Formulation and Evaluation of Pomegranate Peel Extract Buccal Film for Pediatric Dyslipidemia.

S GOPI; CLEVIA MIRANDA F; T BHAVYASHREE; R SHABARAYA A; M S ANUSHA

doi:10.5281/zenodo.18309072

Research Paper | Open Access
Volume 04 | Issue 01 | Article Id IJPS/260401124

Formulation and Evaluation of Pomegranate Peel Extract Buccal Film for Pediatric Dyslipidemia.
S GOPI* CLEVIA MIRANDA F T BHAVYASHREE R SHABARAYA A M S ANUSHA
Department of Pharmaceutics, Srinivas college of pharmacy, Mangalore, Karnataka, 574143

Abstract

Dyslipidemia in children is becoming increasingly common, largely due to the global rise in childhood obesity. While pharmacological treatments are available, their potential impact on the growth and development of children has raised safety concerns. This has prompted the exploration of safer, plant-based alternatives. The current study focuses on the formulation and evaluation of a mucoadhesive buccal film containing pomegranate peel extract, designed specifically for children with dyslipidaemia. Pomegranate peel was collected, dried, and subjected to ethanolic extraction using a Soxhlet apparatus. The extract was then evaluated for phytochemical constituents, including total phenolic and flavonoid contents. The active extract was incorporated into buccal films using the solvent casting method. The formulated buccal films were subjected to comprehensive physicochemical and mechanical evaluation, including: Surface pH, Thickness, Folding endurance, Tack test.

Keywords

Dyslipidemia, Pediatric, Pomegranate Peel Extract, Mucoadhesive Buccal Film, Solvent Casting, Phytochemicals, Bioavailability, Insoluble Drugs, Solid dispersions, rapid disintegration

Introduction

Atherosclerotic cardiovascular disease remains the leading cause of death and a major cause of morbidity. Although clinical outcomes rarely occur in youth, mounting evidence suggests that atherosclerotic lesions can begin in childhood. Childhood dyslipidaemia is one of the main traditional cardiovascular risk factors that initiate and exacerbate the atherosclerotic process.¹ Dyslipidaemia causes a wide spectrum of lipoprotein abnormalities leading to an elevated total cholesterol (TC), low-density lipoprotein-cholesterol(LDL-C),non-high-density lipoprotein cholesterol (HDLC), triglycerides (TG) or decreased HDL-C levels.² Dyslipidaemia has emerged as an increasingly prevalent risk factor in children, concomitant with the worldwide epidemic of obesity. Dyslipidaemia can alter vascular endothelial function and impair some of its profibrinolytic and antithrombotic regulatory properties, as well as initiate the atherosclerotic process.³ Dyslipidaemia, when recognized and treated in childhood, can reduce the risk of premature adverse cardiovascular events and mortality. ⁴ The currently available drugs show certain kind of effects on the growth, maturation and pubertal development of the children. Hence the need of the hour is to pay attention to natural or herbal remedies. Pomegranate (Punica granatum) peel is rich in many compounds such as phenolics, flavonoids, ellagitannins, proanthocyanidin compounds and many minerals including potassium, nitrogen, sodium etc.⁵Polyphenolic compounds such as punicalagin, punicalin, gallic acid and ellagic acid have shown beneficial effects on serum lipid levels and macrophage atherogenic properties.⁶ It has also been found that the pomegranate peel extract has antioxidant activity beside the lipid lowering effects.⁷ The dosage forms that are available in the market are mostly in the form of tablets which is difficult to administer to children. Hence buccal drug delivery can be used to obtain better compliance. The term "buccal drug delivery" pertains to the administration of pharmaceutical substances to the buccal mucosa, situated on the inner side of the cheek within the oral cavity. This route offers the advantage of facilitating both local and systemic drug delivery. By bypassing first-pass metabolism and enzymatic drug degradation, buccal drug delivery enhances the effectiveness of therapy. Moreover, this method proves particularly beneficial for patient cohorts experiencing challenges with swallowing or those who are unable to swallow altogether.⁸ Hence the present study aims to develop a mucoadhesive buccal film containing pomegranate peel extract for treating dyslipidaemia in children.

Objectives of the Study

The primary objective of this study was to formulate and evaluate a novel, safe, and effective buccal film containing pomegranate peel extract rich in phenolics, flavonoids, and polyphenols for the potential management of dyslipidaemia in children.

To collect, identify, and authenticate pomegranate peel.
To extract active constituents from pomegranate peel using the Soxhlet extraction method.
To formulate a buccal film incorporating pomegranate peel extract.
To evaluate the formulated buccal film for anti-lipidemic activity and other relevant parameters.

METHODOLOGY:

Collection of plant material and Morphological study of the Pomegranate.
- Collection of plant material: Punicaceae peel was collected from local farmhouse in the month of January 2025.
- Morphological study of Pomegranate¹⁰.
Method of Preparation of Pomegranate peel extract.^9,10

The pomegranate peels were separated manually and then subjected to Hot air oven for drying at 40°C until it reaches a moisture content of approximately 5% (dry basis). Following this, the dried peels were milled before being subjected to the extraction process. 100 g of the blended peels were extracted with 400ml ethyl acetate in a Soxhlet extractor for 3, 6 and 15 hrs in the dark (4-5 extraction cycles per hour). The obtained solution was evaporated at 40°C in the dark to a dry yellow-brown residue.

Preliminary Phytochemical screening^11,12:

The above extract was studied for their Phytochemical identifications and reported. Qualitative preliminary phytochemical analysis was performed with different chemical reagents to detect the nature of phytoconstituents present in the pomegranate extract. The presence of amino acids, terpenoids, saponins and alkaloids were evaluated by standard qualitative methods.

Table No. 1: Formulation Table

Sl. No.	Ingredients	Weight%
1.	PVP K30	1.4
2.	HPMC K4 M	0.2
3.	Propylene glycol	1
4.	Acetic acid	0.2
5.	Chitosan	0.2
6.	Water	20
7.	Extract	0.5g

Method of preparation of Anti lipidemic Buccal film:¹³

Buccal films of Pomegranate peel extract were prepared by solvent casting technique using film forming mucoadhesive polymers. HPMC was weighed accurately and dissolved in 2ml ethanol. The beaker containing polymer and ethanol was kept aside for 5 min for swelling of the polymer. Further 3ml of ethanol was added to the above polymer solution and the dispersion was stirred. Then one drop propylene glycol was added to the polymer solution. Simultaneously pomegranate peel extract was accurately weighed and then dissolved in 1ml of ethanol in another beaker. The drug solution was added to the polymer solution and was mixed thoroughly with the help of a magnetic stirrer. The whole solution was poured into the glass Petri dish placed over a flat surface and kept for drying.

Evaluation of the Herbal buccal film:

Appearance: The prepared film formulations were checked by visual inspection and surface texture was evaluated by touch or feel of the film.
pH Determination:^13,14,15The pH of the formulation was measured using a pH meter. The film to be tested was placed in a petri dish and dissolved in 10ml distilled water for 30sec and pH was measured.
Thickness:^15,16 The thickness of film was measured by micrometre screw gauge. For evaluation of film thickness, three films (2*2cm²) of each formulation were taken and the film thickness was measured at different strategic locations (at least 5 locations) and the mean thickness of the films was calculated.
Folding endurance:^13,16,17 Folding endurance of the films determines the flexibility of the films. It was determined by repeatedly folding a small strip of the film (2*2cm²) at the same place until it breaks. The number of times strips can be folded at the same place, without breaking gives the value of folding endurance.
Tack test:¹⁸Thumb tack test was performed to determine the tackiness by gently squeezing a thumb on a film for 5sec and then quickly removing it. The parameters which represent the adhesive property of film is expressed by following value ranges: No adhesion (-), Poor adhesion (+), Medium adhesion (++) and Good/excellent adhesion (+++).

RESULTS AND DISCUSSIONS:

- Morphological studies of pomegranate (Punica granatum)

Pomegranate tree is a member of family “Punicaceae” which comprises only one genus “Punica” and two species. The plant is a deciduous, evergreen attractive shrub 6–10 m in height and long lived. The tree bears thorny multiple branches. The leaves of the plant are evergreen, 1–10 cm long and short stemmed are placed as a cluster of 5–6 on branches. The flowers, 3 cm wide, are characterized by red colour, pointed sepals and numerous stamens. Two to seven flowers are present at the periphery of each branch. The hexagonal shaped fruit is 6–12 cm wide and weighs around

200 g. The fruit is crowned by thick tubular calyx. The rind of the fruit is red, hard and leathery. The edible fruit, sarcotesta, is a berry which comprises of 600 arils (a casing for seeds) under a thick skin. The ripe fruit attains red colour. The arils are placed in several bunches separated by membranous walls and white spongy tissue.

Fig No. 01: Soxhlet extractions of Punica granatum

Qualitative preliminary phytochemical analysis

Phytochemical screening was done to detect the phytoconstituents present in ethanolic extract of pomegranate. It revealed that the extract was rich in chemical constituents like phenolics, flavonoids, ellagitannins, proanthocyanidin compounds, many minerals including potassium, nitrogen, sodium and polyphenolic compounds such as punicalagin, gallic acid, ellagic acid.

Fig No. 02: Punica granatum peel extract

Table No. 2: Phytochemical Screening of Punica granatum extract

Phytochemical Specific Test		Pomegranate peel extract
Phenolics	Ferric chloride Test	+++
Polyphenols	Liebermann Test	+++
Anthocyanins	Lead Acetate Test	+
Flavonoids	FeCl3 Lead Acetate	++ +++
Saponins	Froth’s Test	+
Glycosides	Keller Killiani Test	+
Sterols	Salkowski Test	+
Tannins	FeCl₃	++
Fixed oil Saponification Test			-
Reducing sugar Fehling’s Test			-

+++ refer appreciable amounts (+ve within 5min)

++ refer moderate amount (+ve after 5min but within 10 min)

+ refer trace amount: - refer completely absent

Organoleptic characteristics and evaluation of the formulation:

The formulated buccal film was evaluated for physicochemical test to observe its properties. The prepared herbal buccal film showed yellow-brown colour, it exhibited good characteristics in terms of appearance, colour, homogeneity and consistency.

Table no. 3: Physicochemical tests of the prepared buccal film.

Evaluation test	Result
Appearance	Good
Colour	Yellow-brown
Odour	Characteristic

Fig No. 03: Herbal buccal film of pomegranate peel extract

pH, thickness and folding endurance:

The pH of the formulation was determined by using a digital pH meter. From the result it was found that the formulation had desired pH which is compatible with mouth.

Table no. 4: pH of Herbal buccal film

Formulation	Herbal buccal film
pH*	6.6±0.03

*Data expressed as a mean ±SD, n=3

Thickness:

The thickness of Mucoadhesive Buccal films depends on the concentration of polymer and the thickness. The Mucoadhesive Buccal films were measured with micrometre screw gauge. The thickness of Mucoadhesive Buccal Contributed to increase the knowledge in the subject. The increasing prevalence of childhood dyslipidaemia, fueled by the global obesity epidemic, underscores the urgent need for safe, effective, and child-friendly therapeutic options. Conventional lipid lowering agents often carry concerns regarding their impact on growth and development in paediatric patients. In this context, the use of natural remedies, such as pomegranate peel extract, offers promising potential due to its rich profile of polyphenolic compounds known for their films was found to be 0.34±0.0056 mm.

Table no. 5: Thickness

Parameter	Value (mm)
Thickness	0.34±0.0056

Folding endurance:

Folding endurance of the film was found to be 154±1 N/cm².

Tack test:

Thumb tack test was performed to determine the tackiness by gently squeezing a thumb on a film for-5 s and then quickly removing it.

Table no. 6: Tackiness

Test	Tackiness
Tack test	Good (+++)

Lipid-lowering and antioxidant properties:

This study focused on the development of a mucoadhesive buccal film as an innovative drug delivery system to administer pomegranate peel extract for the management of dyslipidaemia in children. Buccal delivery offers a practical alternative to traditional oral dosage forms, especially for children who may have difficulty swallowing tablets. By bypassing hepatic first pass metabolism, this route also enhances bioavailability and therapeutic efficacy.

The formulated buccal film not only aims to improve patient compliance but also leverages the natural bio actives in pomegranate peel to address lipid abnormalities safely. Overall, the study presents a novel, non-invasive approach that aligns with the growing interest in phytopharmaceuticals and paediatric friendly drug delivery systems. Future research should explore clinical validation and long-term safety to support its integration into mainstream pediatric care.

Scope for Further Studies:

The development of a mucoadhesive buccal film containing pomegranate peel extract to manage dyslipidemia in children presents a promising avenue in pediatric drug delivery. However, this research opens up several new possibilities for further exploration. Firstly, future studies can focus on clinical trials involving pediatric populations to assess the efficacy and safety of the formulated buccal film. While in-vitro and in-vivo animal studies offer initial insights, clinical data will be crucial to validate its therapeutic potential and tolerability in children. Secondly, there is scope to explore the long-term effects of polyphenol-rich extracts like those from pomegranate peel on lipid profiles and vascular health. Studies could investigate not just lipid lowering effects but also improvements in endothelial function, oxidative stress markers, and inflammatory mediators over extended periods. Lastly, future investigations could examine the cost-effectiveness, scalability, and shelf-life stability of the formulation, especially with a view to its commercialization and inclusion in public health initiatives targeted at childhood obesity and related metabolic disorders.

SUMMARY AND CONCLUSION:

The objective of this study was to formulate and evaluate a buccal film containing pomegranate peel extract as a natural and child-friendly approach to managing dyslipidemia in children. The research was carried out in the following steps:

Extraction of Pomegranate Peel: Pomegranate peel was dried, powdered, and subjected to Soxhlet extraction to obtain a concentrated extract rich in bioactive compounds with potential lipid lowering effects.
Phytochemical Screening: Preliminary phytochemical analysis confirmed the presence of important constituents such as flavonoids, tannins, saponins, and phenolic compounds, which are known for their antioxidant and antilipidemic properties.
Formulation of Buccal Film: The extract was incorporated into a mucoadhesive buccal film using suitable film-forming agents and plasticizers to ensure flexibility and patient acceptability.

Evaluation of Buccal Film:

The formulated films were evaluated for key physicochemical parameters:

(i) pH: Found to be within the acceptable range for buccal application, ensuring no irritation.

(ii)Thickness: Consistent across samples, indicating uniform drug distribution.

(iii)Folding Endurance: Demonstrated good mechanical strength and flexibility.

(iv)Tack Test: Showed appropriate mucoadhesiveness for buccal retention. The study successfully developed a buccal film containing pomegranate peel extract with acceptable physicochemical properties and phytochemical composition, suggesting potential for treating dyslipidemia in children.

REFERENCES:

Fiorentino R, Chiarelli F. Treatment of dyslipidaemia in children. Biomedicines. 2021;9(9):1078–9.
Gujral J, Gupta P. Pediatric dyslipidemia [Internet]. Treasure Island (FL): StatPearls Publishing; 2024 Jan. Available from: https://www.ncbi.nlm.nih.gov/books/NBK585106
McCrindle BW. Hyperlipidaemia in children. Thromb Res. 2006;118(1):49–58.
Wiegman A, Gidding SS, Watts GF. Familial hypercholesterolaemia in children and adolescents: gaining decades of life by optimizing detection and treatment. Eur Heart J. 2015;36(36):2425–37.
Viuda-Martos M, Fernández-López J, Pérez-Álvarez JA. Pomegranate and its many functional components as related to human health: a review. Compr Rev Food Sci Food Saf. 2010;9(6):635–54.
Lee CJ, Chen LG, Liang WL, Wang CC. Anti-inflammatory effects of Punica granatum Linné in vitro and in vivo. Food Chem. 2010;118(2):315–22.
Haghighian MK, Rafraf M, Moghaddam A, Hemmati S, Jafarabadi MA, Gargari BP. Pomegranate (Punica granatum L.) peel hydro-alcoholic extract ameliorates cardiovascular risk factors in obese women with dyslipidaemia: a double-blind, randomized, placebo-controlled pilot study. Eur J Integr Med. 2016;8(5):676–82.
Alqahtani MS, Kazi M, Alsenaidy MA, Ahmad MZ. Advances in oral drug delivery. Front Pharmacol. 2021;12:618411.
Alessandra M, Andrea C, Eugenio L, Luciana M, Patrizia P, Stefania C. Evaluation of different extraction methods from pomegranate whole fruit or peels and the antioxidant and antiproliferative activity of the polyphenolic fraction. Food Chem. 2016;202:59–69.
Negi PS, Jayaprakasha GK, Jena BS. Antioxidant and antimutagenic activities of pomegranate peel extracts. Food Chem. 2003;80(3):393–7.
Jayaprakash A, Sangeetha R. Phytochemical screening of Punica granatum Linn. peel extracts. J Acad Ind Res. 2015;4(5):160–2.
Farag RS, Abdel-Latif MS, Emam SS, Tawfeek LS. Phytochemical screening and polyphenol constituents of pomegranate peels and leaf juices. Landmark Res J Agric Soil Sci. 2014;1(6):86–93.
Semalty A, Semalty M, Nautiyal U. Formulation and evaluation of mucoadhesive buccal films of enalapril maleate. Indian J Pharm Sci. 2010;72(5):571–5.
Khade A, Gadge G, Mahajan U. An overview on natural polymer-based mucoadhesive buccal films for controlled drug delivery. Int J Pharm Res Technol. 2020;10(1):48–57.
Lynthong B, Bhavyashree T, Kamath K, Shabaraya AR. Oro-dispersible film: an effective approach for quick drug release. Int J Pharm Sci Rev Res. 2022;75(2):180–5.
Haju S, Yadav S, Baig R, Sawant G. Buccal film: a novel approach for oral mucosal drug delivery system. Asian J Pharm Clin Res. 2021;5(1):27–35.
Chauhan I, Yasir M, Verma M. Oral delivery of zolmitriptan loaded fast disintegrating film: formulation development, statistical optimization, in-vitro and in-vivo evaluation. J Appl Pharm Sci. 2019;2(1):13–22.
Dharmasthala S, Shabaraya AR, Andrade GS, Shriram RG, Hebbar S, Dubey A. Fast dissolving oral film of piroxicam: solubility enhancement by forming an inclusion complex with β-cyclodextrin, formulation and evaluation. J Young Pharm. 2019;11(1):1–6.
Lei F, Zhang XN, Wang W. Evidence of anti-obesity effects of pomegranate leaf extract in high-fat diet-induced obese mice. Int J Obes. 2007;31(6):1023–9.