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FORMULATION DEVELOPMENT OF RANITIDINE HYDROCHLORIDE CONTAINING WAFERS AND ITS IN-VITRO KINETIC STUDIES A thesis submitted in partial fulfillment of the requirements for the degree of MASTER OF PHILOSOPHY (Pharmaceutics) By RAUF-UR-REHMAN 04-PHP-S-14 Session: 2014-2016 Department of Pharmacy Bahauddin Zakariya University Multan Pakistan. I II IN THE NAME OF ALLAH THE MOST GRACIOUS, THE MOST MERCIFUL “...Say: ‘Are those equal, those who know and those who do not know? It is those who are endowed with understanding that receive admonition.” (Qur’an, 39:9) Saying of Prophet Muhammad (PBUH) “A believer is never satiated with gainful knowledge; he goes acquiring it till his death and entry into Paradise.” (Tirmidhi 222) Saying of Prophet Muhammad (PBUH) Teach! Make things easy! And do not make them complicated! Be cheerful! And do not be Repulsive.” III Declaration I, Rauf-ur-Rehman S/o Muhammad Siddique M. Phil (Pharmaceutics) Scholar of the Department of Pharmacy, Bahauddin Zakariya University Multan, hereby declare that the research work entitled “Formulation Development of Ranitidine Hydrochloride containing wafers and its invitro kinetic studies” is done by me. I also certify that nothing has been incorporated in this thesis work without acknowledgment and that to the best of my knowledge and belief that it does not contain any material previously published or written by any other person or any material previously submitted for a degree in any university where due reference is not made in the text. Rauf-ur- Rehman S/o Muhammad Siddique IV Supervisor’s Declaration It is hereby certified that work presented by Rauf-ur-Rehman in the thesis entitled “Formulation Development of Ranitidine Hydrochloride containing wafers and its invitro kinetic studies” is based on the result of research study conducted by candidate under my supervision. No portion of this thesis work has formerly been offered for higher degree in this university or any other institute of learning and to best of the author’s knowledge, no material has been used in this thesis which is not his own work, except where due acknowledgement has been made. He has fulfilled all the requirements and qualified to submit this thesis in partial fulfillment for the degree of Master of Philosophy (MPhil. Pharmaceutics) in the Department of Pharmacy, Bahauddin Zakariya University Multan. Dr. Muhammad Hanif Research Supervisor, Assistant Professor, Department of Pharmacy, Bahauddin Zakariya University Multan Pakistan. V Certificate It is hereby certified that work presented by Rauf-ur-Rehman S/o Muhammad Siddique in the thesis entitled “Formulation Development of Ranitidine Hydrochloride containing wafers and its in-vitro kinetic studies” has been successfully presented and is accepted in its present form as satisfying the requirement for the degree of Master of Philosophy (Pharmaceutics) in the Department of Pharmacy, Bahauddin Zakariya University Multan. Candidate Rauf-ur-Rehman Supervisor Dr. Muhammad Hanif Assistant Professor Chairman Department of Pharmacy Bahauddin Zakariya University Multan. VI Dedication To, All family members, especially my Father and Mother, for their efforts in putting their children to the pursuit of knowledge. Throughout my life, they have supported me in my determination to find and realize my potential. Thanks for their unconditional love and support throughout the course of this thesis. My brothers, and Late sister. Finally my most respected teacher Dr. Muhammad Hanif and truly loving friends who have always been a source of love, care and strength for me. VII Acknowledgment Words are bound and knowledge is limited to praise Almighty ALLAH, the Lord of words, the gracious, most beneficent, the compassionate, and the merciful and with the blessings of whom the pavements of my life became smooth. Countless blessings upon Holy Prophet HAZRAT MUHAMMAD ﷺwho have shown the right direction to humanity and thereby enlightening their ways with faith through his eternal teachings. My sincere gratitude to my worthy supervisor Dr. Muhammad Hanif, Assistant Professor of Pharmaceutics, Faculty of Pharmacy Bahauddin Zakariya University, Multan, Pakistan for his professional guidance, supportive attitude, and scientific advice throughout the research work despite of his multifarious engagements. His inspiring help, consistent encouragement and affectionate behavior during the entire study duration will ever be remembered. I am grateful and feel highly privileged in Prof. Dr. Nazar Muhammad Ranjha (Chairman Department of Pharmacy BZU Multan) who maintained discipline that acts as a bridge between my target and accomplishment. Appreciation goes to the teaching and non-teaching staff of the department of Pharmacy whose encouragement contributed significantly to the successful completion of my work. Words are inadequate to express my special thanks to my parents, brothers, sisters, my M.Phil. Seniors, fellows especially my best friend and other junior research fellows whose nice company, prayers for my success, moral support and special care encouraged me to achieve goal. Rauf-ur-Rehman VIII Table of Contents Supervisor’s Declaration .................................................................................................... ……….V Certificate ............................................................................................................................ ………VI Dedication ............................................................................................................................ ……..VII Acknowledgment ................................................................................................................. ……..VIII Table of Contents ................................................................................................................ ………IX List of figures……………………………………………………………………….…..………XIII List of Tables ....................................................................................................................... ……...XVI List of abbreviation ............................................................................................................. ……..XVII Abstract ................................................................................................................................ ……...XIX 1. 2. Introduction ................................................................................................................. …………1 1.1. Types of wafers ................................................................................................................ 2 1.2. Polymers used in Wafers preparations ............................................................................. 4 1.3. Hydroxypropyl methyl cellulose (HPMC) ....................................................................... 5 1.4. Carboxy methylcellulose sodium (CMC) ........................................................................ 5 1.5. Guar gum (GG) ................................................................................................................ 5 1.6. Pectin (PC) ....................................................................................................................... 6 1.7. Polyethylene Glycol (PEG 200) ....................................................................................... 6 Drug profile .................................................................................................................. ……..8 2.1. Description ....................................................................................................................... 8 2.2. Molecular formula ............................................................................................................ 8 2.3. Chemical formula ............................................................................................................. 8 2.4. Chemical name ................................................................................................................. 8 2.5. Characteristics of Ranitidine hydrochloride:.................................................................... 9 2.6. Solubility of Ranitidine .................................................................................................... 9 2.7. Pharmacology ................................................................................................................... 9 2.7.1. Pharmacokinetic ........................................................................................................ 9 IX 2.7.2. Volume of distribution ..................................................................................................... 10 2.7.3. Metabolism and Excretion ...................................................................................... 10 2.7.4. Pharmacodynamics ................................................................................................. 10 2.7.5. Clinical indications ................................................................................................. 11 2.7.6. Adverse effects........................................................................................................ 11 2.7.7. Dosage and administration ...................................................................................... 11 2.7.8. Contraindication ...................................................................................................... 12 2.7.9. Cautions .................................................................................................................. 12 2.7.10. Pregnancy category ................................................................................................. 12 2.7.11. Toxicity signs and symptoms.................................................................................. 12 2.7.12. Management of toxicity .......................................................................................... 13 3. 4. Literature review ......................................................................................................... ……14 3.1. Hydroxypropyl methylcellulose (HPMC) ...................................................................... 14 3.2. Pectin wafers .................................................................................................................. 17 3.3. Carboxy methyl cellulose wafers (CMC)....................................................................... 18 3.4. Guar gum containing wafers .......................................................................................... 19 3.5. Sodium alginate and gelatin wafers ............................................................................... 21 3.6. Polyethylene glycol (PEG) wafers ................................................................................. 24 Material and Method................................................................................................... ……27 4.1 . Chemicals .................................................................................................................... 27 4.2 . Apparatus /equipment .................................................................................................. 27 4.3. Experimental design (Box- Behnken) ............................................................................ 28 4.4. Preparation of wafers ..................................................................................................... 29 4.5. Characterization of wafers ............................................................................................. 30 4.5.1. Physical parameters of wafers ................................................................................ 30 4.5.2. Surface pH .............................................................................................................. 30 4.5.3. Wafers thickness ..................................................................................................... 30 4.5.4. Weight variation test ............................................................................................... 30 4.5.5. Swelling index ........................................................................................................ 31 4.5.6. Percentage moisture absorption (PMA) .................................................................. 31 X 4.5.7. Percentage moisture loss (PML) ............................................................................. 31 4.5.8. In-vitro mucoadhesion test ...................................................................................... 31 4.5.9. Folding endurance ................................................................................................... 31 4.5.10. Disintegration time.................................................................................................. 31 4.5.11. Total dissolving time (TDT) ................................................................................... 32 4.6. Morphological studies .................................................................................................... 32 4.6.1. Scanning Electron Microscopy (SEM) ................................................................... 32 4.6.2. Optical microscopy ................................................................................................. 32 4.7. FTIR spectroscopy ......................................................................................................... 32 4.8. Content uniformity ......................................................................................................... 33 4.9. Stability studies .............................................................................................................. 33 4.10. Dissolution study ........................................................................................................ 33 4.11. Release kinetics .......................................................................................................... 33 4.11.1. Zero order release ................................................................................................... 33 4.11.2. First order release .................................................................................................... 34 4.11.3. Higuchi model ......................................................................................................... 34 4.11.4. Korsmeyer-peppas model ....................................................................................... 34 5. Result and Discussion .................................................................................................. ……35 5.1. Organoleptic evaluation ................................................................................................. 35 5.2. Effects of excipients on various parameters of wafers formulations ............................. 35 5.3. Physicochemical parameters .......................................................................................... 43 5.3.1. Surface pH .............................................................................................................. 43 5.3.2. Wafers thickness ..................................................................................................... 43 5.3.3. Weight variation test ............................................................................................... 45 5.3.4. Swelling index ........................................................................................................ 51 5.3.5. Percentage moisture absorption (PMA) .................................................................. 53 5.3.6. Percentage moisture loss (PML) ............................................................................. 54 5.3.7. In-vitro mucoadhesion test ...................................................................................... 54 5.3.8. Folding endurance ................................................................................................... 57 5.3.9. Disintegration time.................................................................................................. 57 5.3.10. Total dissolving time (TDT) ................................................................................... 59 XI 5.4. Morphological studies .................................................................................................... 64 5.4.1. Microscopic images of wafers ................................................................................ 64 5.4.2. SEM of wafers formulations ................................................................................... 65 5.5. FTIR studies ................................................................................................................... 65 5.6. Content uniformity of wafers ......................................................................................... 68 5.7. Stability studies .............................................................................................................. 68 5.8. Preparation of standard /calibration curve ..................................................................... 69 5.9. Dissolution studies ......................................................................................................... 70 5.9.1. Kinetic models of formulations (FI-FIII) ................................................................ 72 5.9.2. In-vitro kinetics of formulation (FIV-FVI) ............................................................. 78 5.9.3. In-vitro kinetics of formulation (FVII-FIX) ........................................................... 84 5.9.4. Kinetic models of formulation (FX-FXII) .............................................................. 91 5.10. Discussion ................................................................................................................... 98 5.11. Conclusion ................................................................................................................ 100 6. Refrence ........................................................................................................................ …..101 XII List of figures Figure 1: Excipients effect on surface pH of Formulations (FI-FIII) ........................................... 36 Figure 2: Excipients effect on folding endurance of formulations (FI-FIII) ................................ 36 Figure 3: Excipients effect on mucoadhesion time of formulations (FI-FIII) .............................. 36 Figure 4: Excipients effect on percentage moisture loss of formulations (FI-FIII) ..................... 37 Figure 5: Effect of excipients on surface pH of formulations (FIV-FVI)..................................... 37 Figure 6: Effect of excipients on folding endurance of formulations (FIV-FVI) ......................... 38 Figure 7: Excipients effect on in-vitro mucoadhesion time of formulation (FIV-FVI) ................ 38 Figure 8: Excipients effect on percentage moisture loss of formulations (FIV-FVI) ................... 39 Figure 9: Excipients effect on surface pH of formulations (FVII-FIX)........................................ 39 Figure 10: Excipients effects on folding endurance of formulations (FIV-FVI) .......................... 40 Figure 11: Excipients effect on mucoadhesion time of formulations (FIV-FVI) ......................... 40 Figure 12: Excipients effect on moisture loss of formulations (FVII-FIX) .................................. 41 Figure 13: Excipients effect on surface pH of formulations (FX-FXII) ....................................... 41 Figure 14: Excipients effect on folding endurance of formulation (FX-FXII) ............................. 42 Figure 15: Excipients effect on in-vitro mucoadhesion of formulations (FX-FXII) .................... 42 Figure 16: Excipients effect on moisture loss of formulations (FX-FXII) ................................... 43 Figure 17: Thickness of wafers formulations FI-FIII (n=5) ......................................................... 44 Figure 18: Thickness of wafers formulations FIV-FVI (n=5) ...................................................... 44 Figure 19: Thickness of wafers formulations FVII-FIX (n=5) ..................................................... 45 Figure 20: Thickness of wafers formulations (FX-FXII) ............................................................. 45 Figure 21: Weight variation of wafers formulationF1 (n=20) ...................................................... 46 Figure 22: Weight variation of wafers formulation FII (n=20) .................................................... 46 Figure 23: Weight variation of wafers formulation FIII (n=20) ................................................... 47 Figure 24: Weight variation of formulation FIV (n=20) .............................................................. 47 Figure 25: Weight variation of formulation FV (n=20) ................................................................ 48 Figure 26: Weight variation of formulation (FVI) ........................................................................ 48 Figure 27: Weight variation of formulation FVII (n=20) ............................................................. 49 Figure 28: Weight variation of formulation FVIII (n=20) ............................................................ 49 Figure 29: Weight variation of formulation FIX (n=20) .............................................................. 50 Figure 30: Weight variation of formulation FX (n=20) ................................................................ 50 Figure 31: Weight variation of formulation XI (n=20)................................................................. 51 Figure 32: Weight variation of formulation XII (n=20) ............................................................... 51 Figure 33: Percentage radial swelling of formulations FI-FIII (n=5) ........................................... 52 Figure 34: Percentage radial swelling of formulations FIV-FVI .................................................. 52 Figure 35: Percentage radial swelling of formulations FVII-FIX (n=5)....................................... 53 Figure 36: Percentage radial swelling of formulations FX-FXII (n=5) ........................................ 53 Figure 37: In-vitro mucoadhesion time of wafers formulations FI-FIII ....................................... 55 Figure 38: In-vitro mucoadhesion time of wafers formulations FIV-FVI .................................... 55 XIII Figure 39: In-vitro mucoadhesion time of wafers formulations FVII-FIX) ................................. 56 Figure 40: In-vitro mucoadhesion of wafers formulations FX-FXII ............................................ 57 Figure 41: Disintegration time of formulations (FI-FIII) ............................................................. 58 Figure 42: Disintegration time of formulations from FV-FVI...................................................... 58 Figure 43: Disintegration time of formulation (FVII-FIX)........................................................... 59 Figure 44: Total dissolving time of wafers formulations FI-FIII ................................................. 60 Figure 45: Total dissolving time of wafers formulation FIV-FVI ................................................ 60 Figure 46: Total dissolving time of wafers formulation FVII-FIX............................................... 61 Figure 47: Total dissolving time of wafers formulation FX-FXII ................................................ 61 Figure 48: Total dissolving time of wafers formulation FII, FV, FVIII and FXII ....................... 62 Figure 49: Total dissolving time of wafers formulation FV, FVIII .............................................. 62 Figure 50: Total dissolving time with agitation formulation FI-FIII ............................................ 63 Figure 51: Total dissolving time of wafers formulations FIV-FVI .............................................. 63 Figure 52: Total dissolving time of wafers formulations from FVII-FIX .................................... 64 Figure 53: Microscopic images wafers prepared with (HPMC) ................................................... 65 Figure 54: Microscopic images of wafers prepared with Pectin................................................... 65 Figure 55: FTIR spectra (A) Ranitidine HCL (B)Pectin ,(C) Pectin wafers ,(D) CMC wafers and (E) HPMC wafers ......................................................................................................................... 67 Figure 56: Calibration curve of ranitidine .................................................................................... 70 Figure 57: Percentage drug release of wafers formulation FI-FIII ............................................... 70 Figure 58: Percentage drug release of wafers formulations FIV-FVI .......................................... 71 Figure 59: Percentage drug release of wafers formulations FVII-FIX ......................................... 72 Figure 60: Percentage release of wafers formulations FX-FXII ................................................... 72 Figure 61: Zero order release models of formulation FI............................................................... 73 Figure 62 : Higuchi release model of wafers formulation (FI) ..................................................... 74 Figure 63: Korsmayer-peppas model of wafers formulation (FI) ................................................. 74 Figure 64: Zero order release model of wafers formulation (FII) ................................................ 75 Figure 65: First order release model of wafers formulation (FII) ................................................. 75 Figure 66: Higuchi release model of wafers formulation (FII) ..................................................... 76 Figure 67: Korsmeyer-peppas release model of (FII) ................................................................... 76 Figure 68: Zero order release model of wafers formulation (FIII) ............................................... 77 Figure 69: First order release model of wafers formulation (FIII) ............................................... 77 Figure 70: Higuchi release model of wafers formulation (FIII) ................................................... 77 Figure 71: Korsmeyer-peppas model of wafers formulations (FIII) ............................................ 78 Figure 72: Zero order release model of wafers formulation (FIV) ............................................... 79 Figure 73: First order release model of wafers formulation (FV) ................................................ 79 Figure 74: Higuchi release model of wafers formulation (FIV) ................................................... 80 Figure 75: Korsmeyer-Peppas release model of wafers formulation (FIV) ................................. 80 Figure 76: Zero order release model of formulation (FV) ............................................................ 81 Figure 77: First order release model of wafers formulation (FV) ................................................ 82 XIV Figure 78: Higuchi release model of wafers formulation (FV) .................................................... 82 Figure 79: Korsmayer-peppas release model of wafers formulation (FV). .................................. 82 Figure 80: Zero order release model of wafers formulation (FVI) ............................................... 83 Figure 81: First order release model of wafers formulation (FVI) ............................................... 83 Figure 82: Higuchi release model of wafers formulation (FVI) ................................................... 84 Figure 83: Korsmeyer-peppas release model of wafers formulation (FVI) .................................. 84 Figure 84: Zero order release model of wafers formulation (FVII).............................................. 85 Figure 85: First order release model of wafers formulation (FVII) .............................................. 86 Figure 86: Higuchi release model of wafers formulation (FVII) .................................................. 86 Figure 87: Korsmeyer-peppas model of wafers formulations (FVII) ........................................... 87 Figure 88: Zero order release model of FVIII .............................................................................. 87 Figure 89: First order release model of wafers formulation (FVIII) ............................................ 88 Figure 90: Higuchi release model of wafers formulations (FVIII) ............................................... 88 Figure 91: Korsmeyer peppas release model of formulation (FVIII) ........................................... 89 Figure 92: Zero order release model of wafers formulation (FIX) ............................................... 89 Figure 93: First order release model of wafers formulation (FIX) ............................................... 90 Figure 94: Higuchi release model of wafers formulation (FIX) ................................................... 90 Figure 95: Korsmeyer-peppas release model of wafers formulation (FIX) ................................ 91 Figure 96: Zero order release model of wafers formulation (FX) ................................................ 92 Figure 97: First order release model of wafers formulation (FX) ................................................ 92 Figure 98: Higuchi release model of wafers formulation (FX) .................................................... 93 Figure 99: Korsmeyer-peppas release model of wafers formulation (FX) ................................... 93 Figure 100: Zero order release model of wafers formulation (FXI) ............................................. 94 Figure 101: First order release model of wafers formulation (FXI) ............................................. 94 Figure 102: Higuchi release model of wafers formulation (FXI) ................................................. 95 Figure 103: Korsmeyer peppas release model of wafers formulation (FXI) ................................ 95 Figure 104: Zero order release model of wafers formulation (FXII) .......................................... 96 Figure 105: First order release model of wafers formulation (FXII) ............................................ 96 Figure 106: Higuchi release model of wafers formulation (FXII) .............................................. 97 Figure 107: Korsmeyer peppas release model of wafers formulation (XII) ................................. 97 XV List of Tables Table 1: Mucoadhesive strength of natural and synthetic polymers............................................... 4 Table 2: Mucoadhesive strength of natural and synthetic polymers............................................... 7 Table 3: Coded values of the variables of Box-behnken design for wafers formulation ............. 28 Table 4. Composition of formulations of Ranitidine hydrochloride 50mg wafers (All quantities were given in %ages) .................................................................................................................... 29 Table 5: Evaluation of various physical parameters of ranitidine hydrochloride wafers ............. 54 Table 6: Stability studies of Ranitidine hydrochloride wafers...................................................... 69 Table 7: In-vitro drug release kinetics and its model dependent approaches (FI-FIII) ................ 73 Table 8: In-vitro drug release kinetics and its model dependent approaches (FIV-FVI) ............. 78 Table 9: In-vitro drug release kinetics and model dependent approaches (FVII-FIX) ............... 85 Table 10: In-vitro release kinetics model dependent approaches (FX-FXII) ............................... 91 XVI List of abbreviation GRDDS Gastroretantive drug delivery system NDDS Novel drug delivery system GIT Gastrointestinal tract SV Surface to volume ratio HPMC Hydroxypropyl methyl cellulose NaCMC Sodium carboxymethyl cellulose PVP Polyvinyl pyrollidone GG Guar gum PC Pectin PEG Polyethylene glycol FE Folding endurance GIF Gastrointestinal fluid SEM Scanning Electron Microscopy FTIR Fourier Transform Infrared Spectroscopy RNH Ranitidine Hydrochloride NZD Nizatidine XVII ABA Absolute bioavailability SR Sustained release QID Four times a day PCM Paracetamol RSM Response surface methodology UV Ultra Violet TDT Total dissolving time PMA Percentage moisture absorption PML Percentage moisture loss CU Content Uniformity PM Petri plate method XVIII Abstract Aim of study was to develop the sustained release wafers by using solvent casting technique with various concentration of natural and synthetic polymers, plasticizers and surfactant. Box Benkhen design was applied to observe the effect of excipients on wafers formulations. Six formulations of each polymer hydroxy propylmethyl cellulose, carboxymethyl cellulose (HPMC, CMC, Pectin and Guargum) was designed by taking different concentrations of plasticizers and surfactant. On the bases of physicochemical parameters like surface texture, surface pH, tack test, weight variation wafers thickness, folding endurance, radial swelling, moisture loss, percentage moisture absorption, invitro mucoadhesion time, disintegration time, total dissolving time, content uniformity were applied and three among the six formulations of each polymer were optimized. Ranitidine hydrochloride was used as model drug in selected wafer formulations. FTIR analysis and stability studies were carried out to evaluate the excipient interactions and there was no interaction observed. FTIR spectra of drug, polymers and prepared wafers showed peaks at 3300cm-1, 1640cm-1, 1590cm-1 and 1051cm-1 due to –CH2 alkane group stretching, C=O stretching, carboxylic group stretching and stretching vibration of C-0H group respectively. Three months stability showed the stable formulations under 75% humidity and 40 ± 2 0C while the content uniformity was observed up-to 85-98%. Dissolution studies showed that formulation FV, FVIII were found to be most suitable for sustained release delivery out of 12 optimized formulations (FI-FXII) these formulations showed more than 85 % drug released after 8 hrs. In-vitro drug release data was analyzed by applying different kinetic models zero order, first order, higuchi and korsmeyer-peppas to all formulations which showed observe value of R2 in case of first order was very close to 1 indicating concentration dependent release of drug. First order release model results showed that R2 value of formulation FVIII was very close to 1 showed best release pattern out of all formulations. Korsmeyer –peppas model was applied results showed n value of all formulation were observed to be less than 0.45 showed fickian release pattern. XIX Keywords: Wafers; solvent casting method; mucoadhesion; in-vitro drug release; release kinetic models; tackiness. XX 1. Introduction Various routes are used for adminstration of drugs to the human body these routes includes oral parental, intrathecal and subcutaneous, however the oral route has gained much more importance due to its ease of administration [1]. In oral dosage systems various oral dosage forms includes liquids, solids and wafers are given much more importance. Pharmatechnologist have an urge to develop such dosage forms which are used as conventional dosage systems [2]. Conventional dosage system employed for treatment of various diseases, tablet is preferred due to its ease of administration ease of manufacturing process as compared to other dosage forms however is not suitable for elderly patients and children. Most recently pharmaceutical industry is going to divert their research from conventional dosage forms to novel drug delivery system based on changing patient requirements and global development. The novel micropartices, lozenges, wafers and films are being made during the last couple of decades. Wafers are the one of most advance drug delivery system used for the management of diseases. Wafers disintegrate or dissolute when they come in contact with the oral mucosa or mucosa of the gastrointestinal tract. Wafers have porous structure and a relatively more surface area as compared to the other dosage forms therefore they also have more drug loading capacity. There are however certain limitations for the development of wafers formulations which need considerable attention with respect to solubility of drug, polymer mucoadhesive property which achieves efficient targeted delivery and its bioavailability. Wafers as novel drug delivery system shows certain benefits such as rapid disintegration, controlled release, measured (accurate quantity) can be administered, improved patient compliance, efficacy(E) and safety However, as controlled released drug delivery system can also be used either as mucoadhesive wafers formulations or as gastroretantive drug delivery system (GRDDS) thus enhance retention time in gastric fluid. Drugs that are less absorbed through the intestine or those having the narrow therapeutic window. Therefore, to maintain the therapeutic concentration of drug in a suitable range for the management of either acute or chronic diseases, it becomes compulsory to administer drugs multiple times in a day, due to 1 which a significant variations takes place. For all categories of treatment, a major challenge is to define the optimal dose, time, rate, and. Recent developments in drug delivery techniques make it possible to control the rate of drug delivery to sustain the duration of therapeutic activity and or target the delivery of drug to a special organ or tissue. Many investigations are still going on to apply the concepts of controlled delivery for a wide variety of drugs. Changes plasma concentration causes changes in steady state level which results in improved therapeutic efficacy of drugs for sever conditions drug plasma concentration of highly potent drugs improves drugs safety and efficacy. Utilization of maximum concentration of drugs depends upon the administered dose sustained release drug delivery system has several advantages reduction in the frequency toxicity of drugs is reduced there are so many disadvantages of sustained release drug delivery which includes decreased in the retention time, bioavailability is not predicted accurately disadvantages of sustained released drug delivery system can be improved by increasing the retention time of formulations. Novel drug delivery system (NDDS) is gaining interest in advance pharmaceutical techniques. 1.1. Types of wafers Various types of the wafers were prepared with different release characteristics such as mucoadhesive melt away, mucoadhesive sustained release and flash release wafers. Mostly made by the good film forming agents that provides the good release characteristics and make the consumers handling and transportation easily, also the manufacturing is almost easy because it was prepared by simple means. These wafers release drug rapidly and showed rapid onset of action complete drug releases within 30 seconds site of administration and absorption of these types of drugs is oral, local or systemic effect is induced by these types of wafers[3]. These wafers disintegrate rapidly within 1 mint they completely disintegrate and dissolute, the absorption of the released drug occurs in the gastrointestinal tract (GIT), these shows systemic or local effect on the body [4]. These wafers formulations releases drugs within 5-30 mints [5]. Drug released for several hours absorption site is gastric mucosa or gum ,onset of action is delayed, such type of formulations are suitable for prolonged systemic effect in the body [6]. 2 In controlled release drug delivery system the term mucoadhesion was first time introduced in 1980 [7].Therefore, most of researcher start focusing on the phenomena of mucoadhesive polymers with the mucus of the membrane, Mucoadhesive drug delivery system. Bio adhesion was used which indicates attachment of molecules of the drug to the biological surface or tissue of the epithelium, attachment of drug to the mucoadhesive membrane is referred to as mucoadhesion [8]. Mucoadhesive polymers for sustained release drug delivery system provide excessive advantage as compared to non mucoadhesive polymers enhanced absorption and bioavailability of the drugs due to a high surface to volume (S/V) ratio, increase contact time to mucus layer due to increase absorption of drug increases [8, 9]. Wafers due to greater mucoadhesive property at the gastric pH can attach to the surface of tissues of gastric mucosal tissue due to which it provides systemic and local sustained release of drugs, for sustained released formulations administration frequency is decreased and enhanced bioavailability targeted drug delivery using hydrophilic mucoadhesive polymers is increased Mucoadhesion has become an important issue for the various drugs as mucosa delivery system due to its optimal potential of targeted delivery where its desired concentration reached at the target site gastrointestinal mucosa or the buccal mucosa. Mucosal drug delivery system is of significant importance and was widely used for the administration of mucoadhesive formulations to the sites in the body such as buccal, gastric and nasal mucosa, also applicable to the wound surface. The contact between the mucosal surface and the dosage form may be either due to the physicochemical interactions which can plays an important role in the absorption as well as bioavailability of the drugs. The drug containing formulation wafers can bind to the mucosal surface which may be either due to the hydration and swelling which leads to the inter penetration of chains of the polymeric material with the mucin or mucosal surface. This may be suitable method for the administration of drugs as it reduces the adverse effect, toxicity, also reduces the choking and pain caused by the parental dosage forms. Mucoadhesive polymers can be categorized broadly into two classes’ i.e. Hydrophilic polymers and hydrophobic polymers, based on their solubility and chemical nature. Based on their origin polymers was categorized into three types, natural polymers, synthetic polymers, semisynthetic polymers. Hydrophilic polymers which contain carboxylic acid (C=0) group shows efficient adhesion on the mucosal surface e.g., hydroxypropyl cellulose (HPC), sodium carboxymethyl 3 cellulose (SCMC), methyl cellulose (MC), poly vinyl pyrollidone (PVP), and other cellulose derivatives. Polymers which possess greater mucoadhesive strength, hydrophobic polymers showed increased water absorption swelling and epithelial attachment [8]. Table 1: Mucoadhesive strength of natural and synthetic polymers. Polymers Mucoadhesive strength Sodium alginate +++ Carboxymethyl cellulose +++ Tragacanth ++ Gelatin ++ Pectin + Good (+), very good (++), Excellent (+++) During the selection of the mucoadhesive polymers it must be considered that it shows maximum gastrointestinal absorption, degradation by products should be less toxic or nontoxic even in low doses it must be completely eliminated from body within a short duration of time. It should be nonirritant to the mucus membrane. It should preferably form a strong covalent bond with the mucin–epithelial cell surfaces it should adhere quickly to most tissue and should possess some site specificity. It should allow easy incorporation of the drug and should offer no hindrance to its release. Polymers should not decomposed on storage or during the shelf life of the dosage form. Cost of polymer should not be high so that the prepared dosage form remains competitive in market. 1.2. Polymers used in Wafers preparations Various polymers were mostly used in formulation preparation for the wafers. Mucoadhesive polymers can be used either alone or in combination with other polymers. Hydrophilic polymers were commonly used in wafers development. Various hydrophilic polymers includes HPMC, sodium carboxymethyl cellulose, pectin, gelatin, hydroxypropyl ethyl cellulose and polyvinyl pyrrolidone. 4 1.3. Hydroxypropyl methyl cellulose (HPMC) HPMC use was started in the 1960,s as a US patent of Dow chemical company, however its use on large scale was started in 1960,s to 1970,s. Hydroxypropyl methyl cellulose are ethers of cellulose which can be used for manufacturing controlled released formulations. HPMC a white, off-white granular or fibrous powders which is a synthetic modification of natural polymers, it is prepared when wood pulp is treated with sodium hydroxide (NaOH), 18% hydroxypropyl and hydroxymethyl groups are introduced using propylene glycol and methyl chloride, number of the group added to the molecules give unique property to the polymer like soluble in cold water, or hot water. HPMC is widely used in the research work due to its properties such as it cannot interfere with disintegration time and bioavailability of the drug molecules. Solubility in the aqueous ,organic solvent and gastrointestinal fluids (GIF), odorless, tasteless, ability to incorporate other colors in to the films due to lack of interaction with coloring materials [10]. 1.4. Carboxy methylcellulose sodium (CMC) Sodium carboxy methyl cellulose was first prepared in 1918 and introduced commercially in 1920,s by Germany, carboxy methyl cellulose was prepared by treating sodium hydroxide with akali cellulose and sodium salt of monochlorooacetic acid in the presence of isopropanol or ethanol as organic solvent [11]. CMC is a water soluble polymer, its viscosity is high in dilute solutions, it is off-white –white powder having good film forming property, mucoadhesive strength, thickening effect and Excellent colloids protection [12]. 1.5. Guar gum (GG) Guar gum was obtained from the seeds of the guar plants and exactly present in the endosperm of the seeds, plant is cosmopolitan in nature and mostly cultivated in Pakistan and India, it is a polysaccharides composed of β-D-mannopyrunasoyl and galactomannans molecular weight of Guar gum lies in the range of 50,000-80,000,000[13]. Gaur gum powder is with free flowing properties which is odorless yellowish white to white in appearance, polymer insoluble in hot aquous and organic solvents, however it is soluble in cold water and forms a viscous gel, solution 5 has buffering capacity therefore stable at pH 4-10.5.gum is used as emulsifiers, stabilizers and thickening agent in various dosage forms. 1.6. Pectin (PC) Major constituents of plants and citrus fruits, it has very good gelling property pectin chemically composed of alpha 1-4 galacturonic with methylation of carboxylic groups it is structurally divided into branched, smooth and hairy region [14]. Pectin is prepared from the plant cell wall by esterification it was observed that pectin can decreases glucose ,cholesterol level and also possess anticancer property, in adenocarcinoma patient apoptosis can be induced by using pectin [15]. Cheapest as compared to other polymers due to low cost of production, also nontoxic and availability is high, Pectin can be used for delivery of drugs through various routes like nasally, orally and vaginally also patient compliance is good. Pectin is widely used in pharmaceutical and food industry [16]. 1.7. Polyethylene Glycol (PEG 200) Plasticizer reduced the glass transition temperature (GT)of the film forming polymers and thus increases mechanical properties of wafers such as percentage elongation, tensile strength, elastic strength of wafers formulations, it improves the flexibility and thus reduces the brittleness of the wafers formed[16]. Depending upon the nature of the solvent plasticizer was selected some most commonly used plasticizers were Polyethylene glycol, Glycerol, Castor oil, Glycerin. Wafers are not formed if excessive or low quantity of plasticizer is used [17]. FDC approved colors can be used, however the concentration of coloring material should be less than 1%. Drug should not be bitter in taste. Solubility of drug should be maximum in water Penetration in the oral and gastrointestinal tract (GIT) should be Maximum. Stability of drug in water should be Maximum. Drug having extensive first pass metabolism. 6 Table 2: Mucoadhesive strength of natural and synthetic polymers Serial No. Excipients Max % used 1 Polymer 0-45% 2 Plasticizer 0-20% 3 Colorant ˓˃1% 4 Surfactants q.s The main objective of study was development of sustained released (SR) mucoadhesive wafers contain ranitidine hydrochloride as model drug. Due to shortest half-life of Ranitidine hydrochloride and avoid hepatic first pass metabolism novel dosage form was developed by using various concentrations of natural and synthetic hydrophilic polymers. Synthetic hydrophilic polymers used were hydroxypropyl methyl cellulose, sodium carboxymethyl cellulose. Natural hydrophilic polymers used were Pectin and Guar gum. Effect of polyethylene glycols (PEG) different concentrations used in combination with different concentrations of polymers. Effect of surfactant Tween 80 on the disintegration time and dissolution studies of wafers formulations. Various parameters were evaluated such as weight variation, surface pH, folding endurance (FE), total dissolving time disintegration time. In-vitro mucoadhesion, percentage moisture loss, uniformity of percentage moisture absorption. Microscopic images and scanning electron microscopy (SEM). Dissolution studies, kinetic models were applied to study drug release mechanism such as Zero, first order, higuchi, korsmayer-peppas models were applied to study drug release 7 2. Drug profile In 1990 James black introduced selective H2 antagonist Ranitidine hydrochloride for the first time. He got Nobel Prize for introducing selective receptors antagonists (selective H2 receptor antagonist and beta blockers). Cimetidine was first approved H2 antagonist in united states (US) in 1977, after this ranitidine hydrochloride (RNH) in 1983 and Nizatidine (NZD) in 1988. 2.1. Description Ranitidine Hydrochloride is without out the imidazole ring and cyanoguanidine group which is present in famotidine however RNH is an amino alkyl substituted furan 2.2. Molecular formula 350.87 2.3. Chemical formula C13H22N4O3S.HCl Melting point 134c˚ 2.4. Chemical name N-[2-[5-(Dimethylaminomethyl)-2furylsulphanyl]-N-methyl-2-nitro-ethane-1,1-diamine) [18]. 8 2.5. Characteristics of Ranitidine hydrochloride: Ranitidine exist in two forms, crystalline and amorphous. Ranitidine actually showed polymorphism i.e. crystalline nature commonly known as form 2, (F2) form1(F1), and these possess different pseudo polymorphic structure [19]. Polymorphs of ranitidine depends on type and nature of crystallizing solvents, for example, when solvent used is ethyl acetate along with the ethanolic solution crystallized form thus obtained is Form1, However when the isopropanol Hydrochloride is used as for crystallization the crystals thus formed are form 2 [20]. Difference in the solubility as well as the bioavailability of two forms were found [21, 22] ranitidine showed two different forms which were considered to be the tautomeric to each other as concerned to the manufacturing process form 1 is expensive and difficult to manufacture, however, form 2 is easy to manufacture and cheapest GSK most commonly used the form 2 ,two forms of ranitidine cannot be converted to each other [23]. 2.6. Solubility of Ranitidine Ranitidine hydrochloride a pale yellow white, off white crystalline powder having sulphur like odor and slightly bitter taste. Solubility of the RNH in water is 660mg/ml and is considered freely soluble in water, however at pH range 1 to 7.4 and methanol solubility lies upto 550mg/ solubility is at the temperature 37 ◦C in methanol, however it is sparingly soluble in 96% ethanol [24]. 2.7. Pharmacology 2.7.1. Pharmacokinetic Ranitidine administered through oral route, however oral sustained release formulation releases most of the drug at the colon, the drug showed its absorption either in the gastrointestinal tract or colon. In first (initial) segment of small intestine ranitidine absorption occurs, due to which it showed fifty percent (50%) absolute bioavailability (ABA [25]. Ranitidine metabolism occurred in colon which is responsible for the poor bioavailability. Therefore RNH (ranitidine hydrochloride) is not favored by the traditional approaches for the development of sustained release delivery of drug [26]. Hence, conventional technology may not be successful for the preparation of ranitidine hydrochloride. Ranitidine hydrochloride is rapidly absorbed oral, 9 almost 50%-60% bioavailability of ranitidine after single dose administration within 0.5-1.5hrs first peak plasma concentration achieved and within 3-4hrs second peak appears [27]. Ranitidine administered orally only fraction of drug (0.4%) is excreted through biliary pathway (biliary excretion), when solution of drug was administered directly in to the colon. Plasma was analyzed drug shows double peak which is still unclear why it occurs. The bioavailability of ranitidine is relatively lower when administered as a solution directly to the colon instead of stomach, ileum, or jejunum as tight junctions in colon are considerably less permeable than those in the small intestine, main absorption site of ranitidine is small intestine, where absorption takes place through paracellular mechanism, absorption of drug is not effected by food items. Due to its short biological half-life of ranitidine hydrochloride (2.5to 3hrs). Suitable candidate for sustained release (SR) formulation. Delivery at the local site can increase the bioavailability for receptors located on stomach wall, due to which there is tremendous increase in the ability of drug to reduce the secretion of gastric acid. This principal may improve local as well as the systemic delivery of ranitidine hydrochloride, this results in decreased the gastric acid secretions [18]. 2.7.2. Volume of distribution Volume of distribution(Vd) of Ranitidine Hydrochloride (RNH) for the initial phase ranges from 1.16 to 1.187 liter per kilogram (L/kg) [28]. whereas the protein binding of RNH is very less which is almost less than 15% [28]. 2.7.3. Metabolism and Excretion 70-80% of ranitidine remains unchanged and excreted through urine following intravenous administration however 30% of unchanged ranitidine excreted through urine when administered orally. Ranitidine 10% is metabolized administered through intravenous route and 26% is metabolized when administered orally, colonic metabolism of ranitidine is partial due to which availability of drug is poor, however it is unclear whether ranitidine follows linear or nonlinear pharmacokinetics. 2.7.4. Pharmacodynamics Ranitidine is an histamine H2 receptor antagonist, it bind to H2 receptors located on the mucosal surface of gastric parietal (GP) on antiluminal surface of endothelial mucosa, thus it can 10 block secretion and production pathway of gastric acid, due to which it decreases the gastric acid secretion. Important factor for the common use of ranitidine is its selectivity of H2 blockers, it has lesser or negligible effect on H1 receptors, the H1 receptor antagonist are used for the management of allergic reactions but has little effect on H2 receptors [29]. 2.7.5. Clinical indications Ranitidine hydrochloride is one of the most commonly prescribed histamine H2-receptor antagonists [1].It has two polymeric forms which are used in the manufacture of commercial tablets or dosage forms, commonly used in treatment of gastroesophageal reflux disease (GERD) [30]. Peptic ulcer disease (PUD), widely, used in the management of gastric ulcer, erosive esophagitis, Zollinger ellison syndrome and active duodenal ulcers. In management of hives ( allergic skin condition)in combination with antihistamines such as fexofenadine it shows better treatment outcomes [31]. Scleroderma esophagitis an infection of oesophagitis. 2.7.6. Adverse effects Aplastic anemia Cholestasis Pancreatitis Granulocytopenia Alopecia Insomnia Bradycardia Dizziness Depression Hypersensitivity reactions 2.7.7. Dosage and administration Daily recommended doses of ranitidine hydrochloride for adult is 150mg BD or 300mg OD erosive esophagitis can be treated by administering 150mg of ranitidine four times a day (QID). 150mg of ranitidine inhibits the secretion of gastric acid upto 5 hours. 300mg of ranitidine increased plasma drug level therefore ,sustained release formulation of Ranitidine required [32]. 11 For management of Zollinger-Ellison syndrome a recommended dose up to 900 mg can be used without any adverse effects [33]. 2.7.8. Contraindication Patient hypersensitive to H2 antagonist/Ranitidine 2.7.9. Cautions Pediatric patient Elderly patient Patient on ventilation Porphyria Used not for more than 18 months Hepatic impairment Renal impairment Mechanical ventilated patients 2.7.10. Pregnancy category B Animal trials showed no risk to fetus, however no adequate and well controlled studies were carried out on pregnant women. 2.7.11. Toxicity signs and symptoms Meiosis Ear and mouth redness Excessive salivation Mucous membrane pallor Fast respiration Tremors in muscles tachycardia Thrombocytopenia (TCP) Leucopenia Agranulocytopenia Angioedema 12 Elevated serum creatinine 2.7.12. Management of toxicity After ingestion of ranitidine hydrochloride (RNH) administration of Charcoal decreases absorption of (RNH), in severe condition hemodialysis can be done for the removal of drug from patient plasma. 13 3. Literature review 3.1. Hydroxypropyl methylcellulose (HPMC) Kiran Goutam et al., 2015 developed promethazine theoclate wafers, various grades of hydroxypropyl methylcellulose were used for development. Solvent casting method was used for prepration of formulation. Wafers were evaluated an antihistamine drug used in the treatment of vomiting and nausea. Phosphate Buffer of 6.8pH is used as the dissolution medium and drug showed better release pattern [34]. Jagadesh Kumar et al., 2014 prepared the buccal wafers of antihypertensive drug Labetalol hydrochloride. The objective of development of films was to increase the bioavailability of the drug. Polyvinyl pyrollidone (PVP), hydroxypropyl methylcellulose were used in combination for film formation by using solvent casting method. FTIR analysis showed no polymer –drug interaction formulation contain higher percentage of polymer shows good release (94.35)% is considered to be optimized formulation [35] . Pahoa et al., 2014 they developed the transdermal wafers of Glibenclaimide employing cellulose polymers as film forming agent. In-vitro skin permeation study was conducted which showed better results of penetration of EC/HPMC and EC/PVP shows good permeability. Prepared films were smooth, uniform and flexible [36]. Shiaimaa N et al., 2014 prepared wafers of Flurbiprofen as model drug by using solvent casting technique. Objective of formulation development was to relief pain within a short period. Wafers were prepared by using hydroxypropyl methylcellulose as film forming polymers, polyethylene glycol was used as plasticizers. Wafers (films) disintegrate within 30 seconds however, almost 77.5% of the drug released in 2 mints, therefore prepared Flurbiprofen formulation can be used for the relief of pain [37]. Farhana Sultana et al., 2013 developed wafers formulations containing anhydrous Caffeine using various concentrations of film forming polymers such as sodium alginate, white Kollicoat and hydroxypropyl methylcellulose percentage released from formulations containing Kollicoat polymer is 99% in 240 seconds and disintegration time is 12 s comparatively HPMC contain 14 formulation disintegrate within 13 seconds and 100% drug was released in 120 second so the HPMC containing formulations were considered suitable for use [38]. Joshua Boateng et al., 2013 developed solvent casted films of paracetamol and Amoxicillin as combination therapy by using combination of hydrophilic polymers such as carrageenan, carboxymethyl cellulose and sodium alginate. Films were evaluated by various parameters such as mucoadhesion, swelling index, content uniformity, SEM (scanning electron microscopy), XRD. Drug release occurred through erosion and diffusion process the %age for Amoxicillin was 70.59% and paracetamol 84.65% [39]. P.Narayana Raju et al., 2013 prepared fast dissolving films of Loratidine an antihistaminic drug by using film forming polymer hydroxypropyl methylcellulose in organic solvents such as methanol and dichloromethane by using solvent casting technique. Sweetness were added to mask the bitter taste of Loratidine. Casted films were evaluated for various physicochemical parameters like disintegration time, surface pH, content uniformity, thickness and weight variation test, disintegration time of the formulations lies in the range of 24-29 seconds. In-vitro dissolution studies showed drug release for 5 mints completely. Fourier transformed spectroscopy (FTIR) showed no incompatibility between polymer –drug molecules [40]. Patel et al., 2012 they carried out screening of the polymers that are most suitable for formulation of oral fast dissolving film. Different polymers like HPMC- E3, HPMC- E5, HPMC E6, HPMC- E15, Pullulan, Gelatin, Chitosan and PVA were used for the preparation of wafers. Pullulan and HPMC- E series showed transparent appearance, fast disintegration and dissolution time with good folding endurance good tensile strength [41]. Sane et al., 2012 they have developed fast dissolving film of Zolmitriptan by solvent casting method. The effect of different concentration of polymers and plasticizers evaluated for film forming capacity, appearance and disintegration time .Films with HPMC E3 in combination with PVA, were found to be very transparent having least disintegration time and best film forming capacity [9]. Nagar et al., 2011 described oral route considerations, film forming polymers, ideal characteristics of a polymer to be selected. They classified polymers as natural and synthetic. They described advantages and disadvantages of pullulan, its properties, chemical structure, etc. 15 was categorized as the best film forming polymer. Starch and modified starches, pectin, sodium alginate, gelatin, Maltodextrin and description of each. Also described synthetic polymers like HPMC, hydroxypropyl cellulose, sodium carboxymethyl cellulose, polyvinyl alcohol, polyethylene oxide and their properties in detail. Quality control tests for all polymers were discussed. Pathan et al., 2010 they developed the mucoadhesive buccal wafers of salbutamol sulphate for the systemic delivery using the HPMC -K15 and Eudragit –S100 as the film forming polymers no physicochemical interactions were studied when FTIR spectroscopic analysis was carried out. physical and chemical characterization of the drug was carried out for the evaluation of the drug which includes swelling index ,mucoadhesive strength weight variation, pH of the Patch and invitro kinetic studies were performed developed films showed controlled release over a period of 6h and follow the zero order release kinetics [42]. Alagusundaram et al., 2009 employed solvent casting method for the development of mucoadhesive buccal film using HPMC, PVP as film forming polymers. Dichloromethane and ethanol is used as solvent and propylene glycol was used as plasticizers as well as penetration enhancer. Swelling characteristics were determined the drug loaded films showed greater swelling as compared to unloaded films. Content uniformity studies indicates that drug was uniformly dispersed [43]. Mona et al.,2008 developed buccal film of Glipizide using cellulose polymers and Eudragit RL100.Films were evaluated for their weight variation ,thickness, surface pH ,in-vitro residence time and, folding endurance. In-vivo studies and permeation studies showed better release of drug over 6h. Film showed satisfactory swelling and maximum retention time in combination with HPMC and CMC [44]. Gohel et al., 2007 prepared the taste-masked film of valdecoxib for oral use. Films were prepared using eudragit and hydroxypropyl methylcellulose as a polymer. Polymers are soluble in organic solvent and water respectively. The films were prepared by solvent casting method and evaluated by hydration study folding endurance and dissolution. Glycerol played a critical role in imparting flexibility to the film and improving the drug release [14]. 16 Maseru et al., 2005 developed and evaluated a fast dissolving film of salbutamol sulphate. The prepared films were clear, transparent with smooth surface and has an acceptable mechanical characteristics and satisfactory % drug release, tmax for fast dissolving film was lower than that of conventional tablets. A fast dissolving film of Salbutamol sulphate could be helpful for relief of fast onset asthmatic attack and also provides a rapid mean of management of asthma [15]. 3.2. Pectin wafers Vivek Gupta et al., 2013 developed pectin based polymeric wafers for intestinal mucoadhesion using combination of Pectin and other hydrophilic polymers such ethyl cellulose (EC), sodium carboxymethyl cellulose and carbopol. Calcitonin salmon (polypeptide) was used as model drug. Various parameters such as mucoadhesive strength, increased drug absorption and dissolution were carried out, formulation showed 44 fold increased bioavailability as compared to injectable dosage form [16]. Olga et al., 2012 developed the topical wafers of antimicrobials and antibiotic therapy for treatment of wound infections using Pectin as polymeric material and polyethylene glycol was used as plasticizers .Swelling behavior of the wafers was determined by using the analytical methods and reweighed after the period of 24 hours. Contents were assayed using the diffusion disc method [17]. Rubina P shakikh et al., 2012 developed lyophilized wafers of diphenhydramine an antihistaminic agent, comparison of non-crosslinked pectin wafers and non-crosslinked pectin wafers were carried out. Various parameters were evaluated such as porosity ,surface morphology ,vibrational and transitional texture attributes in-vitro and thermal release studies shows that 80% burst release was observed from non-crosslinked wafers, however cross linked wafers showed 40% release in first 30 mints, study showed that crossed linked wafers formulations were suitable for antihistamine drug release [45]. Sevsagi Gangor et al., 2008 prepared transdermal films of hydrophilic polymer Pectin as polymeric material in combination with polyvinyl pyrollidone (PVP) and propylene glycol (PG) used as plasticizer, methane and eucalyptol was added to increase the permeability of skin for rapid drug absorption. Using tail cuff method these patches were applied on rats and systolic 17 blood pressure was measured, within 30 mints a marked decrease in systolic blood pressure was observed [34]. 3.3. Carboxy methyl cellulose wafers (CMC) Mura P et al., 2015 prepared sustained released econazole nitrate antifungal agent by freeze drying method using carboxymethyl cellulose sodium (NaCMC) and pectin as polymeric material developed formulations showed mucoadhesion for 88 mints. Dissolution studies showed drug 5mg drug release for every hour which is suitable for the treatment of candida albicans [46]. Nq SF et al., 2014 prepared exudates absorbent wafers for healing wounds using freeze drying (Lyophilization) method for the development of wafers. Cellulose polymers such as methyl cellulose (MC) and carboxymethyl cellulose sodium (Na CMC) were used purpose of development was to improve healing process and reduced healing time. Sulphacetamide (SCM) silver nitrate (SN) and neomycin sulphate (NS) were used as active ingredient in various formulations, which were evaluated physically such as hydration capacity, swelling index, uniformity of content, %moisture loss, microscopic evaluation, antimicrobial activity and drug release. Sodium carboxymethyl cellulose wafers showed better results compared to methyl cellulose [47]. Noelle H.et al., 2013 developed the wafers formulations for treatment of wound infection by lyophilization method, cationic antimicrobial peptides with natural polymeric combinations were evaluated for their antimicrobial activity using disc diffusion method (DM). Prepared wafers were used against bacteria such as Pseudomonas aeruginosa, Staphylococcus aureus and natural polymers contain formulations showed efficient antimicrobial activity [48]. Stephen CB Lim et al., 2013 developed lyophilized (freeze dried wafers) by using amylogen carboxymethyl cellulose sodium (NaCMC) as film forming polymers. Wafers were evaluated by various parameters such as, percentage moisture content, friability, weight uniformity and dissolution studies were performed. Morphology of wafers were evaluated by X-ray diffraction and SEM (scanning electron microscopy. Wafers showed 53% absolute bioavailability in-vitro dissolution studies showed 90 % drug released within 1 mint from wafers formulation [24]. 18 Boateng JSet al., 2010 developed wafers using solvent casting and lyophilization method using carboxymethyl cellulose and sodium alginate as polymeric material for oral and gastric mucosa as well as wound surface. Paracetamol (PCM) was used as model drug. Hydration capacity and morphology of wafers were evaluated with laser scanning microscopy (LCM) and SEM (scanning electron microscopy). Lyophilized wafers had porous structure, thus high hydration and drug loading capacity as compared to casted wafers, due to different mechanical and physical properties, ease of hydration solvent casted wafers and lyophilized wafers showed different mucosal application time [25]. Amit Khair nar et al., 2009 prepared solvent casted wafers using Aceclofenac as drug of choice. Formulations were prepared by using carboxymethyl cellulose sodium(Na CMC ), polyvinyl alcohol (PVA) and gelatin casted films were evaluated for various physicochemical and chemical properties such as folding endurance (FE),%age entrapment efficiency ,weight variation ,thickness ,mucoadhesion (in-vitro),% age elongation in-vitro dissolution and stability studies were carried out. Formulation F5 was optimized with more than 88% release after 8hrs, all parameters shows that F5 was best formulation [49]. Joshau Boateng et al., 2009 prepared Paracetamol (PCM) wafers for the immediate analgesic effect by using solvent casting, freeze drying (lyophilization) methods. Carboxy methyl cellulose (CMC) as hydrophilic polymer, morphology of prepared films by both methods were evaluated by using scanning electron microscopy (SEM). Exchange cell was used for dissolution studies release of drug was measured by ultraviolet spectrophotometer (UV) at (243 nm),release rate of drug was better from wafers as compared to the films [50] . 3.4. Guar gum containing wafers Ismail sait Dogan et al., 2015 prepared gluten free backed products using natural hydrophilic polymers such as gaur gum, xanthan gum that gluten free product are suitable for healthy living of celiac patients. Products improved health of patients 0.5% guar gum and 0.1% xanthan gum formulations were considered to be optimized for patients who need gluten free products [28]. V Senthil et al., 2015 purpose of current study was development of the taste masked oral wafers of Ambroxol hydrochloride and Levocetrizine dihydrochloride. Various polymers such as, Guar 19 gum, sodium alginate (SA), Pectin, Propylene glycol (PG), polyvinyl pyrollidone, hydroxypropyl methyl cellulose (HPMC k15) and sodium Alginate were used in combinations in different ratios. Wafers were prepared by using solvent casting technique and all the prepared formulations were evaluated for their physical and chemical parameters formulations contain HPMC K15 and pectin showed best release studies [51]. A.Deepthi et al., 2014 prepared Zolmitriptan wafers by solvent casting method using, Guar gum, Sodium alginate, xanthan gum as polymeric material, polyethylene (400) was used as plasticizers. No drug –polymer in compatibility was found by Fourier transformed infrared spectroscopy (FTIR). Wafers were characterized by various physical and chemical parameters such as total disintegration time folding endurance, tensile strength (TS), disintegration time, uniformity of content and dissolution studies maximum drug released within 6 mints is 98.5% [30]. Labvitidia et al., 2012 prepared wafers of cyclohexatriene digluconate for treatment of wounds using Guar gum as film forming polymeric material. Wafers were prepared by using freeze drying method. Wafers heals the wound by absorbing the exudates (fluid )from surface of wound in-vitro kinetic studies showed that dug release through diffusion process which showed drug release follows korsmayer-peppas release model [52]. Noelle H et al., 2012 developed Guar gum containing, antimicrobial peptide containing wafers used for direct supplication to the skin for the potential treatment of skin wound. These wafers were prepared by using the free drying process, it was observed that the topical delivery of drug for the treatment of wounds by using wafers as drug delivery system was considered important dosage system for the eradication of wound infection. Antimicrobial cationic peptide with Guar gum natural polymer was considered to be important formulation for application on skin [32]. Olga Labvitidia et al., 2011 prepared antimicrobial wafers using Guar gum, xanthene gum and sodium alginate, polymers were mixed. Polymeric material and combination of various antimicrobial agents that were silver sulphadiazene (SA), povidone iodine (PI), neomycin sulphate (NS) and cyclohexatrienes, wafers were used against various microbes the results showed [53]. 20 3.5. Sodium alginate and gelatin wafers J Boateng et al., 2015 prepared and characterized wafers containing silver sulphadiazene for treatment of wound infection by using gelatin and sodium alginate based as polymeric materials by changing the concentration of polymers. FTIR spectroscopic studies were used to determined polymer drug interaction. Physical parameters such as surface pH, thickness weight variation, and content uniformity and in-vitro kinetic studies were carried out [34]. Josaine Guanclaves et al.,2015 developed and characterized orally disintegrating films of gelatin and collagen using the ethanol extract of propolis. Films were prepared by using solvent casting technique. Physical parameters such as mucoadhesion, swelling index, in-vitro release kinetics and FTIR were carried out, extract containing films were more resistant as compared to the blank films containing gelatin and collagen alone. Extract containing films showed antimicrobial activity and can be used for the treatment of diseases caused by the Staphylococcus auras [35]. Yeole Shaiaka et al., 2015 they developed the herbal favors by using variety of various plants due to their medicinal effect on the human body. Various plants and their parts used includes Asparagus sativus which contains the steroidal saponins Lepidium sativum has the potential to improves the growth and possess the lactogenic property. Morinaga oliefera can be used for the treatment of malnutrition as it is rich in the constituents having proteins and micronutrients in the excessive quantity [36]. Indranil Gangualy et al., 2014 developed formulation of promethazine hydrochloride an antiemetic drug for the sublingual administration. Various polymers were used in different concentrations such as methylcellulose, gelatin, xanthan gum and Beta-Cyclodextrin was added to mask the bitter taste of promethazine HCL. FTIR analysis was carried out and no interactions were found. Physical and chemical parameters were evaluated such as surface pH, uniformity of contents, moisture uptake, moisture loss, disintegration. In-vitro drug release and Ex-vivo permeation studies were carried out using the porcine membrane model. In-vitro drug release studies were observed to be 98-100% within 6 hours. Stability studies showed that lyophilized drug wafers with good stability and compatibility were obtained [54]. 21 Boating et al., 2014 developed sodium Alginate and chitosan based wafers. Delivery of macromolecules for buccal drug delivery by using the rolling drum method, differential scanning calorimetric, a comparative study of drug loaded wafers were carried out. Various tests were performed like mucoadhesion test, Hardness testing, swelling capacity and dissolution studies. Swelling capacity of drug loaded wafers was observed to be greater as compared to blank wafers [55]. René et al., 2013 they studied the in vitro, ex vivo and in vivo examination of buccal absorption. Metoprolol TR146 cell culture, porcine buccal mucosa and Gottingen minifies, a significant higher absolute bioavailability was obtained after buccal dosing (58–107%). Compared to oral (3%) administration in Gottingen minifies. A very clear level C in vitro in vivo correlation (r2 = 0.98) was obtained between the observed in vitro permeability’s and the bioavailability observed. [39]. Joshua Boateng et al., 2013 developed solvent casted films of paracetamol and Amoxicillin as combination therapy by using combination of hydrophilic polymers such as Carrageenan, Carboxy methyl cellulose and sodium alginate. Films were evaluated by parameters such as mucoadhesion, swelling index, content uniformity ,SEM (scanning electron microscopy ) ,X-RD, release of drugs occurs through erosion and diffusion process the %age for Amoxicillin was 70.59% and paracetamol 84.65% [39]. Gag et al., 2011 described novel approaches in fast dissolving oral drug delivery systems. Authors described need, advantages, and disadvantages of oral films. Formulation considerations included a typical composition of film, description of polymers, plasticizers, surfactants. Authors stated manufacturing methods with advantages and disadvantages of each [41]. Cilurzo et al., (2010) evaluated feasibility of a fast dissolving film made of Maltodextrin containing nicotine hydrogen tartrate(NHT) salt using solvent free hot-melt extrusion technology. The bitterness and astringency intensity of NHT and the suppression effect of several TMA were evaluated by a Taste-Sensing System. The addition of NHT and tastemasking agents affected film tensile properties, however, effect of addition of these components can be counterweighted by modulating the glycerin content and MDX molecular weight [56]. Kunte et al., 2010 they developed a fast dissolving Oral film for the delivery of Verapamil. Thus the patient-friendly dosage form of bitter drugs such as Verapamil can be successfully 22 formulated using this technology and it can be especially useful for geriatric, bedridden and noncooperative patient due to its ease of administration [57]. Vishw et al. 2010 have formulated and evaluated fast dissolving tablet of metoprolol tartrate using semisolid casting. They compared the effect of various superdisintegrants on the drug release. Sodium starch glycolate was found to be best superdisintegrants; however the other superdisintegrants shows the poor property [44]. Sumitha et al., 2009 designed a taste masked films of Ondansetron hydrochloride by rolling drum method. Polymer carrier system and formulation of rapid-disintegrating films. Rapid dissolving film containing taste masked Ondansetron hydrochloride showed acceptable properties. Drug release profile indicated that it could be used for the oral delivery of Ondansetron hydrochloride in chronic and acute post-operative or chemotherapy or radiotherapy induced emesis, so it possesses good antiemetic property. Good correlation between in vitro disintegration behavior and in the oral cavity was recognized, patient friendly dosage form has been accomplished [45]. Verena et al. 2009 prepared films of Caffeine, gelatin, pullulan by solvent casting technique. The film casting solution was prepared by dissolving the excipients in the solvent and heating up to 60C˚. Films thickness, disintegration, dissolution, and texture analysis was carried out thickness of wet films decreases about 86-97%.films also showed good endurance strength [49]. Dinge et al., 2008 formulated and evaluated fast dissolving films of triclosan. Various film forming agents, film modifiers and polyhydric alcohols were evaluated for optimizing the composition of fast dissolving films. The potential of poloxamer 407 and hydroxypropyl-βcyclodextrin (HPBCD) to improve solubility of TC was investigated. Fast dissolving films containing hydroxypropyl methylcellulose (HPMC), xanthan gum, and xylitol were formulated. Use of poloxamer 407 and HPBCD resulted in significant improvement in the solubility of TC. Fast dissolving films containing TC-HPBCD complex and TC-Poloxamer 407 were formulated and were evaluated [50]. 23 3.6. Polyethylene glycol (PEG) wafers Youhan Zhao et al., 2015 prepared meclizine hydrochloride wafers for treatment of motion sickness. Polyethylene glycol (PEG) was used as plasticizer, wafers (films) were prepared by using solvent casting technique. FTIR results showed no polymer drug interaction and dissolution studies were carried out in 0.1N HCL and distilled water. Absorbance was measured by using UV-spectrophotometer results showed more than 80% drug released within 5 mints. Kianfar et al., 2014 prepared lyophilized (freeze dried) wafers using polyethylene glycol (peg600) a water soluble plasticizers, poloxamer and carrageenan as film forming polymeric materials. Wafers were evaluated by scanning electron microscopic, hydration, in-vitro mucoadhesion studies were carried out which showed that hydration and mucoadhesion of poloxamer (4%) carrageenan (2%) and PEG 600 (4.5%) showed wafers with best hydration capacity and porous network for loading of drug properly [52]. Kianfar et al., 2013 developed wafers of ibuprofen and paracetamol (PCM) using Carrageenan and Pouluric acid as film forming polymers. Polyethylene Glycol 1.8%w/w was used as plasticizers to increase the elastic strength, formulation remains stable for up to six month, mucoadhesive strength was observed using mechanical texture analyzer, drugs gradually releases up to 2 hours results showed prepared wafers were suitable for mucosa of buccal cavity [58]. Gorakh et al.,2013 developed solvent casted wafers (oral films ) using polyethylene glycol (PEG) as plasticizer, in combination with various film forming polymers such as hydroxypropyl methyl cellulose (KM100), hydroxypropyl methylcellulose (E5),hydroxypropyl methyl cellulose (cps3). Wafers formulations were evaluated by various parameters such as disintegration time (DT), weight variation, folding endurance (FE), surface pH, dissolution and stability studies, thickness of wafers was in the range of 0.22 to 0.33mm,maximum folding endurance 200 complete disintegration time was within 60seconds. 100% drug release was observed within 2 mints which showed formulation possess best release characteristic and suitable for immediate effect [54]. Kumaragiri Sasi Deepthi et al., 2012 they developed fast dissolving oral wafers (films )using plasticizer polyethylene glycol (PEG 400), sweating agent glycerin (GLY), film forming 24 polymers such as HPMC, tween (80) were used. FTIR results were evaluated no polymer drug interaction was found. Physicl and chemical parameters were evaluated such as content uniformity, disintegration, folding endurance, thickness, surface pH, weight variation and dissolution studies. Tensile strength of all formulations were carried formulation (F8) showed less tensile strength and optimum drug release [59] . Irana Alexendra Paun et el., 2012 developed implantable wafers using indomethacin as model drug. Polyethylene glycol, polylactide-co-glycodie were used as plasticizers and polymeric material. Formulations were prepared using laser evaporation (LP) techniques. Sustained released (SR) wafers of indomethacin for more than three weeks. Kinetic models were applied to study drug release mechanism which showed that drug releases by diffusion mechanism and follow higuchi model when phosphate buffer saline was used as dissolution medium [56]. Farinose et al., 2011 prepared Carrageenan wafers containing ibuprofen by constant stirring at 80 ◦C. Poloxamer 407 and polyethylene Glycol (PEG600) were used as plasticizers. Thick viscous gel is casted for 24hrs in an oven films formed were analyzed for its tensile strength, elasticity and percentage elongation. Drug dissolution and Release profile was determined by using HPLC by using different media as deionized water of pH 5.6. Buffer solution of pH 6.5.XRay powder diffraction was determined to measure the nature of film ,kinetic property of drug release of the film was evaluated by using korsmayer-peppas, Higuchi, zero order and first order release model [60]. Isaac et al., 2011 prepared wafers by lyophilization of aqueous gels of the polymers using various concentration of polyethylene glycol as plasticizers and D- manifold as cytoprotectant. SEM was carried out with stereoscan-s-360. Phase separation study was carried out using the DSC at different speeds. The drug loading capacity of formulation was determined by using the assayed content and the initial loading of drug .Drug loading was observed upto 90%. Mishra et al., 2011 developed wafers of cetirizine hydrochloride using different concentrations of polyethylene glycol 400 as plasticizers and pullulan was used as film forming polymer. Optimized batch of pullulan showed satisfactory thickness and mechanical properties. It showed stability for six months under specified conditions. Films were prepared by using semisolid 25 casting. Prepared films showed better in-vitro and in-vivo disintegration time within 20-30s [61]. Hiroyoshi Shimoda et al., 2009 prepared dexamethasone wafers (films) using polyethylene glycol as plasticizer and hydroxypropyl methylcellulose was used as film forming polymers.at 75% humidity and 40 0C, formulation was stable within 5mints. 90% dexamethasone was released however films disintegrate completely within 15 seconds. Treatment of emesis caused by anticancer drugs dexamethasone oral wafers were used as drug of choice [62]. 26 4. Material and Method 4.1. Chemicals Ranitidine Hydrochloride was gifted from (Ferozsons Pharmaceutical Karachi Pakistan) Hydroxypropyl methylcellulose (HPMC- E5) Sigma Aldrich Germany Carboxy methylcellulose (CMC) sigma Aldrich (Germany) Pectin purchased from (Dae-Jung chemicals and Metals CO.LTD Korea) Guar gum purchased Sigma Aldrich (Germany ) Polyethylene glycol (PEG 200) E-Merck Dermastad (Germany) Amaranth coloring dye ( Dae-Jung chemicals and Metals CO.LTD Korea) Tween 80 (Dae-Jung chemicals and Metals CO.LTD Korea) Potassium dihydrogen phosphate (KH2PO4) Sigma Aldrich (Germany) Sodium hydroxide (NaoH) Sigma Aldrich (Germany) 4.2. Apparatus /equipment Glass petriplates (Pyrex) Electrical weighing balance (Eutect instruments) Hot plate stirrer ( Dahan Lab tech co.ltd) Measuring cylinder Beakers (Pyrex) Hot air Oven (Memmert Germany ) Soni-cator (FAD Instruments Lab Lahore Pakistan) Micrometer screw gauge (Hitech Karachi Pakistan) Optical light microscopy (Germany ) Vernier caliper (Hitech Karachi Pakistan) Disintegration apparatus (Erweka Germany) Dissolution apparatus (USP type II) ( Digiteck instruments Lahore Pakistan ) FTIR spectrophotometer (Agilent Karachi Pakistan) UV-spectrophotometer (Perkin Almer Germany) PH meter (510 Eutech instrument) 27 Aluminium foil (Zamas international FZCO, U.A.E.) 4.3. Experimental design (Box- Behnken) Response surface methodology is the technique used to develop the relationship between quantitative and qualitative data of the experiment. In present study three factors evaluated at 2 levels offer advantages in the form of proposed formulations [63]. A standard (RSM) of boxbehnken design was used to study the variable concentration of polymers, plasticizers and surfactant. Box-behnken design is a three factors .Variable with minimum (-1) and the maximum(+1) was observed [64]. As shown in equation 1, 2 and 3. (1) -1 = (2) 0= (3) Alpha (α) a constant value which is (1.68), min and max are the lowest and highest values of the variables used, +1, 0, -1 are the maximum average and minimum value of variables as shown in table3. Table 3: Coded values of the variables of Box-behnken design for wafers formulation Independent variables Level High Medium Low Code +1 0 -1 Polymer 33.58 30.64 27.7 Plasticizer 20 18.81 17.63 Surfactant 1.79 1.49 1.20 28 4.4. Preparation of wafers Solvent casting method was used for preparation of Ranitidine wafers. Accurately weighed polymer, plasticizer and ranitidine were dissolved in 15-30ml of water and stirred at 500rpm to make clear solution at temperature 55-600C. Solution was poured in petri dish and allowed to dry in an oven. After 2h hours removed the wafers from oven and wrapped in an aluminum foil and stored in air tight glass container [65]. Table 4. Composition of formulations of Ranitidine hydrochloride 50mg wafers (All quantities were given in %ages) Class Polymer Formulation HPMC CMC Pectin Guar gum PEG Tween Water FI 27.7 - - - 17.02 1.20 5.0 FII 33.58 - - - 17.02 1.50 6.76 FIII 33.58 - - - 20 1.20 4.0 FVII - 27.7 - - 17.02 1.20 5.0 FVIII - 33.58 - - 17.02 1.50 6.76 FIX - 33.58 - - 20. 1.20 4.0 FIV - - 27.7 - 17.02 1.20 5.0 FV - - 33.58 - 17.02 1.50 6.76 FVI - - 33.58 - 20 1.20 4.0 FX - - - 27.7 17.02 1.20 5.0 FXI - - - 33.58 17.02 1.50 6.76 FXII - - - 33.58 20 1.20 4.0 Code Synthetic HPMC Polymer CMC Natural Polymer Pectin Guar gum 29 4.5. Characterization of wafers Physicochemical parameters such as surface texture, transparency, tackiness, surface pH, thickness, weight variation, swelling index, percentage moisture absorption and percentage moisture loss were calculated. Mucoadhesion, folding endurance, disintegration time and dissolving time were performed on wafers. Morphological studies of wafers were performed by optical microscopy. Drug and polymer interaction were studied by Fourier transform infrared spectroscopy (FTIR). Cumulative percent drug release was studied by dissolution and in-vitro kinetic were studied by applying different kinetic models. 4.5.1. Physical parameters of wafers Surface texture of wafers were evaluated by organoleptic method. A rectangular piece of each wafer film was taken and placed on the internal side of spectrophotometer and transmittance was measured at 600nm and transparency of film was calculated by using equation 4. Transparency test (t) = (4) Where t is wavelength and b is the film thickness. Three films of each formulation were taken and placed them on the piece of paper and tackiness was measured [65 and 66]. 4.5.2. Surface pH Three films of each formulation were taken at random and allowed to swell in agar plate for about 2 hours. pH of wafers was measured by placing the pH paper on the swollen surface of the wafers [43]. 4.5.3. Wafers thickness Thickness of wafers were measured by using the micrometer screw gauge at five different position of the films i.e. the center and the four corners [67]. 4.5.4. Weight variation test Wafers were weighed accurately on the analytical weighing balance and average weight for each formulation was measured, weight variation should be within the Pharmacopoeial limits ±10% [54 and 68]. 30 4.5.5. Swelling index Swelling index of wafers was determined by measuring initial diameter of each wafer then placed in agar petri dish at 37ºC. Diameter was again measured after 1hour [69]. Radial swelling was calculated by using equation 5. (5) SD% means the percentage radial swelling, Dt is the diameter of swollen wafers and D0 is the diameter of original wafers. 4.5.6. Percentage moisture absorption (PMA) Physical stability was measured by using the 1-2cm weighed wafers in desiccator. Percentage moisture absorption was calculated using following formula. (6) 4.5.7. Percentage moisture loss (PML) 3-4cm of wafers film were cut, accurately weighed and kept in desiccator which contains fused anhydrous calcium chloride. Films were removed after 72 hours and weighed. Average percentage moisture loss of desiccated wafers was calculated using equation 7 [70]. (7) 4.5.8. In-vitro mucoadhesion test In-vitro mucoadhesion test was carried out using the modified disintegration apparatus. A petri dish cover was attached with the wall of disintegration apparatus. Everted sac technique was used to evaluate mucoadhesion time, when it completely attached to the plate then wafers were placed on the surface and move apparatus at 75rpm and time was measured at which wafers detached from membrane [49]. 4.5.9. Folding endurance Folding endurance was evaluated by repeatedly folding wafers at same place until it breaks [71]. 4.5.10. Disintegration time 31 Disintegration was carried out by using non agitation method (petriplate method) and agitation (Beaker) method. 4.5.10.1. Beaker method (agitation method) Three pieces of each wafers film were taken randomly and placed in a beaker containing 250 ml of phosphate buffer 6.8 pH and placed on hot plate magnetic stirrer at 200 rpm and time was noted until wafers start to disintegrate. 4.5.10.2. Petriplate method non-agitation method Wafers samples were placed on a glass petri dish containing 10-20ml of distilled water and time was noted which the wafers disintegrate. 4.5.11. Total dissolving time (TDT) Three wafers of each formulation were placed in the Petriplate contain 15-20ml of phosphate buffer pH 6.8 used as dissolution medium and time was noted at which the wafers completely dissolves and becomes a solution [72]. 4.6. Morphological studies 4.6.1. Scanning Electron Microscopy (SEM) SEM images of wafers were obtained at a magnification of 100um and spot size of 320A◦ carried out to study the surface morphology of wafers [60]. 4.6.2. Optical microscopy Optical microscopy was carried out to study the surface morphology of drug loaded and unloaded wafers by using low and high resolution of optical microscope [47]. 4.7. FTIR spectroscopy FTIR spectra of ranitidine, polymers and drug loaded and unloaded wafers were recorded at 4000-400 cm-1 by using FTIR spectrophotometer (Agilent, USA) [51]. 32 4.8. Content uniformity Content uniformity was determined by dissolving wafers in water to prepare solution of ranitidine. Solution was filtered through 0.45um filter and was analyzed at 323nm by UV-visible spectrophotometer. Each experiment was performed three times (n=3) [42]. 4.9. Stability studies Stability studies were performed at 400C ±0.50C and humidity of about 75% for 3 months. After three months wafers were again evaluated for surface pH, folding endurance, drug content, uniformity of weight and in-vitro drug release [73]. 4.10. Dissolution study Drug release studies were performed by using USP type II dissolution paddle apparatus by using phosphate buffer pH 6.8. Temperature of dissolution medium was maintained at 37± 0.50C and peddle was rotated at 75rpm. Wafers were fixed on glass slide and placed in the dissolution medium. Sink condition was maintained throughout the experiment. Sample was analyzed at 323nm [43 and 74] 4.11. Release kinetics The mechanism drug release kinetics from wafers was analyzed by using various kinetic models such as zero order, higuchi model, first order and korsmeyer-peppas model. 4.11.1. Zero order release When drug release is not dependent on its concentration is referred as zero order release mechanism and is calculated by following equation. (9) Where Ft is fraction of drug release at time t and Ko is rate constant for zero order release. 33 4.11.2. First order release When drug release pattern depends upon concentration, it is referred as first order release mechanism and calculated by following equation. (10) F is drug release in time t and K1 is first order release constant 4.11.3. Higuchi model Higuchi model shows time dependent diffusion process from insoluble matrix. It is based on Fick’s law of diffusion and calculated by following equation. (11) F is fraction of drug release in time t and K2 is Higuchi constant 4.11.4. Korsmeyer-peppas model Korsmeyer-peppas equation is as follows. Mt is amount of drug release at time t, M∞ is amount of drug release at infinity and n is diffusion coefficients. 34 5. Result and Discussion Pakistan is an under developed country where almost 51 % of population is living below poverty line, it is difficult for people to purchase costly medicine. A dosage form, more effective having less cost is much needed. Ranitidine hydrochloride (RNH) is commonly recommended drug for management of erosive esophagitis, gastric ulcer (GU), Zollinger –Ellison’s syndrome, duodenal ulcer, Ranitidine is recommended ranging from 150mg twice daily (BD). A novel drug delivery system (NDDS) sustained released wafers of RNH was prepared by solvent casting technique using various natural and synthetic polymers. 5.1. Organoleptic evaluation Natural polymer (Pectin and Gaur gum) containing wafers showed the smooth surface while the synthetic polymers (HPMC) containing wafers showed slightly rough texture. CMC containing wafers had the crystalline texture. All the prepared formulations showed maximum transparency when observed in the cube cell of the spectrophotometer. Prepared formulations of natural polymer were evaluated for their Stickiness and dryness characteristics at the surface of the paper were found to be some stickiness while the lack stickiness was observed in synthetic polymers containing wafers. 5.2. Effects of excipients on various parameters of wafers formulations Effect of excipients on surface pH and folding endurance of formulation FI-FIII was evaluated as variables using box behnken design which showed that increasing the concentration of polymer showed no marked variations in surface pH, however increasing concentration of polymer and decreasing plasticizer concentration showed increased folding endurance of formulation as shown in figure 1 and 2. 35 Figure 1: Excipients effect on surface pH of Formulations (FI-FIII) Figure 2: Excipients effect on folding endurance of formulations (FI-FIII) Mucoadhesion time and % moisture loss of formulation FI-FIII was study and was found to be increased concentration of polymer enhanced mucoadhesion time and reduces percentage moisture loss as shown in figures 3 and 4 respectively. Figure 3: Excipients effect on mucoadhesion time of formulations (FI-FIII) 36 Figure 4: Excipients effect on percentage moisture loss of formulations (FI-FIII) Effect of Pectin polyethylene concentration was observed on surface pH and folding endurance of formulations FIV-FVI. No significant effect on surface pH was observe, however increasing effect on folding endurance was observed with increasing polymer concentration and decreasing plasticizer concentration as shown in figures 5 and 6 respectively. Figure 5: Effect of excipients on surface pH of formulations (FIV-FVI) 37 Figure 6: Effect of excipients on folding endurance of formulations (FIV-FVI) Effect of excipients on mucoadhesion time and %moisture loss of formulations FIV-FVI was observed which showed that formulations contaning higher polymer concentration with more mucoadhesion and less moisture loss as shown in figures 7 and 8. Figure 7: Excipients effect on in-vitro mucoadhesion time of formulation (FIV-FVI) 38 Figure 8: Excipients effect on percentage moisture loss of formulations (FIV-FVI) Surface pH and folding endurance of wafers formulations (FVII-FIX) contain various concentration of CMC, PEG were evaluated which showed that increasing concentration have no significant effect on surface pH, however significant increase folding endurance was observed as shown in figures 9 and 10 respectively. Figure 9: Excipients effect on surface pH of formulations (FVII-FIX) 39 Figure 10: Excipients effects on folding endurance of formulations (FIV-FVI) CMC possess an excellent mucoadhesive strength due to which it showed increased mucoadhesion time and less %moisture loss as shown in figures 11 and 12 respectively. Figure 11: Excipients effect on mucoadhesion time of formulations (FIV-FVI) 40 Figure 12: Excipients effect on moisture loss of formulations (FVII-FIX) Effect of excipients concentration on surface pH of formulations (FX-FXII) showed changing concentrations had no effect on surface pH of formulations. However increasing polymer concentration results in increasing folding endurance as shown in figure 13 and 14. Figure 13: Excipients effect on surface pH of formulations (FX-FXII) 41 Figure 14: Excipients effect on folding endurance of formulation (FX-FXII) Effect of excipients on percentage moisture loss of formulations containing Guar gum which showed that increasing polymer concentration reduces % moisture loss of formulations however when plasticizers concentration increased moisture loss increases. Polymer concentration showed increased mucoadhesion. Figure 15: Excipients effect on in-vitro mucoadhesion of formulations (FX-FXII) 42 Figure 16: Excipients effect on moisture loss of formulations (FX-FXII) 5.3. Physicochemical parameters 5.3.1. Surface pH Surface pH of wafers formulations was found to be within range of 6.52-6.56 ± 0.1 which was suitable for absorption of drug at intestinal pH as shown in table 5. 5.3.2. Wafers thickness Five wafers of each formulation were taken randomly and thickness of each wafer was measured at five different positions and average thickness of wafers was upto 0.35mm as shown in figures 17, 18 and 19 respectively. 43 Figure 17: Thickness of wafers formulations FI-FIII (n=5) Figure 18: Thickness of wafers formulations FIV-FVI (n=5) Thickness of various formulations contain various concentrations of excipients was evaluated which shows that all formulations FVII-FXII were of uniform thickness which was less than 1mm and was found to be within limit as shown in figures 19 and 20. 44 Figure 19: Thickness of wafers formulations FVII-FIX (n=5) Figure 20: Thickness of wafers formulations (FX-FXII) 5.3.3. Weight variation test Twenty wafers of each formulation were randomly taken and weighed individually. Average weight of twenty wafers was calculated variation of wafers were not found to be more then ±5±10%. Weight variation of each formulations is shown in graphs below. Weight variation of Twenty wafers (films) of FI,FII and FII were taken randomly and weighed individually and average weight was calculated which was found by 647mg ±3.56 and was found to be within pharmacopoeial limits ,maximum weight of wafer was found to be 659 mg and minimum weight 643mg which lies within the limit. As shown in figure 21, 22 and 23 respectively. 45 Figure 21: Weight variation of wafers formulationF1 (n=20) Figure 22: Weight variation of wafers formulation FII (n=20) . 46 Figure 23: Weight variation of wafers formulation FIII (n=20) Weight variation of twenty wafers FIV,FV and VI was determined by taking five wafers of each formulation which was found to be 647.20mg ±3.39 and was found to be within pharmacopoeial limits ,maximum weight of wafer was found to be 656mg and minimum weight 644mg which lies within the limit as shown in figures 24 ,25 and 26 respectively. Figure 24: Weight variation of formulation FIV (n=20) 47 Figure 25: Weight variation of formulation FV (n=20) Figure 26: Weight variation of formulation (FVI) Weight variation of twenty wafers (films) of FVII, FVIII and FIX were taken randomly and weighed individually and average weight was calculated which was found by 647.700mg ±3.74 which was found to be within pharmacopoeial limits, maximum weight of wafer was found to be 656mg and minimum weight 639mg which lies within the limit as shown in figure 27, 28 and 29 48 Figure 27: Weight variation of formulation FVII (n=20) Figure 28: Weight variation of formulation FVIII (n=20) 49 Figure 29: Weight variation of formulation FIX (n=20) Weight variation of twenty wafers of FX, FXI and FXII were taken randomly and weighed individually and average weight was calculated which was found to be 647mg ±3.56 which was found to be within Pharmacopoeial limits, maximum weight of wafer was found to be 656 mg and minimum weight 639 mg which lies within the limit as shown in figures 30, 31and32 respectively. Figure 30: Weight variation of formulation FX (n=20) 50 Figure 31: Weight variation of formulation XI (n=20) Figure 32: Weight variation of formulation XII (n=20) 5.3.4. Swelling index Five wafers of each formulation were randomly taken initial diameter and final was measured percentage radial swelling of formulations FI-FVI was measured. Maximum radial swelling was found to be 15 minutes as shown in figures 33and 34. 51 Figure 33: Percentage radial swelling of formulations FI-FIII (n=5) Figure 34: Percentage radial swelling of formulations FIV-FVI Percentage radial swelling of formulations FVII-FXII was determined and radial swelling of formulations was found to be up to 30 mints as shown in figure 35and 36. 52 Figure 35: Percentage radial swelling of formulations FVII-FIX (n=5) Figure 36: Percentage radial swelling of formulations FX-FXII (n=5) 5.3.5. Percentage moisture absorption (PMA) Percentage moisture absorption of various formulations were carried out by taking three wafers of each formulations initial weighed and then placed initially weighed and then placed in desiccator containing calcium chloride solution after 72 hours these were taken out again weighed , %moisture absorption was calculated for each formulation and shown in the table 5. 53 5.3.6. Percentage moisture loss (PML) Percentage moisture loss of various formulations were carried out by taking three wafers of each formulation initial weighed and then placed in oven for 72 hours after specified time interval these were taken out again weighed %moisture loss was calculated for each formulation and shown in the table 5. Table 5: Evaluation of various physical parameters of ranitidine hydrochloride wafers Class Polymer Formulat ion Surface Folding % moisture % moisture absorption Swelling index Thickness Conent uniformty pH Endurance loss FI 6.51 ±0.08 116.42 ±3.20 1.84 ±0.63 1.08 ±0.36 7.81 ±3.10 0.31 ±0.02 93.52±7.22 FII 6.53 ±0.13 142.64±3.162 1.71 ±0.76 1.18 ±0.39 10.80±3.0 0.34 ±.015 92.19±8.32 FIII 6.56 ±0.11 107.36 ±3.54 1.58 ±0.10 1.060±0.42 11.61±2.5 0.32±0.011 95.38±3.84 FVII 6.59 ±0.25 289.81 ±2.25 1.344 ±0.38 1.50 ±0.39 8.74±1.83 0.35±0.021 95.81±3.07 FVIII 6.56 ±0.12 296.73±1.49 1.540 ±0.54 1.682±0.41 11.77±2.3 0.35±0.022 96.92±3.11 FIX 6.51 ±0.10 288.66 ±2.19 1.295 ±0.80 1.572 ±0.32 15.11±3.6 0.39±0.015 95.59±2.79 FIV 6.51 ±0.11 298.29 ±2.30 1.095 ±0.32 1.520 ±0.61 8.77±1.81 0.26±0.021 95.13±2.87 FV 6.53 ±0.10 298.33 ±1.49 1.56 ±0.46 1.40 ±0.50 12.14±2.4 0.33±0.015 97.22±3.07 FVI 6.53 ±0.12 292.03 ±2.18 1.33 ±0.49 1.23 ±0.13 8.571±2.1 0.352±0.02 95.30±2.90 FX 6.56 ±0.25 213.12 ±3.21 1.339±0.49 1.34 ±0.44 9.34±2.18 0.302±0.01 95.91±2.75 FXI 6.56 ±0.25 235.16 ±2.43 1.24 ±0.46 1.630 ±0.40 15.0±1.24 0.318±0.01 95.51±2.37 FXII 6.53 ±0.10 241.71±2.23 1.82 ±0.49 1.28 ±0.22 5.012±2.4 0.318±0.01 98.50±5.501 Code Synthetic HPMC Polymer CMC Natural Polymer Pectin Guar gum 5.3.7. In-vitro mucoadhesion test Mucoadhesion time of wafers formulations containing different concentrations of polymers, plasticizers and surfactant which showed that formulation FI, FIII were found to be with less mucoadhesion time 19.3 and 22.25 minutes however formulation FII showed maximum mucoadhesion time 24 minutes as shown in figure 37. 54 Figure 37: In-vitro mucoadhesion time of wafers formulations FI-FIII Wafers formulations FIV, FV and FVI were evaluated for mucoadhesion time. Mucoadhesion time of formulations was found to be up to 25 minutes as shown in figure 38. Figure 38: In-vitro mucoadhesion time of wafers formulations FIV-FVI Mucoadhesion time of wafers formulations containing different concentrations of polymers, plasticizers and surfactant which showed that formulation FVII, FIX were found to be with less mucoadhesion time, however formulation FVIII showed maximum mucoadhesion which showed that increasing concentration of polymer and lowering plasticizer results in more mucoadhesive formulations as shown in figure 39. 55 Figure 39: In-vitro mucoadhesion time of wafers formulations FVII-FIX) Mucoadhesion time of wafers formulations containing different concentrations of polymers, plasticizers and surfactant which showed that formulation FX, FXII were found to be with less mucoadhesion time, however formulation FXI shows maximum mucoadhesion which showed that increasing concentration of polymer and lowering plasticizer results in more mucoadhesive formulations. Radial graph showed mucoadhesion time of wafers formulations containing different concentrations of polymers, plasticizers and surfactant which showed that formulation FII, FXI were found to be with less mucoadhesion time, however formulation FV,FVIII shows maximum mucoadhesion which showed that increasing concentration of polymer and lowering plasticizer results in more mucoadhesive formulations as shown in figure 40. 56 Figure 40: In-vitro mucoadhesion of wafers formulations FX-FXII 5.3.8. Folding endurance Three wafers of each formulation were taken and folded at the same place for several periods of time until the films break the maximum limit at which film breaks is folding endurance; folding endurance of various formulation as shown in table 5. 5.3.9. Disintegration time Disintegration of five wafers of each formulation were carried out both by agitation and nonagitation method and average disintegration time of each formulation was calculated it was found that disintegration time without agitation is more as compared to agitation as shown in figures 41, 42,43 and 44 respectively. 57 Figure 41: Disintegration time of formulations (FI-FIII) Figure 42: Disintegration time of formulations from FV-FVI 58 time mints 30 25 20 15 10 5 0 fixna Fviia Fviia Fviina Fviiia fixa Fviina Fviiina Fixa Fix na fviina fviiia Figure 43: Disintegration time of formulation (FVII-FIX) 5.3.10. Total dissolving time (TDT) Three wafers of each formulation were placed in the Petriplate having 15-20ml of phosphate buffer pH 6.8 and the time at which the wafers completely dissolved without agitation were calculated. Solution formation after the agitation were also observed as shown in figure 45. Total dissolving time along a single axis graph showed total dissolving time of three wafers formulations FI-FIII, Figure showed that formulation FI and FIII, showed less total dissolving time however FII showed maximum dissolving time as compared to FI,FIII so FII was considered to be optimized formulation as compared to the other two formulations as shown in figures 45 and 46. 59 Figure 44: Total dissolving time of wafers formulations FI-FIII Total dissolving time along a single axis graph showed total dissolving time of three wafers formulations FIV-FVI ,graph showed that formulation FIV,FVI showed less total dissolving time however FV showed maximum dissolving time as compared to FIV, FVI so FV was considered to be optimized formulation as compared to the other two formulations. Figure 45: Total dissolving time of wafers formulation FIV-FVI Total dissolving time along a single axis graph showed total dissolving time of three wafers formulations FVII-FIX ,graph showed that formulation FVII,FIX showed less total dissolving time however FVIII showed maximum dissolving time as compared to FVII, FIX so FVIII was considered to be optimized formulation as compared to the other two formulations as shown in figures 47 and 48 respectively. 60 Figure 46: Total dissolving time of wafers formulation FVII-FIX Total dissolving time along a single axis graph showed total dissolving time of three wafers formulations FX-FXII ,graph showed that formulation FX ,FII showed less total dissolving time however FXI showed maximum dissolving time as compared to FX,FXII so FXI was considered to be optimized formulation as compared to the other two formulations. Figure 47: Total dissolving time of wafers formulation FX-FXII Total dissolving time along a single axis graph showed total dissolving time of three wafers formulations FII, FV, FVIII, FXI, graph showed that formulation FII,FXI showed less total dissolving time however FV, FVIII showed maximum dissolving time as shown in figures 49 and 50. 61 Figure 48: Total dissolving time of wafers formulation FII, FV, FVIII and FXII Figure 49: Total dissolving time of wafers formulation FV, FVIII 62 Figure 50: Total dissolving time with agitation formulation FI-FIII Total dissolving time with agitation along a single axis graph showed total dissolving time of three wafers formulations FIV-FVI ,graph showed that formulation FIV,FVI showed less total dissolving time however FV showed maximum dissolving time as compared to FIV,FVI so FV was considered to be optimized formulation as compared to the other two formulations. Figure 51: Total dissolving time of wafers formulations FIV-FVI Total dissolving time with agitation along a single axis graph showed total dissolving time of three wafers formulations FVII-FIX ,graph showed that formulation FVII,FIX showed less total 63 dissolving time however FVIII showed maximum dissolving time as compared to FVII,FIX so FVIII was considered to be optimized formulation as compared to the other two formulations as shown in figure 51 and 52. Figure 52: Total dissolving time of wafers formulations from FVII-FIX 5.4. Morphological studies Microscopic image analysis of wafers formulations containing different concentrations of polymers and plasticizer and tween 80 were compared. Wafers formulation (FIII) showed crosslinked network like structure with uniform loading of drug concentration. Formulation FIV contained pectin polymers was wrinkled in shape and showed agglomeration with drug molecules. The difference was observed due to irregular mixing of excipients and drug molecules in formulation. FV formulation containing 17% of plasticizer showed smooth surface in texture and was easily inspected visually due to uniform distribution of drug particles. Wafers formulations containing lesser concentration of polymeric material showed crystalline texture as shown in figures 53 and 54. 5.4.1. Microscopic images of wafers 64 Figure 53: Microscopic images wafers prepared with (HPMC) Figure 54: Microscopic images of wafers prepared with Pectin 5.4.2. SEM of wafers formulations 5.5. FTIR studies FTIR studies of pure Ranitidine hydrochloride (RNH) prepared wafers formulations containing polymers such as HPMC, CMC, Pectin and Guar gum were evaluated ranging from 4000-400 cm. Pure drug showed peaks due to secondary diamine, furan ring alkene and nitro group at2500cm-1 peak observed which is due to the presence of N+-H- bond protonated tertiary amine, 2nd peak was observed at the 1610cm-1which was due to stretching of acetonitrile group and third peak of pure ranitidine was observed at 1460cm-1 for diamine contain group due to vibrational 65 motion of the molecules. Standard FTIR spectra of pure ranitidine was compared with the spectra of formulations containing various polymers in different concentration along with plasticizers and surfactants. FTIR spectra of tween 80 showed peaks at 3350 cm-1 and 1640 cm-1 due to stretching of CH2 alkane group and at 1640cm-1 due to C=O stretching as shown in Figure 56.FTIR spectra of pectin showed peaks at 1750cm-1, 1400cm-1, 1200cm-1,1050cm-1 and 980cm-1 due to ester carbonyl group ( C=O) stretching ,1400 cm-1 due to CH bonding (in –plane) and at 1200 cm-1 due to =C-H out –of –plane bending as shown in Figure55(A). FTIR spectra of pectin wafers showed peaks at 3300cm-1, 1600cm-1, 1400cm-1, 1100cm-1 and 1000cm-1 due to C=O stretching –CH bending and =C-H bending respectively as shown in Figure55(B). FTIR spectra of CMC showed peaks at 3300cm-1,1590cm-1,1390cm-1,1240cm-1,1100cm-1, and 1000cm-1 due to –OH stretching, carboxylic group stretching and stretching vibration of the C-OH group respectively as shown in Figure 55(C). FTIR spectra of drug contain wafers showed peaks at3300cm-1,2100cm-1 1564cm-11421cm-1,1321cm-1 and 1051cm-1 due to –OH stretching, carboxylic group stretching and stretching vibration of C-OH group respectively as shown in Figure60.FTIR spectra of Guar gum drug loaded wafers showed peaks at 2490cm-1,1616cm1 ,1370cm-1,1220cm-1 and 1000cm-1 due to methyl C-H stretching, 1616 cm-1 due to=C- Hstretching,1220cm-1 due to methylene twisting and 1000 cm-1 due to stretching vibration of COH group as shown in Figure55(D). No changes in the peaks were observed which shows that the excipients shows compatibility with the all the excipients.as no changes in these peaks were evaluated so no drug –excipients interactions were found as shown in figure (E). 66 Figure 55: FTIR spectra (A) Ranitidine HCL (B)Pectin ,(C) Pectin wafers ,(D) CMC wafers and (E) HPMC wafers 67 5.6. Content uniformity of wafers Content uniformity test was carried out on various wafers formulation, contents of the formulations were found to be within the limit. Content uniformity values of various formulations were presented in table 5. 5.7. Stability studies Prepared wafers were placed in the stability chamber at humidity of about 75% and temperature of the chamber was maintained at 400C for 3 months. After three months in the stability chamber these formulations were again evaluated for surface pH, folding endurance, drug content, uniformity of weight, results were presented in the table 6. 68 Table 6: Stability studies of Ranitidine hydrochloride wafers Parame ters F1 F2 F3 F4 F5 Color F6 F7 F8 F9 F10 F11 F12 Light red Surface pH 6.51±0.1 1 6.46± 0.11 6.41± 0.13 6.59± 0.25 6.56± 0.13 6.51± 0.10 6.51± 0.12 6.52± 0.10 6.53 ±0.1 2 6.51 ±0.1 4 6.53 ±0.1 1 6.52 ±0.1 3 Folding endura nce 116.4±3. 2 107.4 ±3.5 142.6 ±4.1 296.73 ±2.81 289.8 ±4.25 278.6 ±1.48 292.04 ±2.2 298.3 ±2.3 298. 3±1. 5 262± 2.2 270± 2.7 276 Disinte gration time 10.2±0.5 7 18.7 5± 14.6 ±0.9 6 17.4 ±0.8 9 17.5 ±0.8 9 94.6 8 95.5 4 95.6 15.5± 0.79 17.6± 0.41 12.8± 0.57 18.5± 0.79 18.6± 0.41 20±0. 79 20.65 ±0.78 ±1.5 0.79 Content unifor mity 5.8. 85.30±1 1.12 92.54 ±8.91 89.50 1±17. 993 88.01 ±5.14 94.6± 7.03 95.89 ±12.7 91.12 ±19.4 1 91.70 ±14.5 4 92.6 7 Preparation of standard /calibration curve The stock solution of Ranitidine hydrochloride was prepared in phosphate buffer pH 6.8 and serial dilutions was prepared. Absorbance was measured by using UV-visible spectrophotometer as shown in figure 56. 69 0.93 absorbance(nm) 1 y = 0.2629x + 0.0991 R² = 0.9985 0.8 0.491 0.6 0.4 0.2 0.213 0.156 0.143 0.303 0 0 0.5 1 1.5 2 2.5 3 3.5 Drug percentage Figure 56: Calibration curve of ranitidine 5.9. Dissolution studies In-vitro release of wafers was studied by using modified USP type II dissolution apparatus. Percentage drug release of three wafers formulation FI showed more than 85% drug release within 3hours, FII showed more than 85 % release within 6 hours , however FII showed more than 85% release after 6 hours which as shown in figure 57. Figure 57: Percentage drug release of wafers formulation FI-FIII 70 Percentage drug release of wafers formulation FIV released more than 85% of ranitidine within 2 hours , FV showed more than 85% drug release after 8hours and FVI showed more than 85 % release after 6 hours as shown in figure 58. Figure 58: Percentage drug release of wafers formulations FIV-FVI Percentage drug release of formulation FVII showed more than 85% release within 4 hours, FVIII showed more than 85% drug release after 8hours and FIX showed more than 85% release within 6 hours as shown in figure 59. 71 Figure 59: Percentage drug release of wafers formulations FVII-FIX Radial graph shows %age drug release along a single axis graph shows that %age drug release of formulation FX showed more than 85% release within 6 hours, FXI shows more than 85% drug release after 8hours and FIX showed more than 85 % release within 3 hours as shown in figure 60. Figure 60: Percentage release of wafers formulations FX-FXII Release kinetic models Different kinetic models of release were applied to the 12 formulations of wafers to determine drug follows which type of kinetic models. 5.9.1. Kinetic models of formulations (FI-FIII) Four models zero order, first order, higuchi, and korsmeyer-peppas models were applied to the formulations (FI-FIII) formulations values were shown in table 7. 72 Table 7: In-vitro drug release kinetics and its model dependent approaches (FI-FIII) Kinetic model FI FII FIII - R2 K(h-1)/n R2 K(h-1)/n R2 K(h-1)/n Zero order 0.573 12.21 0.410 11.18 0.379 12.30 First order 0.947 0.402 0.816 0.408 0.809 1.152 Higuchi 0.376 37.44 0.516 33.20 0.556 38.00 Korsmeyer Peppas 0.860 0.255 0.977 0.329 0.923 0.208 Kinetic models i.e. zero order, first order, higuchi model and korsmeyer-peppas were applied to the formulation (FI) values of R2 were upto 0.947 very close to unity(1) which shows that drug release depends on concentration of drug in body as shown in figures 62,63 and 64. Figure 61: Zero order release models of formulation FI 73 Figure 62 : Higuchi release model of wafers formulation (FI) Korsmeyer-peppas release model was applied on formulation (FI) and n value was found to be less than 0.255 which shows that drug release follows Fickian release as shown in figure 65. Figure 63: Korsmayer-peppas model of wafers formulation (FI) Various kinetic models such as zero order, first order and Higuchi were applied successfully on formulation (FII) value of R2 was observed and incase of first order was 0.816 found close to (1) indicating drug dependency release pattern as shown in figure 66,67 and 68. 74 Figure 64: Zero order release model of wafers formulation (FII) Figure 65: First order release model of wafers formulation (FII) 75 Figure 66: Higuchi release model of wafers formulation (FII) Observed n value was less than 0.329 which indicates that drug release follows Fickian kinetic as shown in figure 69. Figure 67: Korsmeyer-peppas release model of (FII) Percentage release data of FIII formulation was fitted in to various kinetic models i.e. zero order, first order and higuchi and observed R2 value in first order model 0.809 which was near to unity which indicates that drug release is concentration dependent as shown in figure 70,71 and 72. 76 Figure 68: Zero order release model of wafers formulation (FIII) Figure 69: First order release model of wafers formulation (FIII) Figure 70: Higuchi release model of wafers formulation (FIII) 77 Korsmeyer peppas model was applied to formulation FIII and n value was observed and found to 0.208 which shows drug release follow Fickian model as shown in figure 73. Figure 71: Korsmeyer-peppas model of wafers formulations (FIII) 5.9.2. In-vitro kinetics of formulation (FIV-FVI) Four models zero order, first order, Higuchi and korsmeyer-peppas were applied to the formulations (FIV-FVI) formulations of wafers as shown in table 8. Table 8: In-vitro drug release kinetics and its model dependent approaches (FIV-FVI) Kinetic model FIV FV FVI - R2 K(h-1)/n R2 K(h-1)/n R2 K(h-1)/n Zero order .704 12.66 0.824 11.17 0.325 11.29 First order 0.882 1.889 0.847 0.577 0.885 0.339 Higuchi 0.361 39.81 0.320 33.801 0.881 32.76 Korsmeyer Peppas 0.796 0.159 0.901 0.244 0.896 0.431 Drug release data of (FIV) was analyzed by Appling zero, first order and higuchi release model and observed R2 of first order 0.882 was very close to 1 in first order which indicates drug release dependents on concentration as shown below in figure 74,75 and 76 respectively. 78 Figure 72: Zero order release model of wafers formulation (FIV) Figure 73: First order release model of wafers formulation (FV) 79 Figure 74: Higuchi release model of wafers formulation (FIV) Korsmeyer-peppas release FIV release was analyzed n value 0.402 which indicates drug release follow Fickian kinetic.as shown in figure 77. Figure 75: Korsmeyer-Peppas release model of wafers formulation (FIV) Kinetic release models i.e. zero, first. Higuchi were applied on drug release data of formulation FV and observed R2 value of first order was 0.847 found to close to unity indicating concentration dependent release pattern as shown in figure 78,79 and 80. 80 Figure 76: Zero order release model of formulation (FV) 81 Figure 77: First order release model of wafers formulation (FV) Figure 78: Higuchi release model of wafers formulation (FV) Observed n value of korsmeyer-peppas was found to be 0.244 which showed Fickian release of FV formulation as shown in figure 81. Figure 79: Korsmayer-peppas release model of wafers formulation (FV). 82 Kinetic models i.e. zero, first and higuchi were applied successfully to FVI formulations. Results of R2 of first order was found to be 0.885 very close to 1 indicating drug concentrating dependent release pattern as shown in figures 82, 83, 84 and 85 respectively. Figure 80: Zero order release model of wafers formulation (FVI) Figure 81: First order release model of wafers formulation (FVI) 83 Figure 82: Higuchi release model of wafers formulation (FVI) Figure 83: Korsmeyer-peppas release model of wafers formulation (FVI) 5.9.3. In-vitro kinetics of formulation (FVII-FIX) Four models zero order, first order, higuchi and korsmeyer-peppas were applied to the formulations (FVII-FIX) formulations of wafers as shown in table 9. 84 Table 9: In-vitro drug release kinetics and model dependent approaches (FVII-FIX) Kinetic model FVII FVIII FIX - R2 K(h-1)/n R2 K(h-1)/n R2 K(h-1)/n Zero order 0.130 11.60 0.476 10.71 0.447 10.84 First order 0.977 0.454 0.964 0.950 0.967 0.312 Higuchi 0.846 34.29 0.945 30.92 0.946 31.36 Korsmeyer Peppas 0.887 0.393 0.949 0.452 0.953 0.454 Drug release data of (FVII) was applied to various kinetic models i.e. zero order ,first order and higuchi and R2 of first order was 0.977 very close to unity (1) indicating concentration depends release as shown in figure 86, 87 and 88. Figure 84: Zero order release model of wafers formulation (FVII) 85 Figure 85: First order release model of wafers formulation (FVII) Figure 86: Higuchi release model of wafers formulation (FVII) Korsmeyer-peppas release model of wafers formulation (VII) was applied and n value was found to be 0.393 shows Fickian release as shown in figure89. 86 Figure 87: Korsmeyer-peppas model of wafers formulations (FVII) Kinetic models zero, first and higuchi were applied successfully R2 value was 0.950 indicating drug release depends on concentration i.e. follow first order release kinetics as shown in figure 90, 91 and 92. Figure 88: Zero order release model of FVIII 87 Figure 89: First order release model of wafers formulation (FVIII) Figure 90: Higuchi release model of wafers formulations (FVIII) Fickian release was observed by applying drug release data in to kinetic models as value of n was 0.452 shown in figure 93. 88 Figure 91: Korsmeyer peppas release model of formulation (FVIII) Percentage drug release data of formulation (FIX) was fitted in to kinetic models and R 2 was observed which showed that first order value 0.967 is close to unity (1) indicates concentration dependent drug release as shown in figure 94, 95 and 96 respectively. Figure 92: Zero order release model of wafers formulation (FIX) 89 Figure 93: First order release model of wafers formulation (FIX) Figure 94: Higuchi release model of wafers formulation (FIX) Korsmeyer peppas release model shows drug release value R2 0.454 follows Fickian release kinetics as shown in figure 97. 90 Figure 95: Korsmeyer-peppas release model of wafers formulation (FIX) 5.9.4. Kinetic models of formulation (FX-FXII) Four models such as zero order, first order, Higuchi and korsmeyer-peppas were applied to formulations (FX-FXII) formulations of wafers as shown in table 10. Table 10: In-vitro release kinetics model dependent approaches (FX-FXII) Kinetic model FX FXI FXII - R2 K(h-1)/n R2 K(h-1)/n R2 K(h-1)/n Zero order 0.456 11.29 0.621 10.15 .5760 11.99 First order 0.971 0.344 0.974 0.245 0.910 0.692 Higuchi 0.922 32.68 0.989 28.96 0.542 36.45 Korsmeyer Peppas 0.926 0.362 0.989 0.399 0.908 0.276 Drug release data of formulations (FX) was fitted in to kinetic models i.e. zero first order and higuchi models observed results are 0.977 showed that drug release is dependent on concentration of drug as shown in figure 98,99 and 100. 91 Figure 96: Zero order release model of wafers formulation (FX) Figure 97: First order release model of wafers formulation (FX) 92 Figure 98: Higuchi release model of wafers formulation (FX) Korsmeyer-peppas release model of wafers formulation (FX) was applied successfully and drug release was observed which shows that n value is 0.393 thus it follows fickian mechanism as shown in figure101. Figure 99: Korsmeyer-peppas release model of wafers formulation (FX) Zero order, first order and higuchi release kinetics models were applied to formulation (FXI) R2 value was observed which was found to be 0.967 and close to unity in first order as compared to zero and higuchi so drug release dependent on drug concentration as shown in figure 102,103 and 104. 93 Figure 100: Zero order release model of wafers formulation (FXI) Figure 101: First order release model of wafers formulation (FXI) 94 Figure 102: Higuchi release model of wafers formulation (FXI) Korsmeyer-peppas release model of wafers formulation (FXI) was successfully applied and it was observed n value was 0.362 that drug release follows fickian mechanism as shown in figure 105 . Figure 103: Korsmeyer peppas release model of wafers formulation (FXI) Percentage drug release data was analyzed by applying drug release mechanism R2 was found to be 0.974and it depends on drug concentration as shown in figure 106,107 and 108. 95 Figure 104: Zero order release model of wafers formulation (FXII) Figure 105: First order release model of wafers formulation (FXII) 96 Figure 106: Higuchi release model of wafers formulation (FXII) Korsmeyer peppas release model of wafers formulation n values was 0.276 (FXII) shows that drug release follows fickian release as shown in figure109. Figure 107: Korsmeyer peppas release model of wafers formulation (XII) 97 5.10. Discussion Ranitidine hydrochloride (RNH) is commonly recommended for the management of erosive esophagitis, gastric ulcer, Zollinger –Ellison’s syndrome and duodenal ulcer. RNH. Various problems are associated with conventional formulations used in the colon [25]. In the present study a novel drug delivery system (NDDS) such as sustained released wafers of RNH was prepared by solvent casting technique by using two natural and two synthetic polymers. Wafers were evaluated for surface texture which was found to be smooth for pectin, guar gum and HPMC, however crystalline texture was observed for CMC wafers formulation was reported by Shaima et al.,2014 [37] . Wafers of 5×4cm2 were placed on piece of paper and was evaluated for tackiness. All prepared wafers showed non-tackiness already reported by Komaragiri et al.,2012 after preparing the fast dissolving wafers of atenolol [59]. Surface pH of all wafers formulations (FI-FXII) was found to be ranging from 6.52-6.62±0.41. The similar studies were already reported by M.Alagus Undrum et al., 2009 [43]. Thickness of wafer was evaluated at five different positions using screw gauge and was observed uniform thickness ranges 0.31-0.35±0.22 mm which was found to be less than 1mm. Pathan et al., 2011 also reported the similar findings [42]. All formulations showed constant values of the thickness due to solubility of exact amount of drug in all the preparations. Garsuch et al.,2013 reported the similar finding after observing the texture, and drug loading of the different compositions of wafers [75]. Twenty films of each formulation were selected and weighed accurately using analytical weighing balance all the formulations showed weight in the range of 647.5-650.5 ±0.025mg which showed that the method used is suitable for development of wafers also reported by Semalty et al.,2008 [44]. Folding endurance of all wafers formulations were calculated by repeatedly folding the wafers at the same place the order of folding endurance is as CMC>Pectin>Gaur gum>HPMC that showed that nature and concentration of polymer effects the folding endurance that was already reported by Devi et al., 2003 that he developed hydroxypropyl methylcellulose oral films using different concentrations of hydroxypropyl methylcellulose [76]. Formulations were evaluated using modified disintegration apparatus. In-vitro mucoadhesion time showed similar results reported by Ayensu et al., 2012 prepared chitosan and thioglycolic acid lyophilized wafers formulations [77]. Percentage moisture loss of prepared wafers formulations was measured by initially weighing the three wafers of each formulation and placed in an oven for 72 hours, after 72 hour’s wafers were reweighed and percentage loss in weight was calculated percentage. Weight loss of 98 formulations was in the order of CMC<Pectin<Guar gum< HPMC showed similar results were reported by Joshu S Boetang et al., 2009, moisture loss was increased by increasing the contents of plasticizers [78]. Disintegration time of various formulations were evaluated using Petriplate and beaker methods showed that disintegration time was more in non-agitation method as compared agitation method reported by N Jummat et al., 2014 [47]. Percentage moisture absorption of wafers formulations were evaluated to check the stability of wafers in the humid conditions which showed moisture absorbance less than 1.06±0.42-1.68±0.62%. Mathews et al., 2008 evaluated the same by using this technique and found that humidity upto 14% could not affect the stability of wafers formulations [79]. Total dissolving time of wafers were determined by agitated and non-agitated method and it was found that wafers dissolve completely in agitated as compared to non-agitation method reported by Sasi Deepthi et al., 2012 [59]. Morphological studies of formulations were evaluated by optical microscopy and some formulations showed smooth appearance however, other formulations showed the wrinkled network like appearance. Gurcsh et al., 2009 evaluated oral films by using same evaluation technique [80]. Scanning electron microscopy of wafers showed that drug was uniformly distributed throughout the wafers Paola Mura et al., 2015 evaluated upper and lower surface morphology of films using SEM [46]. FTIR analysis of pure ranitidine hydrochloride, excipients and prepared wafers showed no incompatibilities or interactions were reported. Dissolution studies were carried out using USP typeII dissolution apparatus using phosphate buffer 6.8pH as dissolution medium absorbance was measured by using spectrophotometer. It was revealed that by increasing the concentration of polymer, release of drug was retarted from formulations. Patil et al., 2010 also found inprepared floating tablets of ranitidine hydrochloride [81]. Rajesh Kaza et al., also reported that increasing concentration of HPMC retards the release of drug which is suitable polymer for the prepration of controlled released formulations [82]. Polyethylene glycol (PEG200) was used as penitratration enhancer and plasticers used ranging from 17-19% study reveiled that formulations containing 17% plasticizers concentration showed best release. Naryan et al., 2014 prepared the Glibinclamide sustained released release transdermal wafers formulations and reported similar findings [36]. Carboxy methylcellulose containg wafers were evaluated for in-vitro kinetic studies which showed that the formulation containing higher CMC concentration showed better results for controlled released as compared to the less quantity of CMC. K Regurum et al., 2003 also reported that increasing concentration of CMC retards the drug release for several hours 99 [83]. Boetnge et al., 2009 also reported that increasing CMC concentration retards release rate of drug [78]. Wafers were prepared using PEG as plasticizers as well as penetration enhancer it was observed that formulations containing increased polymer concentration and 17% plasticizer retard drug release upto several hours. Kinetic models were applied to drug release data of wafers formulations are shown in tables 7, 8, 9 and 10 respectively. R2 values of all the formulations were observed which was found to be 0.977 indicates the formulation followed the first order. Values of n was 0.442 which was less than 0.45 indicates fickian drug release mechanism [84]. 5.11. Conclusion Ranitidine hydrochloride wafers were prepared successfully using solvent casting. Drug release was studied upto 12 hours, results showed that CMC containg formulations showed best sustained release effect up to 12 hours various kinetic models were applied to the drug release data which showed that the drug through the wafers follows first order and fickian drug release mechanism. 100 6. Refrence 1. Raval, K.M., Overview on oral strip. Journal of drug discovery and therapeutics, 2013. 1(03). 2. Jangra, P.K., S. Sharma, and R. Bala oral , International journal of universal Pharmacy and Biosciences fast dissolving oral films , 2014.(3) 1,6 3. Patil, P. and S. Shrivastava, Fast dissolving oral films: An innovative drug delivery system. Structure, 2012. 20(70): p. 50-500. 4. Vibhooti, P. and K. Preeti, wafers technology a new approch to smart drug delivery system. . Indian Journal of Research in Pharmacy and Biotechnology, 2013. 1(3): p. 428. 5. Khatoon, N., N.R. Rao, and B.M. Reddy, Overview on fast dissolving oral films. Int J Chem Pharm Sci, 2013. 1(1): p. 63-75. 6. Bala, R., Parven p,Shushil khana and Redupa R , Orally dissolving strips: A new approach to oral drug delivery system. International journal of pharmaceutical investigation, 2013. 3(2): p. 67. 7. Peggs, K. and S. Mackinnon, Imatinib mesylate-the new gold standard for treatment of chronic myeloid leukemia. New England Journal of Medicine, 2003. 348(11): p. 10481048. 8. Jyotsana, M., B. Sagar, and D. Mahesh, fast dissolving fims of chlorpromazine www. ijrap. net.2009 9. Madan, J.,Avachat A,Bandode S and Dangi M, Formulation and evaluation of a bilayer floating drug delivery system of Nizatidine for nocturnal acid breakthrough. 2012. 10. Ghosal, K., S. Chakrabarty, and A. Nanda, Hydroxypropyl methylcellulose in drug delivery. Der pharmacia sinica, 2011. 2(2): p. 152-68. 11. Heinze, T. and K. Pfeiffer, Studies on the synthesis and characterization of carboxymethylcellulose. Die Angewandte Makromolekulare Chemie, 1999. 266(1): p. 3745. 12. Barba, C. Montane, D. Rinado, M and Ferrori, X. , Synthesis and characterization of carboxymethylcelluloses (CMC) from non-wood fibers I. Accessibility of cellulose fibers and CMC synthesis. Cellulose, 2002. 9(3-4): p. 319-326. 101 13. Bhagat, S., R. Gaikwad, and A. Warade, Guar Gum Products, Its Research & Uses in Chemical Industries. 14. Mishra, R., A. Banthia, and A. Majeed, Pectin based formulations for biomedical applications: a review. Asian Journal of Pharmaceutical and Clinical Research, 2012. 5(4): p. 1-7. 15. Olano-Martin, E. Willat, G. and Mikalson, D. Pectin and pectic-oligosaccharides induce apoptosis in in vitro human colonic adenocarcinoma cells. Anticancer research, 2002. 23(1A): p. 341-346. 16. Yamada, H., Contribution of pectins on health care. Progress in biotechnology, 1996. 14: p. 173-190. 17. Ms, A. and P.M. Amin, Oral Film Technology: Challenges and Future Scope for Pharmaceutical Industry. 18. Patel, D.M., Bhatt A., Floating granules of ranitidine hydrochloride-gelucire 43/01: formulation optimization using factorial design. AAPS PharmSciTech, 2007. 8(2): p. E25-E31. 19. Madan, T. and A. Kakkar, Preparation and characterization of ranitidine-HC1 crystals. Drug development and industrial pharmacy, 1994. 20(9): p. 1571-1588. 20. Crookes, D. Crystalline ranitidine hydrochloride and pharmaceutical composition containing it. in Chem Abstr. 1982. 21. Shen, J., D. Lee, and R. Mc Keag, Bioequivalence of two forms of ranitidine. New Zealand Pharmacy Oct, 1995: p. 24-25. 22. Bawazir, S., et al., Comparative bioavailability of two tablet formulations of ranitidine hydrochloride in healthy volunteers. International journal of clinical pharmacology and therapeutics, 1998. 36: p. 270-274. 23. Wu, V., T. Rades, and D. Saville, Stability of polymorphic forms of ranitidine hydrochloride. Die Pharmazie, 2000. 55(7): p. 508-512. 24. Stosik, A. Patil ,Savile D, Biowaiver monographs for immediate release solid oral dosage forms: Metoclopramide hydrochloride. Journal of pharmaceutical sciences, 2008. 97(9): p. 3700-3708. 102 25. Patel, R., Kumar K, Thakre, K. Vionade, M., Formulation and optimization of floating matrix tablet of ranitidine hydrochloride. International Journal Comprehensive Pharmacy, 2011. 2: p. 36-45. 26. Oates, J.A. Feldman. M,Burtan. M ., Histamine2-receptor antagonists: standard therapy for acid-peptic diseases. New England Journal of Medicine, 1990. 323(24): p. 16721680. 27. Basit, A.W. and L.F. Lacey, Colonic metabolism of ranitidine: implications for its delivery and absorption. International journal of pharmaceutics, 2001. 227(1): p. 157165. 28. Bogues, K.,Roberts C . Pharmacokinetics and bioavailability of ranitidine in humans. in British Journal of Pharmacology. 1981: . 29. Brunton, L.L., Goodman & Gilman's the pharmacological basis of therapeutics. Vol. 12. 2011: McGraw-Hill Medical New York. 30. Yadav, S., K. Kavita, and T. Tamizhamani, Formulation and evaluation of floating tablets of ranitidine hydrochloride using natural and synthetic polymers. Int J pharm Tech Res, 2010. 2(2): p. 1513-1519. 31. Chowdhury, M.E.H. and M. Pathan, Preparation and evaluation of floating matrix tablets of Ranitidine Hydrochloride. The Pharma Innovation, 2012. 1(7). 32. HCL, R., Journal of Drug Discovery and Therapeutics 1 (7) 2013, 42-45. Journal of drug discovery and therapeutics, 2013. 1(7): p. 42-45. 33. Brogden, R. Carmine, A. Avery GS, ., Ranitidine: a review of its pharmacology and therapeutic use in peptic ulcer disease and other allied diseases. Drugs, 1982. 24(4): p. 267-303. 34. Kiran Goutam, R.G., Ajay Sharma, Ajay Singh, Pooja Sharma, Shriya Formulation and evaluation of oral fast dissolving films of promethazine theoclate (25mg):, 2015. 5((8): p. 2536-2544. 35. Kalluri, J.K.Y., Fabrication and Assessment of fast Dissolving Buccal Films of Labetalol Hydrochloride for Hypertension. International Journal of Medicine and Pharmaceutical Research, 2014. 2(1): p. 462-467. 36. Yadav, P.N., P. Bhat, and S. Soni, Glibenclamide fabricated transdermal wafers for therapeutic sustained delivery systems. 2014. 103 37. Abd-Alhammid, S.N. and H.H. Saleeh, Formulation and Evaluation of Flurbiprofen Oral Film. Iraqi J Pharm Sci, 2014. 23(1): p. 53-59. 38. Sultana, F., M. Arafat, and S.I. Pathan, Preparation and evaluation of fast dissolving oral thin film of caffeine. Int. J. Pharm. Biol. Sci, 2013. 3: p. 153-161. 39. Boateng, J., J. Mani, and F. Kianfar, Improving drug loading of mucosal solvent cast films using a combination of hydrophilic polymers with amoxicillin and paracetamol as model drugs. BioMed research international, 2013. 40. Raju, P.N, Kumar. M,Ravishkanar k and Reddy. Ch ., Formulation and Evaluation of Fast Dissolving Films of Loratidine by Solvent Casting Method. The Pharma Innovation, 2013. 2(2). 41. Pallavi Patil.1, S.K.S., Fast Dissolving Oral Films: An Innovative DrugDelivery System. (2012):. 42. Pathan, A.M.,Mahhaduhri.A , Chandewar V and Bakade V ., 5. Development and in vitro evaluation of salbutamol sulphate mucoadhesive buccal patches. Int J Pharm Pharm Sci, 2011. 3(2): p. 39-44. 43. Alagusundaram, M.,Ramkanth. k,Cuhdna.C and Madu. C, ., Formulation and evaluation of mucoadhesive buccal films of ranitidine. International journal of pharmtech research, 2009. 1(3): p. 557-563. 44. Semalty, M., A. Semalty, and G. Kumar, Formulation and characterization of mucoadhesive buccal films of glipizide. Indian journal of pharmaceutical sciences, 2008. 70(1): p. 43. 45. Shaikh, R.P. Vinas. P ,Nadesno. K, Kumar. P, ., The application of a crosslinked pectin‐based wafer matrix for gradual buccal drug delivery. Journal of Biomedical Materials Research Part B: Applied Biomaterials, 2012. 100(4): p. 1029-1043. 46. Mura, P, Kosalec. I, Orlanedi S and Mario. J ., Amidated pectin-based wafers for econazole buccal delivery: Formulation optimization and antimicrobial efficacy estimation. Carbohydrate polymers, 2015. 121: p. 231-240. 47. Ng, S.-F. and N. Jumaat, Carboxymethyl cellulose wafers containing antimicrobials: a modern drug delivery system for wound infections. European Journal of Pharmaceutical Sciences, 2014. 51: p. 173-179. 104 48. Noelle H. O’Driscoll, O.L., K.H.M. T. P. Tim Cushnie, and D.K.M.A.J. Lamb, Production and Evaluation of an Antimicrobial Peptide-Containing Wafer Formulation for Topical Application. 2013. 49. Khairnar, A., Jain. D, Bavishkar. D, Jain. P, , Developmement of mucoadhesive buccal patch containing aceclofenac: in vitro evaluations. Int J Pharm Tech, 2009. 1: p. 978981. 50. Boateng, J.S., Mathews.KH,and Auffret.AD ., In vitro drug release studies of polymeric freeze-dried wafers and solvent-cast films using paracetamol as a model soluble drug. International journal of pharmaceutics, 2009. 378(1): p. 66-72. 51. Senthil, V, Rizwana. B, Venkatta. T, and Rahti v., ., Levocetirizine dihydrochloride and ambroxol hydrochloride oral soluble films: Design, optimization, and patient compliance study on healthy volunteers. International Journal of Health & Allied Sciences, 2013. 2(4): p. 246. 52. Labovitiadi, O., A.J. Lamb, and K.H. Matthews, Lyophilised wafers as vehicles for the topical release of chlorhexidine digluconate—Release kinetics and efficacy against Pseudomonas aeruginosa. International journal of pharmaceutics, 2012. 439(1): p. 157164. 53. Olga Labovitiadia, b., Andrew J. Lamba, Kerr H. Matthewsa, Lyophilised wafers as vehicles for the topical release of chlorhexidine digluconate—Release kinetics and efficacy against Pseudomonas aeruginosa Olga Labovitiadia,b, Andrew J. Lamba, Kerr H. Matthewsa. 2012) p. 157– 164. 54. Ganguly, I.,., Development of Fast Dissolving Sublingual Wafers of Promethazine Hydrochloride. Iranian Journal of Pharmaceutical Sciences, 2014. 10(1): p. 71-92. 55. Boateng, J. and D. Areago, Composite Sodium Alginate and Chitosan Based Wafers for Buccal Delivery of Macromolecules. Austin J Anal Pharm Chem, 2014. 1(5): p. 1022. 56. Cilurzo, F.,Bhurrati.S,Selmini. F, Nicotine fast dissolving films made of maltodextrins: a feasibility study. AAPS PharmSciTech, 2010. 11(4): p. 1511-1517. 57. Kunte, S. and P. Tandale, Fast dissolving strips: A novel approach for the delivery of verapamil. Journal of pharmacy and bioallied sciences, 2010. 2(4): p. 325. 105 58. Kianfar, F., Lyophilized wafers comprising carrageenan and pluronic acid for buccal drug delivery using model soluble and insoluble drugs. Colloids and Surfaces B: Biointerfaces, 2013. 103: p. 99-106. 59. Deepthi, K.S, Goutam.K,Sharma. A, and Khushal. S, , Formulation and Characterization of Atenolol Fast Dissolving Films. Indian Journal of Pharmaceutical Science & Research, 2012. 2(2): p. 58-62. 60. Kianfar, F., ., Formulation development of a carrageenan based delivery system for buccal drug delivery using ibuprofen as a model drug. Journal of Biomaterials and Nanobiotechnology, 2011. 2(05): p. 582. 61. Mishra, R. and A. Amin, Formulation development of taste-masked rapidly dissolving films of cetirizine hydrochloride. Pharmaceutical technology, 2009. 33(2): p. 48-56. 62. Shimoda, H., et al., Preparation of a fast dissolving oral thin film containing dexamethasone: a possible application to antiemesis during cancer chemotherapy. European Journal of Pharmaceutics and Biopharmaceutics, 2009. 73(3): p. 361-365. 63. Ferreira, S.C., Burns R.E,Brando. G,and Suza .A, e, Box-Behnken design: An alternative for the optimization of analytical methods. Analytica chimica acta, 2007. 597(2): p. 179186. 64. Ismail, A.M., ., Early Post Trabeculectomy Hypotony is a Risk Factor for Glaucoma Failure. Liver, 2012. 70: p. 73. 65. Sakhare, A.V., Effect of Glycerin as Plasticizer in Orodissloving Films of Losartan Potassium. 66. Fulzele, S., P. Satturwar, and A. Dorle, Polymerized rosin: novel film forming polymer for drug delivery. International journal of pharmaceutics, 2002. 249(1): p. 175-184. 67. Garsuch, V. and J. Breitkreutz, Novel analytical methods for the characterization of oral wafers. European Journal of Pharmaceutics and Biopharmaceutics, 2009. 73(1): p. 195201. 68. El-Nabarawi, M.,Rehab. A,Taley. s, ., Development and characterization of ketorolac tromethamine (KT) orobuccal films. Int. J. of Pharmacy and Pharm. Sci, 2012. 4(4): p. 186-193. 106 69. Chakraborty, P, Shurjet Dey,Versha. P, ., Design expert supported mathematical optimization and predictability study of buccoadhesive pharmaceutical wafers of loratadine. BioMed research international, 2013. . 70. Kusum Devi, V., ., Design and evaluation of matrix diffusion controlled transdermal patches of verapamil hydrochloride. Drug development and industrial pharmacy, 2003. 29(5): p. 495-503. 71. Semalty, M., Beshant. S, Shree. V , Development of mucoadhesive buccal films of glipizide. International journal of pharmaceutical sciences and nanotechnology, 2008. 1(2): p. 184-190. 72. Bansal, S., M. Bansal, and G. Garg, Formulation and evaluation of fast dissolving film of an antihypertensive drug. Int. J. of Pharmaceutical, Chemical and Biological Sciences, 2013. 3(4): p. 1097-1108. 73. Patel, A.R., D.S. Prajapati, and J.A. Raval, Fast dissolving films (FDFs) as a newer venture in fast dissolving dosage forms. International journal of drug development and research, 2010. 2(2): p. 232-246. 74. Sakuda, Y., .,Yushira.M,Sastu. M, Preparation and evaluation of medicinal carbon oral films. Chemical and Pharmaceutical Bulletin, 2010. 58(4): p. 454-457. 75. Chakraborty, P.,Vercha. P, Goash A, ., Mathematical optimization and characterisation of pharmaceutically developed novel buccoadhesive wafers for rapid bioactive delivery of Loratadine. Journal of Pharmaceutical Investigation, 2013. 43(2): p. 133-143. 76. VK Devi, S.S., GR Maria, PU Deepti. Drug Dev. Ind. Pharm, 2003, 29, 495–503. 77. Ayensu, I. and J. Boateng, Development and evaluation of lyophilized thiolated-chitosan wafers for buccal delivery of protein. Journal of Science and Technology (Ghana), 2012. 32(2): p. 46-55. 78. Boateng, J.S., Mathews K, Development and mechanical characterization of solvent-cast polymeric films as potential drug delivery systems to mucosal surfaces. Drug development and industrial pharmacy, 2009. 35(8): p. 986-996. 79. Matthews, K., Boeatng S.j,., Formulation, stability and thermal analysis of lyophilised wound healing wafers containing an insoluble MMP-3 inhibitor and a non-ionic surfactant. International journal of pharmaceutics, 2008. 356(1): p. 110-120. 107 80. Garsuch, V., Preparation and characterization of fast-dissolving oral films for pediatric use. 2009. 81. Patil, V., Kumar k sunail. V and shunil D., Formulation and evaluation of floating matrix tablet of locally acting h2-antagonist. International Journal of Pharmacy & Technology, 2010. 2(3): p. 528-540. 82. Kaza, R., Arvind. K, Gupta P and Chawla ., Design and Evaluation of Sustained release Floating tablets for the treatment of Gastric Ulcers. J Pharm Sci Res, 2009. 1(4): p. 8187. 83. Reddy, K.R., S. Mutalik, and S. Reddy, Once-daily sustained-release matrix tablets of nicorandil: formulation and in vitro evaluation. AAPS pharmscitech, 2003. 4(4): p. 480488. 84. V.F. Patel, N.M.P.a.P.G.Y., Studies on, f. floating matrix tablets international journal of pharmaceutical sciences 2012 .,(3) 2 108