International Journal of Current Research and Review
ISSN: 2231-2196 (Print)ISSN: 0975-5241 (Online)
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IJCRR - 4(18), September, 2012

Pages: 138-147

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Author: Preeti. R. Meshram, Suvarna P. Gajare, Manoj A. Bhadane

Category: Healthcare

Abstract:Azithromycin is macrolide antibiotic is effective against a wide variety of bacteria such as Hemophilus influenzae, Streptococcus pneumoniae, Mycoplasma pneumoniae, Staphylococcus aureus, mycobacterium avium, and many others. The aim of present work was to formulate sustained released azithromycin microsphere to overcome the problem related to drug like insoluble in water, bitter taste and gastrointestinal side-effects. Microspheres were prepared by varying drug polymer ratio using emulsion solvent evaporation technique. Characterization of microspheres was done by Differential Scanning Calorimetry (DSC), X-Ray diffractometry (XRD), Surface Morphology (SEM), Stability studies.

Keywords: Sustained Released Microsphere, Azithromycin, Solvent evaporation technique, Ethyl Cellulose, Ethyl Acetate

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The objective of any drug delivery system is to provide drug in therapeutic amount to the proper site in the body to achieve immediately and then maintain the desired drug concentration. These idealized objectives are achieved by appropriately developed sustained release drug delivery which also has diverse applicability and merits. There are certain drugs having large dose they may cause GI side effect so to avoid this drug can be formulated in sustained form to release slowly in lower GI tract. The sustained release drug delivery includes the application of physical and polymer chemistry. These polymers slowly release the drug in biosystem and maintain drug blood level within therapeutic range for longer duration. Some of the products characterize the drug permeation through the appropriate biological membrane and any first pass metabolic effects prior to the entry of drug into systemic circulation. The fact that the absorption and release rate of the drug from the dosage form, is one of the interesting and most recent developments in pharmaceutical field1, 2 . Microencapsulation is a process whereby relatively thin coating of polymers are applied to small particles of solid or droplets of liquid and dispersions. The microencapsulation processes produce small particles ranging in size from 1 to 1000 µm. There are different names for these particles: microparticle, microsphere, microcapsule and micromatrix. The microparticle system has become an indispensable part of the sustained drug delivery fields for the past few decades since it can readily be adapted for various administration methods3, 4, 5 .

Procurement of drug and excipients: The drug Azithromycin and polymer Ethyl cellulose (50 cps) were gifted from Zim Laboratories Ltd, Nagpur, where other AR grade excipients like Polyvinyl alcohol, Dichloromethane, Chloroform, Ethyl acetate, Sodium hydroxide and Dibasic sodium phosphate were supplied by Loba chemical, Mumbai. Microspheres were prepared by using mechanical stirrer of Remi Motors Ltd, Mumbai. Evaluation and characterization of microspheres was carried out by using equipments like USP Tablet dissolution apparatus Type II (TDT 08L), UVVisible double beam Spectrophotometer (UV1800), FTIR (8400SCCE), X Ray diffractometer (PW1700), DSC (61000), Scanning Electron Microscope ( JSM 6380 A), Light Microscope and Stage Micrometer. METHODS Preliminary study 6, 7, 8: The Preformulation study like Preliminary identification test (melting point, solubility) characterization of drug and polymer and their interaction study were performed. Spectroscopic studies 9, 10, 11: UV Spectroscopy studies involved Determination of λ max and preparation of standard curve of Azithromycin at 244 nm. The IR spectrum of the drug sample was recorded by IR spectroscopy. The drug polymer interaction was carried out using FT-IR Apparatus. Formulation of microspheres: The formulations were prepared by using solvents ethyl acetate at different drug: polymer ratio (1:1, 1:1.5, and 1:2). Method of preparation 12, 13, 14: In the O/W emulsion solvent evaporation method, the polymer (ethyl cellulose) was dissolved in internal organic phase of ethyl acetate. Accurately weighed quantity 1g of Azithromycin was dispersed or dissolved in the polymer solution. This resulting mixture was poured slowly with stirring into 100 ml of a 0.015% w/v aqueous solution of polyvinyl alcohol. The emulsion was then stirred continuously at 700 rpm for 1 hr to evaporate the solvent. The microspheres were recovered by vacuum filtration, washed with 200 ml of deionized water and dried at room temperature. The compositions of Azithromycin microspheres using ethyl acetate as solvent were given in Table 1. Evaluation of microspheres15, 16: Angle of repose: Angle of repose is defined as the maximum angle possible between the surface of pile of powder and horizontal plane. The angle of repose for the microspheres of each formulation was determined by the funnel method. The microspheres were allowed to flow out of the funnel orifice on a plane paper kept on the horizontal surface. It forms a pile of microspheres on the paper. The angle of repose was calculated by substituting the values of the base radius ‘R’ and pile height ‘H’ in the following equation Bulk density of all batches of microspheres was determined by pouring gently 2 g of sample through a glass funnel into a 10 ml graduated cylinder. The volume occupied by the sample was recorded. Bulk density was calculated as per given formula:


Tapped Density:
The tapped density was determined by pouring 2 g of microspheres through a glass funnel into a 10 ml graduated cylinder. The cylinder was tapped from height of 2 inches until a constant volume was obtained. Volume occupied by the sample after tapping was recorded .The values for tapped density was calculated as per given formula:


Compressibility index:
The compressibility indices of the formulation blends were determined using Carr’s compressibility index formula.


Hausner Ratio:
It provides an indication of the degree of densification which could result from vibration of feed hopper. Lower the Hausner ratio better is the flowability. It was calculated as per given formula.


Determination of particle size 17:
The particle size was determined using stage micrometer. The diameters of about 300 microspheres were measured and the average particle size was determined. Estimation of drug loading18, 19: For determination of drug content, microspheres equivalent to 100 mg were weighed and dissolved in 100ml of acetone. After suitable dilutions were with phosphate buffer (pH 6.0), the resulting solution was analyzed spectrophotometrically at 244 nm.

In-vitro dissolution study20, 21, 22: The study was carried out using dissolution apparatus USP Type-I (Rotating Basket type). Accurately weighed microspheres equivalent to 200 mg of Azithromycin were taken in muslin cloth and it was kept in baskets. Dissolution study was carried out in phosphate buffer pH 6.0 at 50 RPM at temp 37 ± 0.5° c. During dissolution study 10 ml of aliquot was withdrawn at a time intervals of 1 to 12 hr and same was replaced with equal volume of fresh medium. The withdrawn samples were filtered through Whatmann filter paper and absorbances were measured at 244 nm. Drug concentration in the samples was determined from the calibration curve. Characterization of microspheres: Differential Scanning Calorimetry 9, 10, 11, 27:

The DSC measurements were performed on a differential scanning colorimeter with thermal analyzer. All accurately weighed samples (about 10 mg of Azithromycin, ethyl cellulose, physical mixtures, and formulation) were placed in a sealed aluminium pan, before heating under nitrogen flow (10 ml/min) at a scanning rate of 20 ° c per min from 100 to 300 ° c an empty aluminium pan was used as reference. X-Ray diffractometry 13, 14, 15, 23: The powder X-ray diffraction pattern of Azithromycin and polymer were obtained using Phillips X-ray diffractometer with a Ni-filtered CuKα-radiation at a scanning speed of 10 °/min at 2θ.The graph was plotted in 2 theta angle Vs intensity count. Surface Morphology 24 The microspheres were coated with Platinum by ion sputtering using Autofine coater. The microspheres were kept on the sample holder and the scanning electron micrographs were taken. Stability studies 35 The study was carried out by storing the microspheres in glass bottle 40 ° c and 75% RH for 30 days. These samples were collected on 7th, 14th , 21st, 28th day and analyzed for changes drug content and in-vitro dissolution studies. RESULTS Preformulation studies: The melting point of azithromycin was found in the range of 119 – 121 ° c. The wavelength of maximum absorbance (λ max) found was 244 nm. Evaluation of microspheres: All the formulations show angle of repose value in the range of 19.72 to 20.42. The values for bulk density were found in the range of 0.352 to 0.450. The values for tapped density were found to range from 0.414 to 0.510. The compressibility index values were found in the range of 11.76 to 14.99 respectively. Hausner’s ratio was ranging from 1.13 to 1.17. The average particle size of microcapsules is found to be within 151.263 to 171.342 µm. The % encapsulation efficiency is  found to be in the range of 73.50 to 75. 60%. In – vitro dissolution study, % cumulative drug release from formulation in 12 hrs was decreased from 80.13 to 69.98 % represented in Table 2. The dissolution data for formulations C1, C2, and C3 was fitted to various drug release kinetic models like Zero order, First order, Higuchi Matrix and Korsemeyer Peppas, Hixon-Crowel model. Kinetic Treatment to Dissolution Data proved important information about rate constants (K), correlation coefficients (R) obtained for various models were tabulated in Table 3. Characterization of microspheres: The FT-IR spectra of pure drug - Azithromycin, ethyl cellulose, physical mixture and formulation (C1) shows major functional groups of Azithromycin (methyl substituted nitrogen on lactone ring) in spectra of individual drugs as well as in spectra of physical mixture and formulation (C1). FT-IR of Formulation C1 presented in Fig 1. DSC curve presented in fig 2 of Azithromycin shows a single endothermic peak at 121 ° c, due to melting of the drug. Optimized formulation C1 which contains Azithromycin and ethyl cellulose, the thermogram indicates characteristic peaks for melting of Azithromycin at 121 ° c. The XRD pattern presented in fig 3 of formulation exhibited halo pattern with less intense and denser peaks compared to plain Azithromycin, Scanning Electron Microscopy results were presented in fig 4a and 4b. Accelerated stability studies (AST) was carried for optimized batch (C1) by exposing it to 40° c/75% RH for one month and analyzed the samples at the interval of 7,14,21,28 days and the samples was analyzed for drug content and invitro dissolution study. The stability studies show drug content after 28 day was 74.14 where percent cumulative drug release 80.55. DISCUSSION The XRD scan of Azithromycin showed intense peaks of crystallinity. Diffractogram of azithromycin showed high intensity peaks between 2θ of 10-200 values demonstrating the crystalline nature of drug. No intense peaks were observed in diffractogram of ethyl cellulose which indicates amorphous nature. Thus, preformulation studies indicated that there is no interaction between Azithromycin and ethyl cellulose. Physical evaluation of microsphere formulation for angle of repose, Compressibility index and Hausner’s ratio showed that they had good flow properties. The microsphere formulated using ethyl acetate as internal organic phase or solvent in ratio1: 1 (C1) showed better encapsulation efficiency than other formulations. All the formulations showed diffusion exponent (n) varying from 0.524 to 0.668. From the n values of all formulations it can be concluded that as the concentration and viscosity of polymer was increased the value of diffusion exponent also increases. From the dissolution profile of formulations C1, C2 and C3, It can be concluded that formulation C1 shows better drug release (80%in 12 hrs) than C2 and C3. Formulations C1 to C3 showed retardation of Azithromycin as concentration of ethyl cellulose increased. Total release in 12 hrs was decreased from 80% to 70%. The IR data of Azithromycin loaded ethyl cellulose microsphere also showed the peaks same as that the peaks in IR spectrum of pure drug, this suggested that there was no chemical interaction between ethyl cellulose and Azithromycin. The DSC results also support IR spectrometry results indicating absence of drug - polymer interactions. The XRD pattern of formulation C1 exhibited halo pattern with less intense and denser peaks compared to plain Azithromycin. This indicated that Azithromycin is dispersed at the molecular level in the blend of polymeric matrix. The SEM photographs showed that the microspheres were spherical in nature and had a smooth surface. SEM photographs revealed the absence of crystals of drug on the surface of microspheres and uniform distribution of the drug within the microspheres.

The stability study conducted for observing effect of temperature on the formulation showed that there is no significant change in drug content and in-vitro drug release of optimized formulation. CONCUSION From the investigation carried out and results obtained, it was observed that the % of drug released from microspheres was sustained. As the GI side effects of Azithromycin are related to its high dose which can be minimized by formulating as microspheres. It would also helps to improve bioavailability because of absorption window and microencapsulation helps to mask the bitter taste of drug. Hence from this investigation, we can propose that the objective of the study is achieved. ACKNOWLEDGEMENT Authors are grateful to Zim Laboratoris Nagpur and Loba chemical, Mumbai, for providing the gift sample of drug and chemicals.


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Infrared spectroscopy DSC: Differential Scanning Calorimetry XRD: X-Ray diffractometry SEM: Surface Morphology UV: Ultraviolet Visible double beam Spectrophotometer K: Rate constants R: Correlation coefficients AST: Accelerated stability studies


Dr. Pramod Kumar Manjhi joined Editor-in-Chief since July 2021 onwards

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A Study by Varsha M. Shindhe et al. entitled "A Study on Effect of Smokeless Tobacco on Pulmonary Function Tests in Class IV Workers of USM-KLE (Universiti Sains Malaysia-Karnataka Lingayat Education Society) International Medical Programme, Belagavi" is awarded Best article of Vol 12 issue 14, July 2020
A study by Amruta Choudhary et al. entitled "Family Planning Knowledge, Attitude and Practice Among Women of Reproductive Age from Rural Area of Central India" is awarded Best Article for special issue "Modern Therapeutics Applications"
A study by Raunak Das entitled "Study of Cardiovascular Dysfunctions in Interstitial Lung Diseas epatients by Correlating the Levels of Serum NT PRO BNP and Microalbuminuria (Biomarkers of Cardiovascular Dysfunction) with Echocardiographic, Bronchoscopic and HighResolution Computed Tomography Findings of These ILD Patients" is awarded Best Article of Vol 12 issue 13 
A Study by Kannamani Ramasamy et al. entitled "COVID-19 Situation at Chennai City – Forecasting for the Better Pandemic Management" is awarded best article for  Vol 12 issue 12
A Study by Muhammet Lutfi SELCUK and Fatma entitled "Distinction of Gray and White Matter for Some Histological Staining Methods in New Zealand Rabbit's Brain" is awarded best article for  Vol 12 issue 11
A Study by Anamul Haq et al. entitled "Etiology of Abnormal Uterine Bleeding in Adolescents – Emphasis Upon Polycystic Ovarian Syndrome" is awarded best article for  Vol 12 issue 10
A Study by entitled "Estimation of Reference Interval of Serum Progesterone During Three Trimesters of Normal Pregnancy in a Tertiary Care Hospital of Kolkata" is awarded best article for  Vol 12 issue 09
A Study by Ilona Gracie De Souza & Pavan Kumar G. entitled "Effect of Releasing Myofascial Chain in Patients with Patellofemoral Pain Syndrome - A Randomized Clinical Trial" is awarded best article for  Vol 12 issue 08
A Study by Virendra Atam et. al. entitled "Clinical Profile and Short - Term Mortality Predictors in Acute Stroke with Emphasis on Stress Hyperglycemia and THRIVE Score : An Observational Study" is awarded best article for  Vol 12 issue 07
A Study by K. Krupashree et. al. entitled "Protective Effects of Picrorhizakurroa Against Fumonisin B1 Induced Hepatotoxicity in Mice" is awarded best article for issue Vol 10 issue 20
A study by Mithun K.P. et al "Larvicidal Activity of Crude Solanum Nigrum Leaf and Berries Extract Against Dengue Vector-Aedesaegypti" is awarded Best Article for Vol 10 issue 14 of IJCRR
A study by Asha Menon "Women in Child Care and Early Education: Truly Nontraditional Work" is awarded Best Article for Vol 10 issue 13
A study by Deep J. M. "Prevalence of Molar-Incisor Hypomineralization in 7-13 Years Old Children of Biratnagar, Nepal: A Cross Sectional Study" is awarded Best Article for Vol 10 issue 11 of IJCRR
A review by Chitra et al to analyse relation between Obesity and Type 2 diabetes is awarded 'Best Article' for Vol 10 issue 10 by IJCRR. 
A study by Karanpreet et al "Pregnancy Induced Hypertension: A Study on Its Multisystem Involvement" is given Best Paper Award for Vol 10 issue 09

List of Awardees

A Study by Ese Anibor et al. "Evaluation of Temporomandibular Joint Disorders Among Delta State University Students in Abraka, Nigeria" from Vol 13 issue 16 received Emerging Researcher Award

A Study by Alkhansa Mahmoud et al. entitled "mRNA Expression of Somatostatin Receptors (1-5) in MCF7 and MDA-MB231 Breast Cancer Cells" from Vol 13 issue 06 received Emerging Researcher Award

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Disclaimer: International Journal of Current Research and Review (IJCRR) provides platform for researchers to publish and discuss their original research and review work. IJCRR can not be held responsible for views, opinions and written statements of researchers published in this journal.


International Journal of Current Research and Review (IJCRR) provides platform for researchers to publish and discuss their original research and review work. IJCRR can not be held responsible for views, opinions and written statements of researchers published in this journal


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