• Title/Summary/Keyword: cell separation

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Transcriptome Analysis of Streptococcus mutans and Separation of Active Ingredients from the Extract of Aralia continentalis (Streptococcus mutans의 전사체 분석과 독활 추출물로부터 활성 성분 분리)

  • Hyeon-Jeong Lee;Da-Young Kang;Yun-Chae Lee;Jeong Nam Kim
    • Journal of Life Science
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    • v.33 no.7
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    • pp.538-548
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    • 2023
  • The research has been conducted on the isolation of antimicrobial compounds from plant natural extracts and their potential application in oral health care products. This study aimed to investigate the antimicrobial mechanism by analyzing the changes in gene expression of Streptococcus mutans, a major oral pathogen, in response to complex compounds extracted from Aralia continentalis and Arctii Semen using organic solvents. Transcriptome analysis (RNA-seq) revealed that both natural extracts commonly upregulated or downregulated the expression of various genes associated with different metabolic and physiological activities. Three genes (SMU_1584c, SMU_2133c, SMU_921), particularly SMU_921 (rcrR), known as a transcription activator of two sugar phosphotransferase systems (PTS) involved in sugar transport and biofilm formation, exhibited consistent high expression levels. Additionally, component analysis of the A. continentalis extract was performed to compare its effects on gene expression changes with the A. Semen extract, and two active compounds were identified through gas chromatography-mass spectrometry (GC-MS) analysis of the active fraction. The n-hexane fraction (ACEH) from the A. continentalis extract exhibited antibacterial specificity against S. mutans, leading to a significant reduction in the viable cell counts of Streptococcus sanguinis and Streptococcus gordonii among the tested multi-species bacterial communities. These findings suggest the broad-spectrum antibacterial activity of the A. continentalis extract and provide essential foundational data for the development of customized antimicrobial materials by elucidating the antibacterial mechanism of the identified active compounds.

Evaluation of Manufacturing and Biological Properties of Y Zone Care Hydrogel Solution (Y존 케어 하이드로젤솔루션의 제조 및 생물학적 특성 평가)

  • Eun-Ji Kim;In-Kyoung Kim
    • Journal of Advanced Technology Convergence
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    • v.3 no.2
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    • pp.25-31
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    • 2024
  • In recent industrialization and development due to information and communication technology, modern women in modern society are exposed to physical and mental health due to numerous stresses. Popular inflammations are attributable to a decrease in lactic acid bacteria, frequent antibiotic use, and a decrease in immunity. It is necessary to develop products that are helpful and reflected. The inner care gel currently introduced on the market can increase beneficial bacteria and maintain a healthy y-zone. The inner gel contains a hydrogel component. 90% is made up of water, and other components act as support for supporting water and are formed through crosslinking between polymer chains. Hydroxyethyl cellulose (HEC) is a hydroxyethyl ethylenetel of cellulose. The purpose of use is to act as a binder, an emulsion stabilizer, a viscosity enhancer (water-soluble), and a film forming agent. CA (crosslinker) is a crosslinking agent and serves to bind. Hydrogel in the beauty field acts as a film forming agent that gently wraps around the skin by forming a thin film and serves as an emulsion stabilizer that helps to prevent separation of other raw materials. It also acts as a thickener by increasing viscosity in cosmetics. In addition, it is used for glucose monitoring, nursing care, cell transplantation, and wound treatment in the bio field. Currently, it is understood that no products using functional hydrogel have been released, so in this study, a Y zone care hydrogel solution was manufactured to find out the antibacterial properties of the functional hydrogel, and a new solution was developed. As a result, it was confirmed that the appropriate Ph was applied to the Y zone, and after culturing Candida albicans in PDB medium, all three products of the Y zone care hydrogel solution showed an antibacterial effect of 0.5-1.0mm

The Effect of Superoxide Anion Production by PMN on Pneumocyte Injury in Patients with Bronchial Asthma (기관지천식환자에서 다형핵구의 과산화 음이온 생성능이 폐포세포 손상에 미치는 영향)

  • Kim, Young-Kyoon;Park, Sung-Hak
    • Tuberculosis and Respiratory Diseases
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    • v.40 no.3
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    • pp.213-222
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    • 1993
  • Background : Bronchial asthma has been known as an inflmmatory disease. There have been many evidences that polymorphonuclear leukocytes (PMN) might play an important role in the pathogrnesis of asthma. Although many investigators suggested that pneumocyte injury by PMN-derived oxygen radicals may contribute to the pathogenesis of asthma, there has been few report for a direct evidence of oxygen radicals-mediated pneumocyte injury in bronchial asthma. Furthermore the exact mechanism of oxygen radicals-mediated pneumocyte injury is still controversy. This study was designed to establish a direct in vitro evidence and its clinical significance of pneumocyte injury by PMN-derived superoxide anion in bronchial asthma and to elucidate the main mechanism of superoxide anion-mediated pneumocyte injury. Methods : 12 stable asthmatics and 5 healthy volunteers were participated in this study. PMN was separated from peripheral venous blood samples by using dextran sedimentation and Ficoll-Hypaque density gradient separation method. Superoxide anion productions by PMN and plasma SOD activities were measured by spectrophotometric assay using the principle of SOD inhibitable cytochrome c reduction. PMN-mediated pneumocyte injuries were measured by $^{51}Cr$-release assay using A549 pneumocytes and were expressed as percent lysis and percent detachment. Results: 1) PMN from asthmatics produced more amount of superoxide anion compared to PMN from normal subjects ($6.65{\pm}0.58$ vs $2.81{\pm}0.95\;nmol/1{\times}10^6$ cells, p<0.05), and showed an inverse correlation with $FEV_1$(R=-0.63, p<0.05), but no correlation with $PC_{20}$ histamine in asthmatics. 2) Plasma SOD activities were decreased in asthmatics compared to normal subjects but not significant, and showed a positive correlation with $FEV_1$(R=0.63, p<0.05) but no correlation with $PC_{20}$ histamine in asthmatics. 3) There were a positive correlation between plasma SOD activity and superoxide anion production by PMN in normal subjects (R=0.88, p<0.05) but not in asthmatics. 4) PMN-mediated pneumocyte injury was predominantly expressed as cell detachment rather than cell lysis in both groups, and PMN from asthmatics showed more potent cytotoxic effect on A549 pneumocytes compated to PMN from normal subjects. PMN-mediated detachment rather than lysis of A549 pneumocytes was significantly inhibited by in vitro SOD but not by diluted serum. 5) PMN-mediated detachment rather than lysis of A549 pneumocytes showed a good correlation with superoxide anion production by PMN (R=0.90 in normal subjects, R=0.82 in asthmatics, p<0.05) but no correlation with plasma SOD activity. PMN-mediated pneumocyte injuries were not correlated with $FEV_1$ or $PC_{20}$ histamine in asthmatics. 6) There were no significant differences in PMN-mediated pneumocyte injuries between allergic and nonallergic asthmatics. Conclusion : Our results suggest that pneumocyte injury by PMN-derived superoxide anion may partially contribute to the pathogenesis of asthma and that cell detachment rather than cell lysis may be the mechanism of superoxide anion-mediated pneumocyte injury.

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Cytotoxic Effects of Tenebrio molitor Larval Extracts against Hepatocellular Carcinoma (갈색거저리 유충 추출물의 간암세포에 대한 세포독성 효능)

  • Lee, Ji-Eun;Lee, An-Jung;Jo, Da-Eun;Cho, Ju Hyeong;Youn, Kumju;Yun, Eun-Young;Hwang, Jae-Sam;Jun, Mira;Kang, Byoung Heon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.2
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    • pp.200-207
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    • 2015
  • Various natural products or their derivatives, mostly originating from plants, fungi, and bacteria, have been exploited as therapeutic drugs to treat various human diseases. In addition to previously explored organisms, research on natural compounds has now expanded into unexamined living organisms in order to identify novel bioactive substances. Here, we determined whether or not the larval form of the mealworm beetle Tenebrio molitor, a species of darkling beetle, contains cytotoxic substances that exclusively affect cancer cell viability. Ethanol extract and its solvent partitioned fractions, hexane and ethyl acetate fractions, showed anticancer effects against various human cancer cells derived from the prostate (PC3 and 22Rv1), cervix (HeLa), liver (PLC/PRF5, HepG2, Hep3B, and SK-HEP-1), colon (HCT116), lung (NCI-H460), breast (MDA-MB231), and ovary (SKOV3). Cell death induced by the fractions was a mix of apoptosis, necrosis, and autophagy. The hexane fraction was administered intraperitoneally to nude mice bearing a hepatocellular carcinoma SK-HEP-1 and showed inhibition of tumor growth in vivo. Therefore, we concluded that worm extracts contain cytotoxic substances, which can be enriched by proper fractionation protocols, and further separation and purification could lead to the identification of novel molecules to treat human cancers.

Phase Equilibria of Ionic Liquid/Organic Compound/Supercritical CO2 Systems (이온성액체/유기화합물/초임계이산화탄소계의 상평형)

  • Lim, Bang-Hyun;Kim, Jong-Won;Paek, Sang-Min;Son, Bo-Kook;Lee, Yong-Rok;Lee, Chul Soo;Lee, Huen;Ra, Chun-Sup;Shim, Jae-Jin
    • Clean Technology
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    • v.12 no.3
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    • pp.128-137
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    • 2006
  • The volume change of an ionic liquid and the phase separation behavior of room temperature ionic liquid(RTIL)/organic compound mixture in supercritical carbon dioxide were measured in a high pressure view cell. 1-Butyl-3-methylimidazolium hexafluorophosphate ([bmim][$PF_6$]) and 1-butyl-3-methylimidazolium tetrafluoroborate ([bmim][$BF_4$]) was used as ionic liquid(IL). and methanol and dimethyl carbonate were used as organic compound. For a fixed amount of [bmim][$PF_6$] the lower critical endpoint (LCEP) pressure, where the liquid phase is split, decreased as increasing the amount of organic compound. The LCEP pressure became higher as the water content of ionic liquid was higher. However, for water contents above a certain value, no LCEP was formed. LCEP appeared 1.0 MPa higher for a mixture with [bmim][$BF_4$] than with [bmim][$PF_6$]. There was almost no difference in the K-point pressures for different types of ionic liquid and for different amounts of organic liquid. When the concentration of ionic liquid([bmim][$PF_6$]) (IL/(IL+MeOH)) in the initial liquid mixture was larger than 5.9 mol% at the LCEP of the mixture, the volume of $L_1$ because larger than the volume of $L_2$. When it was smaller, however, the volume became smaller, too. The volume change of ionic liquid in the presence of carbon dioxide decreased as increasing the temperature, while it increased as increasing the pressure. For temperatures between 313.15 to 343.15K at 300 bar, it was about 123~125 % of the original volume.

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Animal Experiments on an Antithrombogenic Small-Caliber Vascular Prostheses and Vascualr Patch : Observation in Canine Models (항혈전성 소구경 인조 혈관 및 봉합편에 대한 동물 실험)

  • 김수철;김원곤;유세영
    • Journal of Chest Surgery
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    • v.36 no.2
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    • pp.63-72
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    • 2003
  • Although a variety of synthetic vascular grafts are available in modern vascular surgery, no ideal prosthesis ha,4 yet been developed. Small-caliber vascular grafts with low flow, as used in the lower extremity, continue to become thrombosed at unacceptable rates. We have developed and evaluated the new antithrombogenic blood contacting surfaces in canine model. Material and Method: Two now antithrombogenic blood contacting surfaces(Polyvinylalcohol -Polyurethane(PVA-PU) blend and natural Graphite-polyurethane(G-PU) blend) have been developed and evaluated in canine model, using vascular grafts and patches. The luminal surfaces of the test vascular grafts(5 mm ID) were fabricated by dipping a glass rod in PVA-PU blend solution(50 % PVA) using phase separation method. Mongrel dogs of either sex weighing 18-22 kg were anesthetized by endotracheal intubation using halothane and their lungs were ventilated with a volume-cycled ventilator, Maintenance anesthesia with 0.5-1.0% halothane and supplemental oxygen was used. Two pairs were used for comparison in the bilateral femoral arteries for both vascular grafts(PVA-PU vs. PU) and vascular patches(G-PU vs. PU). Bilateral groin incisions were made and the arteries were exposed and clamped. After an excision of 1 cm of the artery between clamps, a grail of 2.5 cm in length was implanted end-to-end using 6-0 polypropylene suture. The vascular patch was implanted as a form of on-lay patch. Animals were sacrificed at 1, 2, 4, 6, 8 and 16 weeks for vascular grafts and 1, 2. 4 and 6 weeks for vascular patches. Result The vascular grafts of PVA-PU blends showed patent lumina in the 2 and 16 weeks animals, while those of PU showed a patent lumen in 2 weeks animal. PVA-PU graft of 16 weeks showed a fairly clean luminal surface. A light microscopic finding of this graft demonstrated good tissue infiltration through porosity, The animals with vascular patches showed patent arteries in both groups except 2 weeks animal. Scanning electron microscopy of the luminal surfaces of G-PU patches in 4 and 6 weeks animals showed endothelial cell covering with microvilli. PU patches showed qualitatively less endothelial cell covering. Conclusion: In conclusion, PVA-PU and G-PU blends can be a promising blood contacting surfaces for application in a synthetic vascualr graft. However, further animal study is needed to determine the real long-term effects of these methods of surface modifications.

Light and Electron Microscopy of Gill and Kidney on Adaptation of Tilapia(Oreochromis niloticus) in the Various Salinities (틸라피아의 해수순치시(海水馴致時) 아가미와 신장(腎臟)의 광학(光學) 및 전자현미경적(電子顯微鏡的) 관찰(觀察))

  • Yoon, Jong-Man;Cho, Kang-Yong;Park, Hong-Yang
    • Applied Microscopy
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    • v.23 no.2
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    • pp.27-40
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    • 1993
  • This study was taken to examine the light microscopic and ultrastructural changes of gill and kidney of female tilapia{Oreochromis niloticus) adapted in 0%o, 10%o, 20%o, and 30%o salt concentrations, respectively, by light, scanning and transmission electron microscope. The results obtained in these experiments were summarized as follows: Gill chloride cell hyperplasia, gill lamellar epithelial separation, kidney glomerular shrinkage, blood congestion in kidneys and deposition of hyalin droplets in kidney glomeruli, tubules were the histological alterations in Oreochromis niloticus. Incidence and severity of gill chloride cell hyperplasia rapidly increased together with increase of salinity, and the number of chloride cells in gill lamellae rapidly increased in response to high external NaCl concentrations. The ultrastructure by scanning electron microscope(SEM) indicated that the gill secondary lamella of tilapia(Oreochromis niloticus) exposed to seawater, were characterized by rough convoluted surfaces during the adaptation. Transmission electron microscopy(TEM) indicated that mitochondria in chloride cells exposed to seawater, were both large and elongate and contained well-developed cristae. TEM also showed the increased chloride cells exposed to seawater. The presence of two mitochondria-rich cell types is discussed with regard to their possible role in the hypoosmoregulatory changes which occur during seawater-adaptation. Most Oreochromis niloticus adapted in seawater had an occasional glomerulus completely filling Bowman's capsule in kidney, and glomerular shrinkage was occurred higher in kidney tissues of individuals living in 10%o, 20%o, 30%o of seawater than in those living in 0%o of freshwater, and blood congestion was occurred severer in kidney tissues of individuals living 20%o, 30%o of seawater than in those living in 10%o of seawater. There were decreases in the glomerular area and the nuclear area in the main segments of the nephron, and that the nuclear areas of the nephron cells in seawater-adapted tilapia were of smaller size than those from freshwater-adapted fish. Our findings demonstrated that Oreochromis niloticus tolerated moderately saline environment and the increased body weight living in 30%o was relatively higher than that living in 10%o in spite of histopathological changes.

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Study of Polysulfone Membrane for Membrane-covered Oxygen Probe System (산소 전극 시스템에 사용되는 polysulfone막에 대한 연구)

  • Hong, Suk In;Kim, Hyun Joon;Park, Hee Young;Kim, Tae Jin;Jeong, Yong Seob
    • Applied Chemistry for Engineering
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    • v.7 no.5
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    • pp.877-887
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    • 1996
  • The ideal membranes for membrane-covered oxygen probes system should be selectively permeable for oxygen and chemically inert, and have good mechanical strength. Polysulfone(PSf) was selected to develop the membrane for membrane-covered oxygen electrodes system. PSf membranes have properties such as good reproducibility, good mechanical strength, chemical inertness, and high heat resistance. PSf membranes were cast from polymer solution on the glass plate at constant temperature, and casting solvents used were tetrahydrofuran(THF), methylene chloride, and N-methyl-2-pyrrolidone(NMP). Tricresyl phosphate(TCP) as plasicizer was added to PSf to increase the softness of membrane. The permeation characteristics were observed for pure oxygen and nitrogen through pure PSf membranes by variable volume method and membrane-covered electrode system. The permeability coefficients of oxygen and nitrogen measured by variable volume method were slightly decreased with increasing of upstream pressure. The permeation properties of PSf membrane using methylene choride as casting solvent were not affected by the PSf amount of polymer solution. The permeability coefficients of oxygen and nitrogen for PSf membrane containing TCP were very slightly lower than those for pure PSf membrane, but ideal separation factors were slightly higher. The flexibility of PSf membrane containing 2wt% TCP was better than that of pure PSf membrane. It was expected that this increase in flexibility would solve the difficulty of fixing the membrane to the cathode. The membrane-covered oxygen probes system was composed of anode, cathode and electrolyte. The type of the anode was Ag/AgCl half-cell, that of cathode was Ag, and the electrolyte was 4N KCl solution. The result of sampled current voltametry for PSf membrane showed the plateu region at -0.3V~-1.0V. The correlation coefficient of oxygen partial pressure versus current for PSf membrane was relatively high, 0.99949. It was concluded that PSf membrane was the good candidate for the membrane-covered oxygen probes system.

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Detection of Campylobacter jejuni in food and poultry visors using immunomagnetic separation and microtitre hybridization

  • Simard, Ronald-E.
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2000.05a
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    • pp.71-73
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    • 2000
  • Campylobacter jejuni is most frequently identified cause of cause of acute diarrhoeal infections in developeed countries, exceeding rates of illness caused by both salmonella and shigilla(Skirrow, 1990 ; Lior 1994). Previous studies on campylobacter jejuni contamination of commercial broiler carcasses in u.s.(Stern, 1992). Most cases of the disease result from indirect transmission of Campylobactor from animals via milk, water and meat. In addition to Campylobactor jejuni. the closely relates species Campylobactor coli and Campylobactor lari have also been implicated as agents of gastroenteritis in humans. Campylobactor coli represented only approximately 3% of the Campylobactor isolates from patients with Campylobactor enteritis(Griffiths and Park, 1990) whereas Campylobactor coli is mainly isolated from pork(Lmmerding et al., 1988). Campylobactor jejuni has also been isolated from cases of bacteremia, appendicitis and, recently, has been associated with Guillai-Barre syndrome(Allos and Blaser, 1994; von Wulffen et al., 1994; Phillips, 1995). Studies in volunteers indicated that the infectious dose for Campylobactor jejuni is low(about 500 organisms)(Robinson, 1981). The methods traditionally used to detect Campylobactor ssp. in food require at least two days of incubation in an enrichment broth followed by plating and two days of incubation on complex culture media containing many antibiotics(Goossens and Butzler, 1992). Finnaly, several biochemical tests must be done to confirm the indentification at the species level. Therfore, sensitive and specific methods for the detection of small numbers of Campylobactor cells in food are needed. Polymerase chain reaction(PCR) assays targeting specific DNA sequences have been developed for the detection of Campylobactor(Giesendorf and Quint, 1995; Hemandex et al., 1995; Winter and Slavidk, 1995). In most cases, a short enrichment step is needed to enhance the sensitivity of the assay prior to detection by PCR as the number of bacteria in the food products is low in comparison with those found in dinical samples, and because the complex composition of food matrices can hinder the PCR and lower its sensitivity. However, these PCR systems are technically demanding to carry out and cumbersome when processing a large number of samples simutaneously. In this paper, an immunomagnetic method to concentrate Campylobactor cells present in food or clinical samples after an enrichment step is described. To detect specifically the thermophilic Campylobactor. a monoclonal antibody was adsorbed on the surface of the magnetic beads which react against a major porin of 45kDa present on the surface of the cells(Huyer et al., 1986). After this partial purification and concentration step, detection of bound cells was achieved using a simple, inexpensive microtitre plate-based hybridization system. We examined two alternative detection systems, one specific for thermophilic Campylobactor based on the detection of 23S rRNA using an immobilized DNA probe. The second system is less specific but more sensitive because of the high copy number of the rRNA present in bacterial cell($10^3-10^4$). By using specific immunomagnetic beads against thermophilic Campylobactor, it was possible to concentrate these cells from a heterogeneous media and obtain highly specific hybridization reactions with good sensitivity. There are several advantages in using microtitre plates instead of filter membranes or other matrices for hybridization techniques. Microtitre plates are much easier to handle than filter membranes during the adsorption, washing, hybridization and detection steps, and their use faciilitates the simultanuous analysis of multiple sample. Here we report on the use of a very simple detection procedure based on a monoclonal anti-RNA-DNA hybrid antibody(Fliss et al., 1999) for detection of the RNA-DNA hybrids formed in the wells.

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Perfluoropolymer Membranes of Tetrafluoroethylene and 2,2,4Trifluofo- 5Trifluorometoxy- 1,3Dioxole.

  • Arcella, V.;Colaianna, P.;Brinati, G.;Gordano, A.;Clarizia, G.;Tocci, E.;Drioli, E.
    • Proceedings of the Membrane Society of Korea Conference
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    • 1999.07a
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    • pp.39-42
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    • 1999
  • Perfluoropolymers represent the ultimate resistance to hostile chemical environments and high service temperature, attributed to the presence of fluorine in the polymer backbone, i.e. to the high bond energy of C-F and C-C bonds of fluorocarbons. Copolymers of Tetrafluoroethylene (TEE) and 2, 2, 4Trifluoro-5Trifluorometoxy- 1, 3Dioxole (TTD), commercially known as HYFLON AD, are amorphous perfluoropolymers with glass transition temperature (Tg)higher than room temperature, showing a thermal decomposition temperature exceeding 40$0^{\circ}C$. These polymer systems are highly soluble in fluorinated solvents, with low solution viscosities. This property allows the preparation of self-supported and composite membranes with desired membrane thickness. Symmetric and asymmetric perfluoropolymer membranes, made with HYFLON AD, have been prepared and evaluated. Porous and not porous symmetric membranes have been obtained by solvent evaporation with various processing conditions. Asymmetric membranes have been prepared by th wet phase inversion method. Measure of contact angle to distilled water have been carried out. Figure 1 compares experimental results with those of other commercial membranes. Contact angles of about 120$^{\circ}$for our amorphous perfluoropolymer membranes demonstrate that they posses a high hydrophobic character. Measure of contact angles to hexandecane have been also carried out to evaluate the organophobic character. Rsults are reported in Figure 2. The observed strong organophobicity leads to excellent fouling resistance and inertness. Porous membranes with pore size between 30 and 80 nanometers have shown no permeation to water at pressures as high as 10 bars. However high permeation to gases, such as O2, N2 and CO2, and no selectivities were observed. Considering the porous structure of the membrane, this behavior was expected. In consideration of the above properties, possible useful uses in th field of gas- liquid separations are envisaged for these membranes. A particularly promising application is in the field of membrane contactors, equipments in which membranes are used to improve mass transfer coefficients in respect to traditional extraction and absorption processes. Gas permeation properties have been evaluated for asymmetric membranes and composite symmetric ones. Experimental permselectivity values, obtained at different pressure differences, to various single gases are reported in Tab. 1, 2 and 3. Experimental data have been compared with literature data obtained with membranes made with different amorphous perfluoropolymer systems, such as copolymers of Perfluoro2, 2dimethyl dioxole (PDD) and Tetrafluorethylene, commercialized by the Du Pont Company with the trade name of Teflon AF. An interesting linear relationship between permeability and the glass transition temperature of the polymer constituting the membrane has been observed. Results are descussed in terms of polymer chain structure, which affects the presence of voids at molecular scale and their size distribution. Molecular Dyanmics studies are in progress in order to support the understanding of these results. A modified Theodoru- Suter method provided by the Amorphous Cell module of InsightII/Discover was used to determine the chain packing. A completely amorphous polymer box of about 3.5 nm was considered. Last but not least the use of amorphous perfluoropolymer membranes appears to be ideal when separation processes have to be performed in hostile environments, i.e. high temperatures and aggressive non-aqueous media, such as chemicals and solvents. In these cases Hyflon AD membranes can exploit the outstanding resistance of perfluoropolymers.

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