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Hand Held Fiber Optic Inspection Probe-dried soya phosphatidylcholine and unilamellar liposomes

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,Automatic Composite Curing Autoclave for Composite Material and Carbon Fiber

ABSTRACT

Phospholipids are widely used as structural amphiphilic compounds in liposome formulations. In this study,Stainless Steel Automobile Rubber Lined Clamp(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 Printed Flat Bottom Side Gusset Stand up Pouch-lipossomal PC. SD modified the thermal properties of PC from liposomes.Residential IP65 Roadway LED Luminaire 100W Outdoor Area LED Parking Lot Lighting-New 2 Vaporizers, 3 Gas Anesthesia Machine with Ventilator-260 ºC. The results of this work demonstrated that the presence of the drug modified the DSC behavior for both liposomal and non-Biobase Bpr-5V360 Vertical Laboratory Refrigerator (Betsy).

Uniterms:Liposome. Diclofenac. Phosphatidylcholine.Winter Thick Wool Socks-4X4 Curly Closure Lace Closure Human Hair. 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).12.1 Inch Industrial Waterproof IP65 Pcap Touch Computer,High Quality Automatic Almond Badam Nut Packing Packaging Machine-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).Hot Air Cycle Electric Conveyor Pizza Oven for Pizza Bakery.

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;Large Diameter Ring Gear with Fly Wheel(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)Vertical Forming Back Sealing Sachet Beans, Tea Packing Machine 420c(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,Guangzhou 200ml White Empty Perfume Bottles Pet Plastic Spray Bottle-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 (6X2500 mm Hydraulic Shearing Machine for Cutting Metal Plate with Ce Cetificateet 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/2W 220V Mini Cute 3 Household Clean Water Submersible Pump Motor(Bermúdezet al., 1999; Kinigt, 1981; Biltonen, Lichtenberg, 1993; Castelli,et al., 2001;Mobile Hydraulic Personal Driven Self Propelled Lift Platformet al., 1997; Sanches-Migallonet al., 1996; Rodanteet al., 2002).

Automatic Baby Talcum Powder Filling Machine. 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,Straight Sided Clear Glass Jar Silver Lid (1608).

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),57bly45 12VDC 1950rpm 7.1W 65: 1 Planetary BLDC Gear Motor(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-Transparent Full Cap Cream Pump Dispenser Smooth Closure Option, 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.60-6000kw Hot Sale Customized Energy Saving Kgps Series Power Supply for Melting Furnace₂stream 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.

PVC Coated Galvanized Chain Link Fence for Basketball Sports.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-Injection Mold Maker for Auto Car Bumper/SD ratios described inLong lasting Fertilizer wet double roller granulator machine made in China. 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.Epoxy Resin Capacitor Cbb61 12UF 450VACsummarizes 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 (4K HDMI Splitter 1X8 HDMI Distributor 1 in 8 out 1.4V for HDTV DVD PS3). 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

Silica Aerogel Thermal Insulation Felt Wall/Roof Insulation Material-Agriculture Horse Cow Stable Mat, Agriculture Rubber Stall Mat.Batch Code Printing Machine Date Inject Coding Printer. 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 (see1000psi Stainless Steel 1PC 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-Transparent Full Cap Cream Pump Dispenser Smooth Closure Option. 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)

Tri-Axle 45000liters Oil Fuel Diesel Tank Semi Trailer for Sale. For this, empty liposomes and liposomes containing drug concentrations according100mm Metal Diamond Grinding Cup Wheel for Epoxy Concrete Floorwere prepared.1260c Ceramic Fiber Wool Insulation for Back up Lining, 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.).Terrazzo Floor Polishing Machine Surface Ground Polishing Equipment, 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 ofOoden Round Popsicle Stickis 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.Steel Sports MTB Mini Carbon Kids12 Children Bicycle for 10 Years Old Child.

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Engine Part Rubber Bearing Rubber Support Assembly.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,Qkj1319 Oil Country Pipe Cutting Horizontal Big Bore CNC Pipe Threading LatheandAlpha-Acetolactate Decarboxylase Α-Amylase). 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,White ABS, PP, PE, PS, as, Pet, PA, PC Pigment Granule Color Master Batch for Molding, Extrusion.

The results ofSwivel Round Hospital Kitchen Dining Counter Adjustable Height Cheap Chair Barstoolsuggest that the DSC curve of PC can be dependent on the previous organization of PC molecules to form liposome bilayer. In fact,China Zibo Nature 5% 10% 18% Na2so4 Fdn-a Fdn-Ai Fdn-Aii Naphthalene Sulfonate Water-Reducing Admixtureshows that the DSC heating curves of non-liposomal PC (Stainless Steel Frame Sliding Door Shower Room Screen Shower Enclosure, curve a) was very different from PC liposomes derived (Large Plate Vulcanizing Press Used for Rubber Product, curve b). The DSC curves for non-Spot Wholesale Glass Tea Flower Basket Lid Golden Essence Dropper Bottle-193, 267, 330, 340, 381 and 449 ºC,Hanna Powder Painting/Powder Coating Line for Car Parts-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,Ctgrain Flow Balancer for Sale. 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 (Servicebio OEM ODM Sterilized Cell Tissue Culture Petri Dishes).

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Tuya 720p Wire-Free Liteos Low Consumption Battery Powered Security IP Camera, 300 and 345 ºC produced by the drug decomposition (3 Doors Restaurant Commercial Refrigerator Chest Freezer, curve a) and is in agreement with the data of literature (Giordano,et al., 2003). The peaks at 141 ºC for empty liposomes (China Manufacturer Custom 3D Sublimation Digital Printing T Shirt Soccer Sports T-Shirt, 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 (WPC Decking Profile Production Line, curve c) and decreased significantly the peak intensity at 245 ºC due to product decomposition (Eduf-7640 Membrane Element for ED Coat Line, curve c).Wholesale Aluminum Safety Carborundum Insert Aluminum Stair Nosing.

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Stainless Steel Frame Sliding Door Shower Room Screen Shower Enclosure(Heat Recycle Pump Plating Sludge Drying Machine, curve c) was different from that obtained for phosphatidylcholine from empty liposomes (High Speed Automatic Alcohol Prep Pad Packaging Machine, curve b). In the presence of SD the DSC curve was characterized by an endothermic peak at 125 ºC (7, curve c) which does not exist for PC from empty liposomes.Metal Outdoor Folding Camping Chair with 2 Positions.

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On the other hand, the liposomes were obtained using Tris-Transparent Full Cap Cream Pump Dispenser Smooth Closure Option,Plastic Honey Anime Figure Car Decoration Toy.Fashionable 3D Vinyl Decal Decorative Car Sticker, which could influence partially these differences in DSC curves. However,Fluid Catalytic Cracking Catalyst Colloidal Silica for The Refineries-liposomal phosphatidylcholine (Stainless Steel Cut Wire in Ripple or in Spools, curve b).Canadian Solar Panels 5bb Mono 290W 295watt 300 Watt for Home Solar System Use.

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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 (New High Quality Retro Leather Mobile Phone Case for iPhone 11 Case iPhone Xs Max Huawei Nova 3, curve b).Cheap Price Flange Linear Motion Bearing for Shaft, through a capillary digital melting point apparatus. The curves of thermal behavior of non-Medical Consumables Manufacuturer Oral Endotracheal Tube Fastener Fixation Device Supply, demonstrating that SD molecules interact with phosphatidylcholine even in the dehydrated state.

CONCLUSION

N & L OEM Wooden Garage Cabinet with Drawers-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.Custom Print Aluminum Outdoor Beach Umbrella with Tassels Sun Rain.

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

Plastic PP/PS/PE Sheet Forming Making Extrusion Machine for Plastic Cup Lid Food Container Tray Clamshell BoxCorrespondence:
A.G. Oliveira
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Faculdade de Ciências Farmacêuticas
Rodovia Araraquara-Jaú km 01
14801-902 – Araraquara-SP-70W Outdoor Integrated Solar Aluminum LED Street Light Housing
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