this family of herbicides in the environment. In this work, the photodegradation of profoxydim in an amended paddy soil-water system with alperujo compost was studied. Photodegradation experiments were carried out under laboratory conditions using simulated solar light (Suntest equipment) in order to evaluate the reaction kinetics of the active substance. The photochemical behavior of profoxydim was investigated in soil with and without alperujo amendment. Furthermore, due to the rice crop characteristics, profoxydim photodegradation in water in contact with these types of soils was also studied. Determination of profoxydim degradation kinetics was performed by High-Performance Liquid Chromatography with Diode-Array Detection (HPLC-DAD). Furthermore, we followed the evolution of resulting transformation by-products, and their tentative identification was achieved by mass spectrometry. All the experiments allowed us to fit the data of profoxydim photodegradation to a first-order kinetic. Photodegradation of profoxydim was very rapid in all cases. The half-lives in aqueous matrices were in the range of 86±0.3 to 103±0.5 min. The addition of alperujo amendment to the soil produced an increase in the half-life from 62±0.2 min (soil) to 75±0.3 min (amended soil). In addition, a comparison to other organic amendments was also performed. Results showed that the presence of the organic amendment retarded the photodegradation in paddy soil and water. Regarding degradation products, the main process involved was the cleavage of the oxime moiety giving rise to the formation of the corresponding imine compound.

Keywords: by-products, herbicide, organic amendment, photodegradation, profoxydim

Procedia PDF Downloads 44

Authors: Tatjana Jurić, Bojana Blagojević, Denis Uka, Ružica Ždero Pavlović, Boris M. Popović

Abstract:

Natural deep eutectic solvents (NADES) represent a class of modern systems that have been developed as a green alternative to toxic organic solvents, which are commonly used as extraction media. It has been considered that hydrogen bonding is the main interaction leading to the formation of NADES. The aim of this study was phytochemical characterization and determination of the antioxidant and antibacterial activity of Mentha piperita leaf extracts obtained by six choline chloride-based NADES. NADES were prepared by mixing choline chloride with different hydrogen bond donors in 1:1 molar ratio following the addition of 30% (w/w) water. The mixtures were then heated (60 °C) and stirred (650 rpm) until the clear homogenous liquids were obtained. The Mentha piperita extracts were prepared by mixing 75 mg of peppermint leaves with 1 mL of NADES following by the heating and stirring (60 °C, 650 rpm) within 30 min. The content of six phenolics in extracts was determined using HPLC-PDA. The dominant compounds presented in peppermint leaves - rosmarinic acid and luteolin 7-O-glucoside, were extracted by NADES at a similar level as 70% ethanol. The microdilution method was applied to test the antibacterial activity of extracts. Compared with 70% ethanol, all NADES systems showed higher antibacterial activity towards Pseudomonas aeruginosa (Gram -), Staphylococcus aureus (Gram +), Escherichia coli (Gram -), and Salmonella enterica (Gram -), especially NADES containing organic acids. The majority of NADES extracts showed a better ability to neutralize DPPH radical than conventional solvent and similar ability to reduce Fe3+ to Fe2+ ions in FRAP assay. The obtained results introduce NADES systems as the novel, sustainable, and low-cost solvents with a variety of applications.

Keywords: antibacterial activity, antioxidant activity, green extraction, natural deep eutectic solvents, polyphenols

Procedia PDF Downloads 159

Authors: A. K. R. Gobinath, He Jianzhong, Kun-Lin Yang

Abstract:

Fruit waste was used as a feedstock to produce biohydrogen in this study. Fruit waste used in this study was collected from several fruit juice stalls in Singapore. Based on our observation, the fruit waste contained 35-40% orange, 10-20% watermelon, 10-15% apple, 10-15% pineapple, 1-5% mango. They were mixed with water (1:1 ratio based on wet biomass) and blended to attain homogenous mixtures. Later, fruit waste was subjected to one of the following pretreatments: autoclave (121 °C for 20min), microwave (20min) or both. After pretreatment, the total sugar concentration in the hydrolysate was high (>12g/l) when both autoclave and microwave were applied. In contrast, samples without pretreatment measured only less than 2g/l of sugar. While using these hydrolysates as carbon sources, Clostridium strain BOH3 produces 2526-3126 ml/l of hydrogen after 72h of anaerobic fermentation. The hydrogen yield was 295-300 ml/g of sugar which is close to the hydrogen yields from glucose (338 ml/gm) and xylose (330 ml/gm). Our HPLC analysis showed that fruit waste hydrolysate contained oligosugars (25-27%), sucrose (18-23%), fructose (25-30%), glucose (10-15%) and mannose (2-5%). Additionally, pretreatment led to the release of free amino acids (160-512 mg/l), calcium (7.8-12.9 ppm), magnesium (4.32-6.55 ppm), potassium (5.4-65.1 ppm) and sodium (0.4-0.5 ppm) into the hydrolysate. These nutrients were able to support strain-BOH3 to grow and produce high level of hydrogen. Notably, unlike other pretreatment methods (with strong acids and bases), these pretreatment techniques did not generate any inhibitors (e.g. furfural and phenolic acids) to suppress the hydrogen production. Interestingly, strain BOH3 can also ferment pretreated fruit waste slurry and produce hydrogen with a high yield (156-343 ml/gm fruit waste). While fermenting pretreated fruit waste slurry, strain-BOH3 excreted several saccharolytic enzymes majorly xylanase (1.84U/ml), amylase (1.10U/ml), pectinase (0.36U/ml) and cellulase (0.43U/ml). Due to expressions of these enzymes, strain BOH3 was able to directly utilize pretreated fruit waste hydrolysate and produces high-level of hydrogen.

Keywords: autoclave pretreatment, biohydrogen production, clostridial fermentation, fruit waste, and microwave pretreatment

Procedia PDF Downloads 507

Authors: Aghaei Amirkhizi Navideh, Sadjadi Soodeh Sadat, Moghaddam Banaem Leila, Athari Allaf Mitra, Johari Daha Fariba

Abstract:

Dendrimers are good candidates for preparing metal nanoparticles because they can structurally and chemically well-defined templates and robust stabilizers. Poly amidoamine (PAMAM) dendrimer-based multifunctional cancer therapeutic conjugates have been designed and synthesized in pharmaceutical industry. In addition, encapsulated nanoparticle surfaces are accessible to substrates so that catalytic reactions can be carried out. For preparation of dendimer-metal nanocomposite, a dendrimer solution containing an average of 55 Yb+3 ions per dendrimer was prepared. Prior to reduction, the pH of this solution was adjusted to 7.5 using NaOH. NaBH4 was used to reduce the dendrimer-encapsulated Yb+3 to the zerovalent metal. The pH of the resulting solution was then adjusted to 3, using HClO4, to decompose excess BH4-. The UV-Vis absorption spectra of the mixture were recorded to ensure the formation of Yb-G5-NH2 complex. High-resolution electron microscopy (HRTEM) and size distribution results provide additional information about dendimer-metal nanocomposite shape, size, and size distribution of the particles. The resulting mixture was irradiated in Tehran Research Reactor 2h and neutron fluxes were 3×1011 n/cm2.Sec and the specific activity was 7MBq. Radiochemical and chemical and radionuclide quality control testes were carried. Gamma Spectroscopy and High-performance Liquid Chromatography HPLC, Thin-Layer Chromatography TLC were recorded. The injection of resulting solution to solid tumor in mice shows that it could be resized the tumor. The studies about solid tumors and nano composites show that ytterbium encapsulated-dendrimer radiopharmaceutical could be introduced as a new therapeutic for the treatment of solid tumors.

Keywords: nano-radio pharmaceutical, ytterbium, PAMAM, dendrimers

Procedia PDF Downloads 478

Authors: Jamilah Syafawati Yaacob, Muhammad Aiman Ramli

Abstract:

Cili padi or Capsicum frutescens is one of capsicum species from nightshade family, Solanaceae. It is famous in Malaysia and is widely used as a food ingredient. Capsicum frutescens also possess vast medicinal properties. The objectives of this study are to determine the most optimum 2,4-D hormone concentration for callus induction from stem explants C. frutescens and the effects of different 2,4-D concentrations on expression of compounds from C. frutescens. Seeds were cultured on MS media without hormones (MS basal media) to yield aseptic seedlings of this species, which were then used to supply explant source for subsequent tissue culture experiments. Stem explants were excised from aseptic seedlings and cultured on MS media supplemented with various concentrations (0.1, 0.3 and 0.5 mg/L) of 2,4-D to induce formation of callus. Fresh weight, dry weight and callus growth percentage in all samples were recorded. The highest mean of dry weight was observed in MS media supplemented with 0.5 mg/L 2,4-D, where 0.4499 ± 0.106 g of callus was produced. The highest percentage of callus growth (16.4%) was also observed in cultures supplemented with 0.5 mg/L 2,4-D. The callus samples were also subjected to HPLC-MS to evaluate the effect of hormone concentration on expression of bio active compounds in different samples. Results showed that caffeoylferuloylquinic acids were present in all samples, but was most abundant in callus cells supplemented with 0.3 & 0.5 mg/L 2,4-D. Interestingly, there was an unknown compound observed to be highly expressed in callus cells supplemented with 0.1 mg/L 2,4-D, but its presence was less significant in callus cells supplemented with 0.3 and 0.5 mg/L 2,4-D. Furthermore, there was also a compound identified as octadecadienoic acid, which was uniquely expressed in callus supplemented with 0.5 mg/L 2,4-D, but absent in callus cells supplemented with 0.1 and 0.3 mg/L 2,4-D. The results obtained in this study indicated that plant growth regulators played a role in expression of secondary metabolites in plants. The increase or decrease of these growth regulators may have triggered a change in the secondary metabolite biosynthesis pathways, thus causing differential expression of compounds in this plant.

Keywords: callus, in vitro, secondary metabolite, 2, 4-Dichlorophenoxyacetic acid

Procedia PDF Downloads 348