ISSN 0976-111X

INTERNATIONAL JOURNAL OF PHARMA WORLD RESEARCH

(An International Quarterly Published Online Research Journal)

www.ijpwr.com E-mail:editorijpwr@gmail.com

Title:

Development and Characterization of Cefpodoxime Proxetil Niosomes

Pandey Shivanand* Smt. R. B. P. M. Pharmacy College, Atkot-360040, Rajkot, Gujarat. India

Correspondence to: Mr. Shivanand Pandey Tel: (02821) 278-349 Email: dot.shivanand@gmail.com

IJPWR VOL1 ISSUE 3 (Jun-Sep) 2010

ISSN 0976-111X Abstract:

Niosomes are NSVs which act as vehicle for drug delivery. Thus noisome are prepared by using incorporation of synthetic non-ionic surfactant with cholesterol. So, thus niosomes are formed from synthetic non-ionic surfactant which act as a planner bilayer lattices i.e., hydration of surfactant encapsulating the hydrating aqueous solution/non-ionic surfactant which to encapsulate the aqueous solution of solute or surfactant orient with aqueous media but hydrophilic head group of surfactant and hydrocarbon segment of surfactant which align facing with aqueous media without interaction. So their head groups are hydrophilic and their fatty acyl chains are hydrophobic in nature. Niosomes are biodegradable, biocompatible non-immunogenic and exhibit flexibility in their structural characterization. Niosomes have been widely evaluated for controlled release and targeted delivery for the treatment of cancer, viral infections and other microbial diseases. Niosomes can entrap both hydrophilic and lipophilic drugs and can prolong the circulation of the entrapped drug in body. Encapsulation of drug in vesicular system can be predicted to prolong the existence of drug in the systemic circulation and enhance penetration into target tissue, perhaps reduce toxicity if selective uptake can be achieved.

Keywords:

Cefpodoxime

proxetil,

niosomes,

targeted delivery,

biocompatible,

biodegradable, and non-immunogenic

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Introduction: At the beginning of the era of controlled drug delivery systems, a controlled release system utilizes a polymer matrix or pump as a rate-controlling device to deliver the drug in a fixed, predetermined pattern for a desired time period
1, 2, 3

. this basic rationale is behind controlled

drug delivery. A controlled drug delivery system requires simultaneous consideration of several factors, such as the drug property, route of administration, nature of delivery vehicle, mechanism of drug release, ability of targeting, and biocompatibility. Handjani-Vila et al., 5 reported formation of vesicles on hydration of mixture of cholesterol and a single alkyl-chain non-ionic and non-toxic surfactant. A number of non-ionic surfactants have been used to prepare vesicles viz.polyglycerol alkyl ethers6, glucosyl dialkyl ethers7, crown ethers8, ester linked surfactants, poly oxyethylene alkyl ether, brij, and series of spans and tweens9,
10

resultant vesicles have been termed Niosomes. Thus, Niosomes may be defined as unilamellar or multilamellar vesicles where in an aqueous solution is enclosed in highly ordered bilayer made up of non-ionic surfactant with or without cholesterol and dicetylphosphate and exhibit behaviour similar to liposomes in-vivo11. Materials and Methods: Cefpodoxime Proxertil U.S.P Gift sample from Biochem chemicals, Sorbitan mono laurate(Span20), Sorbitan mono palmitate (span 40), Sorbitan mono oleate (Span80), Cholesterol, Chloroform, Potassium dihydrogen phosphate, Triton X100 from Loba chemie, Mumbai, Dialysis membrane bag (2.4 nm), Himedia, Mumbai, Whatman membrane filter paper, Whatman membrane filter paper, N-N- Dimethyl Formamide, Nice chemicals, Mumbai. Instruments and their supplier: Scanning electron microscope(Zeol JSM-5610 ) UV/VISIBLE spectrophotometer(Elico Double beam spectrophotometer), Weighing balance(Schimadzu electrical balance), Magnetic stirrer(Remi equipments, Mumbai), Microscope(Acculab microscope), FTIR(Nexus 670 Thermoelectron corporation), pH meter(Elico L 140 ), Rotary Evaporator(Super fit apparatus, Mumbai ), Sonicator (Bath type PCI, Mumbai), Incubator(Genuine Ltd., Mumbai). Preformulation studies: The preformulation studies were needed for the selection of suitable surfactant for a drug based on the solubility, compatibility and formation of niosomes, stability etc. Preparation of Niosomes: Various batches of niosomes were prepared by slight modification of Thin Film Hydration Technique introduced by Azmin et IJPWR VOL1 ISSUE 3 (Jun-Sep) 2010 3

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al.12 Thin film Hydration Technique the surfactants (Span20, Span 40, span 80) and cholesterol were used as the lipid ingredients. The surfactants and cholesterol were dissolved in about 20 ml chloroform and added drug in the evaporating flask of rotary flash evaporator. Then it was dipped into a water bath at 30o C. The organic solvent was evaporated under reduced pressure at 140 rpm to form a thin, dry film on the wall of flask. The excess of organic solvents were removed by keeping the flask in dessicator for some time. Then the dried lipid film was hydrated with required phosphate buffer saline (20 ml) into the flask and it was heated to 50oC on the water bath, rotating around 120 rpm for complete hydration of layer. Then, the solution was sonicated to form niosomes and stored in the amber coloured bottle, kept in refrigerator. Evaluation of Prepared Niosomes: Particle Size analysis by Optical Microscopy After storing for 24 hours, the various ratio of niosomal formulation prepared from all batches was taken for size analysis. Before to determine the size analysis, first we calculated the correction factor using stage micrometer and eye piece micrometer with compound microscope using 10X magnification. The vesicle size and a size distribution value of various ratios were first done under light microscope at using 45X magnification and later by scanning electron microscopy.13 Shape and Characterization of Niosomes: Large samples for SEM analysis were mounted on the specimen stubs using adhesive. Small samples were mounted directly using a double adhesive tape. A sample of niosomes

suspension were directly placed on the cover glass fixed on the speciment stub. All samples were coated with gold to a thickness of about 100 using Zeol JSM-5610 Evaporator, Model HUS 5GB. Coated samples were viewed in Zeol Scanning Electron Microscope Model JSM5610 operated at 15 kilovolts (kv) with different magnification and then photographed. 13 The entrapment efficiency of the complex within the niosomes was determined by separating the unentrapped drug using dialysis14, 15. Briefly, the prepared niosomes were filled into an open ended glass cylinder to which a Himedia dialysis membrane was securely attached to one side and the free cefpodoxime proxetil dialyzed for 1 hour each time into 100 ml phosphate buffered saline (pH 7.4) which was magnetically stirred at 50 revolutions / min and maintained of 371C. The dialysis of free cefpodoxime proxetil was complete after several changes of saline (8 times), when no cefpodoxime proxetil was detectable in the recipient solution (UV detection at 256 nm). Entrapment efficiency by Indirect method The concentration of cefpodoxime proxetil separated (dialyzate) into 0.9% sodium chloride

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solution was obtained from the calibration curve (it was estimated by UV detection at 256 nm), since the initial amount of cefpodoxime proxetil incorporated in the vesicles was known, thus drug entrapped can be calculated. This is the indirect method of estimation of entrapped drug. Entrapment efficiency by Direct method In the direct method Triton-X100 was used to break the membranes.16 1 ml of 0.1% Triton X100 solution was added to 0.1ml of noisome preparation and made up to 5 ml with PBS. Then it was kept at 37C for 30 minutes for complete break up of the niosome membrane and to release the entrapped material. The sample was filtered through Whatman Filter Paper [no.44] and the filtrate was measured spectrophotometrically at 256nm for cefpodoxime proxetil. The amount of cefpodoxime proxetil was derived from the calibration curve. Invitro Drug Release Studies: A study was done on the release pattern of the drug from the niosomal formulations prepared by Thin film hydration method. After seperating the unentrapped drug, the niosomal suspension containing drug equivalent to drug content was pipetted into the dialysis bag which was previously soaked and washed several times with distilled water. This was placed in 100 ml of phosphate buffer saline (pH 7.4) and kept with constant agitation on a magnetic stirrer maintaining a temperature of 37C. Each periodical time the whole sample were withdrawn and same volume of fresh sample was replaced. Then the sample were assayed spectrophotometrically (Elico Model-SL 164) at 256 nm using medium as blank. The release was compared with pure cefpodoxime proxetil drug solution. Kinetics of Drug Release: In order to understand the mechanism and kinetics of drug release, the drug release data of the in vitro dissolution study were analyzed with various kinetic models like Zero order, First order and coefficient of correlation (r) values were calculated for the linear curves obtained by regression analysis of the plots.17 Compatibility studies by Fourier transform infrared Spectroscopy (FTIR): The interaction between drug substance, surfactants and best formulations were evaluated by comparing the IR spectrum of pure drug sample and the surfactants. Infrared Spectroscopy using Fourier Transform infrared (FTIR Nexus-670) i.e., Compatibility study of drug with the surfactants was determined by I.R. Spectroscopy by using KBr in the ratio of sample to KBr is 1:100, were examined and the spectra of drug and other ingredients in the formulations were compared with that of the original spectra.18Stability Studies of Prepared Niosomes: The formulated niosomes were divided into 3 groups and each one was stored in amber-colored vials, one at 4C, the second portion

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at room temperature 27C and the third at 37C. Every 7 days 0.1ml of the niosome suspension was withdrawn and made upto 1ml with phosphate buffer saline. The amount of drug leached from the niosomes present in the dialysate was estimated spectrophotometrically.18 Results And Discussion Particle size determination by optical microscope Size Distribution/range of Niosomes after storing for 24 hours, the various ratios of all niosomal formulations were taken for size analysis. Measurement of the niosome size was made by using a compound microscope. Before to determine the size analysis, first we calculated the correction factor using stage micrometer and eye piece micrometer with microscope using 10 X magnification. The vesicle size and size distribution values of various ratios of formulation were calculated using 45 X magnification. The diameter of niosomes was found to be in the range of 6.14 10.5 m. Morphology: The niosomes prepared by thin film hydration method when viewed under light microscope at a magnification of 45 X and later under scanning electron microscope (S.E.M) for characterization, shape, size and of niosomes for best formulation were spherical, loaded niosome [Span 40 (1:6:1 ratio)] at magnification of 500X and has a diameter of 0.0035 m, vesicle diameter of 0.0065 m , spherical niosome [Span 40 (1:6:1 ratio)] at a magnification of 500X having a vesicle diameter of 0.058 m and spherical niosomes [Span 20 (1:9:1 ratio)] at a magnification of 500 X and has a diameter of about 0.064 m. The niosomes were mostly spherical in shape and the size range of all ratios formulation which is in the size range of small unilamellar vesicles (0.1m). The vesicles were discrete and separate with no aggregation or agglomeration. The size of the vesicles was uniform and independent of surfactant. Entrapment efficiency by Indirect Method the dialysis method [indirect method] was used for niosomes prepared by Thin film Hydration Technique with various surfactants like Span 20, 40 and 80 (Each 1:3:1, 1:6:1 and 1:9:1 ratio) respectively. Total amount of cefpodoxime proxetil in niosomes used for dialysis studies is 50 mg. In indirect method, the maximum amount drug entrapped in Span 40 and 20 was found to be high when compared with Span 80. In direct method, the entrapment efficiency in Span 20 were found to be 97.74%, 97.67%, 98.97% (F1, F2, F3 respectively) Span 40 were found to be 98.23%, 99.37%,

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98.18% (F4, F5, F6 respectively) and Span 80 were found to be 96.69%, 96.15%, 97.29% (F7, F8, F9 respectively). Compatibility studies by Fourier transform infrared Spectroscopy (FTIR): 1780-1770, 1620-1590, 1075-1020, 1870-1540, 2950-2835, 14851370 wave number for pure drug and 1732.4, 1629.4, 1046.3, 16219, 2922, 1410 for

formulation F5 S 40 (1:6:1) and 1736.5, 1630.1, 1035, 1732.4, 2857, 1462 were found. The similarity in peaks indicates there is no incompatibility between drug and the surfactants used for formulation. Invitro Drug Release Studies: %CDR from formuations F1, F2, F3, F4, F5, F6, F7, F8, and F9 were 61.25, 61.74, 61.56, 51.26, 58.05, 66.33, 63.5 and 57.64 respectively. Kinetics of Drug Release: all the formulation followed zero and first order of kinetics and their r2 were lied in between 0.996 to 0.999. for the best formulation these were 0.998, 0.9825, 0.9956 ( F5) for zero , first and Higuchi plot. Stability Studies of Prepared Niosomes: The formulated niosomes were divided into 3 groups and each one was stored in amber-colored vials, one at 4C, the second portion at room temperature 27C and the third at 37C. Every 7 days 0.1ml of the niosome suspension was withdrawn and made upto 1ml with phosphate buffer saline. The amount of drug leached from the niosomes present in the dialysate was estimated spectrophotometrically. For F3 S20,1:9:1 % drug release 96.589% at 4+1oC same at 25+2oC and 37+2oC. For F5 S40, 1:6:1at 4+1oC, 25+2oC and 37+2oC were 96.864%.

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Graphs and Tables

C prativestudyof diffrent form om ulations

100 90 80 70 F 1 F 2 F 3 F 4 F 5 50 40 30 20 10 0 0 0.5 1 2 3 4 5 6 7 8 9 10 11 F 6 F 7 F 8 F 9

%cu u m latived grelease ru

60

T e inh im rs

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Comparative Dissolution Study of All Formulations Infrared Graph of F5 S 40 (1:6:1) Consolidation Chart of Kinetic Study Formulati on Code F1 F2 F3 F4 F5 F6 F7 F8 F9 Zero order Slope 5.093 5.189 5 4.774 4 5.208 52 4.336 0 4.837 4 5.484 1 5.400 1 4.886 1 First orde r r2 0.99 93 0.99 86 0.99 69 0.99 98 0.99 8 0.99 98 0.99 95 0.99 73 0.99 89 Higuchi plot Slope -0.0335 -0.0343 -0.0301 -0.0346 -0.026 -0.032 -0.0382 -0.0361 -0.0309 r2 0.9772 0.9744 0.9720 0.9848 0.9825 0.9732 0.9731 0.9689 0.9785 Slope 5.0085 5.1009 4.692 5.122 4.2615 4.7572 5.393 5.306 4.8024 r2 0.9593 0.9964 0.9943 0.9986 0.9956 0.9986 0.9982 0.9945 0.9966

Conclusion : The formulations were showing satisfactory particle size, encapsulation efficiency and invitro release studies. The stability studies show the presence of drug in formulation even after storage for some period of time which shows an increase in drug stability by formulating it into niosomes. The incompatibility between the drug and excipients shows that span is suitable for cefpodoxime proxetil to prepare niosomes. The in vitro release studies and kinetic study declared the release is zero order and in controlled manner, so there will be not chances of dose dumping during usage

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References: 1.Aarti Jagtap and Deepali Inamdar Formulation and Evaluation of Niosome Entrapped Pentoxifylline Using in vivo Bronchodilatory Activity in Guinea Pigs. Indian J. Pharm., 2001:63; 49-54. 2. Satturwar PM, Khandare J.N and Nande J.S. Niosomal Delivery of Ketoconazole. Indian Drugs. 2001; 38, 620-624. 3.Langer R. Implantable controlled release systems. J.Pharmacol.Ther. 1983:21; 3551. 4.Adjani-Vila, M., Ribier, A., Rondot, B., and Vanlerberghe, G Dispersions of Lamellar phases of non-ionic lipids in cosmetic products; Int.J.Cosmo.Sci. 1979:1; 303-314. 5.Baillie, A.J., Coombs, G.H., Dolan, T.F., and Laurie J, Non-ionic Surfactant vesicles, J. Pharm. Pharmacol.,1986:38;502. 6.Kiwada H., Nimura H., Kato, Y., Application of synthetic alkyl glycoside vesicles, Chem. Pharm. Bull., 1985:33;2475. 7.Echegoyen, L.E., Hermandez.J.C. Kaifer, A.E., Gokel, G.W., Echegoyen, L. Lariat ether Bolaamphiphiles, J.Chem. Soc. Chem. Comm., 1988:12; 836. 8.Chandraprakash, K.S., Udupa, N., Umadevi, P and Pillai, G.K, Formulation and evaluation of Methotrexate niosomes Ind. J .Pharm. Sci., 1992:54;197. 9.Hunter, C.A., Dolan, T.F., Coombs, L.H., and Baillie A.J., Vesicular Systems for Delivery of Sodium Stilbo gluconate in experimental murine visceral leishmaniasis,

J.Pharm.Pharmacol.,1988:40;161-162. 10.Jain, N.K., Alok Nam Deo, Niosomes as Drug carriers Ind.J.Pharm. Sci., 1996:58; 41-46. 11.Azmin, M.N., Florence, A.T. Handjani Vila, R.M., Stvart, J.F.B., Vanlerberghe.G and Whittazker. J.S.J. Pharm. Pharmacol., 1985:3; 237. 12.Manjusha Malhotra and N.K. Jain, Indian Drugs,1993:31; 81-83.

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13.Himansu M, Varsheny, Tanwar Y.S. Rohit lowalekar and Rathore K.S Designing and Characterization of Verapamil Hydrochloride Niosomes Indian Pharm., 2007:6;77-79. 14.Hardy J.G., Kellaway I.W., Rogers Wilson CG, The distribution and Fate of I131 Labelled Lipsomes, J. Pharm., Pharmacol., 1980:32;309 313. 15.Nidhi N.Dubey, Jolly R. Parikh, Parikh R.H, Solanki A.B and Nitin dubey, Preparation and optimization of Niosomes containing piroxicam, Indian pharm .,2006:5;97-103. 16.Merchant H.A, ShoaibH.M, Tazeen J, Yousuf RI, once dialy tablet formulation and Invitro release evaluation of cefpodoxime proxetil using hydroxypropyl methyl cellulose : A Technical note,AAPS Pharm., Tech., 2006:7; 55-64. 17.Robert M., Silverstein. Francis X.webster., Spectrophotometric identification of organic compounds., 6th edition., Wiley Publishers., 79-81. 18.John R Dyer., Spectroscopy of organic compounds. 12th Edition. 45-54.

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