Green Cosmeceuticals : Formulation and Evaluation of Anti-Aging Cream Formulations from Orange Peel Extract

In recent years, there has been a growing preference for skincare products that are derived from natural sources, driven by concerns over the side effects associated with synthetic ingredients. Herbal  cosmetics are gaining popularity not only for their therapeutic benefits but also for their safety and eco-friendliness. Among the various botanicals explored for skincare, orange peel (Citrus sinensis) stands out due to its rich profile of bioactive compounds, including flavonoids, essential oils, phenolic acids, and antioxidants.

Orange peel, often discarded as industrial waste, has shown promising potential in cosmetic applications, particularly in anti-aging formulations. Its key constituents, such as hesperidin, naringin, and limonene, exhibit strong antioxidant and anti-inflammatory properties, making it effective in combating signs of skin aging like wrinkles, dryness, and loss of elasticity. These compounds work by neutralizing free radicals, protecting collagen and elastin fibers, and supporting the skin’s barrier function.

The aging of skin is influenced by both intrinsic factors (such as genetic aging) and extrinsic factors (such as UV radiation and pollution), which together lead to oxidative stress and the breakdown of the skin’s structural proteins. Natural ingredients like orange peel extract offer a safer, plant-based alternative to chemical anti-aging agents by addressing these underlying causes.

This research focuses on the formulation of a herbal anti-aging cream using orange peel extract and its subsequent evaluation for key quality attributes. The study not only highlights the therapeutic value of natural ingredients in dermatology but also promotes sustainable utilization of agro-industrial byproducts, thereby contributing to the advancement of green cosmeceuticals.

PLANT PROFILE :-

Orange peel, a byproduct of the sweet orange (Citrus sinensis), is valued for its aromatic and medicinal properties. It contains bioactive compounds like essential oils, flavonoids, and antioxidants, contributing to its health benefits. Originally native to Southeast Asia, orange cultivation has spread globally to regions with warm, tropical, and subtropical climates. Major producers of sweet oranges include Brazil, the U.S., Mexico, Spain, and India, making orange peel widely available for use in various industries such as food, cosmetics, and pharmaceuticals. They are given many names depending on the regions; some of the common names are given in Table 1.[1]

Common names and botanical classification of Citrus sinensis:-

Table 1

Table :- 2

 Botanical Description:

The orange tree (Citrus sinensis), of the family Rutaceae, is a medium-sized evergreen tree with dark green, aromatic leaves and fragrant white flowers. The fruit is a hesperidium with a leathery peel composed of two layers: the flavedo, a bright orange, oil-rich outer layer, and the albedo, a white, pectin-rich inner layer. Widely cultivated in tropical and subtropical regions, it is valued for its nutritional and medicinal properties. [1,2]

Fig 1 . orange peel and it’s powder

Geographical Source :-

Orange (Citrus sinensis) are widely cultivated in tropical and subtropical regions across the globe. Major producers include India (Nagpur, Assam, Coorg), Brazil, the United States (Florida, California), Spain, and South Africa. These areas offer the warm climate, well-drained soil, and moderate rainfall essential for optimal growth and fruit production. [3,4]

Fig 2 . Whole Plant of Citrus Sinensis Linn.

Morphology :-

Citrus sinensis, commonly known as the sweet orange, is a small to medium-sized evergreen tree that typically reaches a height of 4 to 10 meters. It has a thick, woody trunk covered with smooth, greyish-brown bark. The branches are extensive and slightly zig-zag, contributing to the tree’s rounded canopy. The leaves are simple, lanceolate in shape, and glossy, with a smooth, leathery texture. They are dark green on the upper side and pale green on the underside, measuring about 5 to 10 cm in length and 2 to 5 cm in width. The petiole is winged and short, and the margins of the leaves are smooth or lightly serrated. These leaves contain numerous oil glands, which release a fragrant citrus aroma when crushed. The sweet orange tree produces white, fragrant flowers that are approximately 2 to 3 cm in diameter. The flowers can appear either solitarily or in small clusters and have five petals. Each flower is hermaphroditic, containing both male and female reproductive organs. The flowers bloom in spring or early summer, attracting pollinators with their strong citrus scent.

The fruit of Citrus sinensis is round or slightly flattened, typically 6 to 10 cm in diameter, with a smooth, orange-colored peel when ripe. The peel is thick and textured, rich in essential oils. Inside, the fruit contains 10 to 12 segments filled with juice sacs, and the pulp is sweet, with a slight tanginess. The high water content of the fruit makes it a popular choice for fresh consumption or juicing. The tree also has a fibrous and shallow root system that is extensive and spreads widely, allowing for efficient absorption of water and nutrients from the soil.

Histology and Microscopical character  :-

Peel: The peel of Citrus sinensis consists of two main layers—flavedo (outer colored layer) and albedo (inner spongy layer).

• Flavedo (Epicarp): The outer surface is lined with a single-layered epidermis covered by a thick cuticle. Oil glands (secretory cavities) are present throughout the flavedo and contain essential oils like limonene, giving the orange its scent. The epidermal cells are compact and polygonal, and few multicellular trichomes may be observed.
• Albedo (Mesocarp): The inner layer is made of loosely packed parenchymatous cells with large intercellular spaces. These cells may contain pectin and mucilage, and vascular bundles are scattered within. Occasionally, calcium oxalate crystals may also be seen.

Powder analysis of citrus sinensis:

The powder of Citrus sinensis peel is yellowish-orange to light brown in color with a pleasant citrus aroma. Microscopically, it shows fragments of polygonal epidermal cells with cuticle, multicellular uniseriate trichomes (3–6 celled), and numerous oil gland remnants. Parenchymatous cells are visible, some containing pigments or mucilage. Scattered calcium oxalate crystals in the form of druses or raphides are present. Occasional spiral and reticulate xylem vessels can be seen, while starch grains are generally absent or very few.

Fig 3 .  Showing microscopy of orange peel and it’s powder

Pharmacological Activities [20]  :-

1. Antiaging and Skin’s Barrier Improvement

Intrinsic skin aging is a natural and gradual process influenced by internal and external factors, often leading to dryness, rough texture, and visible signs such as wrinkles and uneven pigmentation. These changes can affect self-esteem and comfort. A key contributor to this aging is oxidative stress, which damages the skin’s extracellular matrix (ECM) through enzymes like matrix metalloproteinases (MMPs). When overactive, MMPs degrade essential proteins such as collagen and elastin, increasing reactive oxygen species (ROS) and causing structural harm at the cellular level, including DNA damage and lipid peroxidation. This weakens the skin barrier and results in moisture loss. Hesperidin, a flavonoid found in orange peel, can counteract these effects by boosting antioxidant defenses, inhibiting oxidase activity, and regulating MMPs through the expression of tissue inhibitors (TIMPs). Additionally, hesperidin helps protect skin exposed to environmental stressors like pollution and UV radiation by neutralizing harmful free radicals and supporting skin barrier repair, potentially through the Nrf2 signaling pathway. Its integration into skincare products shows promise in delaying signs of skin aging and maintaining healthier skin.

2.   UVA and UVB Radiation-Induced Skin Damage

Prolonged exposure to sunlight is a primary cause of premature skin aging, often referred to as photoaging. This process is primarily triggered by ultraviolet (UV) radiation, particularly UVA (320–400 nm) and UVB (280–320 nm) rays, which penetrate the skin and cause structural and functional damage. While UVA contributes to deeper dermal alterations, UVB is known for its more intense effects, including inflammation, sunburn (erythema), and even cellular apoptosis. Continuous UV exposure results in oxidative stress through the generation of reactive oxygen species (ROS), which in turn leads to DNA damage, protein oxidation (such as carbonylation), and disorganization of the connective tissue, along with abnormal epidermal thickening.

Hesperidin, a natural flavonoid found in orange peel, exhibits strong protective properties against such UV-induced damage. Research suggests that hesperidin helps minimize epidermal thickening, regulates pro-angiogenic and inflammatory signaling pathways like PI3K/Akt, VEGF, HIF-1α, and MMPs, and modulates oxidative stress-related apoptotic proteins. It also plays a role in controlling immune cell responses and reducing inflammatory cytokines, contributing to an overall anti-inflammatory effect on the skin. Moreover, formulations containing hesperidin, especially those with nano-lipid carriers (NLCs), have shown exceptional photoprotective capabilities—blocking up to 99% of UVB and 83% of UVA rays. These properties highlight hesperidin’s potential as an effective sunscreen agent in both cosmetic and therapeutic skin care products designed to combat photoaging.

3. Skin’s Hyperpigmentation and Depigmentation Conditions

Changes in an individual's natural skin tone are often linked to pigmentation disorders such as melasma, solar lentigo, or depigmentary conditions like vitiligo. These conditions are usually visible and can negatively affect a person’s confidence and self-image. Melanin, the pigment responsible for skin and hair color, also plays a crucial role in shielding the skin from harmful UV rays and oxidative stress. Hyperpigmentation typically arises due to an increase in melanin synthesis or its excessive distribution, a process known as melanogenesis, which takes place within organelles called melanosomes in specialized cells known as melanocytes. These melanosomes are eventually transferred to keratinocytes, leading to pigmentation. Several factors influence melanogenesis, including internal regulators like the enzyme tyrosinase and the transcription factor MITF, as well as environmental triggers such as UV radiation. Hence, controlling these pathways—either by inhibiting tyrosinase or downregulating MITF—has become a key approach in managing hyper pigmentary disorders. Conversely, reduced activity of factors like MSH or increased oxidative stress can lead to  hypopigmentation, as seen in vitiligo. Given the importance of melanin regulation in skin health and appearance, there is growing interest in identifying bioactive compounds for skincare applications.

 One such compound is hesperidin, a citrus-derived flavonoid found in orange peels, which shows                   promising effects on pigmentation. Research has revealed that hesperidin can inhibit melanin production by interfering with melanosome transport—specifically, by blocking the Rab27A–melanophilin protein complex. It also reduces melanin synthesis by lowering MITF and tyrosinase expression through the MEK/ERK1/2 signaling pathway. Interestingly, hesperidin doesn’t only inhibit melanin; when applied in suitable formulations, it may also promote re-pigmentation, making it potentially useful for treating conditions like vitiligo. Studies suggest that when hesperidin is combined with compounds like trimethylpsoralen in nanoemulsion gels, it may help restore skin pigmentation, likely through its immunomodulatory properties—though the exact mechanism is still under investigation.

4. Wound healing

Chronic wounds, including ulcers and burns, are characterized by prolonged healing periods—often exceeding three months—or may fail to heal entirely due to the persistent inflammatory state that disrupts the natural wound-repair process. Such wounds are commonly associated with compromised immune responses, typically linked to underlying conditions like diabetes, infections, or metabolic disorders, which significantly diminish the patient's quality of life. These wounds frequently produce excessive exudate, which can harm surrounding healthy tissue and hinder the healing process by maintaining the wound in an inflammatory phase. This is largely attributed to imbalanced expression of molecular mediators, including growth factors, matrix metalloproteinases (MMPs), tissue inhibitors of metalloproteinases (TIMPs), and integrins, along with elevated levels of reactive oxygen species (ROS). As a result, the proliferation and differentiation of key cells such as fibroblasts and keratinocytes are impaired, ultimately obstructing stable extracellular matrix (ECM) formation. If untreated, chronic wounds may escalate into more serious conditions, including infections, hemorrhage, or even gangrene—highlighting their global significance as a public health challenge. Wound healing is a highly regulated and dynamic biological process required for restoring skin integrity following physical or chemical injury. This process typically unfolds in three overlapping phases: (i) hemostasis and inflammation, (ii) proliferation with ECM synthesis, and (iii) re-epithelialization and tissue remodeling . Numerous cellular players, enzymes, and signaling molecules coordinate these stages. Among natural therapeutic agents, hesperidin—a flavonoid predominantly found in orange peel—has attracted attention due to its anti-inflammatory and antimicrobial effects, making it a promising compound for dermatological formulations. Emerging studies suggest that hesperidin aids wound repair by scavenging free radicals, downregulating pro-inflammatory cytokines such as IL-1β, IL-8, and TNF-α, and promoting fibroblast proliferation and tissue regeneration.

5. Skin Cancer and Other Cutaneous Diseases

Skin cancer, especially in fair-skinned individuals, is increasingly common. It includes types like melanoma and basal cell carcinoma. Research aims to target DNA damage, promote cancer cell death, and prevent abnormal cell growth and angiogenesis. Natural compounds like hesperidin, a flavonoid from citrus fruits, have shown anti-cancer effects by reducing oxidative stress and disrupting cancer cell function. Studies suggest hesperidin can slow tumor growth, arrest the cell cycle, and influence proteins linked to cell death in cancerous cells. Additionally, it has been found to support the treatment of skin conditions such as rosacea, psoriasis, and atopic dermatitis by reducing inflammation and abnormal blood vessel formation through pathways like PI3K/Akt and VEGF. Hesperidin also shows promise in aiding hair regrowth by improving blood flow and stimulating hair follicles. Overall, it holds great potential for use in pharmaceutical skincare products aimed at treating various skin disorders and enhancing treatment outcomes.

Table 1.  Phytochemical Screening of orange Extract

Collection and processing of sample

Orange fruit was bought from the local market. The peel was separated from the fruit, washed under running water to remove any dirt, and left to dry in the shade for four days. After drying, the peel was ground into a fine powder, sieved using a No. 60 mesh, and stored in an airtight container for use in research and formulation studies.

Chemicals and Reagents

Orange peel powder, beeswax, almond oil, borax, olive oil, sandalwood, distilled water, ethanol, Molisch’s reagent, concentrated H?SO?, Fehling’s reagents A and B, Wagner’s reagent, lead acetate, ninhydrin solution, dilute H?SO?, FeCl?, DPPH solution, Folin -Ciocalteu’s reagent, gallic acid, E. coli bacteria strain, rose water, 20% sodium carbonate, and ruthenium red dye.

Instruments and Equipments

Sieve no:60 mesh, round conical flask(250ml), whatman filter paper no:41, aluminium foil, test tubes, nutrient agar plates, porcelain dish, thermometer, Buchner funnel, colourimetry, Ph meter, two glass slides of 20 cm × 20 cm, microscopic slide, Brookfield Viscometer.

Preparation of pomegranate extract [20]

a) Preparation of Aqueous Extract of Orange Peel

About 10 g of orange peel powder was placed in a 250 ml conical flask and subjected to a decoction process using 100 ml of boiled distilled water at room temperature for 1 hour. The mixture was then filtered using Whatman filter paper No. 41. The clear extract obtained was used for phytochemical screening and cream formulation.

b) Preparation of Alcoholic Extract of Orange Peel

A total of 150 g of orange peel powder was placed in a conical flask, and 300 ml of ethanol was added and macerated (Orange Peel Powder to ethanol ratio should be 1:10). The flask was sealed and kept for maceration for 8-10 days.

Fig 4 . alcoholic extraction of orange peel

Formulation Of Anti- ageing Cream

Table No. 2: Ingredients and its role.

Table No. 3: Formulation Table. [7]

The right amounts of white beeswax, almond oil, and olive oil were heated together in a porcelain dish at 70°C until melted. In another container, borax was dissolved in water, and the orange peel extract (in alcohol) was added. (For the F3 formulation) the required amount of honey was slowly mixed in. This mixture was also warmed to 70° C.

Next, the water-based mixture was slowly poured into the melted oils while stirring constantly. A small amount of sandalwood was added if required. Stirring continued until a smooth cream was formed. Just before storing the cream in containers, a fragrance was added. Four versions of the cream—F1, F2, F3, and F4—were made by changing the ingredient ratios and were later tested.

F1

F2

F3

F4

Fig no. 4. Formulations of Anti – aging Cream

Evaluation of Herbal Cream [8,9,10]

Organoleptic Evaluation

The physical properties of the cream, such as color, odor, appearance, and homogeneity, were analyzed through visual inspection after storage.

Homogeneity

The homogeneity of the cream was tested by visually observing its appearance after preparation. A small quantity was pressed between the thumb and index finger to check for consistency and ensure the cream was homogeneous.

pH of the Cream

The pH meter was calibrated with a standard buffer solution. About 1 g of the cream was dissolved in 50 ml of distilled water, and the pH of the suspension was measured at 27°C.

 Viscosity

The viscosity of the cream was measured using a rotational viscometer (Brookfield DVII+ Pro, spindle No. 7) at 25 ± 1°C. Measurements were taken in triplicates at 100 rpm, and viscosity values were recorded in centipoise (cp).

Dye Test

The ruthenium red dye was mixed with the cream. A drop of the cream was placed on a microscope slide, covered with a cover slip, and examined under a microscope. If the dispersed globules appeared red and the ground was colorless, the cream was an oil-in-water (o/w) type. The reverse indicated a water-in-oil (w/o) type.

Spreadibility

Place 1gm of sample between two glass slides of 20 cm × 20 cm. A weight of 100 gm was placed on the upper slide so that the cream was pressed uniformly between the two slides to form a thin layer. The weight was removed and then fixed to a stand without slightest disturbance in such a way that the upper slide slides off freely, to the force of weight [30 gm] tied to it. The time taken for the upper slide to travel the distance of 5.0cm and separate away from the lower slide under the direction of the weight was noted.^[14]15]

S = m × L/T

Where

S = Spreadability

M = Weight tied to the upper slide

L = Length of the glass

T = Time taken in seconds.

Stability Studies

Stability studies were conducted to assess the effect of temperature, humidity, and light on the cream. The formulations were stored at room temperature (26°C ± 2) and in a refrigerator (4°C ± 2) for two months. At the end of the

Results and Discussion :

Organoleptic  Evaluation

The organoleptic evaluation revealed that all formulations maintained their appearance, odor, color, and overall elegance throughout the observation period.

Viscosity

Viscosity of cream was in the range of 500-1000 cps which indicates that the cream is readily spreadable by small amount of shear.

Table No. 6: Viscosity values of formulation.

Spreadability

The spreadability of the formulation was in the range of 13 to 18gm.cm/min, which indicated that the spreadability of the formulation was considered high with low spreading time.

Table No. 7: Spreadability value of formulations.

pH of the Cream

The pH of the formulations was found to be in the range of 6.0–6.35, which is close to the natural pH of the skin. This ensures the formulations are non- irritating and suitable for skin application.

Stability testing

The cream filled in the bottle and kept in humidity chamber maintained at 260C ±2 and 40C±2 for 2.5 months. At the end of studies samples were analysed and found to be stable