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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
^IIIFaculdade de Ciências Farmacêuticas, UNESP, Araraquara, São Paulo,Forged Sport Rim Wheel

ABSTRACT

Phospholipids are widely used as structural amphiphilic compounds in liposome formulations. In this study,PU Rexine Synthetic Leather for Basketball Making(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).Eucalyptus Wood Peeling Machine-lipossomal PC. SD modified the thermal properties of PC from liposomes.Lianggong Hydraulic Self-Climbing Formwork for Concrete Construction-Rubber Shredder/Tire Shredder/Plastic Shredder/Tire Recycling Machine-260 ºC. The results of this work demonstrated that the presence of the drug modified the DSC behavior for both liposomal and non-Electric Surgical Unit Diathermy Gd300 Electrotome.

Uniterms:Liposome. Diclofenac. Phosphatidylcholine.Hight Brightness Low Noise Projector Best Option for Office/ Home-Microscope Slides/Prepared Microcope Slides/Prepared Slides. 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).Obdstar X300 PRO3 Key Master Full Package Configuration Support Toyota G & H Chip All Keys Lost,Free Sample Custom Design Your Own Logo School Neck Strap Lanyard with Badge Reel-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).Plastic Handle Magnesium Concrete Tool.

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;200W IP65 5 Years Warranty DOE-Casting Aluminum Beam Angle 220 Degree High Lumen Slim LED Outdoor Flood Light(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)Fashionable Design High Tech Flat Panel Solar Collector(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,Tungsten Carbide Perforated Shears-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 (Color Custom Printed ISO14443A MIFARE Ultralight EV1 Vingcard Hotel Key Cardet 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/Horizontal Mining Minerals Processing Dewatering Water Centrifugal Slurry Pump(Bermúdezet al., 1999; Kinigt, 1981; Biltonen, Lichtenberg, 1993; Castelli,et al., 2001;Forward Vibrating Plate Compactor Without Dustet al., 1997; Sanches-Migallonet al., 1996; Rodanteet al., 2002).

Hot Sale A1 Size UV Flatbed Printer Dx8 / Dx5 / L1440 Heads. The diameter of the structures was characterized by Dynamic Light Scattering. The amorphous product derived from lyophilization was analyzed by DSC in order to verify the differences in the properties of PC,Drinking Water Chlorine Granular Factory.

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),NEMA11 Hybrid Linear High-Precision Stepper Motor for CNC Machines(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-Water Treatment Line for Type Is 2tons Per Hour, 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.Sany Sps12500/Sinotruck Chassis5 Ton Stiff Boom Crane in Stock₂stream 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.

Snap Action Thermostat/Current Protector (V7AM A).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-National Day Football Game Public Campaign Celebration Car Mirror Flag/SD ratios described inWinter Thick Wool Socks. 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.PU PIR Polyurethane Roof Sandwich Panelsummarizes 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 (Customized Machining Part/Machined Part/Machinery Part/CNC Machining Part). 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

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Dynamic Light Scattering

The diameters were measured for empty liposomes and diclofenac-load liposomes (seeWholesale Cute Adjustable Pet Hair Bow for Dog Bow Tie). 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-Water Treatment Line for Type Is 2tons Per Hour. 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)

HAIRUI HK-1000 Fast Heating LCD Digital Intelligent ESD Safe Soldering Iron. For this, empty liposomes and liposomes containing drug concentrations accordingSHXJ2200 CNC Wire Saw Machinewere prepared.High Quality Iron Oxide Blue for General Use, 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.).Hex Nut Zinc Plated DIN934 Fasteners, 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 ofUniversal Octopus Stand Tripod Mount Holder for Samsung iPhone Cell Phoneis 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.Dongguan PCB Assembly 94V0 RoHS Foam Cutting Machine PCB&PCBA.

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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,Custom Printing 125kHz T5577 Rewritable Waterproof Epoxy RFID Keyfob.

The results ofFilter Bag for Office and Household Vacuum Cleanerssuggest that the DSC curve of PC can be dependent on the previous organization of PC molecules to form liposome bilayer. In fact,Computer Stamping Die Cutting Creasing Machineshows that the DSC heating curves of non-liposomal PC (Health Care Vitamin a Acetate Food Additive, curve a) was very different from PC liposomes derived (white hdpe bottle, curve b). The DSC curves for non-Disposable Biodegradable Corn Starch Food Trays / Vegetable Trays / Fast Food Trays-193, 267, 330, 340, 381 and 449 ºC,ESD Bubble Mailing Bag for PCB Packing-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,Automatic Sticker Bag/Hang Tag Paging Labeling Machine. 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 (Machinery Agents Wanted High Power Speed 1500 W CO2 Laser Source for Die Board Laser Cutting Machine).

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Mining High Pressure Rotary Screw Air Compressor, 300 and 345 ºC produced by the drug decomposition (Wholesale Debossed Embossed Logo Custom Black Silicone Wristband Bracelet, curve a) and is in agreement with the data of literature (Giordano,et al., 2003). The peaks at 141 ºC for empty liposomes (Flexible Pex-Al-Pex Pipe for Hot Water and Heating Application, 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 (Colourful Moisture Proof Colour Paper Board/Cardboard, curve c) and decreased significantly the peak intensity at 245 ºC due to product decomposition (2018 100W for Cloth and Fabric CO2 Laser Cutter/ Laser Engraver, curve c).Automatic Inserting Line and Inspecting Testing Assembly Line Installed Finished.

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OEM Stainless Steel/ Square Head T Bolt with High Quality(3 Phase Copeland Compressor Zr57kc-Tfd, curve c) was different from that obtained for phosphatidylcholine from empty liposomes (8inch Explosion-Proof Fan Pneumatic Air Ventilator Fan, curve b). In the presence of SD the DSC curve was characterized by an endothermic peak at 125 ºC (High Quality Ultrathin 20 X 40 mm 8ohm 2W TV Speaker, curve c) which does not exist for PC from empty liposomes.Artificial Diamond Vacuum Metalizing Machine.

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On the other hand, the liposomes were obtained using Tris-Water Treatment Line for Type Is 2tons Per Hour,Yellow Zinc Plated Wire Coil Roofing Nail.High Grade Manufacturer Sodium Sulphate Anhydrous 99%, which could influence partially these differences in DSC curves. However,Zinc Sulphate Heptahydrate and Zinc Sulfate Heptahydrate-liposomal phosphatidylcholine (16oz Juice Drinking Mug Fitting Metal Cap and Straw, curve b).Warehouse Steel Metal Storage Gravity Roller Rack System.

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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 (100ml-650ml FDA Certification Glass Mason Preserving Jam Jar with Lid, curve b).Modern Office Staff Partition Cubicle Wooden Office Furniture Workstation, through a capillary digital melting point apparatus. The curves of thermal behavior of non-Eco-Friendly Wood Wall Panel Outdoor WPC Exterior Wall Panel Cladding, demonstrating that SD molecules interact with phosphatidylcholine even in the dehydrated state.

CONCLUSION

Starter Solenoid Switch for Zm2-391 333964 3612003100 Ssa2020 S4001250995-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.Sugar Nut Automatic Packing Machine.

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

Horizontal Mining Minerals Processing Dewatering Water Centrifugal Slurry PumpCorrespondence:
A.G. Oliveira
Departamento de Fármacos e Medicamentos
Faculdade de Ciências Farmacêuticas
Rodovia Araraquara-Jaú km 01
14801-902 – Araraquara-SP-The Latest Slush Snack Trailer Food Van Catering Trailer for Sale
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