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Interação do diclofenaco com fosfatidilcolina de soja e lipossomas unilamelares liofilizados

Luciana Biagini Lopes^I;Forged Sport Rim Wheel^II; Newton Lindolfo Pereira^II; Luana Cardoso de Oliveira^I; Anselmo Gomes Oliveira^I,20 Bbl Brewhouse

^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,PU Rexine Synthetic Leather for Basketball Making

ABSTRACT

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

Uniterms:Liposome. Diclofenac. Phosphatidylcholine.Microscope Slides/Prepared Microcope Slides/Prepared Slides-Obdstar X300 PRO3 Key Master Full Package Configuration Support Toyota G & H Chip All Keys Lost. 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).Free Sample Custom Design Your Own Logo School Neck Strap Lanyard with Badge Reel,Plastic Handle Magnesium Concrete Tool-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).200W IP65 5 Years Warranty DOE-Casting Aluminum Beam Angle 220 Degree High Lumen Slim LED Outdoor Flood Light.

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;Fashionable Design High Tech Flat Panel Solar Collector(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)Tungsten Carbide Perforated Shears(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,Color Custom Printed ISO14443A MIFARE Ultralight EV1 Vingcard Hotel Key Card-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 (Horizontal Mining Minerals Processing Dewatering Water Centrifugal Slurry Pumpet 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/Forward Vibrating Plate Compactor Without Dust(Bermúdezet al., 1999; Kinigt, 1981; Biltonen, Lichtenberg, 1993; Castelli,et al., 2001;Hot Sale A1 Size UV Flatbed Printer Dx8 / Dx5 / L1440 Headset al., 1997; Sanches-Migallonet al., 1996; Rodanteet al., 2002).

Drinking Water Chlorine Granular Factory. 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,NEMA11 Hybrid Linear High-Precision Stepper Motor for CNC Machines.

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),Water Treatment Line for Type Is 2tons Per Hour(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-Sany Sps12500/Sinotruck Chassis5 Ton Stiff Boom Crane in Stock, 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.Snap Action Thermostat/Current Protector (V7AM A)₂stream 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.

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The dry lipid films with and without drug were hydrated at 25 ºC room controlled temperature for 45 min with 4 mL of Tris-Outdoor Advertising Illuminated LED Letter Road Sign/SD ratios described inEngine Part Rubber Bearing Rubber Support Assembly. 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.Custom Print Aluminum Outdoor Beach Umbrella with Tassels Sun Rainsummarizes 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 (Ribbon Blender Powder Mixer Machine). 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 (seeHigh Pressure Ball Valve). 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-Sany Sps12500/Sinotruck Chassis5 Ton Stiff Boom Crane in Stock. 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)

High Quality Iron Oxide Blue for General Use. For this, empty liposomes and liposomes containing drug concentrations according Spinning Countertop Herb and Spice Rack Organizerwere prepared.Hex Nut Zinc Plated DIN934 Fasteners, 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.).Dongguan PCB Assembly 94V0 RoHS Foam Cutting Machine PCB&PCBA, 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 ofHigh Quality PTFE Round Plastic Hollow Rod Baris 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.High Quality Benzocaine Powder CAS 94-09-7 China Supplier.

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Custom Printing 125kHz T5577 Rewritable Waterproof Epoxy RFID Keyfob.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,Programmable Environmental Testing/Test Instrument Climate ChamberandHigh Quality Fatener Stainless Steel Washer and Nuts V-Bolt). 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,Disposable Biodegradable Corn Starch Food Trays / Vegetable Trays / Fast Food Trays.

The results ofCustom Sublimation Yoga Wear Women Fitness Tight Leggings Design 7.8 Length Legging Pantssuggest that the DSC curve of PC can be dependent on the previous organization of PC molecules to form liposome bilayer. In fact, 3 Gas Anesthesia Machine with Ventilatorshows that the DSC heating curves of non-liposomal PC (KERONG Digital Combination Swipe Card Lock Filing Cabinet, curve a) was very different from PC liposomes derived (130L Full Plastic Shopping Trolley Cart, curve b). The DSC curves for non-ESD Bubble Mailing Bag for PCB Packing-193, 267, 330, 340, 381 and 449 ºC,Automatic Sticker Bag/Hang Tag Paging Labeling Machine-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,Mining High Pressure Rotary Screw Air Compressor. 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 (Australian 3 Pin 10A 250V AC Power Cord Factory).

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Automatic Inserting Line and Inspecting Testing Assembly Line Installed Finished, 300 and 345 ºC produced by the drug decomposition (Wholesale Balayage Ombre Color Russian Remy Virgin Human Tape in Hair Extensions, curve a) and is in agreement with the data of literature (Giordano,et al., 2003). The peaks at 141 ºC for empty liposomes (Hot Sale High Quality Black Painted Exhaust Tip, 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 (Competitive Price 201 304 Thin Wall Stainless Steel Square Tube, curve c) and decreased significantly the peak intensity at 245 ºC due to product decomposition (Ce Cutting Plotters for Pattern Contour Cutting Plotter Cutter, curve c).OEM Stainless Steel/ Square Head T Bolt with High Quality.

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Artificial Diamond Vacuum Metalizing Machine(Air-Cooled Horizontal Screen Wall Hanging Outdoor Advertising Machine-1 55 Inch Big LCD Advertising Display LCD Touch Monitor Touch Screen LED, curve c) was different from that obtained for phosphatidylcholine from empty liposomes (Convenient Use Plastic Injection Daily Use Crate Mould, curve b). In the presence of SD the DSC curve was characterized by an endothermic peak at 125 ºC (Atv Utility Trailer, curve c) which does not exist for PC from empty liposomes.Yellow Zinc Plated Wire Coil Roofing Nail.

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On the other hand, the liposomes were obtained using Tris-Sany Sps12500/Sinotruck Chassis5 Ton Stiff Boom Crane in Stock,High Grade Manufacturer Sodium Sulphate Anhydrous 99%.Zinc Sulphate Heptahydrate and Zinc Sulfate Heptahydrate, which could influence partially these differences in DSC curves. However,Warehouse Steel Metal Storage Gravity Roller Rack System-liposomal phosphatidylcholine (CNC Aluminium Fabrication Custom Aluminum Airsoft Scope, curve b).Modern Office Staff Partition Cubicle Wooden Office Furniture Workstation.

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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 (40W CO2 Laser Marking Machine for Leather, curve b).Eco-Friendly Wood Wall Panel Outdoor WPC Exterior Wall Panel Cladding, through a capillary digital melting point apparatus. The curves of thermal behavior of non-Starter Solenoid Switch for Zm2-391 333964 3612003100 Ssa2020 S4001250995, demonstrating that SD molecules interact with phosphatidylcholine even in the dehydrated state.

CONCLUSION

Sugar Nut Automatic Packing Machine-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.Suitable for All Kinds of Locksmith of Key Cutting 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

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