0Development of Zingiber officinale in oral 2 dissolving films: Effect of polymers on in vitro, ^ in vivo parameters and clinical efficacy
^ Anwar S Daud, Nidhi P Sapkal1, Minal N Bonde
Zim Laboratories Ltd., MIDC, Kalmeshwar, Nagpur, 1Gurunanak College of Pharmacy, Nari, Nagpur, India
ral dissolving drug delivery system offers a solution for those patients having difficulty in swallowing tablets/capsules, etc. Zingiber officinale, has been used for medicinal purpose since antiquity to treat motion sickness, pregnancy, and cancer-chemotherapy-induced vomiting, mild stomach upset, cough, chronic bronchial problems, and low-grade infections of all kinds and anorexia condition. This work investigates the possibility of developing Zingiber officinale oral dissolving films allowing fast, reproducible dissolution in oral cavity; thus bypassing first pass metabolism. The oral dissolving films were prepared by solvent-casting method. Prepared films were evaluated for film-forming properties, physico-mechanical properties, palatability, microbial limit test, accelerated stability studies, and clinical efficacy test. The different polymers such as hydroxypropyl methylcellulose 5cps, maltodextrin, pullulan, and polyvinyl alcohol were explored individually and in combination with each other for the formation of film. Among all polymers, maltodextrin and HPMC 5 cps alone and in combination showed excellent film-forming properties as well as very good physico-mechanical properties. The films resulted into excellent palatability along with least disintegrating time. But at accelerated stability studies, only HPMC 5 cps was found to be stable when compared with other formulations. So, it was concluded that HPMC 5 cps is the best film forming as well as stable polymer with respect to Zingiber officinale oral dissolving film.
Key words: Film-forming polymers, oral thin films, solvent-casting method, Zingiber officinale
INTRODUCTION
From past one decade, there has been an enhanced demand for more patient-friendly and compliant dosage forms. As a result, the demand for developing new technologies has been increased greatly. Oral-dissolving films (ODF) are the latest development in this field. ODFs are the ultrathin films of postage stamp size with an active agent or active pharmaceutical ingredient and other excipients.[1] These dosage forms can rapidly disintegrate and/ or dissolve to release the medicament as soon as they come in contact with saliva thus obviating the need for water during administration, an attribute that makes them highly attractive for pediatric and geriatric patients.[2] ODFs provide accurate dosing in safe and efficacious format, without the need of measuring devices as is the case with liquid oral dosage forms.
Address for correspondence:
Mr. Anwar Siraj Daud, Zim Laboratories Ltd., B-21/22, MIDC,
Kalmeshwar, Nagpur, India. E-mail: anwar_daud@rediffmail.com
Ginger, rhizome of Zingiber officinale, has been used for medicinal purpose since antiquity. In particular, it has been an important plant for the traditional Chinese and Indian pharmacopoeia. Several components of ginger such as 6-gingerol, 6-shogaol and galanolactone have been reported to have anti-5HT3 activity.[3,4] Zingiber officinale is used to treat various types of "stomach problems," including motion sickness, morning sickness, upset stomach, gas, and diarrhea. It is also used for treatment of nausea caused by cancer treatment, nausea, and vomiting after surgery, as well as loss of appetite.[5-7] It is also used as a digestive aid for mild stomach upset, cough, chronic bronchial problems, and low-grade infections of all kinds and anorexic conditions. Currently Zingiber officinale is marketed as extract (Zinaxin), tincture (Bioforce Jan De Vries), capsule and oil for the oral use. But out of these,
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DOI: 10.4103/0973-8398.91995
liquid formulations have the problems such as inaccuracy in dosing, transportation, etc., while the capsules contain powders that contain powders which take time for wetting and, hence, dissolution is slower. Since the main use of ginger is in motion sickness and nausea and vomiting, so it is advisable to formulate it in oral thin films. This dosage form will allow convenient administration even for the patients who are traveling as water is not required for swallowing. During frequent vomiting, oral antiemetic agents are expelled out with the emesis, the sublingual absorption of the ginger through ODF will produce the action faster. There are no ODF of ginger in the market. Hence, there is a need to develop the same.
Development of herbal medicines in novel drug delivery systems is the need of hour. This approach is an interesting blend of therapeutic effectiveness of herbal medicines and advantages of novel drug delivery systems. This increases the patient compliance for the useful herbal drugs, which generally are free from side effects. At present, researchers are working toward developing novel drug delivery systems such as mouth-dissolving tablets, soft gelatin capsules, sustained and extended release formulations, mucoadhesive systems, transdermal dosage forms, microparticles, microcapsules, nanoparticles, implants, etc. of herbs, along with this, many oral retention formulations consisting of gels, pastes, and chewing gums have been developed from the last few years. But poor flowability, poor compressibility of plant extracts, requirement of more functional excipients, etc. poses formulation hurdles for the development of such novel formulations.[8] But as recent market studies indicate that among the different oral marketed dosage forms, ODFs are largely gaining acceptance as the delivery system of choice. Thus there is a great need to deliver this potentially useful herbal drug into the ODF.
The aim of present invention is to formulate and develop ODFs of hydro-alcoholic extract of Zingiber officinale. The ODFs were formulated using wide range of film-forming agents such as hydroxypropyl methylcellulose (HPMC), pullulan, maltodextrin, polyvinyl alcohol (PVA). The polymers
were used alone and in combination to obtain the desired film properties which were later evaluated for film forming properties, physico-mechanical properties, palatability, microbial limit test, accelerated stability studies, and clinical efficacy test.
HPMC is known for its good film-forming properties and has excellent acceptability. Maltodextrin is classified as a complex carbohydrate, but acts such as a simple carbohydrate in the body. It acts as film-forming agent, solubilizer, and imparts sweetness to the formulation. Pullulan is natural water-soluble polysaccharide, produced from starch by fermentation. It is an excellent film-former. For the fabrication of films, polyethylene glycol (PEG) 4000 was used as a plasticizer along with preservative, sweetener, emulsifier, flavoring agent.
MATERIALS AND METHODS Materials
Zingiber officinale extract was procured as a gift sample from Unijules Life Science Limited, Nagpur, India. Maltodextrin was received as generous gift from Gujrat Ambuja export Ltd., India. Pullulan was procured from Hayashibara International, Leatherhead, UK. HPMC 5 cps, tween 80, mentha oil and spearmint oil were procured from S. D. Fine Chemical Lab., A.B. Enterprises, Mumbai, India. All other chemicals and reagents were of analytical grades. Deionized double-distilled water was used throughout the study.
Preparation of Zingiber officinale ODFs
ODFs of Zingiber officinale extract were prepared using solvent casting method. The formulation codes and their respective compositions are given in Table 1. Zingiber officinale extract was mixed with measured amount of water using over headed stirrer for 5 min. The extract was then filtered through the muslin cloth. To this filtered extract of Zingiber officinale, successively measured amounts of polymer, tween 80, bronopol, sweetening and flavoring agents were added, and the solution was stirred for 30 min. The thick viscous solution was degassed to remove air entraps by ultrasonication.
Table 1: Composition of different formulation of ODFs of ginger
Ingredients F1 F2 F3 F4 F5 F6 F7
Zingiber officinale extract 10 10 10 10 10 10 10
HPMC 5cps 30 - - - 15 15 15
Maltodextrin - 30 - - 15 - -
Pullulan - - 30 - - 15 -
PVA - - - 30 - - 15
Polyethylene glycol 10 10 10 10 10 10 10
Polysorbate 80 5 5 5 5 5 5 5
Bronopol 0.1 0.1 0.1 0.1 0.1 0.1 0.1
Sucralose 3 3 3 3 3 3 3
Distilled water Q. S. Q. S. Q. S. Q. S. Q. S. Q. S. Q. S.
Note: quantities given in mg/film, HPMC: Hydroxypropyl methylcellulose, PVA: Polyvinyl alcohol
Measured quantity of solution was casted on a 30 X 45 cm2 glass plate and was kept in hot air oven at about 80°C for 15 min. The film was carefully removed from the glass plate, checked for any imperfections and cut to the required size to deliver the dose equivalent to 100 mg (2 X 3 cm2) per film. The films were stored in airtight plastic containers for further studies. The film samples were also stored for accelerated stability studies as per International Conference on Harmonisation (ICH) guidelines.
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 casted. The films were characterized as very poor, poor, average, good, better, best depending upon their ability to form films.191
Appearance offilms
Appearance of film was evaluated by visual observation. The films were characterized as transparent or translucent.!9"111
Tackiness
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.
Thickness
All the formulations were evaluated for uniformity in thickness by using calibrated digital vernier caliper. Ten films (pieces) from each formulation were taken randomly from different places of the plate. Thickness was measured and mean value was calculated. The uniformity in thickness is directly related to the accuracy of dose in the film.19,101
Folding endurance
The 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 crack in the film was noted as the value of folding endurance.!12,131
Surface pH study
The surface pH of fast dissolving strip was determined in order to investigate the possibility of any side effects in vivo. As an acidic or alkaline pH may cause irritation to the oral mucosa, it was determined to keep the surface pH to neutral as close as possible. A combined pH electrode was used for this purpose. Oral strip was slightly wet with the help of water. The pH was measured by bringing the electrode in contact with the surface of the oral film. The experiments were performed in triplicate, and average values were reported.
Disintegration test
Disintegration test was performed in the USP disintegration
Table 2: Key for evaluation of organoleptic study
Parameters Taste After taste Physical appearance
0 Not good Not bitter Not good
+ Good Slightly bitter Good
++ Very good Bitter Very good
+++ Excellent Very bitter Excellent
apparatus. 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 disintegration time of six films from each formulation was noted.
Organoleptic evaluation
Since the ODFs are intended to disintegrate rapidly in oral cavity, the product needs to have accepted organoleptic palatable characteristics. Organoleptic evaluation of prepared ODFs was carried out on panel of healthy volunteers with sound organoleptic senses, with their prior consents. The ODFs were rated on the basis of taste, mouth feel (grittiness or smoothness) and physical appearance. Table 2 gives the key for the organoleptic evaluation of ODFs.
Stability studies
For stability testing the ODFs were stored under controlled conditions of 40°C±2°C/75%±5% RH, 30°C±2°C/75%±5% RH over a period of 3 months according to the ICH guidelines. During storage the ODFs were checked for their physical appearance, tackiness, disintegration time, active content (6"gingerol) identification.
Analysis of stable formulation (by HPTLC) The formulation performing best in all the above characterization tests was analyzed to estimate the stability of active constituents of ginger extract upon storage. Films containing ginger extract equivalent to 5 g of raw herb from each formulation were picked randomly and weighed individually. Films were agitated in methanol for 45 min and then it was cooled, filtered, and dried on water bath. The dried residue was diluted with 5 ml methanol in volumetric flask. The reference standard was prepared with 10 mg reference standard of 6-Gingerol.
Chromatographic condition
Stationary phase HPTLC precoated, silica gel 60, F254 (Merck) Thickness 0.2 mm
Mobile phase Toluene: ethyl acetate:methanol:formic
acid (5:4:1:1) Scanning 500 nm
wavelength
Visualization aid Through UV-cabinet under 254 nm and
366 nm and under day light also. Spray reagent Anisaldehyde, sulphuric acid reagent
Microbiological limit test
The most stable formulation was subjected to microbiological examination as the herbal products are most susceptible for microbial contamination. This test was designed to determine total aerobic microbial, yeast and mould count including Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa, Salmonella, C. albicans, and A. niger. For ODFs of Zingiber officinale microbiological limit test was performed for the development of safe and effective consumer product. The microbiological limit test was performed according to Indian pharmacopoeia specification.
Clinical efficacy test
Evaluation of clinical effectiveness is a measure of the extent to which a particular formulation works. The most stable formulation was selected for clinical efficacy studies of Zingiber officinale ODFs in given dose of 100 mg of raw herb. These trials were conducted at Clinics of Ayurvedic Physicians with proper ethical consideration for the period of two weeks. All the subjects were completely informed concerning the pertinent details and the purpose of the study. A written consent form was supplied, understood, and signed by each subject prior to dispensing the ODFs of Zingiber officinale. The studies were related to six different
clinical conditions nausea, cough, anorexia, motion sickness, pregnancy-induced vomiting and travel sickness. The studied were conducted on five volunteers from each clinical condition. The effectiveness of Zingiber officinale ODFs, for all the conditions was rated as below:
0% - 25% 25% - 50% 50% - 75% 75% - 100%
Clinically ineffective Comparatively effective Clinically effective Most clinically effective
RESULTS AND DISCUSSION Preliminary Characteristics
Determination of film forming capacity, visual appearance and tackiness for all the formulations are shown in Table 3. Pullulan alone and in combination with HPMC exhibited poor film forming capacity. Pullulan otherwise is good film forming agent but film forming ability of formulation F3 was found to be poor. The reason may be the presence of Zingiber officinale extract that interferes in the linking of pullulan molecules and hence structure flexibility of the films is not achieved. The higher temperatures involved in drying may also cause changes in the mechanical properties of the films, making them brittle and less flexible films. Similarly F6 also did not result in good films. This indicated that in this ratio of HPMC and pullulan the films could not be formed. The reason can be incompatibility between two polymers or the interference of drug extract with the films. Rest of the formulations showed good film forming abilities. F4 also yielded films with average quality. It explained the nature of PVA when used alone as film forming agent. Visual appearance of all films was found to be transparent and free of air bubbles, which is necessary for aesthetic appeal.
Figure 1: Comparative study of folding endurance of ODFs of Zingiber officinale
Table 3: Evaluation of Film forming properties of different polymers with Zingiber officinale
Formulation Film forming Appearance of Tackiness
code capacity films
F1 Very good Transparent Non tacky
F2 Very good Transparent Non tacky
F3 Poor Transparent Slightly tacky
F4 Average Transparent Non tacky
F5 Very good Transparent Non tacky
F6 Poor Translucent Non tacky
F7 Very good Transparent Non tacky
All the formulations were found to be non tacky at ambient conditions except ODFs containing pullulan. This might be because of slightly hygroscopic nature of pullulan. So the formulations F3, F4, and F6 were not studied further as they failed at the first stage only.
Thickness
The thickness of all films was found to be in the range of 0.05-0.07 mm [Table 4]. The difference in the thickness of these formulations might be due to the different viscosities of the polymers which were used to formulate the films. The low standard deviation values for the thickness of these formulations were confirming efficiency of the method that was employed for formulation of films.
Table 4: Evaluation of physical properties of Zingiber officinale
Formulation code Film thickness (mm) Disintegration (s) Surface pH Folding endurance
F1 0.07±0.24 24±0.25 6.91±0.85 258±0.56
F2 0.05±0.28 8±0.47 7±0.25 298±0.25
F5 0.06±0.54 17±0.74 7.1±0.32 351±0.46
F7 0.07±0.42 62±0.37 6.83±0.64 193±0.84
Folding endurance
All the polymers were able to give the acceptable folding endurance values. Figure 1 shows the results for all the formulations, the observed folding endurance was in the order F5 > F6 > F2 > F1. Folding endurance was found to be highest for formulation F5 (351±0.46). Combination of maltodextrin and HPMC 5 cps was found to perform better the folding endurance was increased.[10]
Surface pH study
The surface pH values of the formulations are given in Table 4. All the polymers resulted in the formulations that have neutral surface pH. The surface pH of the strips was ranging from 7.1±0.32 to 6.83±0.64. The neutral values of surface pH of films assured that there will not be any kind of irritation to the mucosal lining of the oral cavity.
In vitro disintegration time
All the films were found to be rapidly disintegrating except F7 [Table 4, Figure 2]. The observed disintegration time was in the order F2 > F5 > F1 > F7 as show in Figure 2. PVA was found to increase the disintegration time of HPMC films. It must be because of the slow solubility of PVA in water that delayed solubility of HPMC. Quickly disintegrating character of maltodextrin was retained even in the presence of other ingredients of film thus allowing F2 to disintegrate rapidly (disintegration time 8±0.47 s). Presence of maltodextrin reduced the disintegration time of HPMC from 24 to 17 s.
Organoleptic evaluation
The results of the organoleptic characterization are shown
in Table 5 [key in Table 2]. All the formulations scored well in physical appearance but formulations F2 and F5 were found to be excellent in palatibity test. This suggested that maltodextrin acts both as film-forming agent and sweetener thus enhances palatability of formulation F2 and F5 [Table 5]. All the formulations also showed no bitter after taste, except F7. It suggested that appropriate amounts of flavors and sweeteners have already been incorporated in the films. The failure of F7 in after taste parameter might be due to the higher disintegration times required which ultimately reduces the palatability of prepared films. Formulation F7 was discarded at this stage and was not studied further.
Accelerated stability studies
The results of accelerated stability studies are given in Table 6. When selected Zingiber officinale ODFs formulations were stored at 40±2°C and 75%±5% RH, 30±2°C and 75%±5% RH changes were observed in formulation F2 and F5 in physical appearance, tackiness, separation of the films. The formulation F1 was found to be stable and subjected to the disintegration test, active content identification and for preservative efficacy test. The formulations F2 and F5 were found to be tacky and difficult to separate at accelerated stability conditions because of very hygroscopic nature of maltodextrin at relative humidities greater than 50%. Thus further studies were carried out only on F1.
Analysis of F1 (by HPTLC)
The active constituent of Zingiber officinale, i.e., 6-gingerol was assessed by comparing the spectra at peak start, peak apex, and peak end positions of the spot. The spectrum of
Table 5: Organoleptic characterization of selected formulations
Formulation Taste Bitter Physical
code aftertaste appearance
F1 ++ 0 +++
F2 +++ 0 +++
F5 +++ 0 +++
F7 0 + +++
Figure 2: Comparative study of disintegration time of ODFs of Zingiber officinale
Table 6: Stability studies of selected formulations
Formulation Physical Tackiness Film Disintegration Active content Preservative
code appearance separation time (s) identification efficacy test
40±2°C, 30±2°C, 40±2°C, 30±2°C,
75%±5% 75%±5% 75%±5% 75%±5%
RH RH RH RH
F1 Very good Non tacky Separates 25±0.35 24±0.96 Present of 6-Gingerol Present of 6-Gingerol Pass as per IP
F2 Good Tacky Difficult to separate NA NA NA NA NA
F5 Good Tacky Difficult to separate NA NA NA NA NA
F1 could be completely overlaid on the spectrum of Zingiber officinale extract [Figure 3]. According to the method when
Figure 3: Overlay spectral display of ODFs (green and yellow color) along with the reference standard (pink color) of Zingiber officinale after scanning under 200 nm-700 nm
Table 7: Identification of bands of ginger raw herb and F1 under daylight
Identification of band Rf F1 Ginger extract
Green spot 0.06 + +
Yellow spot 0.66 + -
6-Gingerols 0.69 + +
Violet spot 0.72 + +
Violet spot 0.81 + +
Violet spot 0.87 + +
Table 8: Identification of bands of ginger raw herb and
F1 at 366 nm
Color Rf F1 Ginger extract
Brown spot 0.06 + +
Light orange spot 0.66 + +
Dark orange spot 0.70 + +
Orange Spot 0.82 + +
Orange spot 0.86 + +
TLC plate was scanned at day light and 366 nm, six spots (Rf values 0.06, 0.66, 0.69, 0.72, 0.81, 0.87) [Table 7] and five spots (Rf value 0.06, 0.66, 0.70, 0.83, 0.86) [Table 8] were observed respectively. The spots obtained in the F1 were compared with those obtained in standard Zingiber officinale extract.
Microbiological limit test
Microbial limit test was performed on the most stable formulation F1. The results of microbial limit test are given in Table 9. From the results obtained, it was concluded that formulation F1 was free from microbial contamination.
Clinical efficacy studies
The studies were related to six different clinical conditions that are nausea, pregnancy-induced vomiting, travel sickness, motion sickness, cough, anorexia. The studies were conducted on the five human volunteers from each clinical condition. The results obtained are given in Table 10. From the clinical efficacy studies, it was found that ODFs of Zingiber officinale are the most promising herbal remedy for these clinical conditions.
CONCLUSION
Among the prepared formulations, formulation F1 was found to have transparent visual appearance, best film-forming capacity, least disintegration time and also found to be stable at accelerated stability studies. In vitro and in vivo evaluation of the films confirmed their potential as an innovative dosage form to deliver Zingiber officinale. Thus, Zingiber officinale ODFs are found to be suitable especially for geriatric, bedridden, and non-cooperative patients due to its ease of administration as well as patient friendly dosage form.
Table 9: Results of microbiological limit tests
Test Limits Results
1 month 2 months 3 months
Total microbial plate count NMT 105 cfu/g of sample 185 cfu/g 186 cfu/g 185 cfu/g
Total yeast and mould NMT 103 cfu/g of sample 19 cfu/g 23 cfu/g 20 cfu/g
Specified Micro-organism
Escherichia coli Absent Absent Absent Absent
Salmonella Absent Absent Absent Absent
Pseudomonas aeruginosa Absent Absent Absent Absent
Staphylococcus aureus Absent Absent Absent Absent
Note: cfu/g (colony forming unit/gram), NMT: not more than
Table 10: Clinical efficacy studies Zingiber officinale ODFs
Formulation F1 Nausea Pregnancy-induced Travel Motion Cough Anorexia
vomiting sickness sickness
Average clinical efficacy 84.2±2.5 85.8±7.2 88.4±3.3 87.6±5.8 87.8±9.6 74.8±10.3
Dosage form acceptability Yes Yes Yes Yes Yes Yes
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How to cite this article: Daud AS, Sapkal NP, Bonde MN. Development of Zingiber officinale in oral dissolving films: Effect of polymers on in vitro, in vivo parameters and clinical efficacy. Asian J Pharm 2011;5:183-9. Source of Support: Nil. Conflict of Interest: None declared.
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