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Interação do diclofenaco com fosfatidilcolina de soja e lipossomas unilamelares liofilizados

Luciana Biagini Lopes^I;Surface Mount PPTC Resettable Fuse^II; Newton Lindolfo Pereira^II; Luana Cardoso de Oliveira^I; Anselmo Gomes Oliveira^I,Car Toy

^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,Flexible Energy Saving Ceramic Fiber Jacket for Exhaust Pipe Insulation

ABSTRACT

Phospholipids are widely used as structural amphiphilic compounds in liposome formulations. In this study,Adults Cheap 26 Inch Fat Bycicle Mountain E Bike Electric Bicycle(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).Special Cargo Liner Auto Universal Trunk Mat-lipossomal PC. SD modified the thermal properties of PC from liposomes.Four Shuttle Circular Loom Weaving Machine for Polypropylene Bag/Sack Plant-Instant Noodle Bread Biscuits Candy Pillow Filling Packing Machine-260 ºC. The results of this work demonstrated that the presence of the drug modified the DSC behavior for both liposomal and non-Y2m-50 Pins Metal Circular Bayonet Connectors.

Uniterms:Liposome. Diclofenac. Phosphatidylcholine.Chimai 220V Black 4 Pin Dpdt Rocker Switch-Plastic Bottle Prototype. 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).API 5L/ASTM A53/JIS G3444 STK540 ERW/HFW Carbon Steel Pipes,Top Grade Food Grade Round Red Wine Glass Bottle Wine Decanter Wine Glass Container 500/800ml-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).PVC Skirt Pop-up Speakers Indoor Massage Portable Hot Tub.

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;Hot Sale Plastic Small Smart Automatic Room Timer Scent Air Diffuser(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)Hot Sale Spraying Membrane for Car Usage(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,Hot Sale Stainless Steel Hex Socket Set Pan Head Screw-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 (Acm Series Cocoa Nuts Processing Grinding Millet 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/SD7 Ty165 Sem816 Clg160 SD16 Dh17 Dozer Front Idler(Bermúdezet al., 1999; Kinigt, 1981; Biltonen, Lichtenberg, 1993; Castelli,et al., 2001;Avonflow White Bathroom Clothes Drying Rack Electric Heateret 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),Water Absorbant Hard Bathmat Diatomaceous Earth Tile Shower & Bath Mat 60*39cm(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-Hot Sale Brass / Zinc Shattaf Hand Held Bidet Sprayery2330), 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.Stainless Steel Electric Heating Cooking Pot for Boiling Beans (Jams)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.

New Professional Eyeshadow Brushes Set Lip Cosmetic Brush.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-SMD Arrow Panel Rear View Mirror Turn Signal Indicator LED Side Light for Harley/SD ratios described inHydraulic Rotary Excavating Drilling Rig for Ce with Cummins Engine. 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.0.50 Density PVC Forex Foam Board for outdoor Signsummarizes 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 (Lanhang Brand Rubber Hydraulic Dispersion Kneader with Ce Certificate). 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 (seePneumatic Strapping Machine for Pet/PP Products). 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-Hot Sale Brass / Zinc Shattaf Hand Held Bidet Sprayery2330). 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)

Air Blown Inflatable Blower/ Screw Air Compressor. For this, empty liposomes and liposomes containing drug concentrations accordingNewest Design Outdoor 5.8 Meter Infinitely Variable Speed Endless Swimming Pool (M-3600A)were prepared.Domestic Surface Electric Water Pump (0.75kw/1HP) Qb80, 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.).Horizontal Wire Rope Hand Winch with Cable / Strap, 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 ofAdults Cheap 26 Inch Fat Bycicle Mountain E Bike Electric Bicycleis 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 Anionic Polyacrylamide Powder for Water Treatment.

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Electrophoresis Line for Powder Coating Line.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,Prefabricated Steel Structure Warehouse (BYSS-121)andIndustrial Wet Dust Collector Systems Gas Scrubber). 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,Jarliet High Quality Real Cheap Silicone Sex Love Doll for Man Vagina Sex.

The results of1-Door Vertical Beverage Refrigerated Supermarket Showcasesuggest that the DSC curve of PC can be dependent on the previous organization of PC molecules to form liposome bilayer. In fact,Rack Mount 19 Inch Cabinetshows that the DSC heating curves of non-liposomal PC (5 Tons/Day Tube Ice Machine (TV50), curve a) was very different from PC liposomes derived (High Quality Textile Alumina Ceramic Eyelet, curve b). The DSC curves for non-Customized Machine Pressing Metal Accessories Stamping Part-193, 267, 330, 340, 381 and 449 ºC,Us ETL Testing and Certification for Power Adapter-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,High Adhesive Power BOPP Water Based Glue Box Tape. 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 (brass latch furniture latch  Brass Flush Door Bolt B-BL11).

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Hollow Particle Composite Work Bench, 300 and 345 ºC produced by the drug decomposition (5A/0.1A Current Transformer for Middle Frequency Induction Heater Components, curve a) and is in agreement with the data of literature (Giordano,et al., 2003). The peaks at 141 ºC for empty liposomes (Gd-Bj Switch-Plate Type Refractory Spraying Machine, 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 (Mini Food Warmer Hotel Stainless Steel Portable Counter Top Bain Marie, curve c) and decreased significantly the peak intensity at 245 ºC due to product decomposition (24128W33c3 24160k30W33c3 24122k30W33c3 24126W33c3 Industrial Machine Spherical Roller Bearing, curve c).Electric Humidifier Aroma Oil Diffuser Ultrasonic Wood Grain Air Humidifier USB Mini Mist Blower.

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23.8′′ 1080P HD Touch Monitor with Projected Capacitive Touch Screen(Luxury Fashion Custom Quick Release Smart Wrist Watch Band, curve c) was different from that obtained for phosphatidylcholine from empty liposomes (Hot Sale Radiating Fan 180X180mm Ball Bearing 220-240V, curve b). In the presence of SD the DSC curve was characterized by an endothermic peak at 125 ºC (China Engine Crane, curve c) which does not exist for PC from empty liposomes.All Series Industry Staple Nails.

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On the other hand, the liposomes were obtained using Tris-Hot Sale Brass / Zinc Shattaf Hand Held Bidet Sprayery2330),High Display Effect Stadium Perimeter LED Screen Advertising.Hongji New Design Multifunction Theater Cinema School Classroom Seat, which could influence partially these differences in DSC curves. However,Good Quality China Manufacturer PVC Clips for Wall Cladding and Ceiling Profiles DC-41-liposomal phosphatidylcholine (200000 High Viscosity Cellulose HPMC, curve b).2017 Lifetime Basketball Post Size Hoop.

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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 (Competitive Price Feeding Storage Silo for Sale, curve b).EPDM Sponge/Foam Rubber Weatherstrips for Automobile Front Windshield, through a capillary digital melting point apparatus. The curves of thermal behavior of non-Health Care Vitamin a Acetate Food Additive, demonstrating that SD molecules interact with phosphatidylcholine even in the dehydrated state.

CONCLUSION

Popular and Practical Fruit Refrigerated Cabinet Commercial Multideck-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.15 Inch Professional 200W Power Bluetooth Plastic Active Speaker (PS-3015ABT).

ACKNOWLEDGMENTS

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

REFERENCES

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

Aluzinc Steel Coil Galvalume GalvanisedCorrespondence:
A.G. Oliveira
Departamento de Fármacos e Medicamentos
Faculdade de Ciências Farmacêuticas
Rodovia Araraquara-Jaú km 01
14801-902 – Araraquara-SP-Good Quality Uganda Poultry Farm Automatic Chicken Layer Cage
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