The aim of the current investigation is to design oral once daily modified release dosage forms of amoxicillin trihydrate for treatment of H. Pylori infection, which release the drug for 24 hours and match with theoretical drug release profile. The tablets and capsules were prepared by the different method using different polymers in different concentrations. The interference of the polymers was ruled out by FT-IR spectroscopy studies. The powder-blends of tablets and drug were evaluated for their physical properties like angle of repose, bulk density, compressibility index, and Hausner ratio and found to be satisfactory. The manufactured tablets were evaluated for in process and finished product quality control tests including appearance, Swelling study, Drug entrapment efficiency, mucoadhesive of microsphere, Particle size analysis, drug content, and in vitro drug release. All formulations showed appearance, thickness, weight variation, hardness, friability and drug content in specified limit. All formulations showed acceptable pharmacotechnical properties and complied with in-house specifications for tested parameters. The formulated amoxicillin trihydrate tablets followed zero order release kinetics and Higuchi diffusion was the dominant mechanism of drug release, resulting in regulated and complete release within 24 hours. Formulations were subjected to short term stability studies as per ICH guidelines and were found stable. Microsphere formulations 8 were evaluated for weight uniformity, drug content and in vitro drug release.
Amoxicillin an acid stable, semisynthetic drug belongs to a class of antibiotics called the Penicillin’s (‐ lactam antibiotics). It is shown to be effective against a wide range of infections caused by wide range of Gram ‐positive and Gram‐ negative bacteria in both human and animals.
It is a congener of ampicillin (a semi‐ synthetic amino‐ penicillin) differing from the parent drug only by hydroxylation of the phenyl side chain.
It has found a niche in the treatment of ampicillin-responsive infection infections after oral administration4‐6. Chemically amoxicillin (2S, 5R, 6R) 6[[(2R) 2Amino2(4hydroxyphenyl) acetyl] amino] 3,3dimethyl-7-oxo-4-thia-1-aza-bicylo [3.2.0] heptane2carboxylic acid.
Amoxicillin is white or almost white (amoxicillin trihydrate‐off white crystalline, and amoxicillin‐sodium white or slight pink, amorphous, very hygroscopic) powders, with slight sulphurous odour, compatible with citrate, phosphate and borate buffers [7-9, 25]. Amoxicillin sodium is very soluble in water, sparingly soluble in anhydrous ethanol, very slightly soluble in acetone, while Amoxicillin trihydrate is slightly soluble in water, very slightly soluble in ethanol (96 per cent), practically insoluble in fatty oils. It dissolves in dilute acids and dilute solutions of alkali hydroxides.
Degradation of amoxicillin trihydrate as well as sodium, in sealed and open containers were found to show two step degradation at various temperatures. Under controlled humidity conditions, both the amoxicillin trihydrate and amoxicillin sodium showed first order degradation.
Literature data indicated that amoxicillin in dilute aqueous solution, was found to follow first order or pseudo first order degradation rate at constant pH with a minimum rate at about pH 6. Amoxicillin degradation was subject to catalysis by phosphate and citrate buffers with a 10-fold increase in rate with phosphate. Increasing ionic strength was reported to have positive effect on degradation rate in alkali and a negative effect in acid.
At higher concentration amoxicillin gave non first order degradation kinetics, indicative of dimerization reaction and the rate of dimerization of amoxicillin at pH 9 is greater than that of other aminopenicillins. Degradation of amoxicillin (sodium salt) at higher concentration became faster in the presence of carbohydrates (dextrose, dextran, and sorbitol) and alcohols. Amoxicillin showed a pH dependent stability with the stability increasing with decrease in pH 26. The acid‐base catalysis was seemed to be the mechanism of degradation of amoxicillin in buffered solutions25. Amoxicillin was reported with pKa values of 2.67, 7.11 and 9.55 at 370C26, 27 found to have the lowest solubility at a pH range of 4 to 6 28. The relative solubility (the pH‐apparent solubility profile) of amphoteric penicillin (amoxicillin) was determined under physiological temperature conditions for amoxicillin and the results indicated U‐shaped pH‐solubility curve, with the minimum solubility at the pH near the isoelectric point.
Amoxicillin is bactericidal against susceptible micro‐organisms through the inhibition of biosynthesis of cell wall mucopeptide during bacterial multiplication. It acts by binding to penicillin‐ binding protein 1A (PBP‐1A) located inside the bacterial cell well. The penicillin’s (amoxicillin), acylate the penicillin‐sensitive transpeptidase C‐terminal domain by opening the lactam ring causing inactivation of the enzyme, prevents the formation of a cross‐link of two linear peptidoglycan strands, inhibiting the third and last stage of bacterial cell wall synthesis, which is necessary for cell division and cell shape and other essential processes; and thus, the lethality of penicillin for bacteria involves both lytic and non‐lytic mechanisms. Cell lysis is than mediated by bacterial cell wall autolytic enzymes such as autolysins; it is possible that amoxicillin interferes with an autolysin inhibitor. The imperfect cell wall synthesis makes bacterial cells to absorb water by osmosis; as gram positive & gram-negative bacteria have 10‐30 & 3‐5 times intracellular osmotic pressure than external environment. Amoxicillin is more effective against gram positive than gram negative micro‐organisms and it demonstrates greater efficacy to penicillin, penicillin V and comparable to other antibiotics, e.g. ampicillin, azithromycin, clarithromycin cefuroxime and doxycycline, in treatment of various infections/ diseases.
In the past decade, amoxicillin has been reported to be useful in the management of many indications and is used to treat infections of the middle ear (otitis media), tonsils (tonsillitis & tonsillopharyngitis), throat, larynx (laryngitis), pharynx (pharyngitis), bronchi (bronchitis), lungs (pneumonia), urinary tract (UTI), skin and to treat gonorrhoea. Published reports also suggest amoxicillin as a potential candidate in treatment of Chlamydia trachomatis, typhoid fever, early Lyme disease, erythema migrans & erythema migrans borreliosis, mucopurulent cervicitis, acute maxillary sinusitis, gastritis & peptic ulcers and meningitis conditions. Recently American Heart Association (AHA), American Dental Association (ADA) and new recommendation by American Academy of Orthopaedic Surgeons (AAOS) changed prophylaxis protocols suggested that it can be used as prophylaxis against bacterial endocarditis, in patients with prosthetic joint replacements and in dentistry. Amoxicillin is susceptible to degradation by β‐lactamase‐producing bacteria, and so may be given with β‐lactamase inhibitor such as clavulanic acid.
Amoxicillin is well absorbed (at different rate and extent from various regions of gut) from GIT. It enjoys widespread clinical use, not only because of its broad antibacterial.
Oral bioavailability : 95 % by oral route
Protein binding : 20 %
Volume of distribution (V/ F) : 0.2-0.4 L/kg
Half- life : 60 min (average)
Route of administration : Especially oral
Fatty meal significantly interferes with amoxicillin, the time above MIC (T>MIC) was prolonged by administration with food. Mean unbound T>MIC of 0.06 µg/ml (minimum required for the inhibition of S. pyogenes) increased from 11.0 hours under fasting conditions to 12.2 hours with a low‐fat meal and 14.6 hours with a high‐fat meal.
Amoxicillin was received fromDSM Anti-infective India Limited, India and other excipient received from Oxford laboratory,Loba. Chemicals Bombay Becta Laboratories Surat Qualigens Fine Chemical
Dissolution of polymer in a suitable organic solvent. Ex. Dichloromethane, acetone Dispersion of drug in the polymer solution. Under high homogenization speed. Atomization of dispersion in a stream of hot air Formation of small droplets due to atomization Solvent evaporation instantaneously. Formation of microspheres in size range of 1-100 Separation of microparticles from the hot air by means of cyclone separator. Vacuum drying is used to remove traces of solvent
The pure drug amoxicillin was scanned by UV Spectrophotometer at 200-400nm to determine λmax .The peak was observed at 228nm for amoxicillin microsphere in 1.2 pH.
The standard solution (100µg/ mL) of pure drug (Amoxicillin) was prepared in freshly prepared 0.1 N hydrochloric acid (pH-1.2). The standard calibration curve of Amoxicillin was obtained by plotting Absorbance V/s. Concentration. Table 1 shows the absorbance values of amoxicillin. The standard curve is shown in Fig. no.1.
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| 1. | 3 | 0.1851 |
| 2 | 6 | 0.2621 |
| 3. | 9 | 0.3590 |
| 4. | 12 | 0.4372 |
| 5. | 15 | 0.5576 |
| 6. | 18 | 0.6841 |
Angle of repose is defined as the maximum angle possible between the surface of the pile of powder and horizontal plane. The angle of repose were found between 160.54-210.47.
It may be defined as the mass of powder divided by the bulk volume. The bulk density were found between 0.158 -0.278 g/cc.
It may be defined as the mass of the powder divided by tapped volume. The tapped density were found between 0.225 -0.401 g/cc.
It is used to measure the porosity of the powder to be compressed to evaluate the inter particulate interactions. It were found between 27.58 -42.80 which revealed that all powders had excellent to passable flow properties.
It is used to know ease of flow of powder. Hausner’s ratio were found between 1.21-1.74 which revealed that all powder blends had good flow properties.
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| B1 | 180.22’ ± 0.22 | 0.189 |
0.286 |
33.62 ± 2.3 | 1.5112 ± 0.1 |
| B2 | 190.06’± 0.80 | 0.218 |
0.374 |
38.59 ± 2.7 | 1.7148 ±0.08 |
| B3 | 200.89’± 1.23 | 0.228 |
0.400 |
42.80 ± 1.2 | 1.7489 ± 0.03 |
| B4 | 160.54’± 0.99 | 0.278 |
0.401 |
30.07± 4.6 | 1.4341 ± 0.09 |
| B5 | 220.05’± 1.29 | 0.209 |
0.326 |
35.70 ± 3.1 | 1.5578 ± 0.07 |
| B6 | 170.11’± 0.90 | 0.274 |
0.379 |
27.58 ± 0.4 | 1.3809 ± 0.009 |
| B7 | 180.00’± 0.5 | 0.263 |
0.319 |
28.55 ± 3.0 | 1.2140± 0.04 |
| B8 | 210.47’± 0.24 | 0.158 |
0.225 |
30.45 ± 1.4 | 1.4384 ± 0.02 |
The percentage yield of different formulations was determined by weighing the microspheres after drying. The percentage yield was found between 15-36.20%.
The particle size analysis was carried out to found the particle size of microspheres. The particle size was found between 585.6-852.7.
Efficiency of drug entrapment was calculated in terms of entrapment efficiency (EE). The drug entrapment efficiency was found between 53.91-89.43.
Mucoadhesive properties of the microspheres were evaluated by the
A known weight of (50 mg) microspheres was placed in a glass vial containing 10 ml of distilled water at 37±0.5°C in with occasional shaking. It was found between 0.623-1.269.
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| 1 | B1 | 30.00 | 12.8 | 75.28 | 0.969 | 73.16 |
| 2 | B2 | 36.20 | 14.2 | 53.07 | 0.7276 | 75.28 |
| 3 | B3 | 16.72 | 19.5 | 53.91 | 0.623 | 68.86 |
| 4 | B4 | 28.50 | 13.4 | 89.43 | 1.36 | 91.14 |
| 5 | B5 | 22.40 | 12.4 | 66.13 | 1.263 | 81.85 |
| 6 | B6 | 50.32 | 15.6 | 54.12 | 1.1428 | 90.69 |
| 7 | B7 | 35.28 | 18.7 | 87.18 | 0.8822 | 71.32 |
| 8 | B8 | 15.00 | 17.5 | 67.35 | 1.2694 | 89.25 |
Release of amoxicillin from microspheres was evaluated in 0.1 N HCl (pH1.2).
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| 1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
| 2 | 1 | 21.69±1.64 | 39.72±0.68 | 44.75±1.37 | 17.09±2.20 | 12.73±0.69 | 19.38±0.39 | 16.85±2.46 | 15.78±0.14 |
| 3 | 2 | 31.16±0.14 | 43.69±0.91 | 51.31±3.83 | 24.45±0.55 | 21.02±0.10 | 25.09±0.15 | 29.05±1.36 | 25.35±0.06 |
| 4 | 3 | 34.90 ± 1.14 | 55.40 ± 1.86 | 59.74 ± 2.46 | 33.80 ± 2.57 | 32.52 ± 0.79 | 32.12 ± 1.19 | 35.17± 0.01 | 31.40± 0.09 |
| 5 | 4 | 40.95 ± 0.12 | 59.19 ± 0.46 | 61.35 ± 2.93 | 46.17± 0.38 | 43.26 ± 0.18 | 40.54 ± 0.24 | 44.34 ± 0.55 | 38.50 ± 0.12 |
| 6 | 5 | 47.16±0.18 | 64.28± 0.52 | 63.76± 1.84 | 63.48± 1.80 | 62.65±0.18 | 44.23±0.27 | 57.39±0.82 | 46.95±0.11 |
| 7 | 6 | 51.14± 0.11 | 66.98 ± 0.63 | 70.76 ± 0.00 | 78.00 ± 1.37 | 76.14 ± 1.43 | 48.44 ± 0.19 | 75.14 ± 0.55 | 59.76 ± 0.08 |
| 8 | 7 | 59.82 ± 1.26 | 72.16 ± 2.38 | 75.88± 7.28 | 86.06 ± 1.56 | 85.59 ± 2.10 | 55.61 ± 1.39 | 86.77 ± 1.36 | 67.47 ± 0.11 |
| 9 | 8 | 65.22±0.09 | 74.72±1.06 | 77.77±6.56 | 97.00±1.29 | 89.68±0.47 | 67.41±0.52 | 93.74±0.96 | 80.21±0.35 |
Drug release kinetics of B4 (best formulation) formulation.
Drug release kinetics studies for best formulation (B4) were done using software KinetDS3 and it was observed that B4 formulation followed Zero Order, Higuchi, Korsmeyer-peppas, Hixon Crowell drug kinetic model for drug release.
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| 1 | Zero Order | 0.988 |
| 3 | Higuchi | 0.972 |
| 4 | Korsemayer – Peppas | 0.963 |
| 5 | Hixon Crowell | 0.964 |
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| Appearance | Yellowish White | Yellowish White | Yellowish White | Yellowish White | Yellowish White |
| Entrapment Efficiency (%) | 44.51 ± 0.02 | 44.51 ± 0.02 | 43.39 ± 0.04 | 44.26 ± 0.01 | 43.63 ± 0.04 |
| Percent Release (%) | 92.74 ± 0.3 | 92.74 ± 0.3 | 91.89 ± 0.67 | 91.37 ± 0.73 | 90.65 ± 0.88 |
Amoxicillin with its comparable clinical efficacy to other antibacterial and favourable dosage, pharmacokinetic profile and tolerability is an excellent candidate to treat various infectious diseases. As it is less effective against gram negative organisms and bacterial resistance develop to the drug candidate, it is the one area where major development is required. Progression in work is also required to investigate new routes of administration and dosage forms with more efficacies to reduce the dose and associated side effects.
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