Search results for: high perforamance liquid chromatography
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 21448

Search results for: high perforamance liquid chromatography

21328 High Acid-Stable α-Amylase Production by Milk in Liquid Culture

Authors: Shohei Matsuo, Saki Mikai, Hiroshi Morita

Abstract:

Objectives: Shochu is a popular Japanese distilled spirits. In the production of shochu, the filamentous fungus Aspergillus kawachii has traditionally been used. A. kawachii produces two types of starch hydrolytic enzymes, α-amylase (enzymatic liquefaction) and glucoamylase (enzymatic saccharification). Liquid culture system is a relatively easy microorganism to ferment with relatively low cost of production compared for solid culture. In liquid culture system, acid-unstable α-amylase (α-A) was produced abundantly, but, acid-stable α-amylase (Aα-A) was not produced. Since there is high enzyme productivity, most in shochu brewing have been adopted by a solid culture method. In this study, therefore, we investigated production of Aα-A in liquid culture system. Materials and methods: Microorganism Aspergillus kawachii NBRC 4308 was used. The mold was cultured at 30 °C for 7~14 d to allow formation of conidiospores on slant agar medium. Liquid Culture System: A. kawachii was cultured in a 100 ml of following altered SLS medium: 1.0 g of rice flour, 0.1 g of K2HPO4, 0.1 g of KCl, 0.6 g of tryptone, 0.05 g of MgSO4・7H2O, 0.001 g of FeSO4・7H2O, 0.0003 g of ZnSO4・7H2O, 0.021 g of CaCl2, 0.33 of citric acid (pH 3.0). The pH of the medium was adjusted to the designated value with 10 % HCl solution. The cultivation was shaking at 30 °C and 200 rpm for 72 h. It was filtered to obtain a crude enzyme solution. Aα-A assay: The crude enzyme solution was analyzed. An acid-stable α-amylase activity was carried out using an α-amylase assay kit (Kikkoman Corporation, Noda, Japan). It was conducted after adding 9 ml of 100 mM acetate buffer (pH 3.0) to 1 ml of the culture product supernatant and acid treatment at 37°C for 1 h. One unit of a-amylase activity was defined as the amount of enzyme that yielded 1 mmol of 2-chloro-4-nitrophenyl 6-azide-6-deoxy-b-maltopentaoside (CNP) per minute. Results and Conclusion: We experimented with co-culture of A. kawachii and lactobacillus in order to get control of pH in altered SLS medium. However, high production of acid-stable α-amylase was not obtained. We experimented with yoghurt or milk made an addition to liquid culture. The result indicated that high production of acid-stable α-amylase (964 U/g-substrate) was obtained when milk made an addition to liquid culture. Phosphate concentration in the liquid medium was a major cause of increased acid-stable α-amylase activity. In liquid culture, acid-stable α-amylase activity was enhanced by milk, but Fats and oils in the milk were oxidized. In addition, Tryptone is not approved as a food additive in Japan. Thus, alter SLS medium added to skim milk excepting for the fats and oils in the milk instead of tryptone. The result indicated that high production of acid-stable α-amylase was obtained with the same effect as milk.

Keywords: acid-stable α-amylase, liquid culture, milk, shochu

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21327 One-Pot Synthesis of 5-Hydroxymethylfurfural from Hexose Sugar over Chromium Impregnated Zeolite Based Catalyst, Cr/H-ZSM-5

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

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21326 Electricity Production from Vermicompost Liquid Using Microbial Fuel Cell

Authors: Pratthana Ammaraphitak, Piyachon Ketsuwan, Rattapoom Prommana

Abstract:

Electricity production from vermicompost liquid was investigated in microbial fuel cells (MFCs). The aim of this study was to determine the performance of vermicompost liquid as a biocatalyst for electricity production by MFCs. Chemical and physical parameters of vermicompost liquid as total nitrogen, ammonia-nitrogen, nitrate, nitrite, total phosphorus, potassium, organic matter, C:N ratio, pH, and electrical conductivity in MFCs were studied. The performance of MFCs was operated in open circuit mode for 7 days. The maximum open circuit voltage (OCV) was 0.45 V. The maximum power density of 5.29 ± 0.75 W/m² corresponding to a current density of 0.024 2 ± 0.0017 A/m² was achieved by the 1000 Ω on day 2. Vermicompost liquid has efficiency to generate electricity from organic waste.

Keywords: vermicompost liquid, microbial fuel cell, nutrient, electricity production

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21325 Anti-Melanogenesis and Anti-Inflammatory Effects of Opuntia humifusa

Authors: Yonghwa Lee, Yoon Suk Kim, Yongsub Yi

Abstract:

This study was to confirm the effects of anti-melanogenesis and anti-inflammatory effects from Opuntia humifusa fruit and stem extracts. A potent anti-oxidant activity was shown from the leaf extract at IC50 value of 38.33±1.07 μg/mL and fruit extract at IC50 value of 40.23±2.21 μg/mL by 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay. Also, phenolic contents were confirmed total phenolic assay by high performance liquid chromatography (HPLC). Fraction of taxifolin from leaf extract was identified using HPLC and gas chromatography/mass spectrometry. The extracts of Opuntia humifusa fruit and stem were confirmed about toxicity effect in B16 F1 by cell viability. Melanin contents were decreased. Opuntia humifusa fruit and stem extracts had a positive effect of melanin synthesis inhibition for skin whitening. In investigating the anti-inflammatory activities of Opuntia humifusa, the results of cell viability indicated that taxifolin did not show cytotoxicity on RAW264.7 cells at 500 μM of concentration. The results show that taxifolin inhibited lipopolysaccharide (LPS)-induced production of Nitrite oxide (NO). In addition, taxifolin indicated the inhibition of lipopolysaccharide (LPS)-induced tumor necrosis factor (TNF) -α and interleukin (IL) -6 productions by cytokine assay and cyclooxygenase (COX)-2 expression by western blot analysis, meaning that taxifolin had a significant anti-inflammatory effect. Our results suggested that taxifolin from Opuntia humifusa has anti-melanogenesis and anti-inflammatory activities.

Keywords: anti-melanogenesis, anti-inflammatory, Opuntia humifusa, taxifolin

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21324 Phase Diagrams and Liquid-Liquid Extraction in Aqueous Biphasic Systems Formed by Polyethylene Glycol and Potassium Sodium Tartrate at 303.15 K

Authors: Amanda Cristina de Oliveira, Elias de Souza Monteiro Filho, Roberta Ceriani

Abstract:

Liquid-liquid extraction in aqueous two-phase systems (ATPSs) constitutes a powerful tool for purifying bio-materials, such as cells, organelles, proteins, among others. In this work, the extraction of the bovine serum albumin (BSA) has been studied in systems formed by polyethylene glycol (PEG) (1500, 4000, and 6000 g.mol⁻¹) + potassium sodium tartrate + water at 303.15°K. Phase diagrams were obtained by turbidimetry and Merchuk’s method (1998). The experimental tie-lines were described using the Othmer-Tobias and Bancroft correlations. ATPSs were correlated with the nonrandom two-liquid (NRTL) model. The results were considered excellent according to global root-mean-square deviations found which were between 0,72 and 1,13%. The concentrations of the proteins in each phase were determined by spectrophotometry at 280 nm, finding partition efficiencies greater than 71%.

Keywords: aqueous two phases systems, bovine serum albumin , liquid-liquid extraction, polyethylene glycol

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21323 Giant Achievements in Food Processing

Authors: Farnaz Amidi Fazli

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After long period of human experience about food processing from raw eating to canning of food in the last century now it is time to use novel technologies which are sometimes completely different from common technologies. It is possible to decontaminate food without using heat or the foods are stored without using cold chain. Pulsed electric field (PEF) processing is a non-thermal method of food preservation that uses short bursts of electricity, PEF can be used for processing liquid and semi-liquid food products. PEF processing offers high quality fresh-like liquid foods with excellent flavor, nutritional value, and shelf-life. High pressure processing (HPP) technology has the potential to fulfill both consumer and scientific requirements. The use of HPP for over 50 years has found applications in non-food industries. For food applications, ‘high pressure’ can be generally considered to be up to 600 MPa for most food products. After years, freezing has its high potential to food preservation due to new and quick freezing methods. Foods which are prepared by this technology have more acceptability and high quality comparing with old fashion slow freezing. Thus, quick freezing has further been adopted as a widespread commercial method for long-term preservation of perishable foods which improved both the health and convenience of everyone in the industrialised countries. Above parameters are achieved by Fluidised-bed freezing systems, freezing by immersion and Hydrofluidisation on the other hand new thawing methods like high-pressure, microwave, ohmic, and acoustic thawing have a key role in quality and adaptability of final product.

Keywords: quick freezing, thawing, high pressure, pulse electric, hydrofluidisation

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21322 Thermodynamic Modelling of Liquid-Liquid Equilibria (LLE) in the Separation of p-Cresol from the Coal Tar by Solvent Extraction

Authors: D. S. Fardhyanti, Megawati, W. B. Sediawan

Abstract:

Coal tar is a liquid by-product of the process of coal gasification and carbonation. This liquid oil mixture contains various kinds of useful compounds such as aromatic compounds and phenolic compounds. These compounds are widely used as raw material for insecticides, dyes, medicines, perfumes, coloring matters, and many others. This research investigates thermodynamic modelling of liquid-liquid equilibria (LLE) in the separation of phenol from the coal tar by solvent extraction. The equilibria are modeled by ternary components of Wohl, Van Laar, and Three-Suffix Margules models. The values of the parameters involved are obtained by curve-fitting to the experimental data. Based on the comparison between calculated and experimental data, it turns out that among the three models studied, the Three-Suffix Margules seems to be the best to predict the LLE of p-Cresol mixtures for those system.

Keywords: coal tar, phenol, Wohl, Van Laar, Three-Suffix Margules

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21321 Investigating the Sloshing Characteristics of a Liquid by Using an Image Processing Method

Authors: Ufuk Tosun, Reza Aghazadeh, Mehmet Bülent Özer

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This study puts forward a method to analyze the sloshing characteristics of liquid in a tuned sloshing absorber system by using image processing tools. Tuned sloshing vibration absorbers have recently attracted researchers’ attention as a seismic load damper in constructions due to its practical and logistical convenience. The absorber is liquid which sloshes and applies a force in opposite phase to the motion of structure. Experimentally characterization of the sloshing behavior can be utilized as means of verifying the results of numerical analysis. It can also be used to identify the accuracy of assumptions related to the motion of the liquid. There are extensive theoretical and experimental studies in the literature related to the dynamical and structural behavior of tuned sloshing dampers. In most of these works there are efforts to estimate the sloshing behavior of the liquid such as free surface motion and total force applied by liquid to the wall of container. For these purposes the use of sensors such as load cells and ultrasonic sensors are prevalent in experimental works. Load cells are only capable of measuring the force and requires conducting tests both with and without liquid to obtain pure sloshing force. Ultrasonic level sensors give point-wise measurements and hence they are not applicable to measure the whole free surface motion. Furthermore, in the case of liquid splashing it may give incorrect data. In this work a method for evaluating the sloshing wave height by using camera records and image processing techniques is presented. In this method the motion of the liquid and its container, made of a transparent material, is recorded by a high speed camera which is aligned to the free surface of the liquid. The video captured by the camera is processed frame by frame by using MATLAB Image Processing toolbox. The process starts with cropping the desired region. By recognizing the regions containing liquid and eliminating noise and liquid splashing, the final picture depicting the free surface of liquid is achieved. This picture then is used to obtain the height of the liquid through the length of container. This process is verified by ultrasonic sensors that measured fluid height on the surface of liquid.

Keywords: fluid structure interaction, image processing, sloshing, tuned liquid damper

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21320 How Addictive Are They: Effects of E-Cigarette Vapor on Intracranial Self-Stimulation Compared to Nicotine Alone

Authors: Annika Skansberg

Abstract:

Electronic cigarettes (e-cigarettes) use vapor to deliver nicotine, have recently become popular, especially amongst adolescents. Because of this, the FDA has decided to regulate e-cigarettes, and therefore would like to determine the abuse liability of the products compared to traditional nicotine products. This will allow them to determine the impact of regulating them on public health and shape the decisions they make when creating new laws. This study assessed the abuse liability of Aroma E-juice Dark Honey Tobacco compared to nicotine using an animal model. This e-liquid contains minor alkaloids that may increase abuse liability compared to nicotine alone. The abuse liability of nicotine alone and e-juice liquid were compared in rats using intracranial self-stimulation (ICSS) thresholds. E-liquid had less aversive effects at high nicotine doses in the ICSS model, suggesting that the minor alkaloids in the e-liquid allow users to use higher doses without experiencing the negative effects felt when using high doses of nicotine alone. This finding could mean that e-cigarettes have a higher abuse liability than nicotine alone, but more research is needed before this can be concluded. These findings are useful in observing the abuse liability of e-cigarettes and will help inform the FDA while regulating these products.

Keywords: electronic cigarettes, intra-cranial self stimulation, abuse liability, anhedonia

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21319 Polyclonal IgG glycosylation in Patients with Pediatric Appendicitis

Authors: Dalma Dojcsák, Csaba Váradi, Flóra Farkas, Tamás Farkas, János Papp, Béla Viskolcz

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Background: Appendicitis is a common acute inflammatory condition in both children and adults, but current laboratory markers such as C-reactive protein (CRP), white blood cell count (WBC), absolute neutrophil count (ANC), and red blood cell count (RNC) lack specificity in detecting appendicitis-related inflammation. N-glycosylation, an asparagine-linked glycosylation process, plays a vital role in cellular interactions, angiogenesis, immune response, and effector functions. Altered N-glycosylation impacts tumor growth and both acute and chronic inflammatory processes. IgG, the second most abundant glycoprotein in serum, shows altered glycosylation patterns during inflammation, suggesting that IgG glycan modifications may serve as potential biomarkers for appendicitis. Specifically, increased levels of agalactosylated IgG glycans are a known feature of various inflammatory conditions, potentially including appendicitis. Identifying pediatric appendicitis remains challenging due to the absence of specific biomarkers, which makes diagnosis reliant on clinical symptoms, imaging such as ultrasound, and nonspecific lab indicators (e.g., CRP, WBC, ANC). In this study, we analyzed the IgG derived N-glycome in pediatric patients with appendicitis compared with healthy controls. Methodology: The N-glycome was analyzed by high-performance liquid-chromatography combined with mass spectrometry. IgG was isolated from serum samples by Protein G column. The IgG derived glycans were released by enzymatic deglycosylation and fluorescent tags were attached to each glycan moiety, which made necessitates the sample clean-up for further reliable quantitation. Overall, 38 controls and 40 serum samples diagnosed with pediatric appendicitis were analyzed by HILIC-MS methods. Multivariate statistical tests were performed with area percentage under the peak data derived from the integrated peaks, which were obtained from the chromatograms. Conclusions: Our results represented the altered N-glycome of IgG in pediatric appendicitis is similar with other observations. The glycosylation pattern reported so far for IgG is characterized by decreased galactosylation and sialylation, and an increase in fucosylation.

Keywords: N-glycosylation, liquid chromatography, mass spectrometry, inflammation, appendicitis, immunoglobulin G

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21318 Waste Management in a Hot Laboratory of Japan Atomic Energy Agency – 2: Condensation and Solidification Experiments on Liquid Waste

Authors: Sou Watanabe, Hiromichi Ogi, Atsuhiro Shibata, Kazunori Nomura

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As a part of STRAD project conducted by JAEA, condensation of radioactive liquid waste containing various chemical compounds using reverse osmosis (RO) membrane filter was examined for efficient and safety treatment of the liquid wastes accumulated inside hot laboratories. NH4+ ion in the feed solution was successfully concentrated, and NH4+ ion involved in the effluents became lower than target value; 100 ppm. Solidification of simulated aqueous and organic liquid wastes was also tested. Those liquids were successfully solidified by adding cement or coagulants. Nevertheless, optimization in materials for confinement of chemicals is required for long time storage of the final solidified wastes.

Keywords: condensation, radioactive liquid waste, solidification, STRAD project

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21317 Detection of Some Drugs of Abuse from Fingerprints Using Liquid Chromatography-Mass Spectrometry

Authors: Ragaa T. Darwish, Maha A. Demellawy, Haidy M. Megahed, Doreen N. Younan, Wael S. Kholeif

Abstract:

The testing of drug abuse is authentic in order to affirm the misuse of drugs. Several analytical approaches have been developed for the detection of drugs of abuse in pharmaceutical and common biological samples, but few methodologies have been created to identify them from fingerprints. Liquid Chromatography-Mass Spectrometry (LC-MS) plays a major role in this field. The current study aimed at assessing the possibility of detection of some drugs of abuse (tramadol, clonazepam, and phenobarbital) from fingerprints using LC-MS in drug abusers. The aim was extended in order to assess the possibility of detection of the above-mentioned drugs in fingerprints of drug handlers till three days of handling the drugs. The study was conducted on randomly selected adult individuals who were either drug abusers seeking treatment at centers of drug dependence in Alexandria, Egypt or normal volunteers who were asked to handle the different studied drugs (drug handlers). An informed consent was obtained from all individuals. Participants were classified into 3 groups; control group that consisted of 50 normal individuals (neither abusing nor handling drugs), drug abuser group that consisted of 30 individuals who abused tramadol, clonazepam or phenobarbital (10 individuals for each drug) and drug handler group that consisted of 50 individuals who were touching either the powder of drugs of abuse: tramadol, clonazepam or phenobarbital (10 individuals for each drug) or the powder of the control substances which were of similar appearance (white powder) and that might be used in the adulteration of drugs of abuse: acetyl salicylic acid and acetaminophen (10 individuals for each drug). Samples were taken from the handler individuals for three consecutive days for the same individual. The diagnosis of drug abusers was based on the current Diagnostic and Statistical Manual of Mental disorders (DSM-V) and urine screening tests using immunoassay technique. Preliminary drug screening tests of urine samples were also done for drug handlers and the control groups to indicate the presence or absence of the studied drugs of abuse. Fingerprints of all participants were then taken on a filter paper previously soaked with methanol to be analyzed by LC-MS using SCIEX Triple Quad or QTRAP 5500 System. The concentration of drugs in each sample was calculated using the regression equations between concentration in ng/ml and peak area of each reference standard. All fingerprint samples from drug abusers showed positive results with LC-MS for the tested drugs, while all samples from the control individuals showed negative results. A significant difference was noted between the concentration of the drugs and the duration of abuse. Tramadol, clonazepam, and phenobarbital were also successfully detected from fingerprints of drug handlers till 3 days of handling the drugs. The mean concentration of the chosen drugs of abuse among the handlers group decreased when the days of samples intake increased.

Keywords: drugs of abuse, fingerprints, liquid chromatography–mass spectrometry, tramadol

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21316 Liquid Phase Catalytic Dehydrogenation of Secondary Alcohols to Ketone

Authors: Anıl Dinçer, Dilek Duranoğlu

Abstract:

Ketones, which are widely used as solvent and chemical intermediates in chemical process industry, are commercially produced by using catalytic dehydrogenation of secondary alcohols at higher temperature (300-500ºC), and pressure (1-5 bar). Although it is possible to obtain high conversion values (60-87%) via gas phase catalytic dehydrogenation, working high temperature and pressure can result in side reactions and shorten the catalyst life. In order to overcome these challenges, catalytic dehydrogenation in the presence of an appropriate liquid solvent has been started to use. Hence, secondary alcohols can be converted to respective ketones at relatively low temperature (150-200ºC) under atmospheric pressure. In this study, methyl ethyl ketone and acetone was produced via catalytic dehydrogenation of appropriate secondary alcohols (isopropyl alcohol and sec-butyl alcohol) in the presence of liquid solvent at 160-190ºC. Obtained methyl ethyl ketone and acetone were analyzed by using FTIR and GC spectrometer. Effects of temperature, amount of catalyst and solvent on conversion and reaction rate were investigated. Optimum process conditions, which gave high conversion and reaction rate, were determined. According to GC results, 70% of secondary butyl alcohol and 42% of isopropyl alcohol was converted to related ketone (methyl ethyl ketone and acetone, respectively) at optimum process conditions. After distillation, 99.13% methyl ethyl ketone and 99.20% acetone was obtained. Consequently, liquid phase dehydrogenation process, which can compete with commercial gas phase process, was developed.

Keywords: dehydrogenation, liquid phase, methyl ethyl ketone, secondary alcohol

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21315 Slug Initiation Evaluation in Long Horizontal Channels Experimentally

Authors: P. Adibi, M. R. Ansari, S. Jafari, B. Habibpour, E. Salimi

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In this paper, the effects of gas and liquid superficial inlet velocities and for the first time the effect of liquid holdup on slug initiation position are studied experimentally. Empirical correlations are also presented based on the obtained results. The tests are conducted for three liquid holdups in a long horizontal channel with dimensions of 5cmx10cm and 36m length. Usl and Usg rated as to 0.11m/s to 0.56m/s and 1.88m/s to 13m/s, respectively. The obtained results show that as αl=0.25, slug initiation position is increasing monotonically with Usl and Usg. During αl=0.50, slug initiation position is almost constant. For αl=0.75, slug initiation position is decreasing monotonically with Usl and Usg. In the case of equal void fraction of phases, generated slugs are weakly (low pressure). However, for the unequal void fraction of phases strong slugs (high pressure) are formed.

Keywords: liquid holdup, long horizontal channel, slug initiation position, superficial inlet velocity

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21314 The Effect of Heating-Liquid Nitrogen Cooling on Fracture Toughness of Anisotropic Rock

Authors: A. Kavandi, K. Goshtasbi, M. R. Hadei, H. Nejati

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In geothermal energy production, the method of liquid nitrogen (LN₂) fracturing in hot, dry rock is one of the most effective methods to increase the permeability of the reservoir. The geothermal reservoirs mainly consist of hard rocks such as granites and metamorphic rocks like gneiss with high temperatures. Gneiss, as a metamorphic rock, experiences a high level of inherent anisotropy. This type of anisotropy is considered as the nature of rocks, which affects the mechanical behavior of rocks. The aim of this study is to investigate the effects of heating-liquid nitrogen (LN₂) cooling treatment and rock anisotropy on the fracture toughness of gneiss. For this aim, a series of semi-circular bend (SCB) tests were carried out on specimens of gneiss with different anisotropy plane angles (0°, 30°, 60°, and 90°). In this study, gneiss specimens were exposed to heating–cooling treatment through gradual heating to 100°C followed by LN₂ cooling. Results indicate that the fracture toughness of treated samples is lower than that of untreated samples, and with increasing the anisotropy plane angle, the fracture toughness increases. The scanning electron microscope (SEM) technique is also implemented to evaluate the fracture process zone (FPZ) ahead of the crack tip.

Keywords: heating-cooling, anisotropic rock, fracture toughness, liquid nitrogen

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21313 Intensifying Approach for Separation of Bio-Butanol Using Ionic Liquid as Green Solvent: Moving Towards Sustainable Biorefinery

Authors: Kailas L. Wasewar

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Biobutanol has been considered as a potential and alternative biofuel relative to the most popular biodiesel and bioethanol. End product toxicity is the major problems in commercialization of fermentation based process which can be reduce to some possible extent by removing biobutanol simultaneously. Several techniques have been investigated for removing butanol from fermentation broth such as stripping, adsorption, liquid–liquid extraction, pervaporation, and membrane solvent extraction. Liquid–liquid extraction can be performed with high selectivity and is possible to carry out inside the fermenter. Conventional solvents have few drawbacks including toxicity, loss of solvent, high cost etc. Hence alternative solvents must be explored for the same. Room temperature ionic liquids (RTILs) composed entirely of ions are liquid at room temperature having negligible vapor pressure, non-flammability, and tunable physiochemical properties for a particular application which term them as “designer solvents”. Ionic liquids (ILs) have recently gained much attention as alternatives for organic solvents in many processes. In particular, ILs have been used as alternative solvents for liquid–liquid extraction. Their negligible vapor pressure allows the extracted products to be separated from ILs by conventional low pressure distillation with the potential for saving energy. Morpholinium, imidazolium, ammonium, phosphonium etc. based ionic liquids have been employed for the separation biobutanol. In present chapter, basic concepts of ionic liquids and application in separation have been presented. Further, type of ionic liquids including, conventional, functionalized, polymeric, supported membrane, and other ionic liquids have been explored. Also the effect of various performance parameters on separation of biobutanol by ionic liquids have been discussed and compared for different cation and anion based ionic liquids. The typical methodology for investigation have been adopted such as contacting the equal amount of biobutanol and ionic liquids for a specific time say, 30 minutes to confirm the equilibrium. Further, biobutanol phase were analyzed using GC to know the concentration of biobutanol and material balance were used to find the concentration in ionic liquid.

Keywords: biobutanol, separation, ionic liquids, sustainability, biorefinery, waste biomass

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21312 Thermo-Physical and Morphological Properties of Pdlcs Films Doped with Tio2 Nanoparticles.

Authors: Salima Bouadjela, Fatima Zohra Abdoune, Lahcene Mechernene

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PDLCs are currently considered as promising materials for specific applications such as creation of window blinds controlled by electric field, fog simulators, UV protective glasses, high data storage device etc. We know that the electrical field inside the liquid crystal is low compare with the external electric field [1,2]. An addition of high magnetic and electrical, properties containing compounds to the polymer dispersed liquid crystal (PDLC) will enhance the electrical, optical, and magnetic properties of the PDLC [3,4]. Low Concentration of inorganic nanoparticles TiO2 added to nematic liquid crystals (E7) and also combined with monomers (TPGDA) and cured monomer/LC mixture to elaborate polymer-LC-NP dispersion. The presence of liquid crystal and nanoparticles in TPGDA matrix were conformed and the modified properties of PDLC due to doped nanoparticle were studied and explained by the results of FTIR, POM, UV. Incorporation of nanoparticles modifies the structure of PDLC and thus it makes increase the amount of droplets and decrease in droplet size. we found that the presence of TiO2 nanoparticles leads to a shift the nematic-isotropic transition temperature TNI.

Keywords: nanocomposites, PDLC, phases diagram, TiO2

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21311 Behavior of an Elevated Liquid Storage Tank under Near-Fault Earthquakes

Authors: Koushik Roy, Sourav Gur, Sudib K. Mishra

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Evidence of pulse type features in near-fault ground motions has raised serious concern to the structural engineering community, in view of their possible implications on the behavior of structures located on the fault regions. Studies in the recent past explore the effects of pulse type ground motion on the special structures, such as transmission towers in view of their high flexibility. Identically, long period sloshing of liquid in the storage tanks under dynamic loading might increase their failure vulnerability under near-fault pulses. Therefore, the behavior of the elevated liquid storage tank is taken up in this study. Simple lumped mass model is considered, with the bilinear force-deformation hysteresis behavior. Set of near-fault seismic ground acceleration time histories are adopted for this purpose, along with the far-field records for comparison. It has been demonstrated that pulse type motions lead to significant increase of the responses; in particular, sloshing of the fluid mass could be as high as 5 times, then the far field counterpart. For identical storage capacity, slender tanks are found to be more vulnerable than the broad ones.

Keywords: far-field motion, hysteresis, liquid storage tank, near fault earthquake, sloshing

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21310 Isolement and Identification of Major Constituents from Essential Oil of Launaea nudicaulis

Authors: M. Yakoubi, N. Belboukhari, A. Cheriti, K. Sekoum

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Launaea nudicaulis (L.) Hook.f. is a desert, spontaneous plant and endemic to northem Sahara, which belongs to the Asteraceae family. This species exists in the region of Bechar (Local name; El-Rghamma). In our knowledge, no work has been founded, except studies showing the antimicrobial and antifungal activity of methalonic extract of this plant. The present paper describes the chemical composition of the essential oil from Launaea nudicaulis and qualification of isolation and identification of some pure products by column chromatography. The essential oil from the aerial parts of Launaea nudicaulis (Asteraceae) was obtained by hydroditillation in 0.4% yield, led to isolation of four several new products. The isolation is made by column chromatography and followed by GC-IK and GC-MS analysis.

Keywords: Launaea nudicaulis, asteraceae, essential oil, column chromatography, GC-FID, GC-MS

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21309 Spectrophotometric Detection of Histidine Using Enzyme Reaction and Examination of Reaction Conditions

Authors: Akimitsu Kugimiya, Kouhei Iwato, Toru Saito, Jiro Kohda, Yasuhisa Nakano, Yu Takano

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The measurement of amino acid content is reported to be useful for the diagnosis of several types of diseases, including lung cancer, gastric cancer, colorectal cancer, breast cancer, prostate cancer, and diabetes. The conventional detection methods for amino acid are high-performance liquid chromatography (HPLC) and liquid chromatography-mass spectrometry (LC-MS), but they have several drawbacks as the equipment is cumbersome and the techniques are costly in terms of time and costs. In contrast, biosensors and biosensing methods provide more rapid and facile detection strategies that use simple equipment. The authors have reported a novel approach for the detection of each amino acid that involved the use of aminoacyl-tRNA synthetase (aaRS) as a molecular recognition element because aaRS is expected to a selective binding ability for corresponding amino acid. The consecutive enzymatic reactions used in this study are as follows: aaRS binds to its cognate amino acid and releases inorganic pyrophosphate. Hydrogen peroxide (H₂O₂) was produced by the enzyme reactions of inorganic pyrophosphatase and pyruvate oxidase. The Trinder’s reagent was added into the reaction mixture, and the absorbance change at 556 nm was measured using a microplate reader. In this study, an amino acid-sensing method using histidyl-tRNA synthetase (HisRS; histidine-specific aaRS) as molecular recognition element in combination with the Trinder’s reagent spectrophotometric method was developed. The quantitative performance and selectivity of the method were evaluated, and the optimal enzyme reaction and detection conditions were determined. The authors developed a simple and rapid method for detecting histidine with a combination of enzymatic reaction and spectrophotometric detection. In this study, HisRS was used to detect histidine, and the reaction and detection conditions were optimized for quantitation of these amino acids in the ranges of 1–100 µM histidine. The detection limits are sufficient to analyze these amino acids in biological fluids. This work was partly supported by Hiroshima City University Grant for Special Academic Research (General Studies).

Keywords: amino acid, aminoacyl-tRNA synthetase, biosensing, enzyme reaction

Procedia PDF Downloads 285
21308 Bacillus thuringiensis CHGP12 Uses a Multifaceted Strategy to Suppress Fusarium Wilt of Chickpea and to Enhance the Total Biomass of Chickpea Plants

Authors: Muhammad Naveed Aslam, Rida Fatima, Anam Moosa, Muhammad Taimoor Shakeel

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Bacillus strains produce antifungal secondary metabolites making them potential candidates for suppressing Fusarium wilt of chickpea disease. In this study, eighteen Bacillus strains were evaluated for their antagonistic effect against Fusarium oxysporum f. sp. ciceris causing Fusarium wilt of chickpea disease. In a direct antifungal assay, thirteen strains showed significant inhibition zones while the remaining five strains did not produce inhibition zones of FOC. Bacillus thuringiensis CHGP12 was the most promising strain exhibiting the highest inhibition of FOC. Antifungal lipopeptides were extracted from CHGP12 strain which showed significant inhibition of the pathogen. Liquid chromatography mass spectrometry (LCMS) analysis revealed that CHGP12 was positive for the presence of iturin, fengycin, surfactin, bacillaene, bacillibactin, plantazolicin, and bacilysin. CHGP12 was tested for biochemical determinants in an in vitro qualitative test where it showed the ability to produce lipase, amylase, cellulase, protease, siderophores, and indole 3-acetic acid (IAA). Furthermore, in a greenhouse experiment CHGP12 also showed a significant decrease in the disease severity in treated plants compared to control. Moreover, CHGP12 also exhibited a significant increase in plant growth parameters viz, root and shoot growth parameters, stomatal conductance, and photosynthesis rate. Conclusively, our findings present the promising potential of Bacillus strain CHGP12 to suppress Fusarium wilt of chickpea and to promote plant growth.

Keywords: liquid chromatography mass spectrometry, growth promotion, antagonism, hydrolytic enzymes, inhibition, lipopeptides.

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21307 Research Developments in Vibration Control of Structure Using Tuned Liquid Column Dampers: A State-of-the-Art Review

Authors: Jay Gohel, Anant Parghi

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A tuned liquid column damper (TLCD) is a modified passive system of tuned mass damper, where a liquid is used in place of mass in the structure. A TLCD consists of U-shaped tube with an orifice that produces damping against the liquid motion in the tube. This paper provides a state-of-the-art review on the vibration control of wind and earthquake excited structures using liquid dampers. Further, the paper will also discuss the theoretical background of TCLD, history of liquid dampers and existing literature on experimental, numerical, and analytical study. The review will also include different configuration of TLCD viz single TLCD, multi tuned liquid column damper (MTLCD), TLCD-Interior (TLCDI), tuned liquid column ball damper (TLCBD), tuned liquid column ball gas damper (TLCBGD), and pendulum liquid column damper (PLCD). The dynamic characteristics of the different configurate TLCD system and their effectiveness in reducing the vibration of structure will be discussed. The effectiveness of semi-active TLCD will be also discussed with reference to experimental and analytical results. In addition, the review will also provide the numerous examples of implemented TLCD to control the vibration in real structures. Based on the comprehensive review of literature, some important conclusions will be made and the need for future research will be identified for vibration control of structures using TLCD.

Keywords: earthquake, wind, tuned liquid column damper, passive response control, structures

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21306 Preparation and Electro-Optic Characteristics of Polymer Network Liquid Crystals Based On Polymethylvinilpirydine and Polyethylene Glycol

Authors: T. D. Ibragimov, A. R. Imamaliyev, G. M. Bayramov

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The polymer network liquid crystals based on the liquid crystals Н37 and 5CB with polymethylvinilpirydine (PMVP) and polyethylene glycol (PEG) have been developed. Mesogene substance 4-n-heptyoxibenzoic acid (HOBA) is served for stabilization of obtaining composites. Kinetics of network formation is investigated by methods of polarization microscopy and integrated small-angle scattering. It is shown that gel-like states of the composite H-37 + PMVP + HOBA and 5CB+PEG+HOBA are formed at polymer concentration above 7 % and 9 %, correspondingly. At slow cooling, the system separates into a liquid crystal –rich phase and a liquid crystal-poor phase. At this case, transition of these phases in the H-37 + PMVP + HOBA (87 % + 12 % + 1 %) composite to an anisotropic state occurs at 49 оС and и 41 оС, accordingly, while the composite 5CB+PEG+HOBA (85% +13 % +2%) passes to anisotropic state at 36 оС corresponding to the isotropic-nematic transition of pure 5CB. The basic electro-optic parameters of the obtained composites are determined at room temperature. It is shown that the threshold voltage of the composite H-37 + PMVP + HOBA increase in comparison with pure H-37 and, accordingly, there is a shift of voltage dependence of rise times to the high voltage region. The contrast ratio worsens while decay time improves in comparison with the pure liquid crystal at all applied voltage. The switching times of the composite 5CB + PEG + HOBA (85% +13 % +2%) show anomalous behavior connected with incompleteness of the transition to an anisotropic state. Experimental results are explained by phase separation of the system, diminution of a working area of electro-optical effects and influence of areas with the high polymer concentration on areas with their low concentration.

Keywords: liquid crystals, polymers, small-angle scattering, optical properties

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21305 Evaluation of Oxidative Changes in Soybean Oil During Shelf-Life by Physico-Chemical Methods and Headspace-Liquid Phase Microextraction (HS-LPME) Technique

Authors: Maryam Enteshari, Kooshan Nayebzadeh, Abdorreza Mohammadi

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In this study, the oxidative stability of soybean oil under different storage temperatures (4 and 25˚C) and during 6-month shelf-life was investigated by various analytical methods and headspace-liquid phase microextraction (HS-LPME) coupled to gas chromatography-mass spectrometry (GC-MS). Oxidation changes were monitored by analytical parameters consisted of acid value (AV), peroxide value (PV), p-Anisidine value (p-AV), thiobarbituric acid value (TBA), fatty acids profile, iodine value (IV), and oxidative stability index (OSI). In addition, concentrations of hexanal and heptanal as secondary volatile oxidation compounds were determined by HS-LPME/GC-MS technique. Rate of oxidation in soybean oil which stored at 25˚C was so higher. The AV, p-AV, and TBA were gradually increased during 6 months while the amount of unsaturated fatty acids, IV, and OSI decreased. Other parameters included concentrations of both hexanal and heptanal, and PV exhibited increasing trend during primitive months of storage; then, at the end of third and fourth months a sudden decrement was understood for the concentrations of hexanal and heptanal and the amount of PV, simultaneously. The latter parameters increased again until the end of shelf-time. As a result, the temperature and time were effective factors in oxidative stability of soybean oil. Also intensive correlations were found for soybean oil at 4 ˚C between AV and TBA (r2=0.96), PV and p-AV (r2=0.9), IV and TBA (-r2=0.9), and for soybean oil stored at 4˚C between p-AV and TBA (r2=0.99).

Keywords: headspace-liquid phase microextraction, oxidation, shelf-life, soybean oil

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21304 First Approach on Lycopene Extraction Using Limonene

Authors: M. A. Ferhat, M. N. Boukhatem, F. Chemat

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Lycopene extraction with petroleum derivatives as solvents has caused safety, health, and environmental concerns everywhere. Thus, finding a safe alternative solvent will have a strong and positive impact on environments and general health of the world population. d-limonene from the orange peel was extracted through a steam distillation procedure followed by a deterpenation process and combining this achievement by using it as a solvent for extracting lycopene from tomato fruit as a substitute of dichloromethane. Lycopene content of fresh tomatoes was determined by high-performance liquid chromatography after extraction. Yields obtained for both extractions showed that yields of d-limonene’s extracts were almost equivalent to those obtained using dichloromethane. The proposed approach using a green solvent to perform extraction is useful and can be considered as a nice alternative to conventional petroleum solvent where toxicity for both operator and environment is reduced.

Keywords: alternative solvent, d-limonene, extraction, lycopene

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21303 Contribution to the Production of Phenazine Antibiotics Effect Type Compounds by Some Strains of Pseudomonas spp.fluorescent

Authors: Nacéra Benoussaid, Lehalali Meriem, Benchabane Messaoud

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Our work focuses on the production of compound antibiotic effect of volatile nature namely hydrogen cyanide and the production and identification of molecules phénazinique by some strains of fluorescent Pseudomonas spp isolated from the rhizosphere of some trees for a possible use as bio pesticides antifungal effect and/or antibiotic. We tested the production of hydrogen cyanide of 21 strains of Pseudomonas spp. fluorescent among them 19 strains (90, 47%) showed a positive cyanogenesis.The antagonism test executed in vitro showed that Pseudomonas strains have a higher anti fungal effect relative to their antibacterial effect with diameters of inhibition zones up to 3, 9 cm recorded by the strain F48 against Coleosporiumsp compared with recorded results against bacteria with a maximum inhibition of 1, 26 cm among this antagonistic strain.Three strains were selected by testing for producing phénazines namely PI9, BB9 and F20. The effect of the antimicrobial activity was performed on different culture media (GN, King B, ISP2 and PDA). The results of our study allowed us to retain the King B medium as ideal medium for the production of secondary metabolite. The produced phenazinique compounds was extracted from various organic solvents, and after the results of antibiographie against germs - targets, the extracts of ethyl acetate gave the best results compared to dichloromethane and hexane.The Analysis of these compounds of antibiotic phenazinique effect within layer chromatography (CCM) and high performance liquid chromatography( HPLC) indicate that both strains PI9 and F20 are productive of phenazine-1-carboxylic acid (PCA). The BB9 strain is suspected to be productive of another phenazinique compound.

Keywords: Pseudomonas ssp. fluorescents, antagonism in vitro, secondary metabolite, phenazines, biopesticide.

Procedia PDF Downloads 511
21302 Prediction of Ionic Liquid Densities Using a Corresponding State Correlation

Authors: Khashayar Nasrifar

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Ionic liquids (ILs) exhibit particular properties exemplified by extremely low vapor pressure and high thermal stability. The properties of ILs can be tailored by proper selection of cations and anions. As such, ILs are appealing as potential solvents to substitute traditional solvents with high vapor pressure. One of the IL properties required in chemical and process design is density. In developing corresponding state liquid density correlations, scaling hypothesis is often used. The hypothesis expresses the temperature dependence of saturated liquid densities near the vapor-liquid critical point as a function of reduced temperature. Extending the temperature dependence, several successful correlations were developed to accurately correlate the densities of normal liquids from the triple point to a critical point. Applying mixing rules, the liquid density correlations are extended to liquid mixtures as well. ILs are not molecular liquids, and they are not classified among normal liquids either. Also, ILs are often used where the condition is far from equilibrium. Nevertheless, in calculating the properties of ILs, the use of corresponding state correlations would be useful if no experimental data were available. With well-known generalized saturated liquid density correlations, the accuracy in predicting the density of ILs is not that good. An average error of 4-5% should be expected. In this work, a data bank was compiled. A simplified and concise corresponding state saturated liquid density correlation is proposed by phenomena-logically modifying reduced temperature using the temperature-dependence for an interacting parameter of the Soave-Redlich-Kwong equation of state. This modification improves the temperature dependence of the developed correlation. Parametrization was next performed to optimize the three global parameters of the correlation. The correlation was then applied to the ILs in our data bank with satisfactory predictions. The correlation of IL density applied at 0.1 MPa and was tested with an average uncertainty of around 2%. No adjustable parameter was used. The critical temperature, critical volume, and acentric factor were all required. Methods to extend the predictions to higher pressures (200 MPa) were also devised. Compared to other methods, this correlation was found more accurate. This work also presents the chronological order of developing such correlations dealing with ILs. The pros and cons are also expressed.

Keywords: correlation, corresponding state principle, ionic liquid, density

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21301 Preparation and Characterization of Dendrimer-Encapsulated Ytterbium Nanoparticles to Produce a New Nano-Radio Pharmaceutical

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

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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

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21300 In vitro Evaluation of Capsaicin Patches for Transdermal Drug Delivery

Authors: Alija Uzunovic, Sasa Pilipovic, Aida Sapcanin, Zahida Ademovic, Berina Pilipović

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Capsaicin is a naturally occurring alkaloid extracted from capsicum fruit extracts of different of Capsicum species. It has been employed topically to treat many diseases such as rheumatoid arthritis, osteoarthritis, cancer pain and nerve pain in diabetes. The high degree of pre-systemic metabolism of intragastrical capsaicin and the short half-life of capsaicin by intravenous administration made topical application of capsaicin advantageous. In this study, we have evaluated differences in the dissolution characteristics of capsaicin patch 11 mg (purchased from market) at different dissolution rotation speed. The proposed patch area is 308 cm2 (22 cm x 14 cm; it contains 36 µg of capsaicin per square centimeter of adhesive). USP Apparatus 5 (Paddle Over Disc) is used for transdermal patch testing. The dissolution study was conducted using USP apparatus 5 (n=6), ERWEKA DT800 dissolution tester (paddle-type) with addition of a disc. The fabricated patch of 308 cm2 is to be cut into 9 cm2 was placed against a disc (delivery side up) retained with the stainless-steel screen and exposed to 500 mL of phosphate buffer solution pH 7.4. All dissolution studies were carried out at 32 ± 0.5 °C and different rotation speed (50± 5; 100± 5 and 150± 5 rpm). 5 ml aliquots of samples were withdrawn at various time intervals (1, 4, 8 and 12 hours) and replaced with 5 ml of dissolution medium. Withdrawn were appropriately diluted and analyzed by reversed-phase liquid chromatography (RP-LC). A Reversed Phase Liquid Chromatography (RP-LC) method has been developed, optimized and validated for the separation and quantitation of capsaicin in a transdermal patch. The method uses a ProntoSIL 120-3-C18 AQ 125 x 4,0 mm (3 μm) column maintained at 600C. The mobile phase consisted of acetonitrile: water (50:50 v/v), the flow rate of 0.9 mL/min, the injection volume 10 μL and the detection wavelength 222 nm. The used RP-LC method is simple, sensitive and accurate and can be applied for fast (total chromatographic run time was 4.0 minutes) and simultaneous analysis of capsaicin and dihydrocapsaicin in a transdermal patch. According to the results obtained in this study, we can conclude that the relative difference of dissolution rate of capsaicin after 12 hours was elevated by increase of dissolution rotation speed (100 rpm vs 50 rpm: 84.9± 11.3% and 150 rpm vs 100 rpm: 39.8± 8.3%). Although several apparatus and procedures (USP apparatus 5, 6, 7 and a paddle over extraction cell method) have been used to study in vitro release characteristics of transdermal patches, USP Apparatus 5 (Paddle Over Disc) could be considered as a discriminatory test. would be able to point out the differences in the dissolution rate of capsaicin at different rotation speed.

Keywords: capsaicin, in vitro, patch, RP-LC, transdermal

Procedia PDF Downloads 227
21299 Analysis of Evaporation of Liquid Ammonia in a Vertical Cylindrical Storage Tank

Authors: S. Chikh, S. Boulifa

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The present study addresses the problem of ammonia evaporation during filling of a vertical cylindrical tank and the influence of various external factors on the stability of storage by determining the conditions for minimum evaporation. Numerical simulation is carried out by solving the governing equations namely, continuity, momentum, energy, and diffusion of species. The effect of temperature of surrounding air, the filling speed of the reservoir and the temperature of the filling liquid ammonia on the evaporation rate is investigated. Results show that the temperature of the filling liquid has little effect on the liquid ammonia for a short period, which, in fact, is function of the filling speed. The evaporation rate along the free surface of the liquid is non-uniform. The inlet temperature affects the vapor ammonia temperature because of pressure increase. The temperature of the surrounding air affects the temperature of the vapor phase rather than the liquid phase. The maximum of evaporation is reached at the final step of filling. In order to minimize loss of ammonia vapors automatically causing losses in quantity of the liquid stored, it is suggested to ensure the proper insulation for the walls and roof of the reservoir and to increase the filling speed.

Keywords: evaporation, liquid ammonia, storage tank, numerical simulation

Procedia PDF Downloads 288