To study the effect of solvent, viscosity, and temperature on the...

[Full title: To study the effect of solvent, viscosity, and temperature on the mouth-dissolving film of Withania somnifera Linn]

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Asian Journal of Pharmaceutics - July-September 2012 212 INTRODUCTION Herbal medicine is the oldest form of health care known to mankind. Herbs have been used by all cultures throughout history. They have been an integral part in the development of the modern civilization. Many drugs commonly used today are of herbal origin. Indeed, about 25% of the prescription drugs dispensed in the United States contain at least one active ingredient derived from plant material. The World Health organization (WHO) reports that 80% of the world’s population relies on drugs of natural origin. Less toxicity, better therapeutic effect, and cost effectiveness are the reasons for choosing drugs of natural origin. But all the formulations of these compounds are either available in liquid forms like arishtas and asavs or the solid forms like powders that have poor palatability and hence poor patient compliance. There is a need to formulate these compounds into novel drug delivery systems so that the therapeutic effectiveness of these compounds can be clubbed with the novel advantages of these delivery systems W. somnifera (WS), also known as Ashwagandha, Indian ginseng, or winter cherry, has been an important herb in the Ayurvedic and indigenous medical systems for over 3,000 years. The roots of the plant are categorized as Rasayanas , and are described to promote health and longevity by augmenting defenses against disease, arresting the aging process, revitalizing the body in To study the effect of solvent, viscosity, and temperature on the mouth-dissolving film of Withania somnifera Linn Anwar Daud, Minal Bonde 1 , Nidhi Sapkal 2 , Naresh Gaikwad 3 Department of Pharmaceutical Science, NIMS University, Jaipur, 1 Department of Research and Development, Zim Laboratories Ltd, 2 Department of Pharmaceutical Chemistry, Gurunanak College of Pharmacy, Nari, 3 Department of Pharmaceutical Science, RTM Nagpur University, Nagpur, India A n oral-dissolving drug delivery system offers a solution for those patients having difficulty in swallowing tablets/ capsules, and so on. Withania somnifera has been used to promote health and longevity by augmenting defenses against disease, arresting the aging process, revitalizing the body in debilitated conditions, and thus creating a sense of well-being. Commercially, it is available either in powder or liquid dosage forms that do not offer patient compliance. In the present study, an attempt has been made to formulate W. somnifera into thin oral films. An attempt was also made to study factors like the effect of the type of solvent used for casting of the film, effect of drying temperature, and viscosity of the solution on the mouth-dissolving film of W. somnifera Linn. The mouth-dissolving films were prepared by the solvent-casting method. Prepared films were evaluated for film-forming capacity, appearance of film, tack test, thickness, in vitro disintegration time, folding endurance, tensile strength, and percentage elongation. This study illustrated that selected process variables have an influence on the physicomechanical properties of the mouth-dissolving film of W. somnifera Linn. Water was found to be an excellent solvent for casting when hydroxypropylmethylcellulose was used as a film former. When mouth-dissolving film of W. somnifera Linn was dried at lower temperatures, it showed optimum physicomechanical performances. Viscosity of the solution plays an important role in physicomechanical properties of the film; as the viscosity of solution increases, there is an increase in folding endurance, tensile strength, and percentage elongation Keywords: Drying temperature, nature of solvent, process variables, viscosity of solvent, Withania somnifera Linn ORIGINAL ARTICLE Address for correspondence: Mr. Anwar S. Daud, Zim Laboratories Ltd, B-21/22, MIDC Area, Kalmeshwar, Nagpur - 441 501, India. E-mail: md@zimlab.in Access this article online Quick Response Code: Website: www.asiapharmaceutics.info DOI: 10.4103/0973-8398.104838

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Asian Journal of Pharmaceutics - July-September 2012 213 Duad, et al .: Mouth-dissolving film of Withania somnifera Linn debilitated conditions, and thus creating a sense of wellbeing. [1] W. somnifera contains alkaloids (withanine, withasomnin) and steroidal lactones and glycosides also called as withanoloids and sitoindosides, and the extract of W. somnifera has analgesic, mildly sedative, anti-inflammatory, and anabolic activities, [2] and it is useful in stress, strain, fatigue, pain, skin diseases, diabetes, gastrointestinal disease, rheumatoid arthritis, and epilepsy, [3] chronic fatigue syndrome, [4] and even during pregnancy without any side effects [5] It is also used as a general tonic to increase energy and improve health and longevity [6] Clinical studies of W. somnifera suggest that it may promote growth in children, and improve hemoglobin level, red blood cell count, and physical performance in adults [7] The mouth-dissolving film belongs to novel drug delivery systems that offer unique advantages like better patient compliance, lesser excipient load, and faster onset of action. These films can be prepared either by hot melt extrusion method or solvent-casting technique. In this study, the solvent-casting method was used The main objective of the present research work was to study three process variables, namely, nature of the solvent used for casting of the films, effect of drying temperature, and viscosity of the solution [Figure 1]. These process parameters may affect physicomechanical properties of films. The effects of these parameters were studied on film-forming capacity, appearance of film, tack test, thickness, in vitro disintegration time, folding endurance, tensile strength, and percentage elongation MATERIALS AND METHODS Materials W. somnifera Linn extract was obtained from Unijules Life Science Limited, Nagpur, India. Hydroxypropylmethylcellulose (HPMC) 5 cps (cps: centipoise) (Shin-Etsu Chemical Co. Ltd, China), polyethylene glycol (PEG) 6000 (IBIS Chemie International, Mumbai), polysorbate 80 (Vinamax organics Pvt. Ltd., Mumbai), bronopol (A. B. ENTERPRISES, Mumbai), and aspartame (Aarti Pharma, Mumbai) were used as film base materials. All other chemicals and reagents were of analytical grades. Deionized double-distilled water was used throughout the study Methods The effect of the nature of the solvent on the physicochemical properties of mouth-dissolving films of W. somnifera Linn was studied using two solvents and their combinations. The formulations were prepared according to the formula given in Table 1. The detail film forming process including variables studies was explained in the Schematic diagram as shown in Figure 1. The best formulation was selected to study the effect of viscosity of the casting solution on the properties of the film. These formulations were prepared according to the formula given in Table 2. The formulation showing desirable properties was selected for studying the effect of drying temperature on the physicomechanical properties Table 1: Composition to study the effect of the solvent on mouth-dissolving films of Withania somnifera Linn Ingredients Percent composition F 1 F 2 F 3 F 4 F 5 Withania somnifera Linn extract 4 4 4 4 4 HPMC 5 cps 15 15 15 15 15 PEG 6000 3.6 3.6 3.6 3.6 3.6 Polysorbate 80 1.2 1.2 1.2 1.2 1.2 Bronopol 0.01 0.01 0.01 0.01 0.01 Aspartame 1.2 1.2 1.2 1.2 1.2 Distilled water 75 – – – – Isopropyl alcohol – 75 – – – Distilled water: Isopropyl alcohol – – 25: 50 – – Distilled water: Isopropyl alcohol – – – 37.5: 37.5 – Distilled water: Isopropyl alcohol – – – – 50: 25 PEG: Polyethylene glycol, HPMC: Hydroxypropylmethylcellulose Table 3: To study the effect of drying temperature on mouth-dissolving films of Withania somnifera Linn Formulation code Drying temperature (°C) F 1 bi 60–65 F 1 bii 80–85 F 1 biii 100–105 F 1 biv 120–125 Table 2: To study the effect of viscosity on mouthdissolving films of Withania somnifera Linn Formulation code Viscosities of solution (in cps) F 1 a 6500–7000 F 1 b 8500–9000 F 1 c 10500–11000 F 1 d 12000–12500 of films. The experiment was carried out according to the conditions given in Table 3 Evaluation methods Preliminary characteristics Film-forming capacity: It is the ability of a polymer to form films that can be separated from the surface on which they are cast The films were characterized as very poor, poor, average, good, better, and best depending upon their ability to form films Appearance of films: Appearance of film was evaluated by visual observation. The films were characterized as smooth or rough Tack test: Upon stacking, the films should not stick to each other. This is a criterion which a film should possess for better dispensing of dosage form [6,7]

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Asian Journal of Pharmaceutics - July-September 2012 214 Duad, et al .: Mouth-dissolving film of Withania somnifera Linn Figure 1: Schematic diagram showing film-forming process including variable studies (solvent-casting method); evaluation of films Thickness All the formulations were evaluated for uniformity in thickness by using calibrated digital electronic digital micrometer (IP 65, Mitutoyo Co., Japan). Ten films (pieces) were taken randomly from each formulation from different places of the plate. Thickness was measured and mean value was calculated [8] In vitro disintegration test Disintegration test was performed in the united state pharmacopoeia (USP) disintegration apparatus (Electrolab, Mumbai). Simulated salivary fluid (pH 6.8) was used as the medium. The films were placed in the tubes of the container and the discs were placed over it. The average in vitro disintegration time of six films from each formulation was noted [9] Folding endurance Folding endurance was measured manually for the prepared films. A film was cut and firmly folded through the middle. The number of folds on the same crease required to produce a crack in the film was noted as the value of folding endurance [10] Tensile strength Tensile strength is the maximum stress applied to a point at which the strip specimen breaks. It is calculated by the

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Asian Journal of Pharmaceutics - July-September 2012 215 Duad, et al .: Mouth-dissolving film of Withania somnifera Linn applied load at rupture divided by the cross-sectional area of the strip as given in the equation below: Tensile strength load at failure 100 Strip thickness strip = × × width (1) Percent elongation When stress is applied, a strip sample stretches and this is referred to as strain. Strain is basically the deformation of the strip divided by the original dimension of the sample. Percentage elongation was calculated by the equation as given in the equation below: [11] Percentage elongation Increase in length 100 Original le = × nngth (2) RESULTS AND DISCUSSION Determination of film-forming capacity, visual appearance, and tackiness for all the formulations are shown in Table 4. From the physical appearance of the film it was noted that all formulations showed rough appearance except the formulation F 1, which appeared to be smooth and translucent. The formulation F 1 was also found to be thin and flexible. It was observed that isopropyl alcohol (IPA) alone and in combination with distilled water exhibited poor film-forming capacity and rough surface. HPMC 5 cps is otherwise a good film-forming agent but the film-forming ability of formulations F 2, F 3, F 4, and F 5 was found to be poor; this might be because of the insolubility of HPMC 5 cps in IPA. All the films were found to be nontacky to each other The thickness of the formulations varied from 0.07±0.002 to 0.08±0.002 mm. The difference in the thickness of these formulations might be because of the different solvents used for casting of the film. When water was used as a solvent, thickness was found to be at a minimum of 0.07±0.002 mm and with increase in the amount of IPA, the thickness of film was found to increase. The low standard deviation values for the thickness of these formulations confirmed the efficiency of the method that was employed for formulation of the films All the films were found to be rapidly disintegrating within 60 seconds [Table 6]. The observed disintegration time was in the order F 2>F 3>F 4>F 5>F 1. It was observed that the in vitro disintegration time was directly proportional to the thickness of the film and nature of the solvent From the results of mechanical properties, as given in Table 4, it was observed that formulation F 1 was found to have acceptable mechanical properties as compared with formulations F 2, F 3, F 4, and F 5. This may be because of the HPMC 5 cps which got completely dissolved in the water and showed hard and tough characteristics, but in the presence of IPA, alone or in combination, it showed soft and weak characteristics. A suitable mouth-dissolving film requires moderate folding endurance, tensile strength, and percentage elongation. So the formulations F 2, F 3, F 4, and F 5 were not studied further as they failed at the first stage itself From the data given in Table 5, it is observed that all the formulations got separated from the surface and were found to be nontacky. From the visual appearance of the surface of the film, it was noted that formulations F 1 a, F 1 b, and F 1 c were found to have a smooth surface and formulation F 1 d had a rough surface; this might be because of the low solubility of HPMC 5 cps in water which imparted roughness to the film The thickness of all formulations was found to be in the range of 0.063±0.025 mm to 0.09±0.002 mm [Table 5]. The difference in the thickness of these formulations might be due to the different viscosities of the solution which were used to formulate the films. The low standard deviation values for the thickness of these formulations confirmed the efficiency of the method that was employed for formulation of the films In vitro disintegration test was performed for all the formulations and they were found to be disintegrated within 60 seconds. The best disintegrating time was observed for formulations F 1 a and F 1 b, whereas formulation F 1 d was ranked last, where disintegration time was concerned. Therefore, from the results obtained it might be concluded that formulations having a higher viscosity and thickness, that si, F 1 c and F 1 d had a comparatively high disintegration time as given in Table 5 Films of size 3×10 mm 2 were taken for the study. From the results, it was clear that when the viscosity increased, Table 4: Evaluation of mouth-dissolving films of Withania somnifera Linn to study the effect of solvent Properties of films F 1 F 2 F 3 F 4 F 5 Film-forming capacity Very good Poor Average Average Average Appearance of film Smooth, translucent Rough Rough Rough Rough Tack test Nontacky Nontacky Nontacky Nontacky Nontacky Thickness (mm) 0.07±0.002 0.08±0.002 0.075±.002 0.072±0.002 0.07±0.002 Disintegration time (sec) 18±0.43 56±0.29 42±0.83 27±0.14 25±0.71 Folding endurance 20.12±0.41 2.78±0.49 7.23±0.84 10.87±0.91 15.54±0.37 Tensile strength (N/mm 2 ) 18.72±0.93 1.98±0.43 4.84±0.29 6.26±0.47 10.37±0.86 Percentage elongation (%) 85±0.41 12±0.75 16±0.07 37±0.20 54±0.92

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Asian Journal of Pharmaceutics - July-September 2012 216 Duad, et al .: Mouth-dissolving film of Withania somnifera Linn the tensile strength of film also increased. Formulation F 1 c showed the maximum folding endurance, tensile strength, and percentage elongation. Table 5 shows the results for all the batches. Formulation F 1 a, with lower viscosity, was found to have folding endurance of 8±0.45 and as the viscosity of solution was increased, the folding endurance was improved to a certain extent as shown in formulations F 1 a, F 1 b, and F 1 c, and it again decreased in the later formulation F 1 d. Similar observations were noted for tensile strength; in formulations F 1 a, F 1 b, and F 1 c, tensile strength was found to be 8.28±0.49, 12.63±0.27, and 16.98±0.46, respectively, and for formulation F 1 d it was 13.09±0.46 N/mm 2 . The percent elongation for all formulations was found to be in the range of 23.55±0.40% to 45.12±0.26%. The percentage elongation of the films increased linearly with increase in viscosity of the solution used for casting of the film As the formulation F 1 b had adequate physical properties, moderate folding endurance, tensile strength, and percentage elongation with minimum in vitro disintegration, it was selected for further studies of drying temperature From Table 6, it is noted that drying temperature was directly proportional to drying time Determination of film-forming capacity, visual appearance, and tackiness for all the formulations are shown in Table 6. It was observed that all the formulations were found to be nontacky to each other, except formulation F 1 bi. This might be because of the presence of moisture in it which may not completely evaporate when film-dried at 60–65°C The thickness of the formulation was 0.07±0.002 mm. The low values for standard deviation indicate physical uniformity of the membranes All the films were found to be rapidly disintegrating within a span of 18 to 20 seconds [Table 6]. The observed in vitro disintegration time was in the order of F 1 bii >F 1 biii = F 1 biv>F 1 bi. It was observed that the drying temperature did not have much influence on in vitro disintegration time of the film From the results of folding method, tensile strength, and percentage elongation given in Table 6, it was found that increase in temperature was inversely proportional to folding endurance, tensile strength, and percentage elongation. Therefore, the film dried at 60–65°C had excellent mechanical properties with folding endurance (56±0.68), tensile strength (20.21±0.37 N/mm 2 ), percentage elongation (83.08±0.28%), and in vitro disintegration (18±0.35 sec) as compared with all other formulations. But the time required to dry the film was more, that is, 60 minutes and the film was found to be slightly tacky when stacked upon each other. Therefore, the formulation F 1 bii was found to be the most acceptable having comparatively less drying time (40 min), with desired physical properties having folding endurance (49±0.25), tensile strength (18.74±0.48 N/mm 2 ), percentage elongation (72.08±0.29%), and in vitro disintegration time (20±0.84 sec) CONCLUSION The main objective of this research work was to study the effect of the nature of the solvent used for casting of the film, effect of drying temperature, and viscosity of the solution on mouth-dissolving film of W. somnifera Linn. From the Table 6: Evaluation of mouth-dissolving films of Withania somnifera Linn when dried at different temperatures Formulation code F 1 bi F 1 bii F 1 biii F 1 biv Drying time (min) 60 40 30 15 Separation from the surface Good Very good Very good Very good Appearance of film Smooth film Smooth film Smooth film Smooth film Tack test Tacky Nontacky Nontacky Nontacky Thickness (mm) 0.07±0.002 0.07±0.002 0.07±0.002 0.07±0.002 Disintegration time (sec) 18±0.35 20±0.84 19±0.38 19±0.15 Folding endurance 56±0.68 49±0.25 27±0.89 20±0.24 Tensile strength (N/mm 2 ) 20.21±0.37 18.74±0.48 10.00±0.23 5.84±0.28 Percentage elongation (%) 83.08±0.28 72.08±0.29 59.78±0.12 42.76±0.49 Table 5: Evaluation of mouth-dissolving films of Withania somnifera Linn at different viscosities Formulation code F 1 a F 1 b F 1 c F 1 d Film-forming capacity Very good Very good Very good Very good Appearance of film Smooth Smooth Smooth Rough Tack test Nontacky Nontacky Nontacky Nontacky Thickness (mm) 0.063±0.025 0.07±0.0002 0.082±0.0002 0.09±0.002 Disintegration time (sec) 18±0.3 18±0.28 24±0.37 30±0.48 Folding endurance 8±0.45 15±0.36 24±0.54 19±0.43 Tensile strength (N/mm 2 ) 8.28±0.49 12.63±0.27 16.98±0.98 13.09±0.46 Percentage elongation (%) 23.55±0.40 30.87±0.81 37.87±0.29 45.12±0.26

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Asian Journal of Pharmaceutics - July-September 2012 217 Duad, et al .: Mouth-dissolving film of Withania somnifera Linn physicomechanical and in vitro dissolution data obtained, it was confirmed that selected process variables have a potential influence on these properties. When HPMC 5 cps was used as a film former, water was found to be an excellent solvent for casting of the solution. From the present study, it was also observed that 8,500–9,000 cps of viscosity was required to formulate the best-quality film with minimum variations in thickness. When mouth-dissolving film of W. somnifera Linn was dried at 80–85°C, it showed the most favorable physicomechanical proprties REFERENCES 1. Weiner MA, Weiner J. Herbs that Heal. Ashwagandha (Indian ginseng). Mill Valley, CA: Quantum Books; 1994. p. 70-2 2. Mishra LC, Singh BB, Dagenais S. Scientific basis for the therapeutic use of Withania somnifera (ashwagandha): A review. Altern Med Rev 2000;5:334-46 3. Prakash J, Gupta SK, Dinda AK. Withania somnifera root extract prevents DMBA-induced squamous cell carcinoma of skin in Swiss albino mice. Nutr Cancer 2002;42:91-7 4. Singh A, Naidu PS, Gupta S, Kulkarni SK. Effect of natural and synthetic antioxidants in a mouse model of chronic fatigue syndrome. J Med Food 2002;5:211-20 5. Sharma S, Dahanukar S, Karandikar SM. Effects of long-term administration of the roots of ashwagandha and shatavari in rats. Indian Drugs 1985;29:1339 6. Sward G. Drying time. In: Sward G, editor. Paint Testing Manualphysical and chemical examination of paints varnishes lacquers, and colors. 13 th ed. West Conshohocken, Pennsylvania: American Society for Testing and Materials; 1972. p. 268 7. Felton L, O’Donnell P, McGinity J. In: McGinity J, Felton L, editors. Mechanical properties of polymeric films prepared from aqueous dispersions. In: Aqueous polymeric coatings for pharmaceutcal dosage forms. 3 rd ed. Vol. 176. Drugs and the pharmaceutical sciences. Informahealthcare, New York, USA. 2008. p. 108 8. Fulzele SV, Sattuwar PM, Dorle AK. Polymerized rosin: Novel film forming polymer for drug delivery. Int J Pharm 2002;249:175-84 9. American Standard of Testing and Materials, ASTM D 1004 - 08 Standard Test Method for Tear Resistance (Graves Tear) of Plastic Film and Sheeting. ASTM, International West Conshohocken, PA, USA, 2000 10. Shinde AJ, Garala KC, More HN. Development and characterization of transdermal therapeutics system of tramadol hydrochloride. Asian J Pharm 2008;2:265-9 11. Nishimura M, Matsuura K, Tsukioka T, Yamashita H, Inagaki N, Sugiyama T, et al . In vitro and in vivo characteristics of prochlorperazine oral disintegrating film. Int J Pharm 2009;368:98-102 How to cite this article: Daud A, Bonde M, Sapkal N, Gaikwad N. To study the effect of solvent, viscosity, and temperature on the mouth-dissolving film of Withania somnifera Linn. Asian J Pharm 2012;6:212-7 Source of Support: Nil. Conflict of Interest: None declared Author Help: Reference checking facility The manuscript system (www.journalonweb.com) allows the authors to check and verify the accuracy and style of references. The tool checks the references with PubMed as per a predefined style. Authors are encouraged to use this facility, before submitting articles to the journal • The style as well as bibliographic elements should be 100% accurate, to help get the references verified from the system. Even a single spelling error or addition of issue number/month of publication will lead to an error when verifying the reference. • Example of a correct style Sheahan P, O’leary G, Lee G, Fitzgibbon J. Cystic cervical metastases: Incidence and diagnosis using fine needle aspiration biopsy. Otolaryngol Head Neck Surg 2002;127:294-8. • Only the references from journals indexed in PubMed will be checked. • Enter each reference in new line, without a serial number • Add up to a maximum of 15 references at a time • If the reference is correct for its bibliographic elements and punctuations, it will be shown as CORRECT and a link to the correct article in PubMed will be given • If any of the bibliographic elements are missing, incorrect or extra (such as issue number), it will be shown as INCORRECT and link to possible articles in PubMed will be given.

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