CHAPTER - 5 DEVELOPMENT AND EVALUATION OF ALFUZOSIN ER TABLETS

Transcription

1 55 CHAPTER - 5 DEVELOPMENT AND EVALUATION OF ALFUZOSIN ER TABLETS 5.1 MATERIALS AND EQUIPMENTS Table 5.1: List of materials used in research work Name of the Material Manufacturer Alfuzosin Hydrochloride Dr. Reddys laboratory, Hyderabad, Ph.Eur India. Doxazosin Mesylate Ph.Eur Clearsynth labs,india. Microcrystalline Cellulose FMC Bio Polymer, Ireland. ( Avicel PH-101) USP/NF Ammonio Methacrylate co-polymers (Eudragit RLPO) USP/NF Povidone(PVP K-30) USP/NF Partially Pregelatinized Starch (Starch 1500) USP/NF Hypromellose (Methocel K100M) USP/NF Gaur Gum 8000 cps Colloidal Silicon Dioxide USP/NF Magnesium Stearate USP/NF Acetonitrile (AR Grade) Triethylamine (AR Grade) Potassium dihydrogen Orthophosphate (ARGrade) Hydrochloric acid (ARGrade) Orthophosphoric acid (AR Grade) Evonik industries, Germany. ISP Sales (UK) Limited. Colorcon,Goa Dow chemical company, USA Lucid Colloids, Mumbai Evonik industries, Germany. Ferro corporation, Cleveland. Merck Specialties Pvt. Ltd, Mumbai. Merck Specialties Pvt. Ltd, Mumbai. Merck Specialties Pvt. Ltd, Mumbai. Merck Specialties Pvt. Ltd, Mumbai. Merck Specialties Pvt. Ltd, Mumbai.

2 56 Potassium coated EDTA Tubes Rabbits weighing Kg Merck specialities Pvt.Ltd, Saastra College of Pharmaceutical Education and Research, Nellore Table 5.2: List of equipments used in the research work Equipment Name Electronic Weighing Balance Manufactured By Mettler Toledo (AB104), Germany. Rapid mixer granulator Diasona, Bombay. Hot air oven Innovative instruments, Delhi. Double cone blender Shakthi engnering, Ahemadabad Sieves Jayant Scientific Ind., Bombay Compression Machine (8 Station) Cadmach, Ahemadabad Digital Vernier Calipers Mitutoyo (CD-8CSX), China. Friability Apparatus Electrolab EF 2W, Mumbai. Hardness Tester Dr. Schleuniger (6D), Germany. Dissolution Apparatus TDT-08L, Electrolab, Mumbai. Sonicator Power sonic 505, India. HPLC Waters, USA. FT-IR Perkin Elmer,Japan. Vortex Mixer Spinex, India. Stability Chamber Mack, Mumbai. DSC DSC21, Mettler Toledo, USA. UV visible spectrophotometer Analytikjena Specord 210 Overhead 3-blade medium duty stirrer Remi stirrer, Mumbai, India. Cyclomixer Remi Instruments, Mumbai, India. Multifuge Centrifuger Heraus, Germany.

3 ANALYTICAL METHODS There are several reported methods for the estimation of alfuzosin available in the literature, those are UV, Colorimetry, HPLC &LC-MS methods. In the present investigation we have develop a modified Ultraviolet spectroscopic method for the estimation of alfuzosin hydrochloride for dissolution samples*.hplc method was developed for estimation of drug content (Assay)*.The analytical methods (i.e dissolution and assay) of alfuzosin hydrochloride extended release tablets not published in official pharmacopeia.(i.e IP, BP&USP) * The methods was developed on the basis of development and validation of UV spectrophotometric method for estimation of alfuzosin by Adsule Prajakta V et al., 40 for dissolution samples and New RP HPLC method development and validation of assay for alfuzosin in tablet dosage form by K.S.Bharathkumar et al., 44 for uniformity of content and assay samples. A new High performance liquid Chromatographic method was developed and validated for the estimation of alfuzosin in rabbit plasma Method development The solubility of the Alfuzosin was tested in different dissolution media like 0.01N HCl, ph 4.5 Acetate buffer, ph 6.8 phosphate buffers, ph 10.0 phosphate buffer and Purified water. Based on the solubility data dissolution media 0.01N HCl was selected as media with 900mL of volume, maintained at 37 ± 0.5 C and USP Apparatus- II (Paddle). Samples were collected at appropriate time intervals from

4 58 dissolution vessels and diluted the samples and measured the absorbance at 245nm using UV-Visible spectrophotometer and calculated using standard calibration curve Standard calibration curve of Alfuzosin Preparation of 0.01N HCl 8.5mL of Hydrochloric Acid was diluted with water to 10 Litres Preparation of standard stock solution 25.0 mg of Alfuzosin HCl weighed in to 50ml volumetric flask and made up the volume with 0.01N HCl. 2ml of this solution further diluted to 100ml with 0.01N HCl Preparation of standard Calibration Curve From the standard stock solution serial dilution were done to obtain solutions ranging from 0.5µg/mL to 6.0µg/mL, i.e. from 10% to 120% with respect to sample concentration. The absorbance of above solutions was measured at wavelength of 245nm using UV-Visible spectrophotometer (Analytikjena Specord 210), against dissolution media as blank. The absorbance values of standard curve was represented in Table 5.3 and a graph was plotted of concentration v/s absorbance which was shown in Fig. 5.1

5 Absorbance 59 Table 5.3: Standard Calibration curve values of Alfuzosin Concentration in µg/ml Absorbance Slope Intercept Correlation Coefficient The linear equation was y = x Where x is concentration and y is the peak absolute area. The correlation coefficient was r = 1.000, indicating good linearity y = x R² = Concentration (µg/ml) Figure:5.1 Standard Calibration curve of Alfuzosin

6 Assay by HPLC Method development Different columns, mobile phases, flow and column temperatures were tested in the development of the analytical method. C-8 and C-18 columns of the same length, different lengths and diameters were also tested and ph of buffer variations from 3.0 to 6.5 were also tested by keeping all parameters and conditions were constant (0.8 ml/min., injection volume of 20μL,temperature at 25 C). Then the mobile phases with different buffer concentrations and organic content were also tested by keeping the all parameters and conditions were constant. Finally we got the good chromatographic peak with more than 5000 theoretical plates, tailing factor of less than 2.0 and Relative standard deviation of less than 2.0% for six replicate standard injections.

7 Preparation of Dilute Orthophosphoric acid 9.3mL of 82%-Ortho phosphoric acid was diluted to 100mL with water Preparation of Buffer 2.72g of Potassium dihydrogen phosphate weighed and transferred into a beaker containing 1000mL of water. Sonicated to dissolve and 2.0mL of Triethylamine was added and mixed well. ph of the solution was adjusted to 3.0 ± 0.05 with diluted orthophosphoric acid. Solution was filtered through 0.45μ membrane filter Preparation of mobile phase Prepare and degassed the mixture of buffer and acetonitrile in the ratio of 75:25%v/v. Diluent: Mobile phase was used as diluent Chromatographic conditions HPLC System Column Flow rate Detection : Waters Alliance 2695 with empower software : C18, mm 5µ or equivalent : 0.8ml/min : 245nm Injection volume : 20µL Column temperature : 25 C Run time : 10min

8 Standard preparation 50.0mg of Alfuzosin Hydrochloride standard weighed accurately and transfer into a 200.0ml volumetric flask, dissolved and diluted with diluent.5.0ml of this solution was transferred in to 100ml of volumetric flask, dilute to volume with diluent and mixed well Method validation 76 The system suitability linearity, accuracy and precision of the method were validated. The specificity of test method by HPLC demonstrated that the excipients from tablets do not interfere with the analytic peak. The linearity of the method was tested in the concentration range 1.26µg/mL to 15.08µg/mL (10.0% to 120.0%). For accuracy of the method, standard drug was spiked from 70.0% to 130.0% and recovery was found to be 99.2% to 100.9% and RSD 0.8%. The precision of the method was checked were found to be relative standard deviation 0.9%.

9 Peak area 63 Table 5.4: Linearity Concentration in % Concentration in µg/ml Peak Area Slope Intercept Correlation Coefficient y = x R² = Concentration (µg/ml) Figure 5.2: Alfuzosin assay Linearity graph

10 64 Level Table 5.5: Accuracy/recovery Actual weight added in mg % in mg recovery % recovery 70% % % Mean 99.9 SD 0.8 RSD 0.8 Table 5.6: Precision S.No. % Assay Avg 99.4 SD 0.86 % RSD 0.9 Figure 5.3: Alfuzosin Assay Standard

11 PRE-FORMULATION Solubility Analysis The solubility of Alfuzosin Hydrochloride was determined in different media as follows N HCl 2. p H 4.5 Acetate buffer 3. p H 6.8 phosphate buffer 4. p H 10.0 phosphate buffer 5. Purified Water 1.0 gm amount of the drug was weighed and transfer to 25 ml volumetric flasks. To each of the volumetric flasks above mentioned media were added and shaken well. The volume was made up to volume with same media the samples were kept in constant water bath shaker for 24 hours at temperature of 37 C. After 24 hours the samples were removed from bath, equilibrated for 1 hr. then the samples were filtered through 0.45 μm filter. The dissolved drug was measured using UV visible spectrophotometer at 245 nm after suitable dilutions Compatibility Studies Differential Scanning Calorimetry Differential Scanning Calorimetry of active ingredient and polymers were studied to investigate the compatibility of the both materials when mixed together by observing any changes occur in melting points of the drug. The test was performed at a rate of 5 C

12 66 min -1 from 25 C to 300 C temperature range under nitrogen flow of 25 ml min-1 using differential scanning calorimeter Fourier Transform Infra-Red (FT-IR) spectral analysis Fourier Transformed Infrared (FT IR) spectrums of Alfuzosin with HPMC K100M,Guar gum 8000cP, Eudragit RLPO and Povidone K-30 performed individually and in combinations at range of 400 to 4000 cm -1 and the resolution was 1 cm -1 using Fourier Transform Infrared (FTIR) spectrophotometer, (Perkin Elmer, spectrum-100, Japan )using the KBr disk method (2 mg sample in 200 mg KBr). This spectral test was used to check the compatibility of Alfuzosin Hydrochloride with the selected polymers. The spectrums were shown in Fig to 5.31.

13 PREPARATION OF MATRIX TABLETS Preparation of matrix tablets containing Alfuzosin The wet granulation technique was chosen to prepare matrix tablets. The compositions of the formulations were given in Table 5.7. Matrix tablets were prepared using below technique. Step1.Required quantity of Alfuzosin, retardant (HPMC K100 M or Guar gum 8000 cps or Eudragit RLPO) and other excipients (Microcrystalline Cellulose (AVCEL PH 101, Pregelatinized Starch and Povidone) were weighed and sifted through 40# sieve. Step 2. Step1 material was mixed in rapid mixing granulator (RMG) for 15min and blend was granulated using purified water as the granulating agent. Step 3. The wet granules were sifted through 14 # sieve and dried in hot air oven at inlet temperature of 60 ± 5 C till the moisture comes below 3%w/w. Step 4. The dried granules were sieved through 20 # sieve and lubricated with Magnesium stearate and colloidal silicon dioxide (previously shifted through # 40 mesh) for about 5 min in a double cone blender. Step 5. The lubricated granules were compressed into tablets using 8.8 mm round shaped with standard concave punches.(compression machine 8 station ).

14 68 Table5.7: Composition of matrix tablet containing Alfuzosin mg/tablet Name of ingredient ALF/01 ALF/02 ALF/03 ALF/04 ALF/05 ALF/06 ALF/07 ALF/08 ALF/09 ALF/10 ALF/11 ALF/12 Alfuzosin HCL Microcrystalline Cellulose Avicel PH Eudragit RLPO Povidone K Starch Hydroxypropyl methyl cellulose K100M Guar gum 8000 cp Purified Water QS QS QS QS QS QS QS QS QS QS QS QS Collodial silicone dioxide Magnesium Stearate Tablet Weight

15 EVALUATION OF TABLETS Evaluation of physical parameters for granules Flowability 78,79 Flowablility of lubricated granule were tested by using Bulk density, Tap density, Compressibility, Hausner s ratio and Angle of repose. Formulas are as below. The bulk density of prepared granules were determined by three-tap method. Weighed quantity (15gm of granules) was carefully introduced in to a 100mL graduated cylinder. The cylinder was dropped on to a hard wood surface 3 times from a height of 2.5cm at an interval of 2sec. The bulk density was obtained by dividing weight of the sample by volume of the sample. The tap density is the ratio of weight of the dry its tapped volume. The above weighed quantity of granules was placed on tapped density tester (Electrolab Model:ETD-1020) and subjected to USP Type II method i.e, 250 drops per minute and drop height is 3 mm ± 10%. The volume of the powdered weight is measured after increment of 250 drops until the difference of last two values mean is zero. Bulk Density = Tapped Density = Compressibility Index (%) and Hausner s Ratio Can calculated by using the following formulas. Compressibility Index (%) = 100 Hausner s Ratio =

16 70 Angle of repose Take funnel stand with smooth base, keep the funnel and adjust the funnel height such a way that the distance between the powder pile and funnel should be approximately 2-4 cm. keep the graph paper on base, hold the funnel orifice, pore the powder and leave the orifice to fall down. Find the height (H) of the cone of powder and circle of the powder carefully. Find out the angle of repose using following equation: Where, α is angle of the repose. H is height of the powder cone R is radius of the circle. Table 5.8: Scale of flowability 79 Flow Property Compressibility Hausner Angle of Index Ratio Repose Excellent Good Fair Passable Poor Very poor Very, Very poor >38 >1.60 >66

17 Evaluation of physical parameters for Tablets Uniformity of Weight 80 Ten tablets were selected randomly from each batch and weighed individually and determine the average weight, then check for weight variation. The average weight of tablet with % deviation as per Indian pharmacopeia was represented in table Table 5.9: Average weight of tablet with % deviation as per Indian Pharmacopeia. Average weight of Tablet % Deviation 80 mg or less 10 < 80 mg and < 250 mg 7.5 < 250 mg Thickness Thickness of the tablets was checked using digital vernier calipers by placing the tablet in between the two jaws Hardness 81 Hardness is main criteria for tablets and should have enough to withstand mechanical stress like coating, packaging, shipment, and handling by the consumer. The crushing strength test of tablet diametrically was performed on 10 tablets from each formulation by using Dr.Schleuniger, Hardness tester.

18 Friability 82 The friability test is to evaluate the ability of the tablet to withstand abrasion in coating, packaging, handling and shipping. Friability of each formulation tested using 20 tablets was determined using a Roche type friability tester.20 tablets were weighed, transferred to friabilator and performed the test with 100 rotations at speed of 25 rpm. After completion of rotations tablets were removed, dedusted and weighed. Friability of tablet should not be more than 1.0 %. Friability percentage was calculated using the following equation: Drug content by HPLC Accurately weighed 20 tablets and determined the average tablet weight in mg. Tablets were crushed into fine powder. Weighed the sample equivalent to 10.0mg of Alfuzosin and transferred it into individual 200ml volumetric flask with the aid of 120ml diluent. Solution was sonicated for 15min, dissolved and diluted to volume with diluent. Portion of sample was centrifuged for about 15min prior to dilution. 5.0ml of sample solution was transferred into individual 20.0ml volumetric flask, made up the volume with diluent and mix well. Alfuzosin was estimated by HPLC using developed method step:5.2.2.

19 Uniformity of drug content test 77 Ten tablets were selected randomly, weigh the tablet individually and place it into individual 200ml volumetric flask with the aid of 120ml diluent. Solution was sonicated for 15min, dissolved and diluted to volume with diluent. Portion of sample was centrifuged for about 15min prior to dilution. 5.0ml of sample solution was transferred into individual 20.0ml volumetric flask, made up the volume with diluent and mix well and inject to the HPLC System as followed as per assay method. (5.2.2) In-vitro Dissolution Study: (By UV) Dissolution Parameters Medium Volume Apparatus : 0.01N HCl : 900mL : USP-II (Paddle) Revolutions(RPM) : 100 Temperature Time Points Lambda : 37 ± 0.5 C : 1,2,3,6,12 and 20 hours : 245nm Test Solution All the bowls were filled with 900 ml of dissolution medium and maintained at 37±0.5 C. Dropped one tablet in to each dissolution vessel and start the dissolution test ml of aliquot were withdrawn at specific time intervals and same quantity of fresh dissolution media was replaced. Aliquots were filtered through 0.45 μ (Millipore) nylon membrane filter.5ml of this solution was diluted to

20 74 10ml with dissolution medium.samples were estimated by developed method step: Effect of Hardness on dissolution To study the effect of hardness on the tablet formulation, we have compressed the formulation (B.No: ALF/10) at different hardness levels i.e low hardness (8.1 to 9.3kp), optimum hardness (10.7 to 12.2 kp), high hardness (14.4 to 15.7 kp). The dissolution studies were performed arrive the effect of the hardness on the drug release. All the samples were analysing for in-vitro drug release by using same mentioned method (5.2.1). For comparison, marketed samples also analysed by using same method (5.2.1) Statistical approach to difference and similar factor 83 The model independent method is most suitable for dissolution profile comparison when 3 to 4 or more dissolution time points are available. Statistical models such as Difference factor (f1) and similar factor (f2) both were constructed for optimized batch and marketed product dissolution profile by using following equations. Difference factor measures the % difference between 2 curves at each time point and the relative error between the two curves, similarity factor is a measurement of % dissolution similarity between the two curves. Difference Factor f1={[ t=1 n (Rt-Tt)]/[ t=1 n Rt]} x 100 Similar Factor f2=50xlog{[1+(1/n) t=1 n (Rt-Tt) 2 ] -0.5 x 100

21 75 Where, n is number of time points. R(t) is the mean % drug dissolved of Marketed product at time t. T(t) is the mean % drug dissolved of test product. f1 value should be close to 0 (0 to 15) to prove the both the formulations are not different. f2value should be between 50 to 100 to prove the both the formulations are similar Kinetic modeling system for In-vitro release Zero Order Drug dissolution from dosage forms that do not disaggregate and release the drug slowly (a constant release rate) can be represented by zero order equation. To study the release kinetics, in vitro data of drug release studies were plotted as cumulative amount of percentage drug released versus time. It describes the rate of drug release is independent of the concentration of dissolved substance. C = Kot Where, Ko is zero-order rate constant expressed in units of concentration/time and t is the time. Application: This equation can be used to describe the drug dissolution of matrix tablets with low soluble drugs, osmotic systems andtransdermal systems, First Order This model useful in the determination of drug absorption and/or elimination. Drug release depending on the concentration.

22 76 LogC=LogCo-kt /2.303 Where, Co = The initial concentration of drug and K is first order constant. Application: This equation can be used to describe the drug release in porous matrices those containing water-soluble drugs Erosion model 85 This equation defines the drug release based on erosion alone. Q = 1-(1-k3t) 3 Where, Q is the fraction of drug released at time t, k3 is the release rate constant. Thus, a plot between [1-(1-Q) 1/3] against time will be linear if the release obeys erosion equation Korsmeyer-Peppas model To find the drug release mechanism first 60% drug release data were fitted in Korsmeyer-Peppas model, which described drug release from a polymeric system equation. To study release kinetics, in vitro drug release data was plotted as log cumulative % drug release versus log time. Mt / M = Kt n Where Mt / M =a fraction of drug released at time t, K = The release rate constant and n is the release exponent. The n value is used to characterize different mechanism of drug release for cylindrical shaped matrices Higuchi s Model The first mathematical model which describes drug release from a matrix system proposed by Higuchi in It is applicable for

23 77 planar systems initially; it was then extended to different geometrics and porous systems. Q=KH x T1/2 KH = The Higuchi dissolution constant The values of cumulative percentage drug release versus square root of time. Application: This can be used to describe the drug release from matrix tablets with water soluble drugs and transdermal systems. 5.6 Stability Studies 86 Stability study of selected formulation was tested according to international conference of harmonization guidelines. The tablets was stored in Alu-Alu blister for 3 months in stability chamber at 40 C ± 2 C & 75% ± 5 % RH. Stability samples were tested for Physical, drug content and in vitro dissolution.

24 mg/ml RESULTS Solubility Analysis Solubility of Alfuzosin in different media at 37 C ± 0.5 C. Table 5.10: Solubility of Alfuzosin Media p H N HCL p H 4.5 Acetate buffer p H 6.8 Phosphate buffer p H 10.0 Phosphate buffer Purified Water Solubility mg/ml 192( mg/ml) 172( mg/ml) 159( mg/ml) 123( mg/ml) 123( mg/ml) Solubility mg/ml ph N HCL ph 4.5 Acetate buffer ph 6.8 Phosphate buffer Media ph 10.0 Phosphate buffer Purified Water Figure 5.4: Solubility Study of Alfuzosin in Different media

25 Compatibility Studies Differential Scanning Calorimetry Figure 5.5:DSC of Alfuzosin Figure 5.6: DSC of Alfuzosin with HPMC K100M

26 80 Figure 5.7: DSC of Alfuzosin with Guargum 8000 cp Figure 5.8:DSC of Alfuzosin with Eudragit RLPO

27 81 Table 5.11: DSC characteristics of pure drug and Combination with Polymers Parameters Alfuzosine HCl (API) API+HPM C K100M API+Gaurgum API+Eudragit RLPO On Set ( C) Peak ( C) Delta H (J/g) Fourier Transform Infra-Red (FT-IR) spectral analysis Figure 5.9: IR spectrum of Alfuzosin

28 82 Figure 5.10: IR spectrum of HPMC K100M Figure 5.11: IR spectrum of Guargum 8000 cp

29 83 Figure 5.12: IR spectrum of Eudragit RLPO Figure 5.13: IR spectrum of Alfuzosin with HPMC K100M

30 84 Figure 5.14: IR spectrum of Alfuzosin with Guargum 8000 cp Figure 5.15: IR spectrum of Alfuzosin with Eudragit RLPO

31 85 Table 5.12: Characteristic peaks of Alfuzosin Frequency (cm -1 ) Functional Group 1503,1530,1552 C-C stretching 3183 C-H stretching 1213 C-O stretching 2954 Aromatic ring attached to C-H stretching 3345 NH2 stretching 1307 C-N Table 5.13: Characteristic peaks of HPMC K100M Frequency (cm -1 ) Functional Group 1039 C-C stretching 2923 C-H stretching 1279 C-O stretching 3500 CH2CH(OH) CH3 Stretching value -OH stretching Table 5.14:Characteristic peaks of Guargum 8000 cp Frequency (cm -1 ) Functional Group 870 to1022 C-C stretching CH stretching 1154 C-O stretching 3400 OH stretching Table 5.15: Characteristic peaks of Eudragit RLPO Frequency (cm -1 ) Functional Group 1262 C-C stretching 2925 C-H stretching 1020 C-O stretching 1734 C=O stretching

32 86 Table 5.16: Characteristic peaks of Alfuzosin + HPMC K100M Frequency (cm -1 ) Functional Group Alfuzosin HCl 1530 C-C stretching 1240 C-O stretching 2924 Aromatic ring attached to C-H stretching 3400 NH2 stretching 1395 C-N stretching HPMC K100 M 1067 C-C stretching 2920 C-H stretching 1277 C-O stretching 3400 CH2CH(OH) CH3 Stretching value -OH stretching Table 5.17: Characteristic peaks of Alfuzosin+Guargum 8000 cp Frequency (cm -1 ) Functional Group Alfuzosin HCl 1504,1531 C-C stretching 1241 C-O stretching 2924 Aromatic ring attached to C-H stretching 3400 NH2 stretching 1395 C-N stretching Guargum 8000 cps 872 C-C stretching CH stretching 1155 C-O stretching 3400 OH stretching

33 87 Table 5.18: Characteristic peaks of Alfuzosin+ Eudragit RLPO Frequency (cm -1 ) Functional Group Alfuzosin HCl 1529 C-C stretching 2925 C-H stretching 1245 C-O stretching 2853 Aromatic ring attached to C-H stretching 3400 NH 2 stretching 1348 C-N stretching Eudragit RLPO 1277 C-C stretching 2925 C-H stretching 1020 C-O stretching 1731 C=O stretching Evaluation of Matrix Tablets Evaluation of physical parameters for granules Flowability Table 5.19: Bulk properties of blend Parameter B.No Bulk Density Tapped Density Carr s Index Hausner s ratio Angle of repose ( ) ALF/ ALF/ ALF/ ALF/ ALF/ ALF/ ALF/ ALF/ ALF/ ALF/ ALF/ ALF/

34 Evaluation of physical parameters for tablets Physical parameters of tablets Table 5.20: Physical parameters of Tablets B.No Weight of Tablet (in mg) Hardness (in kp) Parameters Thickness (in mm) Friability (%) ALF/ ALF/ ALF/ ALF/ ALF/ ALF/ ALF/ ALF/ ALF/ ALF/ ALF/ ALF/

35 89 Figure 5.16: Photograph of prepared ALFUZOSIN extended release Tablets (B.No: ALF/10) Figure 5.17: Shows the swelling of prepared ALFUZOSIN extended release Tablets at 12 hr (B.No: ALF/10)

36 Evaluation of Chemical parameters for Tablets Drug Content Table 5.21: Chemical parameters of tablets Parameter Drug content ( %) B.No ALF/01 ALF/02 ALF/03 ALF/04 ALF/05 ALF/ ALF/07 ALF/08 ALF/09 ALF/10 ALF/11 ALF/ In-vitro Dissolution Study Table 5.22: Dissolution values B.No Cumulative % Drug Release 1hr 2hr 3hr 6hr 12hr 20hr ALF/ ALF/ ALF/ ALF/ ALF/ ALF/ ALF/ ALF/ ALF/ ALF/ ALF/ ALF/

37 % Drug Release % Drug Release ALF/ Figure5.18: In-vitro drug release of formulations contains Eudragit RLPO In-vitro drug release of formulations contains Guargum 8000cps ALF/02 ALF/03 ALF/ Time (hr.) Figure 5.19: In-vitro drug release of formulations contains Guargum 8000 cp In-vitro drug release of formulations contains HPMC K100M Time (hr.) ALF/05 ALF/06 Figure 5.20: In-vitro drug release of formulations contains HPMC K100M

38 % Drug Release In-vitro drug release of formulations contains HPMC K100M and Guargum Time (hr.) ALF/07 ALF/08 ALF/09 ALF/10 ALF/11 ALF/12 Figure 5.21: In-vitro drug release of formulations contains HPMC K100M and Guargum 8000 cp Figure 22: Photograph shows dissolution of Alfuzosin ER Tablets Table 5.23: Dissolution profile at different hardness of tablets (B.No: ALF/10) Time in (hr.) Low Hardness ( ) Medium Hardness ( ) High Hardness ( )

39 %Drug Release In-vitro drug release of optimised formula at diffrent hardness Time (hr.) Optimum Hardness ( ) High Hardness ( ) Low Hardness ( ) Figure5.23: Dissolution profile at different Hardness of Alfuzosin ER tablets Table 5.24: Comparison of in-vitro release of ALF/10 with Marketed Product Time points Marketed Product (Uroxatral) B.No: ALF/10 1 hour hour hour hour hour hour F2 value F1 value 1.20

40 %Drug Release 94 Table 5.25: Alfuzosin f1& f2 values S.No B.No Dissimilarity factor ( f1) Similarity factor(f2 ) 1. ALF/ ALF/ ALF/ ALF/ ALF/ In-vitro drug release of ALF/10 and Marketed Product ALF/10 Reference Time (hr.) Figure5.24: In-vitro drug release of ALF/10 and Marketed Product

41 Kinetic modeling system for In-vitro release Table 5.26: In vitro release kinetics of Alfuzosin ER tablets Correlation Coefficient (r2) B.No: Zero First Higuchi Erosion Peppas k (/h) n value Uroxatral ALF/ ALF/ ALF/ ALF/ ALF/ ALF/ ALF/ ALF/ ALF/ ALF/ ALF/ ALF/

42 Comulative percent drug release (%) 96 Selected formulation Kinetic models plots (B.NO: ALF/10) Uroxatral ALF/10 Linear (Uroxatral) Linear (ALF/10) Time (hr) Figure 5.25: Zero order plot for B. No: ALF/10

43 Log percent remaining to be released Uroxatral 1 ALF/10 Linear (Uroxatral) Linear (ALF/10) Time (hr) Fig 5.26: First order plot for B.No: ALF/10

44 Comulative percent drug release (%) Uroxatral ALF/10 Linear (Uroxatral) Linear (ALF/10) Square root time (hr) Fig 5.27: Higuchi plot for B.No: ALF/10

45 (1-Q)1/ Uroxatral ALF/10 Linear (Uroxatral) Linear (ALF/10) Time (hr) Fig 5.28: Erosion plot for B.No: ALF/10

46 log fraction release Uroxatral ALF/ Log Time (hr) Fig 5.29: Korsmeyer-Peppas plot for B.No: ALF/10

47 Stability Studies: Stability studies for FormulationB.N0:ALF/10 revealed that there was no significant change in appearance, assay, and drug release profile at 40 C± 2 & 75% RH till 3 months. Table 5.27: Stability study parameters Parameters Time period Initial 1 Month 2 Month 3 Month Description Complies Complies Complies Complies Drug content (%) Uniformity of dosage units ( %) Tablet Tablet Tablet Tablet Tablet Cumulative % Drug Release 1Hr Hr Hr Hr Hr Hr

48 DISCUSSION The solubility results are shown in Table Alfuzosin Hydrochloride was soluble in p H N HCl, p H 4.5 Acetate buffer, p H 6.8 Phosphate buffer, p H 10 Phosphate buffer and purified water. Comparatively all the media alfuzosin was more soluble in 0.01NHcl Differential Scanning Calorimetry (DSC) results are shown in Table The DSC thermograph for Alfuzosine showed melting peak starts at C ending at C and the mixer of the Alfuzosin + HPMC K100M, Alfuzosin + Guar gum & Alfuzosin + Eudragit RLPO showed melting points starting at C, C & C ending at C, C & C respectively. The endothermic energy of Alfuzosine was J/g and the mixer of the Alfuzosin + HPMC K100M, Alfuzosin + Guar gum & Alfuzosin + Eudragit RLPO were -7.60J/g, J/g& J/g respectively. From the obtained results concluded that there is no interaction between the selected polymers and drug substance Fourier Transform Infra-Red (FT-IR) spectral results are shown in Table The FT-IR spectra of pure drug showed characteristic peaks at 1503 cm -1, 1530 cm -1, 1552 cm -1,3183 cm -1, 1213 cm -1,2954 cm -1, 3345 cm -1 & 1307 cm -1. The mixer of drug substance and polymers of the selected in the formulations also showed same characteristic peaks with linear and shift so these values supported that there is no interaction between the selected polymer and the drug substance.

49 The results of bulk properties are shown in Table The bulk density of the prepared formulations are in the range of 0.325g/mL to 0.399g/mL. The tapped density was found to be in the range of 0.457g/mL to 0.500g/mL. The Carr s index and Hausner s ratio varies in the range of %, to %, respectively. Angle of repose was found to be in the range of 33.6 to The bulk properties of B.No: ALF/10 was showed flow properties (15.28 %,1.18, 33.6 )respectively. So the results clearly indicate that the preferred blend have good flowability and compressibility Results of tablet weight variation are shown in Table All batches were found within specified range ±5.0% as per Indian Pharmacopeia Tablet Thickness results are shown in Table5.20. Thickness of tablet containing total weight 300 mg was found in the range of 5.41 to 5.54 mm. Thickness of tablet containing total weight 350 mg was found in the range of 5.68 to 5.76 mm Hardness results are shown in Table Hardness of all tablets were found in the range of 9.0 to 12.2 kp. Selected batch (B.No: ALF/10) found with 10.7 to 12.2kp.

50 Friability results are shown in Table Friability of all tablets was found in the range of % w/w, which found within the specified limit. Friability of selected batch (B.No: ALF/10) was found to be 0.13% The results of the assay shown in Table5.21. The assay results are found within the pharmacopeial limits which indicate uniformity in drug content for all the prepared formulations The results of Dissolution studies are shown in Table5.22. The formulation prepared with Eudragit RLPO (40 % w/w concentration) alone (B.No: ALF/01) able to control drug release 99.8% at 3 hours. Those formulations containing guar gum alone (B.No: ALF/02, ALF/03, ALF/04) contain in the concentration of 13.33, 26.67& % w/w respectively are able to control drug release 98.2% at 3 hours, 99.2% at 6 hours and 99.1% at 20 hours respectively. The formulations contain HPMC K100M alone (B.No: ALF/05, ALF/06) in the 33.33%, 50.0% concentrations released the drug in 96.2% at 6 hours, 95.4% at 12 hours respectively. The formulation containing the combination of HPMC K 100M and Guar gum 8000 cp (B.No: ALF/07, ALF/08, ALF/09, ALF/10, ALF/11 & ALF/12) at the concentrations of & 8.57, 30 & 10, & 11.49, & , & , & respectively controls drug release 95.7% at 20 hours, 95.5% at 20 hours, 94.2% at 20 hours, 91.2% at 20 hours, 89.1% at 20 hours and 92.3% at 20 hours respectively.

51 105 Among all the formulations B.No: ALF/10 are shown able to drug release over 24 hours. Marketed formulation also evaluated for the comparison of the drug release which shows 97.4% released in 20 hours and found that the B.No: ALF/10 is shows similar release of the drug as that of the marketed formulation The similarity and dissimilarity factor values are shown in Table5.25. The f1, f2 are calculated by using the equation proposed by More et al, the results of formulation (B.No: ALF/10) f1= 1.20, f2= so, it indicates the proposed formulation is similar to that of the marketed formulation The results of kinetic profile are shown in Table5.26. The release profile was studied for kinetics of the drug release by zero order and first order kinetics. The correlation coefficient value (r2) of zero order and first order for all formulation were found to be in the range of and These results indicate that the release follows first order kinetics. The mechanism of drug release was studied by using Higuchi s, Erosion and Peppas model. The correlation r 2 values found to be ( ),( ) and ( ) respectively, this results indicates the release mechanism follows with super case II transport.(as the n value more than1) From the observed results indicate the drug release follows first order kinetics with diffusion mechanism.

52 The results of stability studies are shown in Table No: The stability studies were conducted on the selected formulation (B.No: ALF/10) at 40 C/75% RH for 3 months. The tablets were evaluated for Description, Assay, uniformity of dosage units and Dissolution. There is no significance difference between the initial and final exposed samples with required physical stability and chemical stability like assay and dissolution. So this indicates the formulation is stable.