Search results for: pyroglutamic acid
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 3350

Search results for: pyroglutamic acid

2930 Surface Modified Polyamidoamine Dendrimer with Gallic Acid Overcomes Drug Resistance in Colon Cancer Cells HCT-116

Authors: Khushbu Priyadarshi, Chandramani Pathak

Abstract:

Cancer cells can develop resistance to conventional therapies especially chemotherapeutic drugs. Resistance to chemotherapy is another challenge in cancer therapeutics. Therefore, it is important to address this issue. Gallic acid (GA) is a natural plant compound that exhibits various biological properties including anti-proliferative, anti-inflammatory, anti-oxidant and anti-bacterial. Despite of the wide spectrum biological properties GA has cytotoxic response and low bioavailability. To overcome this problem, GA was conjugated with the Polyamidoamine(PAMAM) dendrimer for improving the bioavailability and efficient delivery in drug-resistant HCT-116 Colon Cancer cells. Gallic acid was covalently linked to 4.0 G PAMAM dendrimer. PAMAM dendrimer is well established nanocarrier but has cytotoxicity due to presence of amphiphilic nature of amino group. In our study we have modified surface of PAMAM dendrimer with Gallic acid and examine their anti-proliferative effects in drug-resistant HCT-116 cells. Further, drug-resistant colon cancer cells were established and thereafter treated with different concentration of PAMAM-GA to examine their anti-proliferative potential. Our results show that PAMAM-GA conjugate induces apoptotic cell death in HCT-116 and drug-resistant cells observed by Annexin-PI staining. In addition, it also shows that multidrug-resistant drug transporter P-gp protein expression was downregulated with increasing the concentration of GA conjugate. After that we also observed the significant difference in Rh123 efflux and accumulation in drug sensitive and drug-resistant cancer cells. Thus, our study suggests that conjugation of anti-cancer agents with PAMAM could improve drug resistant property and cytotoxic response to treatment of cancer.

Keywords: drug resistance, gallic acid, PAMAM dendrimer, P-glycoprotein

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2929 Determination of Hydrolisis Condition in the Extraction of Fatty Acids from Pinchagua's (Opisthonema libertate) Heads, a By-Product of Sardine Industry

Authors: Belen Carrillo, Mauricio Mosquera

Abstract:

Fatty acids are bioactive compounds widely used as nutritional supplements in the food and pharmaceutical industry. Bluefish such as sardines have a large variety of these fatty acids in their composition. The objective of this project is to extract these compounds from fishing wastes, to do this, heads of known species as Pinchagua (Opistonema libertate) were used. The conducted study represents a simplified alternative for obtaining and simultaneous saponification of oil through basic hydrolysis, which separates lipids from protein and saponifies sample all the same time to isolate the fatty acid accurately through salts formation. To do these different concentrations of sodium hydroxide were used, it was demonstrated at a concentration of 1 M the highest yield of saponified oil recovery corresponding a value of 3,64% was obtained. Subsequently, the saponified oil was subjected to an acid hydrolysis in which fatty acids were isolated. Different sulfuric acid concentrations and temperatures for the process were tested. Thus, it was shown that the great fatty acids variety were obtained at a 60 °C temperature and sulfuric acid concentration of 50% v/v. Among the obtained compounds the presence of acids such as palmitic, lauric, caproic and myristic are highlighted. Applications of this type of elements are varied and widely used in the nutritional supplements development. Thus, the described methodology proposes a simple mechanism in the revaluation of fishing industry wastes that allow directly generate high added value elements.

Keywords: fatty acids, hydrolysis, Pinchagua, saponification

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2928 Biosorption of Methylene Blue and Acid Red-88 from Wastewater by Using Cypress Cones

Authors: Onur Yel

Abstract:

This study represents the removal of harmful dye substances from wastewaters by using waste and cheap adsorbents. Rapid population growth and industrialization occasion anthropogenic pollution which gives irreversible damage to the environment. One of the ways in which water pollution occurs is caused by the release of the dyestuffs in the textile industry. The release of dyestuffs to the environment directly damages the living creatures that have acquired water habitat. Especially, wastewater cannot be used for nutritional purposes. In addition, some adsorbents have mutagenic and/or carcinogenic effects. By blocking photosynthesis, it hinders the inhibition of photosynthetic bacteria in the water, which damages the ecological balance and also causes the formation of malodorous compounds. Moreover, the lack of oxygen can pose a serious danger to the lives of other living organisms that need oxygen. In recent years, some physical and chemical methods are preferred for the removal of dyestuffs. However, the utilization of these methods is expensive. For this reason, the availability of new and cheap adsorbents becomes the more significant issue. In this study, an investigation of various variables on the removal of Methylene Blue and Acid Red-88 dyestuffs from wastewaters by the usage of pulverized cypress cones has been carried out. Thus, various masses of absorbent (0.1-0.25-0.5-1-2-4-5 grams) are used in 50, 100, 150, 200, 300 ppm concentrations of Methylene Blue and Acid Red-88 dyestuffs’ solutions, and with a variety of the interaction time (0.25-0.5-1-2-4-5 hours). The mixtures were centrifuged and the absorbance of the filtrates was measured on a UV spectrophotometer to determine their remaining concentrations. In the study, the highest removal ratio of Acid Red-88 dyestuff was found to be 81% at 200 ppm of dyestuff with 2 grams of adsorbent at 300 minutes. For Methylene Blue experiments, the removal percentage was found as 98% where 2 grams of adsorbent is used in 200 ppm dyestuff solution at 120 minutes of interaction.

Keywords: acid red-88, biosorption, methylene blue, cypress cones, water pollution

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2927 Cellular Uptake and Endocytosis of Doxorubicin Loaded Methoxy Poly (Ethylene Glycol)-Block-Poly (Glutamic Acid) [DOX/mPEG-b-PLG] Nanoparticles against Human Breast Cancer Cell Lines

Authors: Zaheer Ahmad, Afzal Shah

Abstract:

pH responsive block copolymers consist of mPEG and glutamic acid units were syntheiszed in different formulations. The synthesized polymers were structurally investigated. Doxorubicin Hydrocholide (DOX-HCl) as a chemotherapy medication for the treatment of cancer was selected. DOX-HCl was loaded and their drug loading content and drug loading efficiency were determined. The nanocarriers were obtained in small size, well shaped and slightly negative surface charge. The release study was carried out both at pH 7.4 and 5.5 and it was revealed that the release was sustained and in controlled manner and there was no initial burst release. The in vitro release study was further carried out for different formulations with different glutamic acid moieties. Time dependent cell proliferation inhibition of the free drug and drug loaded nanoparticles against human breast cancer cell lines MCF-7 and Zr-75-30 was observed. Cellular uptakes and endocytosis were investigated by confocal laser scanning microscopy (CLSM) and flow cytometery. The biocompatibility, optimum size, shape and surface charge of the developed nanoparticles make the nanoparticles an efficient drug delivery carrier.

Keywords: doxorubicin, glutamic acid, cell proliferation inhibition, breast cancer cell

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2926 Fabrication of Gold Nanoparticles Self-Assembled Functionalized Improved Graphene on Carbon Paste Electrode for Electrochemical Determination of Levodopa in the Presence of Ascorbic Acid

Authors: Mohammad Ali Karimi, Hossein Tavallali, Abdolhamid Hatefi-Mehrjardi

Abstract:

In this study, an electrochemical sensor based on gold nanoparticles (AuNPs) functionalized improved graphene (AuNPs-IGE) was fabricated for selective determination of L-dopa in the presence of ascorbic acid by a novel self-assembly method. The AuNP IGE modified carbon paste electrode (AuNPs-IGE/CPE) utilized for investigation of the electrochemical behavior of L-dopa in phosphate buffer solution. Compared to bare CPE, AuNPs-IGE/CPE shows novel properties towards the electrochemical redox of levodopa (L-dopa) in phosphate buffer solution at pH 4.0. The oxidation potential of L-dopa shows a significant decrease at the AuNPs-IGE/CPE. The oxidation current of L-dopa is higher than that of the unmodified CPE. AuNPs-IG/CPE shows excellent electrocatalytic activity for the oxidation of ascorbic acid (AA). Using differential pulse voltammetry (DPV) method, the oxidation current is well linear with L-dopa concentration in the range of 0.4–50 µmol L-1, with a detection limit of about 1.41 nmol L-1 (S/N = 3). Therefore, it was applied to measure L-dopa from real samples that recoveries are 94.6-106.2%. The proposed electrode can also effectively avoid the interference of ascorbic acid, making the proposed sensor suitable for the accurate determination of L-dopa in both pharmaceutical preparations and human body fluids.

Keywords: gold nanoparticles, improved graphene, L-dopa, self-assembly

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2925 Exploration Study of Civet Coffee: Amino Acids Composition and Cup Quality

Authors: Murna Muzaifa, Dian Hasni, Febriani, Anshar Patria, Amhar Abubakar

Abstract:

Coffee flavour is influenced by many factors such as processing techniques. Civet coffee is known as one of premium coffee due to its unique processing technique and its superior cupping quality. The desirable aroma of coffee is foremost formed during roasting step at a high temperature from precursors that are present in the green bean. Sugars, proteins, acids and trigonelline are the principal flavor precursors compounds in green coffee bean. It is now widely accepted that amino acids act as precursors of the Maillard reaction during which the colour and aroma are formed. To investigate amino acids on civet coffee, concentration of 20 amino acids (L-Isoleucine, L-Valine, L-Proline, L-Phenylalanine, L-Arginine, L-Asparagine, L-Threonine, L-Tryptophan, L-Leucine, L-Serine, L-Glutamine, L-Methionine, L-Histidine, Aspartic acid, L-Tyrosine, L-Lysine, L-Glutamic acid, and L-Cysteine, L-Alanine and Glycine) were determined in green and roasted bean of civet coffee by LCMS analysis. The cup quality of civet coffee performed using professional Q-grader followed SCAA standard method. The measured parameters were fragrance/aroma, flavor, acidity, body, uniformity, clean up, aftertaste, balance, sweetness and overall. The work has been done by collecting samples of civet coffee from six locations in Gayo Higland, Aceh-Indonesia. The results showed that 18 amino acids were detected in green bean of civet coffee (L-Isoleucine, L-Valine, L-Proline, L-Phenylalanine, L-Arginine, L-Asparagine, L-Threonine, L-Tryptophan, L-Leucine, L-Serine, L-Glutamine, L-Methionine, L-Histidine, Aspartic acid, L-Tyrosine, L-Lysine, L-Glutamic acid, and L-Cysteine) and 2 amino acids were not detected (L-Alanine and Glycine). On the other hand, L-Tyrosine and Glycine were not detected in roasted been of civet coffee. Glutamic acid is the amino acid with highest concentration in both green and roasted bean (21,02 mg/g and 24,60 mg/g), followed by L- Valine (19,98 mg/g and 20,22 mg/g) and Aspartic acid (14,93 mg/g and 18,58 mg/g). Civet coffee has a fairly high cupping value (cup quality), ranging from 83.75 to 84.75, categorized as speciality coffee. Moreover, civet coffee noted to have nutty, chocolaty, fishy, herby and watery.

Keywords: amino acids, civet coffee, cupping quality, luwak

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2924 Improving the Ability of Constructed Wetlands to Treat Acid Mine Drainage

Authors: Chigbo Emmanuel Ikechukwu

Abstract:

Constructed wetlands are seen as a potential means of ameliorating the poor quality water that derives from coal and gold mining operations. However, the processes whereby a wetland environment is able to improve water quality are not well understood and techniques for optimising their performance poorly developed. A parameter that may be manipulated in order to improve the treatment capacity of a wetland is the substrate in which the aquatic plants are rooted. This substrate can provide an environment wherein sulphate reducing bacteria, which contribute to the removal of contaminants from the water, are able to flourish. The bacteria require an energy source which is largely provided by carbon in the substrate. This paper discusses the form in which carbon is most suitable for the bacteria and describes the results of a series of experiments in which different materials were used as substrate. Synthetic acid mine drainage was passed through an anaerobic bioreactor that contained either compost or cow manure. The effluent water quality was monitored with respect to time and the effect of the substrate composition discussed.

Keywords: constructed wetland, bacteria, carbon, acid mine drainage, sulphate

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2923 Physicochemical Analysis of Soxhlet Extracted Oils from Selected Northern Nigerian Seeds

Authors: Abdulhamid Abubakar, Sani Ibrahim, Fakai I. Musa

Abstract:

The aim of the present study is to investigate the potential use of the selected seed oils. The oil was extracted using Soxhlet apparatus and the physicochemical characteristics of the oil determined using standard methods. The following results were obtained for the physicochemical parameters analysed: for Egusi seed oil, Oil yield 53.20%, Saponification value 178.03±1.25 mgKOH/g, iodine value 49.10±0.32 g I2/100 g, acid value 4.30±0.86 mgKOH/g, and Peroxide value 5.80±0.27 meq/kg were obtained. For Pawpaw seed oil, Oil yield 40.10%, Saponification value 24.13±3.93 mgKOH/g, iodine value 24.87±0.19 g I2/100g, acid value 9.46±0.40 mgKOH/g, and Peroxide value 3.12±1.22 meq/kg were obtained. For Sweet orange seed oil, oil yield 43.10%, Saponification value 106.30±2.37 mgKOH/g, Iodine value 37.08±0.04 g I2/100g, acid value 7.59±0.77 mgKOH/g, and Peroxide value 2.21±0.46 meq/kg were obtained. From the obtained values of the determined parameters, the oils can be extracted from the three selected seeds in commercial quantities and that the egusi and sweet orange seed oils may be utilized in the industrial soap production.

Keywords: Carica papaya, Citrus sinensis, physicochemical, iodine value, peroxide value

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2922 Effects of Four Dietary Oils on Cholesterol and Fatty Acid Composition of Egg Yolk in Layers

Authors: A. F. Agboola, B. R. O. Omidiwura, A. Oyeyemi, E. A. Iyayi, A. S. Adelani

Abstract:

Dietary cholesterol has elicited the most public interest as it relates with coronary heart disease. Thus, humans have been paying more attention to health, thereby reducing consumption of cholesterol enriched food. Egg is considered as one of the major sources of human dietary cholesterol. However, an alternative way to reduce the potential cholesterolemic effect of eggs is to modify the fatty acid composition of the yolk. The effect of palm oil (PO), soybean oil (SO), sesame seed oil (SSO) and fish oil (FO) supplementation in the diets of layers on egg yolk fatty acid, cholesterol, egg production and egg quality parameters were evaluated in a 42-day feeding trial. One hundred and five Isa Brown laying hens of 34 weeks of age were randomly distributed into seven groups of five replicates and three birds per replicate in a completely randomized design. Seven corn-soybean basal diets (BD) were formulated: BD+No oil (T1), BD+1.5% PO (T2), BD+1.5% SO (T3), BD+1.5% SSO (T4), BD+1.5% FO (T5), BD+0.75% SO+0.75% FO (T6) and BD+0.75% SSO+0.75% FO (T7). Five eggs were randomly sampled at day 42 from each replicate to assay for the cholesterol, fatty acid profile of egg yolk and egg quality assessment. Results showed that there were no significant (P>0.05) differences observed in production performance, egg cholesterol and egg quality parameters except for yolk height, albumen height, yolk index, egg shape index, haugh unit, and yolk colour. There were no significant differences (P>0.05) observed in total cholesterol, high density lipoprotein and low density lipoprotein levels of egg yolk across the treatments. However, diets had effect (P<0.05) on TAG (triacylglycerol) and VLDL (very low density lipoprotein) of the egg yolk. The highest TAG (603.78 mg/dl) and VLDL values (120.76 mg/dl) were recorded in eggs of hens on T4 (1.5% sesame seed oil) and was similar to those on T3 (1.5% soybean oil), T5 (1.5% fish oil) and T6 (0.75% soybean oil + 0.75% fish oil). However, results revealed a significant (P<0.05) variations on eggs’ summation of polyunsaturated fatty acid (PUFA). In conclusion, it is suggested that dietary oils could be included in layers’ diets to produce designer eggs low in cholesterol and high in PUFA especially omega-3 fatty acids.

Keywords: dietary oils, egg cholesterol, egg fatty acid profile, egg quality parameters

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2921 Development and Total Error Concept Validation of Common Analytical Method for Quantification of All Residual Solvents Present in Amino Acids by Gas Chromatography-Head Space

Authors: A. Ramachandra Reddy, V. Murugan, Prema Kumari

Abstract:

Residual solvents in Pharmaceutical samples are monitored using gas chromatography with headspace (GC-HS). Based on current regulatory and compendial requirements, measuring the residual solvents are mandatory for all release testing of active pharmaceutical ingredients (API). Generally, isopropyl alcohol is used as the residual solvent in proline and tryptophan; methanol in cysteine monohydrate hydrochloride, glycine, methionine and serine; ethanol in glycine and lysine monohydrate; acetic acid in methionine. In order to have a single method for determining these residual solvents (isopropyl alcohol, ethanol, methanol and acetic acid) in all these 7 amino acids a sensitive and simple method was developed by using gas chromatography headspace technique with flame ionization detection. During development, no reproducibility, retention time variation and bad peak shape of acetic acid peaks were identified due to the reaction of acetic acid with the stationary phase (cyanopropyl dimethyl polysiloxane phase) of column and dissociation of acetic acid with water (if diluent) while applying temperature gradient. Therefore, dimethyl sulfoxide was used as diluent to avoid these issues. But most the methods published for acetic acid quantification by GC-HS uses derivatisation technique to protect acetic acid. As per compendia, risk-based approach was selected as appropriate to determine the degree and extent of the validation process to assure the fitness of the procedure. Therefore, Total error concept was selected to validate the analytical procedure. An accuracy profile of ±40% was selected for lower level (quantitation limit level) and for other levels ±30% with 95% confidence interval (risk profile 5%). The method was developed using DB-Waxetr column manufactured by Agilent contains 530 µm internal diameter, thickness: 2.0 µm, and length: 30 m. A constant flow of 6.0 mL/min. with constant make up mode of Helium gas was selected as a carrier gas. The present method is simple, rapid, and accurate, which is suitable for rapid analysis of isopropyl alcohol, ethanol, methanol and acetic acid in amino acids. The range of the method for isopropyl alcohol is 50ppm to 200ppm, ethanol is 50ppm to 3000ppm, methanol is 50ppm to 400ppm and acetic acid 100ppm to 400ppm, which covers the specification limits provided in European pharmacopeia. The accuracy profile and risk profile generated as part of validation were found to be satisfactory. Therefore, this method can be used for testing of residual solvents in amino acids drug substances.

Keywords: amino acid, head space, gas chromatography, total error

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2920 Green Technologies Developed by JSC “NIUIF”

Authors: Andrey Norov

Abstract:

In the recent years, Samoilov Research Institute for Mineral Fertilizers JSC “NIUIF”, the oldest (established in September 1919) industry-oriented institute in Russia, has developed a range of sustainable, environment-friendly, zero-waste technologies that ensure minimal consumption of materials and energy resources and fully consistent with the principles of Green Chemistry that include: - Ecofriendly energy and resource saving technology of sulfuric acid from sulfur according to DC-DA scheme (double conversion - double absorption); - Improved zero-waste technology of wet phosphoric acid (WPA) by dihydrate-hemihydrate process applicable to various types of phosphate raw materials; - Flexible, efficient, zero-waste, universal technology of NP / NPS / NPK / NPKS fertilizers with maximum heat recovery from chemical processes; - Novel, zero-waste, no-analogue technology of granular PK / PKS / NPKS fertilizers with controlled dissolution rate and nutrient supply into the soil, which allows to process a number of wastes and by-products; - Innovative resource-saving joint processing of wastes from the production of phosphogypsum and fluorosilicic acid (FSA) into ammonium sulfate with simultaneous neutralization of fluoride compounds with no lime used. - New fertilizer technology of increased environmental and agrochemical efficiency (currently under development). All listed green technologies are patented with Russian and Eurasian patents. The development of ecofriendly, safe, green technologies is ongoing in JSC “NIUIF”.

Keywords: NPKS fertilizers, FSA, sulfuric acid, WPA

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2919 Flavonoids and Phenolic Acids from the Aerial Parts of Alyssum alyssoides

Authors: Olga St. Tsiftsoglou, Diamanto M. Lazari, Eugene L. Kokkalou

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Most of Alyssum species of Brassicaceae family have been mainly studied for their contribution in ecology. In this study, A. alyssoides was examined for its chemical substitutes. The methanol extract of its aerial parts was fractionated with liquid-liquid extraction (distribution) with four different solvents of increasing polarity: diethyl ether, ethyl acetate, 1-butanol and water. The diethyl ether and ethyl acetate extracts were further studied for their chemical composition. So far, secondary metabolites which belong to phenolics were isolated by using several chromatographic methods (C.C. and HPLC) and were identified by using spectroscopic methods (UV/Vis, NMR and MS): two phenolic acids (p-hydroxy-benzoic acid and 3-methoxy-4-hydroxy-benzoic acid (vanillic acid)), and five flavonoids, which are derivatives of flavonol: kaempferol 3-O-β-D-glucopyranoside (astragalin), kaempferol 3-O-(6′′-α-L-rhamnopyranosyl)-β-D-glucopyranoside (nicotiflorin), quercetin 3-O-β-D-glucopyranoside (isoquercetin), isorhamnetin-3-O-β-D-glucopyranoside, and isoramnetin 3-O-(6′′-α-L-rhamnopyranosyl)-β-D-glucopyranoside (narcissin).

Keywords: Alyssum, chemical substitutes, flavonoids, phenolic acids

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2918 Technological Characterization of Lactic Acid Bacteria Isolated from Algerian's Goat's Milk

Authors: A. Cheriguene, F. Chougrani

Abstract:

A total of 153 wild lactic acid bacteria were isolated from goat’s milk collected from different areas in Western Algeria. The strains were identified using phenotypical, biochemical and physiological properties. API system and SDS-PAGE technique was also used in identification of the strains. Six genera were found Enterococcus (41.83%), Lactobacillus (29.40%), Lactococcus (19.60%), Leuconostoc (4.57%), Streptococcus thermophilus (3.26%) and Pediococcus (1.30%). The most abundant species were Enterococcus faecium (24 isolates), Enterococcus durans (22 isolates), Lactococcus lactis subsp. lactis (25 isolates), Lactobacillus rhamnosus (09 isolates) and Lactobacillus delbrueckii subsp. bulgaricus (07 isolates). The strains were screened for production and technological properties such as acid production, aminopeptidase activity, autolytic properties, antimicrobial activity and exopolysaccharide production. In general most tested isolates showed a good biomass separation when collected by centrifugation; as for the production of the lactic acid, results revealed that our strains are weakly acidifying; nevertheless, lactococci showed a best acidifying activity compared to lactobacilli. Aminopeptidase activity was also weak in most strains; but, it was generally higher for lactobacilli compared to lactococci, where we recorded 30 units for Lactobacillus delbrueckii subsp. bulgaricus M14. Autolytic activity was generally higher for most strains, more particularly lactobacilli where we recorded values of 71.13% and 70% of autolysis rate respectively in Lactobacillus rhamnosus strains 9S10 and 9S7. Antimicrobial activity was detected in 50% of the isolates, particularly in lactobacilli where 80% of strains tested were able to inhibit the growth of other strains. Two strains could produce exopolysaccharides, E. faecium 8M6 and E. durans 7S8. Some strains were able to maintain two or three technological characteristics together.

Keywords: lactic acid bacteria, technological properties, acidification, aminopeptidase acivity (AP), autolysis

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2917 The Solvent Extraction of Uranium, Plutonium and Thorium from Aqueous Solution by 1-Hydroxyhexadecylidene-1,1-Diphosphonic Acid

Authors: M. Bouhoun Ali, A. Y. Badjah Hadj Ahmed, M. Attou, A. Elias, M. A. Didi

Abstract:

In this paper, the solvent extraction of uranium(VI), plutonium(IV) and thorium(IV) from aqueous solutions using 1-hydroxyhexadecylidene-1,1-diphosphonic acid (HHDPA) in treated kerosene has been investigated. The HHDPA was previously synthesized and characterized by FT-IR, 1H NMR, 31P NMR spectroscopy and elemental analysis. The effects contact time, initial pH, initial metal concentration, aqueous/organic phase ratio, extractant concentration and temperature on the extraction process have been studied. An empirical modelling was performed by using a 25 full factorial design, and regression equation for extraction metals was determined from the data. The conventional log-log analysis of the extraction data reveals that ratios of extractant to extracted U(VI), Pu(IV) and Th(IV) are 1:1, 1:2 and 1:2, respectively. Thermodynamic parameters showed that the extraction process was exothermic heat and spontaneous. The obtained optimal parameters were applied to real effluents containing uranium(VI), plutonium(IV) and thorium(IV) ions.

Keywords: solvent extraction, uranium, plutonium, thorium, 1-hydroxyhexadecylidene-1-1-diphosphonic acid, aqueous solution

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2916 Comparative Chromatographic Profiling of Wild and Cultivated Macrocybe Gigantea (Massee) Pegler & Lodge

Authors: Gagan Brar, Munruchi Kaur

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Macrocybe gigantea was collected from the wild, growing as pure white, fleshy, robust fruit bodies in caespitose clusters. Initially, the few ladies collecting these fruiting bodies for cooking revealed their edibility status, which was later confirmed through classical and molecular taxonomy. The culture of this potential wild edible taxa was raised with an aim of domesticating it. Various solid and liquid media were evaluated for their vegetative growth, in which Malt Extract Agar was found to be the best solid medium and Glucose Peptone medium as the best liquid medium. The effect of different temperatures as well as pH was also evaluated for the vegetative growth of M. gigantea, and it was found that it shows maximum vegetative growth at 30° and pH 5. For spawn preparation, various grains viz. Wheat grains, Jowar grains, Bajra grains and Maize grains were evaluated, and it was found that wheat grains boiled for 30 minutes gave the maximum mycelial growth. Mother spawn was thus prepared on wheat grains boiled for 30 minutes. For raising the fruiting bodies, different locally available agro-wastes were tried, and it was found that paddy straw gives the best growth. Both wilds as well as cultivated M. gigantea were compared through HPLC to evaluate the different nutritional and nutraceutical values. For the evaluation of different sugars in wild and cultivated M. gigantea, 15 sugars were taken for analysis. Among these Melezitose, Trehalose, Glucose, Xylose and Mannitol were found in the wild collection of M. gigantea; in the cultivated sample, Melezitose, Trehalose, Xylose and Dulcitol were detected. Among the 20 different amino acids, 18 amino acids were found, except Asparagine and Glutamine in both wild as well as cultivated samples. Among the 37 tested fatty acids, only 6 fatty acids, namely Palmitic acid, Stearic acid, Cis-9 Oleic acid, Linoleic acid, Gamma-Linolenic acid and Tricosanoic acid, were found in both wild and cultivated samples, although the concentration of these fatty acids was more in the cultivated sample. From the various vitamins tested, Vitamin C, D and E were present in both wild and cultivated samples. Both wild as well as cultivated samples were evaluated for the presence of phenols; for this purpose, eleven phenols were taken as standards in HPLC analysis, and it was found that Gallic acid, Resorcinol, Ferulic acid and Pyrogallol were present in the wild mushroom sample whereas in the cultivated sample Ferulic acid, Caffeic Acid, Vanillic acid and Vanillin are present. The flavonoid analysis revealed the presence of Rutin, Naringin and Quercetin in wild M. gigantea, while 5 Naringin, Catechol, Myrecetin, Gossypin and Quercetin were found in cultivated one. From the comparative chromatographic profiling of both wild as well as cultivated M. gigantea, it is concluded that no nutrient loss was found during its cultivation. An increase in percentage of secondary metabolites (i.e., phenols and flavonoids) was found in cultivated one as compared to wild M. gigantea. Thus, from future perspective cultivated species of M. gigantea can be recommended for the commercial purpose as a good food supplement.

Keywords: culture, edible, fruit bodies, wild

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2915 Poly(L-Lactic Acid) Scaffolds for Bone Tissue Engineering

Authors: Aleksandra BužArovska, Gordana Bogoeva Gaceva

Abstract:

Biodegradable polymers have received significant scientific attention in tissue engineering (TE) application, in particular their composites consisting of inorganic nanoparticles. In the last 15 years, they are subject of intensive research by many groups, aiming to develop polymer scaffolds with defined biodegradability, porosity and adequate mechanical stability. The most important characteristic making these materials attractive for TE is their biodegradability, a process that could be time controlled and long enough to enable generation of a new tissue as a replacement for the degraded polymer scaffold. In this work poly(L-lactic acid) scaffolds, filled with TiO2 nanoparticles functionalized with oleic acid, have been prepared by thermally induced phase separation method (TIPS). The functionalization of TiO2 nanoparticles with oleic acid was performed in order to improve the nanoparticles dispersibility within the polymer matrix and at the same time to inhibit the cytotoxicity of the nanofiller. The oleic acid was chosen as amphiphilic molecule belonging to the fatty acid family because of its non-toxicity and possibility for mediation between the hydrophilic TiO2 nanoparticles and hydrophobic PLA matrix. The produced scaffolds were characterized with thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), scanning electron microscopy (SEM) and mechanical compression measurements. The bioactivity for bone tissue engineering application was tested in supersaturated simulated body fluid. The degradation process was followed by Fourier transform infrared spectroscopy (FTIR). The results showed anisotropic morphology with elongated open pores (100 µm), high porosity (around 92%) and perfectly dispersed nanofiller. The compression moduli up to 10 MPa were identified independent on the nanofiller content. Functionalized TiO2 nanoparticles induced formation of hydroxyapatite clusters as much as unfunctionalized TiO2. The prepared scaffolds showed properties ideal for scaffold vascularization, cell attachment, growth and proliferation.

Keywords: biodegradation, bone tissue engineering, mineralization, PLA scaffolds

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2914 Antioxidant Properties, Ascorbic Acid and Total Carotenoids Values of Sweet and Hot Red Pepper Paste: A Traditional Food in Turkish Diet

Authors: Kubra Sayin, Derya Arslan

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Red pepper (Capsicum annum L.) has long been recognized as a good source of antioxidants, being rich in ascorbic acid and other phytochemicals. In Turkish cuisine red pepper is sometimes consumed raw in salads and baked as a garnish, but its most wide consumption type is red pepper paste. The processing of red pepper into pepper paste includes various thermal treatment steps such as heating and pasteurizing. There are reports demonstrating an enhancement or reduction in antioxidant activity of vegetables after thermal treatment. So this study was conducted to investigate the total phenolics, ascorbic acid and total carotenoids as well as free radical scavenging activity of raw red pepper and various red pepper pastes obtainable on the market. The samples were analyzed for radical-scavenging activity (RSA) and total polyphenol (TP) content using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and Folin-Ciocalteu methods, respectively. They were also evaluated for ascorbic acid content (AsA) by HPLC. Total carotenoids content was determined spectrophotometrically. Results suggest that there is no significant (P > 0.05) difference in RSA, TP, AsA and total carotenoids content between various red pepper paste products. However, red pepper paste showed marked differences (P < 0.05) in the RSA, TP and AsA contents compared with raw red pepper. It is concluded that the red pepper paste, that has a wide range of consumption in Turkish cuisine, presents a good dose of phenolic compounds and antioxidant capacity and it should be regarded as a functional food.

Keywords: red pepper paste, antioxidant properties, total carotenoids, total phenolics

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2913 In vitro Analysis of the Effect of Supplementation Oils on Conjugated Linoleic Acid Production by Butyvibrio Fibrisolvense

Authors: B. D. Ravindra, A. K. Tyagi, C. Kathirvelan

Abstract:

Some micronutrients in food (milk and meat), called ‘functional food components’ exert beneficial effects other than their routine nutrient function and conjugated linoleic acid (CLA) is an unsaturated fatty acid of ruminant origin, an example of this category. However, recently the fear of hypercholesterolemia due to saturated fats has led to the avoidance of dietary fat especially of animal origin despite its advantages such as lowering blood cholesterol, immuno-modulation and anticarcinogenic property due to the presence of CLA. The dietary increase of linoleic acid (LA) and linolenic acid (LNA) is one of the feeding strategies for increasing the CLA concentration in milk. Butyrivibrio fibrisolvens is the one potential rumen bacteria, which has high potential to isomerize LA to CLA. The study was conducted to screen the different oils for CLA production, selected based on their LA concentration. Butyrivibrio fibrisolvens culture (strain 49, MZ3, 30/10) were isolated from the rumen liquor of fistulated Buffalo (age ≈ 3 years; weight ≈ 250 kg) were used in in-vitro experiments, further work was carried out with three oils viz., sunflower, mustard and soybean oil at different concentration (0.05, 0.1, 0.15, 0.2, 0.25 and 0.3 g/L of media) to study the growth of bacteria and CLA production at different incubation period (0, 8, 12, 18, 24, 48, 72 h). In the present study, growth of the bacteria was decreased linearly with increase in concentration of three oils. However, highest decrease in growth was recorded at the concentration of 0.30 g of three oils per litre of the media. Highest CLA production was 51.96, 42.08 and 25.60 µg/ml at 0.25 g and it decreased to 48.19, 39.35 and 23.41 µg/ml at 0.3 g supplementation of sunflower, soybean, and mustard oil per litre of the media, respectively at 18 h incubation period. The present study indicates the Butyrivibrio fibrisolvens bacteria involves in the biohydrogenation process, and LA rich sunflower meal can be used to improve the CLA production in rumen and thereby increasing the CLA concentration of milk.

Keywords: Butyrivibrio fibrisolvens, CLA, fatty acids, sunflower oil

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2912 Anti-Aging Effects of Retinol and Alpha Hydroxy Acid on Elastin Fibers of Artificially Photo-Aged Human Dermal Fibroblast Cell Lines

Authors: Mohammed Jarrar, Shalini Behl, Nadia Shaheen, Abeer Fatima, Reem Nasab

Abstract:

Skin aging is a slow multifactorial process influenced by both internal as well as external factors. Ultra-violet radiations (UV), diet, smoking and personal habits are the most common environmental factors that affect skin aging. Fat contents and fibrous proteins as collagen and elastin are core internal structural components. The direct influence of UV on elastin integrity and health is crucial on aging of skin by time. The deposition of abnormal elastic material is a major marker in a photo-aged skin. Searching for compounds that may protect against cutaneous photo-damage is highly valued. Retinoids and Alpha Hydroxy Acids protective and or repairing effects of UV have been endorsed by some researchers. For consolidating a better understanding of anti and protective effects of such anti-aging agents, we evaluated the combinatory effects of various dosages of lactic acid and retinol on the dermal fibroblasts elastin levels exposed to UV. The UV exposed cells showed significant reduction in the elastin levels. A combination of drugs with a higher concentration of lactic acid (30-35 mM) and a lower concentration of retinol (10-15mg/mL) showed to work better in enhancing elastin concentration in UV exposed cells. We assume this enhancement could be the result of increased tropo-elastin gene expression stimulated by retinol and lactic acid probably repaired the UV irradiated damage by enhancing the amount and integrity of the elastin fibers.

Keywords: alpha hydroxy acid, elastin, retinol, ultraviolet radiations

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2911 Studies of Carbohydrate, Antioxidant, Nutrient and Genomic DNA Characterization of Fresh Olive Treated with Alkaline and Acidic Solvent: An Innovation

Authors: A. B. M. S. Hossain, A. Abdelgadir, N. A. Ibrahim

Abstract:

Fresh ripen olive cannot be consumed immediately after harvest due to the excessive bitterness having polyphenol as antioxidant. Industrial processing needs to be edible the fruit. The laboratory processing technique has been used to make it edible by using acid (vinegar, 5% acetic acid) and alkaline solvent (NaOH). Based on the treatment and consequence, innovative data have been found in this regard. The experiment was conducted to investigate biochemical content, nutritional and DNA characterization of olive fruit treated with alkaline (Sodium chloride anhydrous) and acidic solvent (5% acetic acid, vinegar). The treatments were used as control (no water), water control, 10% sodium chloride anhydrous (NaOH), vinegar (5% acetic acid), vinegar + NaOH and vinegar + NaOH + hot water treatment. Our results showed that inverted sugar and glucose content were higher in the vinegar and NaOH treated olive than in other treatments. Fructose content was the highest in vinegar + NaOH treated fruit. Nutrient contents NO3 K, Ca and Na were found higher in the treated fruit than the control fruit. Moreover, maximum K content was observed in the case of all treatments compared to the other nutrient content. The highest acidic (lower pH) condition (sour) was found in treated fruit. DNA yield was found higher in water control than acid and alkaline treated olives. DNA band was wider in the olive treated water control compared to the NaOH, vinegar, vinegar + NaOH and vinegar + NaOH + Hot water treatment. Finally, results suggest that vinegar + NaOH treated olive fruit was the best for fresh olive homemade processing after harvesting for edible purpose.

Keywords: olive, vinegar, sugars, DNA band, bioprocess biotechnology

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2910 Synthesis and Anti-Inflammatory Activity of Pyrazol-3-yl Thiazole 4-Carboxylic Acid Derivatives Targeting Enzyme in the Leukotriene Pathway

Authors: Shweta Sinha, Mukesh Doble, Manju S. L.

Abstract:

Pyrazole scaffold is an important group of compound in heterocyclic chemistry and is found to possess numerous uses in chemistry. Pyrazole derivatives are also known to possess important biological activities including antitumor, antimicrobial, antiviral, antifungal, anticancer and anti-inflammatory. Inflammation is associated with pain, allergy and asthma. Leukotrienes are mediators of various inflammatory and allergic disorders. 5-Lipoxygenase (5-LOX) is an important enzyme involved in the biosynthesis of leukotrienes and metabolism of arachidonic acid (AA) and thus targeted for anti-inflammation. In vitro inhibitory activity of pyrazol-3-yl thiazole 4-carboxylic acid derivatives is tested against enzyme 5-LOX. Most of these compounds exhibit good inhibitory activity against this enzyme. Binding mode study of these compounds is determined by computational tool. Further experiments are being done to understand the mechanism of action of these compounds in inhibiting this enzyme. To conclude, these compounds appear to be a promising target in drug design against 5-LOX.

Keywords: inflammation, inhibition, 5-lipoxygenase, pyrazole

Procedia PDF Downloads 245
2909 Kinetics of Phytochemicals and Antioxidant Activity during Thermal Treatment of Cape Gooseberry (Physalis peruviana L)

Authors: Mary-Luz Olivares-Tenorio, Ruud Verkerk, Matthijs Dekker, Martinus A. J. S. van Boekel

Abstract:

Cape gooseberry, the fruit of the plant Physalis peruviana L. has gained interest in research given its contents of promising health-promoting compounds like contents. The presence of carotenoids, ascorbic acid, minerals, polyphenols, vitamins and antioxidants. This project aims to study thermal stability of β-carotene, ascorbic acid, catechin and epicatechin and antioxidant activity in the matrix of the Cape Gooseberry. Fruits were obtained from a Colombian field in Cundinamarca. Ripeness stage was 4 (According to NTC 4580, corresponding to mature stage) at the moment of the experiment. The fruits have been subjected to temperatures of 40, 60, 80, 100 and 120°C for several times. β-Carotene, ascorbic acid, catechin and epicatechin content were assessed with HPLC and antioxidant activity with the DPPH method. β-Carotene was stable upon 100°C, and showed some degradation at 120°C. The same behavior was observed for epicatechin. Catechin increased during treatment at 40°C, at 60°C it remained stable and it showed degradation at 80°C, 100°C and 120°C that could be described by a second order kinetic model. Ascorbic acid was the most heat-sensitive of the analyzed compounds. It showed degradation at all studied temperatures, and could be described by a first order model. The activation energy for ascorbic acid degradation in cape gooseberry was 46.0 kJ/mol and its degradation rate coefficient at 100 °C was 6.53 x 10-3 s-1. The antioxidant activity declined for all studied temperatures. Results from this study showed that cape gooseberry is an important source of different health-promoting compounds and some of them are stable to heat. That makes this fruit a suitable raw material for processed products such as jam, juices and dehydrated fruit, giving the consumer a good intake of these compounds.

Keywords: goldenberry, health-promoting compounds, phytochemical, processing, heat treatment

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2908 Cold Plasma Surface Modified Electrospun Microtube Array Membrane for Chitosan Immobilization and Their Properties

Authors: Ko-Shao Chen, Yun Tsao, Chia-Hsuan Tsen, Chien-Chung Chen, Shu-Chuan Liao

Abstract:

Electrospun microtube array membranes (MTAMs) made of PLLA (poly-L-lactic acid) have wide potential applications in tissue engineering. However, their surface hydrophobicity and poor biocompatability have limited their further usage. In this study, the surface of PLLA MTAMs were made hydrophilic by introducing extra functional groups, such as peroxide, via an acetic acid plasma (AAP). UV-graft polymerization of acrylic acid (G-AAc) was then used to produce carboxyl group on MTAMs surface, which bonded covalently with chitosan through EDC / NHS crosslinking agents. To evaluate the effects of the surface modification on PLLA MTAMs, water contact angle (WCA) measurement and cell compatibility tests were carried out. We found that AAP treated electrospun PLLA MTAMs grafted with AAc and, finally, with chitosan immobilized via crosslinking agent, exhibited improved hydrophilic and cell compatibility.

Keywords: plasma, EDC/NHS, UV grafting, Chitosan, microtube array membrane (MTAMs)

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2907 Fabrication and Characterization of Dissolvable Microneedle Patches Using Different Compositions and Ratios of Hyaluronic Acid and Zinc Oxide Nanoparticles

Authors: Dada Kolawole Segun

Abstract:

Transdermal drug delivery has gained popularity as a non-invasive method for controlled drug release compared to traditional delivery routes. Dissolvable transdermal patches have emerged as a promising platform for delivering a variety of drugs due to their ease of use. The objective of this research was to create and characterize dissolvable transdermal patches using various compositions and ratios of hyaluronic acid and zinc oxide nanoparticles. A micromolding technique was utilized to fabricate the patches, which were subsequently characterized using scanning electron microscopy, atomic force microscopy, and tensile strength testing. In vitro drug release studies were conducted to evaluate the drug release kinetics of the patches. The study found that the mechanical strength and dissolution properties of the patches were influenced by the hyaluronic acid and zinc oxide nanoparticle ratios used in the fabrication process. Moreover, the patches demonstrated controlled delivery of model drugs through the skin, highlighting their potential for transdermal drug delivery applications. The results suggest that dissolvable transdermal patches can be tailored to meet specific requirements for drug delivery applications using different compositions and ratios of hyaluronic acid and zinc oxide nanoparticles. This development has the potential to improve treatment outcomes and patient compliance in various therapeutic areas.

Keywords: transdermal drug delivery, characterization, skin permeation, biodegradable materials

Procedia PDF Downloads 91
2906 The Production of B-Group Vitamin by Lactic Acid Bacteria and Its Importance in Food Industry

Authors: Goksen Arik, Mihriban Korukluoglu

Abstract:

Lactic acid bacteria (LAB) has been used commonly in the food industry. They can be used as natural preservatives because acidifying carried out in the medium can protect the last product against microbial spoilage. Besides, other metabolites produced by LAB during fermentation period have also an antimicrobial effect on pathogen and spoilage microorganisms in the food industry. LAB are responsible for the desirable and distinctive aroma and flavour which are observed in fermented food products such as pickle, kefir, yogurt, and cheese. Various LAB strains are able to produce B-group vitamins such as folate (B11), riboflavin (B2) and cobalamin (B12). Especially wild-type strains of LAB can produce B-group vitamins in high concentrations. These cultures may be used in food industry as a starter culture and also the microbial strains can be used in encapsulation technology for new and functional food product development. This review is based on the current applications of B-group vitamin producing LAB. Furthermore, the new technologies and innovative researches about B vitamin production in LAB have been demonstrated and discussed for determining their usage availability in various area in the food industry.

Keywords: B vitamin, food industry, lactic acid bacteria, starter culture, technology

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2905 Characterization of (GRAS37) Gibberellin Acid Insensitive (GAI), Repressor (RGA), and Scarecrow (SCR) Gene by Using Bioinformatics Tools

Authors: Yusra Tariq

Abstract:

The Grass 37 gene is presently known in tomatoes, which are the source of healthy substances such as ascorbic acid, polyphenols, carotenoids and nutrients. It has a significant impact on the growth and development of humans. The GRASS 37 gene is a plant Transcription factor group assuming significant parts in various reactions of different Abiotic stresses such as (drought, salinity, thermal stresses, temperature, and bright waves) which could highly affect the growth. Tomatoes are very sensitive to temperature, and their growth or production occurs optimally in a temperature range from 21 C to 29.5 C during the daytime and from 18.5 C to 21 C during the night. This protein acts as a positive regulator of salt stress response and abscisic acid signaling. This study summarizes the structure characterized by molecular formula and protein-binding domains by different bioinformatics tools such as Expasy translate tool, Expasy Portparam, Swiss Prot and Inter Pro Scan, Clustal W tool regulatory procedure of GRASS gene components, also their reactions to both biotic and Abiotic stresses.

Keywords: GRAS37, gene, bioinformatics, tool

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2904 A Mathematical Investigation of the Turkevich Organizer Theory in the Citrate Method for the Synthesis of Gold Nanoparticles

Authors: Emmanuel Agunloye, Asterios Gavriilidis, Luca Mazzei

Abstract:

Gold nanoparticles are commonly synthesized by reducing chloroauric acid with sodium citrate. This method, referred to as the citrate method, can produce spherical gold nanoparticles (NPs) in the size range 10-150 nm. Gold NPs of this size are useful in many applications. However, the NPs are usually polydisperse and irreproducible. A better understanding of the synthesis mechanisms is thus required. This work thoroughly investigated the only model that describes the synthesis. This model combines mass and population balance equations, describing the NPs synthesis through a sequence of chemical reactions. Chloroauric acid reacts with sodium citrate to form aurous chloride and dicarboxy acetone. The latter organizes aurous chloride in a nucleation step and concurrently degrades into acetone. The unconsumed precursor then grows the formed nuclei. However, depending on the pH, both the precursor and the reducing agent react differently thus affecting the synthesis. In this work, we investigated the model for different conditions of pH, temperature and initial reactant concentrations. To solve the model, we used Parsival, a commercial numerical code, whilst to test it, we considered various conditions studied experimentally by different researchers, for which results are available in the literature. The model poorly predicted the experimental data. We believe that this is because the model does not account for the acid-base properties of both chloroauric acid and sodium citrate.

Keywords: citrate method, gold nanoparticles, Parsival, population balance equations, Turkevich organizer theory

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2903 Modifiable Poly Methacrylic Acid-Co-Acrylonitrile Microgels Fabricated with Cu and Co Nanoparticles for Simultaneous Catalytic Reduction of Multiple Compounds

Authors: Muhammad Ajmal, Muhammad Siddiq, Nurettin Sahiner

Abstract:

We prepared poly(methacrylic acid-co-acrylonitrile) (p(MAc-co-AN)) microgels by inverse suspension polymerization, and converted the nitrile groups into amidoxime groups to obtain more hydrophilic amidoximated poly(methacrylic acid-co-acrylonitile) (amid-p(MAc-co-AN)) microgels. Amid-microgels were used as microreactors for in situ synthesis of copper and cobalt nanoparticles. Cu (II) and Co (II) ions were loaded into microgels from their aqueous metal salt solutions and then converted to corresponding metal nanoparticle (MNP) by treating the loaded metal ions with sodium borohydride (NaBH4). The characterization of the prepared microgels and microgel metal nanoparticle composites was carried out by SEM, TEM and TG analysis. The amounts of metal nanoparticles within microgels were estimated by AAS measurements by dissolving the MNP entrapped within microgels by concentrated HCl acid treatment. Catalytic performances of the prepared amid-p(MAc-co-AN)-M (M: Cu, Co) microgel composites were investigated by using them as catalyst for the degradation of cationic and anionic organic dyes such as eosin Y (EY), methylene blue (MB) and methyl Orange (MO), and for the reduction of nitro aromatic pollutants like 2-nitrophenol (2-NP) and 4-nitrophenol (4-NP) to their corresponding amino phenols. Here, we also report for the first time, the simultaneous degradation/reduction of MB, EY, and 4-NP by amid-p(MAc-co-AN)-Cu microgel composites. Different parameters affecting the reduction rates such as metal types, amount of catalysts, temperature and the amount of reducing agent were investigated.

Keywords: microgels, nanoparticles, catalyst, pollutants

Procedia PDF Downloads 358
2902 Improving Dyeability of Cotton Fabric with Juglans regia L. Natural Dyestuff

Authors: M. Heysem Arslan, Ikilem Gocek, U. Kivanc Sahin

Abstract:

Natural dyestuff, extracted from Juglans Regia L., a kind of walnut, was used to dye 100% cotton gabardine fabric. The main goal of this study was to enhance dyeing process of cotton fabric with Juglans Regia L. dyestuff in terms of color fastness values by designing and developing a mordant application process. Within the context of this study, different mordants such as tannic acid, gallic acid, ascorbic acid, potassium sodium tartrate tetrahydrate, calcium carbonate, iron (II) sulphate heptahydrate, aluminum potassium sulphate dodecahydrate and their combinations were applied in the mordanting processes. Spectrophotometric analysis, color fastness to washing and color fastness to light tests were carried out on the fabric samples. In this study, it was shown that by using the right combination of mordants with a proper application process, it is possible to improve color fastness values of cotton fabric samples dyed with natural dyestuff.

Keywords: extraction, Juglans Regia L., mordanting process, natural dyestuff

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2901 A Nucleic Acid Extraction Method for High-Viscosity Floricultural Samples

Authors: Harunori Kawabe, Hideyuki Aoshima, Koji Murakami, Minoru Kawakami, Yuka Nakano, David D. Ordinario, C. W. Crawford, Iri Sato-Baran

Abstract:

With the recent advances in gene editing technologies allowing the rewriting of genetic sequences, additional market growth in the global floriculture market beyond previous trends is anticipated through increasingly sophisticated plant breeding techniques. As a prerequisite for gene editing, the gene sequence of the target plant must first be identified. This necessitates the genetic analysis of plants with unknown gene sequences, the extraction of RNA, and comprehensive expression analysis. Consequently, a technology capable of consistently and effectively extracting high-purity DNA and RNA from plants is of paramount importance. Although model plants, such as Arabidopsis and tobacco, have established methods for DNA and RNA extraction, floricultural species such as roses present unique challenges. Different techniques to extract DNA and RNA from various floricultural species were investigated. Upon sampling and grinding the petals of several floricultural species, it was observed that nucleic acid extraction from the ground petal solutions of low viscosity was straightforward; solutions of high viscosity presented a significant challenge. It is postulated that the presence of substantial quantities of polysaccharides and polyphenols in the plant tissue was responsible for the inhibition of nucleic acid extraction. Consequently, attempts were made to extract high-purity DNA and RNA by improving the CTAB method and combining it with commercially available nucleic acid extraction kits. The quality of the total extracted DNA and RNA was evaluated using standard methods. Finally, the effectiveness of the extraction method was assessed by determining whether it was possible to create a library that could be applied as a suitable template for a next-generation sequencer. In conclusion, a method was developed for consistent and accurate nucleic acid extraction from high-viscosity floricultural samples. These results demonstrate improved techniques for DNA and RNA extraction from flowers, help facilitate gene editing of floricultural species and expand the boundaries of research and commercial opportunities.

Keywords: floriculture, gene editing, next-generation sequencing, nucleic acid extraction

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