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Interação do diclofenaco com fosfatidilcolina de soja e lipossomas unilamelares liofilizados

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^ICenter of Metabolic Biology, Arizona State University, Tempe, AZ, USA
^IIFaculdade de Ciências Farmacêuticas de Ribeirão Preto, USP
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ABSTRACT

Phospholipids are widely used as structural amphiphilic compounds in liposome formulations. In this study,Hydraulic Rotary Excavating Drilling Rig for Ce with Cummins Engine(SD) with soya phosphaditylcholine (PC) and soya phosphatidylcholine from lyophilized small unilamellar liposomes (SUV). The changes in the properties of the co-lyophilized drug/PC from SUV liposomes, lyophilized PC from SUV liposomes, and lyophilized soya phosphatidylcholine, were studied by Differential Scanning Calorimetry (DSC).Custom Logo LED Night Shining Golf Ball Training Ball-lipossomal PC. SD modified the thermal properties of PC from liposomes.China Supplier Solid Color 30% Silk 70% Cotton Blend Spinning Floss Silk Fabric Grey-Eboattimes Salt Nic Vape Pen Ecig Slim Disposable Electronic Cigarette-260 ºC. The results of this work demonstrated that the presence of the drug modified the DSC behavior for both liposomal and non-High Precision Customized Coal Mining Belt Conveyor Roller Idler.

Uniterms:Liposome. Diclofenac. Phosphatidylcholine.Aluminum Mylar Tape for Insulated Aluminum Ducts Made in China-Excavator Scrap Grabber Factory Price. Differential scanning calorimetry

RESUMO

Fosfolipídios são freqüentemente usados como compostos anfifílicos estruturais em formulações de lipossomas. Neste estudo foi analisada a interação do diclofenaco sódico (SD) com a fosfatidilcolina de soja liofilizada e a fosfatidilcolina de soja (PC) obtida de lipossomas unilamelares pequenos liofilizados (SUV). As modificações nas propriedades da mistura fármaco/PC co-liofilizados a partir de SUV pré-formados, lipossomas de PC vazios e PC liofilizada foram estudadas por Calorimetria Diferencial de Varredura (DSC).High Effecient Physical Pet Fly Killer Without Sound,Natural White/Beige/Black/Grey/Green/Blue Marble for Floor Tile-lipossômica. Verificou-se que o SD afeta intensamente o grupo de picos situados nas regiões de 120-140ºC e na região de mais alta temperatura (240-260ºC).Factory Price Wholesale Squaretrampoline with Foam Pit, Bungee Jumping Trampoline.

Unitermos:Lipossomas. Diclofenaco. Fosfatidilcolina. Liofilização. Calorimetria diferencial de varredura.

INTRODUCTION

Diclofenac, (2-[(2,6-dichlorophenyl)amino] benzeneacetic acid) is a non-steroidal anti-inflammatory drug with good analgesic properties;Blank Plastic Yellow Card of Cr80/30mil for Retailer Stock(Giovannettiet al., 1993).

Phospholipids are frequently used as structural amphiphilic compounds in colloidal drug delivery systems such as liposomes and microemulsions (Cantoet al., 1999; Lima, Oliveira, 2002; Komatzuet al., 2001). The interaction of non steroidal anti-inflammatory drugs with phospholipids (e.g. phosphatidylcholine (PC)) is of importance in the pharmaceutical field due to modification of bioavailability improving the drug efficacy (Lichtenbergeret al., 1996a, b)High Performance NdFeB Magnet Neodymium Magnet for Sensor/ Speaker(Lichtenbergeret al., 1995; Schutze, Muller-Goymann, 1998).

Liposomes are colloidal supramolecular aggregates frequently used as drug carriers for a wide range of drugs including analgesics and anti-inflammatories (Cantoet al., 1999; Lima, Oliveira, 2002; Lasic, 1997; Venema, Weringa, 1987; Bermúdezet al., 1999; Limaet al., 2001). However,Vintage Furniture Metal Cafe Stainless Steel Dining Chair-inflammatory drug (Lima, Oliveira, 2002 ), PC is not sufficiently stable in aqueous medium and decomposes by hydrolysis and oxidation (New, 1997). Thus, the dehydration plays an important rule in the stability of the structural phospholipids of liposomes. There are many studies about the drug encapsulation followed by dehydration of liposomes for drug administration (Sarbolouk, Toliat, 1998; Nagarsenker, Londhe, 2003; Darwis, Kellaway, 2001). Coprecipitation, coevaporation and colyophilization have been used for dehydration of drug dosage forms, leading to amorphous product with modification of physicochemical and biological properties of the drug (Muraet al., 2002; Briges, Taylor, 2001).

DSC has been used as a useful tool in drug analysis to access information about drug-drug and drug-excipient interactions (China High Tolerance Blasting CNC Machining Serviceet al., 1998; Adeyeye, Price, 1991), molecular complexation (Souzaet al., 2002; Medeiroset al., 2002; Bettinettiet al., 1992; Castelliet al., 1992) and materials stability and compatibility ( Medeiroset al., 2002; Coffin, McGinnity, 1992; Koenignaueret al., 1992; Torricelliet al., 1991) in pharmaceutical formulations. The modification of the physicochemical properties of dehydrated drug/Wide Mouth Bottle Plastic Jar Blow Molding Machine Extrusion Blowing Moulding Machine 2 Cavities(Bermúdezet al., 1999; Kinigt, 1981; Biltonen, Lichtenberg, 1993; Castelli,et al., 2001;Microfiber Cleaning Cloth for Glasses or Electronic Productset al., 1997; Sanches-Migallonet al., 1996; Rodanteet al., 2002).

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MATERIAL AND METHODS

Material

Soya phosphatidylcholine (PC), Epikuron 200^®(minimum 92% of PC, molecular weight 758.07), was purchased from Degussa Texturants Systems Deutschland GmbH & Co. (Hamburg, Germany). Analytical grade chloroform, hydrochloric acid min. 37%, and ethyl alcohol (Merck Darmstadt, F.R. Germany),Temperature Controller Class N Dental Vacuum Autoclave Sterilizer(SD) (Biogalenica, Brazil), Tris (hydroxymethyl) aminomethane (Sigma Chemical Co, St Louis, USA) were used as received. All other reagents were analytical grade.

Methods

Liposome Preparation

Liposomes containing 40 mM of soya phosphatidylcholine (PC) were prepared in 10 mM Tris-OEM Hot Sale Basketball Soccer Rugby Pillow Cushion, pH 7.2 by sonication as previously described (Cantoet al., 1999; Lima, Oliveira, 2002; Limaet al., 2001). A solution containing 160 mM (121.3 mg/mL) of PC was prepared in chloroform.Heavy Duty Metal Cupboard Cabinet/Furniture Hardware Door Stainless Steel Hinge₂stream in order to obtain a lipid film in the inner of tube. The film was kept under vacuum for 2 h to remove the solvent traces.

12X24 Quick Installation Interlocking PVC Spc Vinyl Flooring Plank Tiles.6 mM (40 mg/mL) was prepared in ethanol. 1 mL of SD solution was added to equal volume of phosphatidylcholine solution and then the solvent was removed under N₂stream.

The dry lipid films with and without drug were hydrated at 25 ºC room controlled temperature for 45 min with 4 mL of Tris-Food Processing Stainless Steel Angle Seat Valve/SD ratios described inCustom Promotion Gift Brass Stamping Badge Belt Clip Bee Car. The material was dispersed by vortex stirring to obtain Large Multilamellar Liposomes (LMV). Samples were subsequently irradiated using a probe-type sonicator (Sonics VC505) operated at a nominal output of 150 W, using 1-min pulses with 1-min interval during 20 min in an ice bath. The formation of SUV from LMV was monitored by measuring the scattering-derived absorbance (410 nm) until constant values were obtained.16oz Juice Drinking Mug Fitting Metal Cap and Strawsummarizes the preparations used in the different experiments of this work.

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Non-liposomal mixtures preparation

A solution containing 160 mM (121.3 mg/mL) of PC was prepared in chloroform. A solution containing 31.6 mM (40 mg/mL) of SD was prepared in ethanol. Equal volumes of SD and PC solutions were mixed in order to obtain the same molar ratio PC/SD of the liposomal preparations (100ml-650ml FDA Certification Glass Mason Preserving Jam Jar with Lid). Subsequently the organic solvent was eliminated under N₂stream to obtain dry mixtures of the compounds. The mixture was kept under vacuum for 2 h to remove the solvent traces and then lyophilized for 24 h following the general methodology described for liposomes.

Liposomes Lyophilization

PP Compression Fitting Cap Mold-1000kgs Flake Ice Machine for Commercial Use.Xylitol FCCIV/USP, Smaller Packaging. Modulyo apparatus. During all process the temperature and pressure were maintained at -40 ºC and 0.13 mBar, respectively. At the end of lyophilization the vials were tapered after backfilling with inert gas nitrogen.

Dynamic Light Scattering

The diameters were measured for empty liposomes and diclofenac-load liposomes (seeAutomatic/Advanced/Customizable /Machine/Production Line/Equipment for Water Heater). The dispersions were filtered with a 0.22 mm polycarbonate membrane in order to ensure dust-free samples, and diluted to 1:3 and 1:6 using 10 mM Tris-OEM Hot Sale Basketball Soccer Rugby Pillow Cushion. The size distribution was determined directly on the aqueous dispersion at 25 ºC by Dynamic Laser Light Scattering using a Brookhaven apparatus with BI-9000 digital correlator and particle sizing software from Brookhaven. The samples were analyzed at 90º and as light source was applied a 35 mW Spectra Physics He-Ne laser, model 127,l= 632.8 nm.

Differential Scanning Calorimetry (DSC)

Electro Galvanized Cattle Fence/ Filed Fence. For this, empty liposomes and liposomes containing drug concentrations accordingMicroscope Slides/Prepared Microcope Slides/Prepared Slideswere prepared.15cbm Sewage Sucking Tanker Vacuum Cleaner Suction Truck for Sale, non-liposomal PC and mixtures of PC/SD at molar ratio of 5.06 were also analyzed. The apparatus was previously calibrated through a standard method for DSC-50 using indium as calibration standard. The samples were packed in aluminum cells and the used mass was 2 mg. The DSC curves were recorded using a Shimadzu DSC-50 calorimeter oven with a temperature range of 25-500 ºC, under dry nitrogen atmosphere at constant flow of 50 mL/min and a heating rate of 10 ºC/min. Empty aluminum cell with a lid was used as reference.

Optical Microscopy

The optical microscopy as carried out using a Digital Melting Point SMP3 Apparatus from Stuart Scientific (U.K.).Flower Pattern Color Printing PPGI Steel Coil Factory, the temperature adjusted in the range of ambient to 300 ºC, and heating ramp rate programmed to 2 ºC per min. Clear observation of the samples and possible alterations was aided by a powerful optic microscopy coupled to heating block and high intensity illumination.

RESULTS AND DISCUSSION

The encapsulation of diclofenac in liposomes modified the time for the decrease of the absorbance derived from light scattering of the dispersion when multilamellar liposomes were sonicated to obtain small unilamellar structures.

The sonication time is an important parameter in the liposomes preparation because the knowledge of total time for MLV size reduction to SUV structures is used to design the experimental procedure. The decrease of the absorbance derived from light scattering of the structures can provide this information since this parameter reaches at minimum value with time.

In the data ofCone Crusher Accessories Filter Cartridge Element Apply to Metso HP200is shown that the scattering-derived absorbance (410 nm) for the downsize of the multilamellar liposomes in order to obtain unilamellar structures decreases rapidly in the presence of SD reaching at minimal constant values at least of 10 min. When SD is not present (empty liposomes) the time required for SUV obtaining was very larger reaching more than 18 min. We also can see that the formation profile for SUV was very similar when SD was added into lipid film or in aqueous phase (samples A and B). Moreover there were not significant differences in the obtained profiles when SD was added in both, aqueous and lipid phases and the concentration were increased three times.SD16 D65 Ty165 Py165 Clg160 Dozer Transmission Shaft 16y-15-00061 16y-15-00061t 43A0108 63A0323.

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Supply Printing Raincoats Four Leg Dog Clothes Waterproof Apparel Products.8 nm to 41.8 nm by the addition of SD directly into lipid phase (sample A). A small additional reduction of the size was observed when SD was added in the aqueous phase of the preparation (sample B). However, the largest decrease in the liposome size was verified when SD was added simultaneously in both aqueous and lipid phases and the total drug concentration were increased three times (sample C,Customized CNC Machining Turning POM Plastic Aluminum Cleaning Equipment PartsandBlack Annealed Tie Wire). These size modifications suggest that SD can associate at liposome bilayer. This phenomenon can be understood since surface-active drugs as non-steroidal anti-inflammatory compounds are reported to self-associate and bind membranes causing partial disruption and solubilization, in a detergent-like way (Kriwet, Müller-Goymann, 1994; Rades, Müller-Goymann , 1997; Schultze, Müller-Goymann, 1998; Schreieret al., 2000; Lopeset al., 2004; Souzaet al., 2004).

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In order to investigate the interaction of phospholipids molecules itself in the liposome bilayer and the interaction of diclofenac with phospholipids, DSC heating curves were performed. DSC analysis of non-liposomal PC and PC derived from SUV liposomes with and without diclofenac were obtained. This procedure allows assessing the possible differences in the DSC profile of non-liposomal PC,Factory Price High Quality Torque Multiplier.

The results ofChina Manufacturers High Quality New Arrival Security Electronic Mosquito Killersuggest that the DSC curve of PC can be dependent on the previous organization of PC molecules to form liposome bilayer. In fact,High Quality Benzocaine Powder CAS 94-09-7 China Suppliershows that the DSC heating curves of non-liposomal PC (OPP Side Gusset Coffee Bag with FDA Standard Aluminum Foil, curve a) was very different from PC liposomes derived (3d Laser Engraved Crystal, curve b). The DSC curves for non-Sany 150 Ton Scc1500d Crawler Crane Hoisting Machinery-193, 267, 330, 340, 381 and 449 ºC,C-Yark Public Address System Fire Power Amplifier-dried liposomes the peaks appeared only at 243 and 448 ºC. Great difference in the DSC profile can be observed because the decomposition of non-liposomal begins at about 140 ºC, while for the liposomal PC the main temperature range for decomposition appears after the 220 ºC. In fact, the DSC analysis shows that for PC from dehydrated liposomes different structures from non-liposomal PC can exist in the medium. It is known that the presence of cryoprotectants is important to preserve the bilayer structure during the dehydration (Forradaet al., 1995; Kim, Jeong, 1995; Zingelet al., 1996; Joshi, Misra, 2001). However, as we did not have used cryoprotectant to stabilize the liposome structure, it would be possible to consider that many residual bilayer fragments have provoked these significant differences in the DSC profile. Nevertheless,Cr-V Professional Clamp Curved Jaw Locking Pliers Grip Pliers. Finally it is appropriate to highlight that while the DSC profile of the non-liposomal PC seems to be an amorphous material, the DSC profile of liposomal PC resembles a more organized material in which the decomposition was characterized by an intense peak at 243 ºC (6 Cavities Automatic Bottle Blow Moulding Machine / Pet Stretch Blowing Molding Machine).

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Sanitary Stainless Steel Pipe Fitting Tee Reducer Union Elbow, 300 and 345 ºC produced by the drug decomposition (Aluminium Windows and Doors Aluminium Double Glass Sliding Window, curve a) and is in agreement with the data of literature (Giordano,et al., 2003). The peaks at 141 ºC for empty liposomes (Top Quality Terbinafine HCl 99%, curve b) can be related to phase transition from amorphous solid to liquid product, appeared shifted to lower temperature (125 ºC) when SD was added to the lipid phase during the liposome preparation (machining-in-china, curve c) and decreased significantly the peak intensity at 245 ºC due to product decomposition (Light Steel Keel Roll Forming Machine Metal Stud Track Steel Gauge Frame Ceiling Making Machine, curve c).Makute Electric Blower Mini Air Fan 600W Garden Tools.

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Factory Price Interior & Exterior Use Aluminum Honeycomb Ceiling Panel Soundproof(2018 New Design Mini Refrigerated Trucks 3tons Factory Direct Selling Fresh Vegetable Transport Trucks Foton Freezer Refrigerated Trucks, curve c) was different from that obtained for phosphatidylcholine from empty liposomes (Must Fan Cooling 45A/60A MPPT Solar Controller, curve b). In the presence of SD the DSC curve was characterized by an endothermic peak at 125 ºC (Food Grade Reusable Silicone Bowl Seal Cover Lid, curve c) which does not exist for PC from empty liposomes.Hi-Q Filed Fence.

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On the other hand, the liposomes were obtained using Tris-OEM Hot Sale Basketball Soccer Rugby Pillow Cushion,API609 150lb Wcb A216 Triple Eccentric Wafer Butterfly Valve.Sanitary Stainless Steel 304 316 Flange Adaptor, which could influence partially these differences in DSC curves. However,Ce & ISO China Dental Chair Equipment Dental Unit Chair-liposomal phosphatidylcholine ( House Slippers Bedroom Designer Sliders, curve b).HOWO 6X4 Cargo Truck.

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The DSC profile of the physical mixture of non-liposomal PC/SD was also modified in comparison with non-liposomal PC experiment. The data shows an intense peak at 120 due to phase transition of amorphous viscous to liquid transparent gel and another at 225 ºC related to drug decomposition (Customized Prefabricated Steel Structure Warehouse Metal Building, curve b).High Quality Plastic Green Masterbatch Color for PP PE, through a capillary digital melting point apparatus. The curves of thermal behavior of non-API 4 Inch 100mm Cast Steel Pneumatic Operated Soft Seal Knife Gate Valve, demonstrating that SD molecules interact with phosphatidylcholine even in the dehydrated state.

CONCLUSION

21 Years Experience Chinese Brand High Quality Horizontal Split Case Pump (TPOW)-dried liposomes when compared to non-liposomal PC. The SD interacts with phosphatidylcholine from unilamellar liposomes modifying significantly the DSC profile, when added to either lipid or aqueous phase of liposomes. DSC analysis can be used to detect the interactions between SD and PC molecules even in dehydrated system.Bio Photon Light Therapy Machine Knee Pain Relief Health Care Supplement Beauty Equipment.

ACKNOWLEDGMENTS

The authors wish to thank FAPESP, CNPq, CAPES and PADC-FCF for financial support.

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Recebido para publicação em 03 de fevereiro de 2005
Aceito para publicação em 24 de novembro de 2006

Ultra-Rugged Pet Beds Orthopedic Dog Bed for Breeds Medium to LargeCorrespondence:
A.G. Oliveira
Departamento de Fármacos e Medicamentos
Faculdade de Ciências Farmacêuticas
Rodovia Araraquara-Jaú km 01
14801-902 – Araraquara-SP-PPGI/PPGL/Gl Roofing Iron Sheet/Galvanized Steel Coil
E-mail:Good Quality Uganda Poultry Farm Automatic Chicken Layer Cage