FORMULATION EVALUATION AND OPTIMIZATION OF AN ORAL IMMEDIATE RELEASE ANTIBIOTIC FORMULATION OF AMOXICILLINE

e-issn 2249 7706 print-issn 2249 7714 International Journal of Advanced Pharmaceutics www.ijapjournal.com FORMULATION EVALUATION AND OPTIMIZATION OF AN ORAL IMMEDIATE RELEASE ANTIBIOTIC FORMULATION OF AMOXICILLINE Tanuja N 1, Vasanth PM* 1, Suresh K 2, Ramesh T 3, Ramesh Malothu 3 1 Department of Pharmacy, UCEV-JNTUK, Vizianagaram, Andhra Pradesh, India. 2 Department of Pharmaceutics, Vignan Inst of Pharm Sciences, Vishakapatnam, Andhra Pradesh, India. 3 Department of Biotechnology, UCEV-JNTUK, Vizianagaram, Andhra Pradesh, India. ABSTRACT The aim of the study was formulation Evaluation and optimization of an oral immediate release antibiotic formulation of Amoxicilline using magnesium stearate sodium starch glycolate povidone microcrystalline cellulose carboxy methylcellulose sodium either alone or combinations tablets were prepared by direct compression wet granulation and dry granulation method and evaluated for various physical parameters. Amoxicilline is the antibiotic, it is used in gram+ve and gram-ve bacteria the drug release studies were performed using USP apparatus type11, using o.1nhcl and ph 6.2 phosphate buffer as dissolution medium. The drug release was dependent on the type and concentration of the polymer dosage forms were characterized for powder properties like angle of repose, bulk density tapped density, and tablet properties like content uniformity, weight variation and hardness and friability and thickness and in-vitro dissolutions studies. Nine formulations were taken. Grade A the wet granulation, direct compression, dry granulation method were found good the compressibility index showed in the flow powder is possible range. Grade B practically the flow of blend for both trails was found good as per angle of repose and hausner s ratio. Grade C according to the angle of repose the flow blen all trails good except trail 1. In selection of API the Grade C API was selected for the formulation by using dry granulation is showed poor flow by direct compression it also reduce the process loss and increase the productivity the selection of the granulation technique dry granulation showed the satisfactory result in this method no flow problem occurred and all the physical parameters observed. Keywords: Immediate release amoxicilline sodium starch glycolate, Microcrystalline cellulose, Povidone, Magnesium stearate. INTRODUCTION Oral drug delivery has been known for decades as the most widely utilized route of administration among all the routes that have been explored for the systemic delivery of drugs via various pharmaceutical products of different dosage forms. The Immediate Release Dosage Form was Compressed tablets may be defined as drug delivery system, intended to be taken by mouth and release the drug in gastrointestinal track, made up of a single solid unit body. It may be compressed with or without addition of diluents. Some of the most common types of tablets include, Immediate release, coated or uncoated (swallow able), Modified release, coated or uncoated (swallow able), Effervescent (dissolved/suspended in water before drinking), Chewable (chewed before swallowing), Lozenge (dissolved slowly in the mouth) [1]. Process optimization is required in the environment of ever increasing domestic and international market, where we have better our competitors in case of quality and lower production cost and at the same time satisfying the demand of the market. The Optimization technology, there are two types of optimization problems. They are Constrained optimization Unconstrained optimization [2]. Corresponding Author:- Vasanth PM Email:- vasanthpharma@gmail.com 37 P a g e

The Amoxicilline used for the gram +ve and gram ve bacteria that kills bacteria or inhibits its growth. The objective of the present study was to develop formulation, evaluate and optimize of an oral immediate release antibiotic formulation of amoxicilline using magnesium stearate sodium starch glycolate povidone microcrystalline cellulose carboxy methylcellulose sodium either alone or combinations tablets were prepared and the manufacturing process selected was the wet granulation dry granulation and directly compressed method. The mechanism of action of Amoxicilline interferes with the construction of the bacterial cell wall, the synthesis of protein, or the replication and transcription of DNA. Relatively few clinically useful agents act at the level of the cell membrane or by interfering with specific metabolic processes of the bacterial cell [3-6]. MATERIALS AND METHODS Materials Amoxicilline was obtained from Oman Chemicals Avicel PH101, Avicel PH102, Avicel RC 591, obtained from FMC Biopolymer Povidone K-30 obtained ISP Magnesium Stearate were obtained from Ferro, Portugal. Methods Direct compression Amoxicilline, Avicel PH102, Povidone K- 30,sodium starch glycolate are accurately weighed then sieved through #18 mesh size, magnesium stearate were individually sifted through #40 mesh size, then sifted material was transferred through octagonal blender, and blended for 12min at 15 rpm then add sifted magnesium stearate and transferred into the blender and blended for 3 min at 15 rpm, then lubricated material was compressed using 8 station compressed machine using 21 x 10mm oblong standard punches. Dry granulation Amoxicilline, Avicel PH102, Povidone K- 30,sodium starch glycolate are accurately weighed then sieved through #18 mesh size, magnesium stearate were individually sifted through #40 mesh size. All the materials were mixed 5 to 8 min other than magnesium stearate, then dry the intra granular material, compact it by using roller Compactor, and the contacted material was milled through 1.5mm screen by using multi mill sifted material was transferred through octagonal blender to blend for 12min at 15 rpm and then add sifted magnesium stearate were lubricated for 3min at 15rpm then compression of the lubricated material was done by using 8 station compressed machine using 21 x 10mm oblong standard punches [7-10]. Wet Granulation Amoxicilline, Avicel PH102, Povidone K- 30,sodium starch glycolate are accurately weighed then sieved through #18 mesh size, magnesium stearate were sifted through #40 mesh. all the materials were mixing 5 to 8 min other than magnesium stearate then the mixing material was granulated by hand granulation or in RMG using purified water as a granulating fluid granulating fluid, wet mass is obtained and then wet mass is dried in the tray drier or rapid drier at 60 0 C for 1hr 50 min for aqueous granulation and for non-aqueous granulation air drying of the milled wet mass. Dried material was sifted through #20 mesh size and add sifted magnesium stearate were lubricated for 3min at 15rpm, compression of the lubricated material was done by using 8 station compressed machine using 21 x 10mm oblong standard punches [11-14]. Evaluation of pre-compression parameters (Blend analysis) Powder flow properties can be evaluated by measurements of angle of repose bulk density, Hausner s ratio and Compressibility index. Friability Test Friability is performed to assess the effect of friction and shocks, which may often cause tablet to chip, cap or break. Roche friabilator was used for this purpose. This device subjects a number of tablets to the combined effect of abrasion and shock by utilizing a plastic chamber that revolves at 25 rpm dropping the tablets at a distance of 6 inches with each revolution. Preweighed sample of 10 tablets were placed in the friabilator and tumbled in a friabilator (modesl, Roche, Bombay) for 4 min at 25 rpm. The tablets then were dedusted, and the loss in weight caused by fracture or abrasion was recorded as % friability [15]. Compressed tablets should not lose more than 1% of their weight. The percentage friability was measured using the formula, F = W (initial) W (final) x 100 W (initial) Dissolution Test: Dissolution of immediate release tablet is done as procedure specified in OGD guidelines. Test samples of dissolution was analysed by using UV Spectroscopy [14]. Dissolution parameters Medium : Water Volume : 900 ml Apparatus : USP type II (paddle) Agitation : 75 RPM Time points : 5min, 10 min, 15 min, 20 min, 30 min, 45 min. Temperature : 37 0 C 0.5 Volume withdrawn: 10 ml Preparation of Std. Stock solution: Accurately weigh and transfer about 51.8 mg of Amoxicilline into a 200 ml volumetric flask, to this add up to 100 ml water and Sonicate for 5-6 min and make up the volume. 38 P a g e

The absorbance of std. Solution and sample preparation were measured at 272 nm. Using dissolution medium as blank. Bulk density and Tap density An accurately weighed quantity of the granules was carefully poured into the graduated cylinder and the volume (V o ) was measured. Then the graduated cylinder set into the density determination apparatus. The density apparatus was set for 100 taps and after that, the volume (V F ) was measured and the operation is continued till the two consecutive readings were equal. The bulk density and tapped density were calculated using the following formula Bulk density=w/ V o Tapped Density= W/V F Where, W = weight of the powder V o = Initial volume V F = Final volume. Hausner s ratio It indicates the flow properties of the powder and the ratio of Tapped density to bulk density of the granules is called Hausner s ratio. It is expressed in percentage and is expressed by- H= Dt / Db Where, Dt = Tapped density of the granules Db = Bulk density of the granules RESULTS AND DISCUSSION In the present investigation it is aimed to formulation, evaluate and optimize of an oral immediate release antibiotic formulation of amoxicilline development was to made to increase the rate of dissolution by increasing the release rate of drug form the solid oral dosage form by using magnesium stearate sodium starch glycolate povidone microcrystalline cellulose carboxy methylcellulose sodium Tablets with various compositions were formulated by direct compression dry granulation wet granulation using high shear granulator. The effect of moisture on the strength of bulk solids was the target of many investigations. After granulation, the extra granular excipients are mixed to provide a smooth surface on the tablet and to increase the flow and compressibility of granules and to reduce the risk of capping and lamination. The ultimate evaluation of tablet formulation lies in its tablet characteristics. Accordingly, tablets prepared by wet granulation mode were compressed at compression force of 21 x 10mm. oblong standard concave punches, flow properties of the powders can be judged from the bulk density and tapped density and hausner s ratio. The hausner s ratio 1.00-1.11 indicates free flowing and 1.60 with poor flow properties. The results of the parameters are given below The prepared tablets were evaluated for weight variation, thickness, hardness, friability and disintegration time and in-vitro drug release. The prepared tablets had acceptable variations in weight and thickness. Since mechanical integrity is importance in successful compressed formulation, the hardness of tablets were determined and found to be in the range of 120-140N.Friability was observed between 0.29-0.46%, which was below 1% indicating the sufficient mechanical integrity and strength of the prepared tablets. The prepared tablets were disintegrated within 2-3 min. Disintegrating agent was added different ratios at the both processing steps for all the formulations. The Dissolution results showed that more than 80% drug release at 10min 100% drug release occurred within 30 min dissolution profile of optimized trail II showed the similar release pattern like innovator product with F2 value 70.02. the trails III,IV,V. are the optimization trails the F2 value observed for III was 51.49 when lubricant concentration was increased by 5% the release was observed slower than innovator product. The F2 value IV was observed 54.93, when the process of granule preparation was changed similar result observed like trail III. The trail V value 69.1, which was similar for the optimization trails, in trail V showed maximum F2 value in optimization wet granulation trails significant change in F2 value observed but most of the trails F2 value was found to be greater than 50, small changes in excipient concentrations in the process shows effect on the dissolution profile of the drug from the tablet hence trail V was found more optimized trail by wet granulation process of amoxicilline grade A. trail II shows similar release with innovator profile with F2 value 74.05%. trail II was the optimized trail, III, IV, V, are the optimized trails III F2 value observed 77.86%, when lubricant concentration was reduced by 5% the release was observed faster or similar to innovator product the F2 value IV 62.36,when the lubricant concentration was increase by 5% the release was slower than innovator product the trail V observed with F2 value 73.39, which was similar as for the optimized trail, the process optimization trail II was taken with lower thickness the F2 value was observed 79.69, which was greater than the optimized trail V, from these results it is confirmed that the greater release observed with lower thickness. In-Vitro Dissolution studies Dissolution profile of innovator product The dissolution results showed that more than 80 % drug released at 10 min, and 100 % drug release occurred within 30 min. 39 P a g e

Table 1. Formulation trials for tablet of Grade A API I II III IV V VI VII VIII IX amoxicilline 1173.84 1173.84 1173.84 1173.84 1173.84 1173.84 1173.84 1173.84 1173.84 Avicel PH 101 - - 64.11 57.56 - - - - - Avicel PH 102 77.31 77.31 - - 24.81 18.26 11.71 18.26 18.26 Avicel RC 591 - - - - 39.3 39.3 39.3 39.3 39.3 Povidone K-30 - - 13.1 6.55 - - - - - Sodium Starch Glycolate 32.75 32.75 52.4 65.5 65.5 65.5 65.5 65.5 65.5 Magnesium stearate 26.2 26.2 6.55 6.55 6.55 13.1 19.65 13.1 13.1 Total weight (mg). 1310 1310 1310 1310 1310 1310 1310 1310 1310 Table 2. Formulation trials for tablet of Grade B API I II Amoxicilline 1168.33 1168.33 Avicel PH 102 82.72 82.72 Sodium Starch Glycolate 32.75 32.75 Magnesium stearate 26.2 26.2 Table 3. Formulation trials for tablet of Grade C API I II III IV V VI Amoxicilline 1169.21 1169.21 1169.21 1169.21 1169.21 1169.21 Avicel PH 102 81.84 81.84 75.29 81.84 69.29 75.29 Sodium Starch Glycolate 32.75 32.75 39.3 39.3 39.3 39.3 Magnesium Stearate 26.2 26.2 26.2 19.65 32.2 26.2 Total Weight (mg) 1310 1310 1310 1310 1310 1310 Table 4. Process optimization trials I II amoxicilline 1169.21 1169.21 MCC PH 102 75.29 75.29 Sodium Starch Glycolate 39.3 39.3 Magnesium Stearate 26.2 26.2 Total Weight (mg) 1310 1310 Table 5. Results of blend analysis for amoxicilline (Grade A) Angle of repose Bulk density Tapped density Compressibiliy index Hausner s ratio I 37.21 0.2780 0.525 47.04 1.88 II 33.69 0.55 0.88 33 1.66 III 29.28 0.4912 0.6231 21.16 1.26 IV 29.29 0.4910 0.6233 21.22 1.26 V 29.3 0.4912 0.6243 21.31 1.26 VI 29.1 0.4912 0.6161 20.27 1.25 VII 29.28 0.4912 0.6160 20.25 1.25 VIII 29.1 0.4914 0.6143 20 1.25 IX 29.0 0.4911 0.61602 20.23 1.25 40 P a g e

Table 6. Results of blend analysis for amoxicilline(grade B) Trial No. Angle of repose Bulk density Tapped density Compressibility index Hausner ratio I 28.1 0.6634 0.8242 16.08 1.24 II 28.30 0.6630 0.8240 19.53 1.24 Table 7. Results of blend analysis amoxicilline (Grade C) Trial No. Angle of repose Bulk density Tapped density Compressibility index Hausner ratio I 37.2 0.55 0.88 33 1.88 II 29.3 0.626 0.7981 21.55 1.26 III 29.3 0.629 0.797 21.07 1.25 IV 29.32 0.625 0.810 22.83 1.28 V 29.3 0.629 0.797 21.68 1.25 VI 29.29 0.627 0.7980 21.42 1.27 Table 8. Results of blend analysis for amoxicilline (Grade C) with changed physical parameter (change in thickness and lubrication time) Trial No. Process change parameters Angle of repose Bulk density Tapped density Compressibility index Hausner ratio VII Thickness 29.7 0.4910 0.6230 21.17 1.26 (7.65mm) VIII Lubrication Time(5min) 28.4 0.4920 0.6221 20.91 1.26 Table 9. Evaluation of Tablet (Grade A) Weight (mg)* Thickness Hardness Friability D.T (mm)* (N) (min) I - - - - - II 1301.3 7.65 150-210 0.41 2-3 III 1312.2 8.22 240-273 0.15 8-9 IV 1312.1 8.14 195-260 0.21 3-4 V 1309.9 7.81 190-245 0.29 3-4 VI 1311.2 7.82 185-240 0.25 3-4 VII 1312.3 7.81 190-240 0.24 3-4 VIII 1312.4 7.82 220-260 0.21 4-5 IX 1311.3 7.82 190-240 0.23 3-4 Table 10. Evaluation of tablet (Grade B) Weight (mg)* Thickness Hardness Friability DT (mm)* (N) (min) I 1312 7.65 90-130 2.31 1-2 II 1309 7.69 160-205 0.79 1-2 Table 11. Evaluation of tablet (Grade C) Weight (mg)* Thickness Hardness Friability DT (mm)* (N) (min) I 1303.3 7.71 180-220 0.46 2-Jan II 1307.1 7.77 190-230 0.41 3-Feb III 1309.2 7.76 185-230 0.29 3-Feb IV 1312.12 7.77 180-230 0.35 3-Feb V 1311.2 7.75 230-250 0.29 3-Feb VI 1309.3 7.76 190-220 0.31 3-Feb 41 P a g e

Table 12. % drug release for innovator sample Time (min) 5 65.70 10 88.30 15 95.80 20 99.50 30 102.20 45 103 Dissolution profile for optimization Trials of amoxicilline Grade A: Table 13. % drug release for Trial II to V Time(min) Innovator Trial II Trial III Trial IV Trial V 5 65.7 69.1 55.1 59.1 64.1 10 88.3 84.9 72.7 79.1 85.8 15 95.8 91.8 89.1 87.7 92.8 20 99.5 95.3 84.1 92 96.2 30 102.2 98.1 101.1 97.6 100.4 45 103 100.2 101.9 99.7 101.2 F2 70.02 51.49 54.93 69.1 Table 14. Dissolution Profile For n trails of amoxicilline Grade C Time(min) Innovator Trial II Trial III Trial IV Trial V 5 65.7 62.3 61.4 64.11 69,2 10 88.3 83.3 91.11 80.14 82.3 15 95.8 96.1 95.86 91.13 96.2 20 99.5 98.1 98.92 92.89 99.1 30 102.2 99.5 102.13 97.66 100.1 45 103 100.2 102.4 99.19 102.2 F2 74.05 77.86 62.36 73.39 Dissolution profile for process optimization Trials: Table 16. % drug release for Trial VII, VIII Time (min) Innovator Trial VII Trial VIII 5 65.7 61.6 56.1 10 88.3 90.3 80.3 15 95.8 96.83 89.2 20 99.5 99.2 95.8 30 102.2 101.1 98.6 45 103 102.1 99.8 F2 79.69 58.37 Figure 1. Drug release profile for Innovator Figure 2. Comparative drug release profile for Trial VII, VIII, IX and innovator 42 P a g e

Figure 3. Comparison of release profile for all Trials Figure 4. Comparative drug release profile for Trials VII, VIII & Innovator CONCLUSION In selection of the API the Grade C API was selected for the formulation by using dry granulation as it showed poor flow by direct compression. It also reduces the process loss and increase the productivity. In selection of the granulation technique dry granulation showed the satisfactory result. In this method no flow problem occurred and all the physical parameters observed as per innovator product. No significant effect of optimization trial was found on granule properties, disintegration time but significant change was observed on release profile of drug. For Trial III and its reproducible Trial V highest F2 value was observed with satisfactory stability when charged at accelerated condition. During stability no changes in tablet properties was observed Hence Trial V was selected as optimized formulation. Hence finally Trial V was selected as reproducible, stable and in-vitro equivalent formulation for innovator formulation of Amoxicilline and can be used for further studies. REFERENCES 1. Chien YW. Novel Drug Delivery Systems, 2nd Ed. (Revised & Expanded), Dekker, New York, 1992, 160-164, 96-139. 2. Singh BN. Oral route- Drug Delivery. In: Swarbrick, J. (3rd Ed), Encyclopedia of Pharmaceutical Technology, Informa Healthcare, New York, 2, 2007, 1242, 1248. 3. Chien, Y.W., Potential developments and new approaches in oral controlled release drug delivery system. Inf. Health, 9(7), 1983, 1294-1330. 4. Ghosh TK. Drug Delivery to the Oral Cavity. In: Swarbrick, J. Drugs and the Pharmaceutical Sciences, New York, 2005, 1-3. 5. Brahmankar PM, Jiaswal SB. Biopharmaceutics and Pharmacokinetics a Treatise, 9th Ed., Vallabh Prakashan, Delhi, 2000, 1-3. 6. Lachman L, Liberman H, Kaning J. The Theory and Practice of Industrial Pharmacy, 3rd Ed., Varghese publishing House, Bombay, 1987, 293-294, 298, 335, 372, 711, 714. 7. Singh SK. Tablet Dosage forms: Non- Parenterals. In: Swarbrick, J. (3rd Ed), Encyclopedia of Pharmaceutical Technology, Informa Healthcare, New York, 2, 2007, 988-989. 8. Aulton ME. Pharmaceutics the Science of Dosage Form Design, 2nd Ed., Churchill Livingstone, 410, 2010, 133-134. 9. Bandelin FJ. Compressed Tablets by Wet Granulation. In: Lieberman, H. A. (2nd Ed), Pharmaceutical Dosage Forms: Tablets, Dekker, New York, 1, 1989, 131-151, 151-154. 10. Armstrong NA. Selection of excipients for direct compression tablet formulations. Pharm. Techn. Eur., 1997, 9(8), 24-30. 11. Armstrong NA. Tablet Manufacture. In: Swarbrick, J.(3rd Ed), Encyclopedia of Pharmaceutical Technology, Informa Healthcare, New York, 6, 2007, 3654, 3655-3657, 3661-3660, 3662, 3659-3661. 12. Banker G. and Rhodes C. Modern Pharmaceutics, 4th Ed., Dekker, New York, 2002, 291-292, 310-314, 315. 13. Parikh D. Handbook of Pharmaceutical Granulation Technology, 2nd Ed., Taylor and Francis, New York, 2005, 99-104. 14. Michael DT. The granulation process 101: Basic technologies for tablet making. Pharm. Techn, 2002, 8-13. 15. Jogani PD, Gohel MC. A review of co-processed directly compressible excipients. J. Pharm. Pharmaceut. Sci, 8(1), 2005, 76-93. 43 P a g e