Formulation and Evaluation of a Peel-Off Face Mask Using Butterfly Pea Flower (Clitoria ternatea Linn.) Extract

Human skin is constantly directly exposed to the air, solar radiation, environmental pollutants, or other mechanical and chemical insults, which causes the generation of free radicals as well as reactive oxygen species (ROS) of our own metabolism. Ionizing radiation, extreme physical and mental stress, alcohol consumption, poor diet, overeating, and pollution in the environment are some of the causes of extrinsic skin damage ^[1]. Among these is UV radiation, and repeated exposure to ultraviolet B (UV-B) is causing damage to DNA and the skin's function and structure, resulting in the skin aging process known as photoaging. Meanwhile, ultraviolet A (UV-A) has a longer wavelength and can penetrate deeper into dermic skin but is not causing damage to DNA. Photoaging is characterized by the appearance of a wrinkle due to collagen damage caused by the formation of ROS. Uncontrolled free radicals can also cause cellular tissue damage, resulting in symptoms such as redness, pigmentation, and an increased risk of cancer ^[2]. Therefore, the addition of antioxidants is necessary to address this insufficiency. Antioxidants are widely used to reduce and protect the skin from harmful ultraviolet radiation. Natural antioxidants from plant origin are well known to have fewer side effects than the manufactured ones ^[3,4]. Butterfly-pea's botanical name is Clitoria ternatea Linn. and belongs to the Fabaceae (Papilionaceae) family. It is known by different names as bluebell vine, blue pea, cordofan pea, Darwin pea, and many more, and is found in several countries worldwide, including Thailand, Malaysia, India, Kenya, Australia, Sri Lanka, and others ^[5]. Clitoria ternatea L. is rich in phenolic content and contains terpenoid compounds, anthocyanin, tannins, phenols, and flavonoids ^[6,7]. Due to these constituents, the butterfly pea (Clitorea ternatea L.) has potential as an anti-aging treatment for facial skin. The presence of phenolic compounds and flavonoids in Clitoria ternatea L. as antioxidants can ward off free radicals to inhibit oxidation, thereby slowing photooxidation from exposure to UV lights ^[8]. Studies have reported its other pharmacological activities, including nootropic, anticonvulsant, antidepressant, antianxiety, antistress, analgesic, anti-inflammatory, antihyperglycemic, antihyperlipidemic, antidiabetic, and platelet aggregation inhibitory actions ^{[9, 10]}. The peel-off face mask is one of the prominent forms of topical applications used to enhance the quality of the facial skin. The skin face mask has the virtue of being easily peeled off or removed as an elastic membrane ^[11]. Once it is completely dried, the liquid film peels off from the face, leaving behind a thin, plasticized film without any residue. It effectively removes skin impurities and tightens, rejuvenates, and heals skin. The peel-off face mask is useful to treat the facial skin and can be used to minimize pores. Additionally, peel-off masks may provide slight moisturization while enhancing the occlusive effect, which promotes increased blood flow, activates skin cells, and facilitates impurity removal. In addition, it is also useful for soothing the muscles of the face and as a cleanser or freshener ^[12]. With these considerations, this study was aimed at developing and testing a peel-off face mask from butterfly-pea flowers.

Pharmacognosy of Butterfly-pea flower:

1. Plant profile:

Botanical Name: Clitoria ternatea Linn.

Common Name: Butterfly pea, blue pea and Cordofan pea

Synonyms: Asphota, Girikarni, Vishnukranta, Sankhapushpi, Sephanda, Sveta, Maha

Family: Papilionaceae

2. Taxonomic hierarchy:

Kingdom: Plantae

Phylum: Angiosperms

Order: Fabales

Family: Fabaceae

Genus: Clitoria

Species: Clitoria Ternatea Linn^.[13]

3. Other species of Clitoria:

Clitoria albiflora Mattei

Clitoria amazonum Benth

Clitoria Andrei Fantz.

Clitoria angustifolia Kunth ^[14]

MATERIAL AND METHOD:

Material:

The dried flowers of butterfly pea were purchased from the online market store, Flipkart E-commerce site, and were stored in airtight conditions till use. The Clitorea ternatea Linn blue flowers  has been identified with the number MIT0227  at Mithibai College Of Arts, Chauhan Institute Of Science & Amrutben Jivanlal College Of Commerce And Economics. The techniques used for authentication are microscopy and morphology. Additional materials were used such as  Polyvinyl alcohol (PVA), glycerine, hydroxypropyl methylcellulose (HPMC), gelatin, polyethylene glycol, citric acid, methanol,  sulfuric acid and Levender oil. 

METHODOLOGY:

1.  Procedure for Extraction of butterfly-pea flower:

For the extraction of butterfly-pea flowers,  it was dried at 40? for 15-20 minutes to remove moisture from flowers and powdered using a mixer. The powder is then sifted using sieves to get fine powder for maximum extraction. According to a literature survey, alcoholic extract of butterfly-pea flower is prominent for antioxidant activity^[15,16]. Butterfly-pea flower extract is made by macerating the powdered drug in acidified methanol for 24 hours and then filtering it out using Whatman filter paper^[17,18]. Then the filtrate solvent is evaporated at 40? in a hot air oven to get the extract ^[19].

2. Phytochemical Screening of Butterfly-Pea Flower Extract:

The alcoholic extract of butterfly-pea (Clitorea ternatea L.) was tested for the presence of alkaloid, carbohydrate, terpenoid compounds, anthocyanin, glycoside, tannins, phenols, and flavonoids ^[20,21].

3. Antioxidant Activity Test of Butterfly-Pea Flower Extract:

Antioxidant activity was tested using the DPPH (1,1-diphenyl-2-picrylhydrazyl) method. As much as 10.0 mg of butterfly-pea flower extract stock solution was then put into a 10.0 ml volumetric flask, and methanol was added until the limit was reached and shaken homogeneously. Stocks of methanol extract were taken in amounts of 100, 200, 300, 400, and 500 μl. A DPPH solution of 3.0 ml and methanol up to the limit mark were added in a 5.0 ml volumetric flask. The solution was incubated for 30 min, and then the absorbance was measured at a wavelength of 516 nm using a UV-VIS spectrophotometer (LABINDIA Analytical UV 3200). The final concentrations of flower extract were 20, 40, 60, 80, and 100 ppm. The absorbance data was used to calculate the inhibition percentage of extract against DPPH free radical using the equation below:

Percentage inhibition (%) = A₀-A₁/ A₀ x 100

Where, A₀ was the absorbance of the blank solution, while A1 was the absorbance of the test solution. The IC50 (Inhibition concentration 50%) was determined using the linear regression equation y = bx + a, where x is the concentration (μg/ml) and y is the percentage of inhibition ^[22-24].

4. Formulation of peel-off gel mask base:

The base of the formulation was made with Polyvinyl alcohol (PVA) and with increasing concentrations of gelatin to form the best base for the peel-off mask with appropriate properties ^[25]. The formulation table of peel-off mask base is listed below:

Table No. 01: Formulation table for peel-off base

4.1 Physical evaluation of peel-off mask base:

The base of the formulation was evaluated by various organoleptic properties and physically by performing tests like pH, drying time, spreadability, and film-forming capacity ^[26]. And on the basis of test results, the most suitable base is selected for further trials.

5. Preparation of Peel-Off Mask:

All ingredients were weighed carefully using an analytical balance. Polyvinyl alcohol (PVA) and HPMC dispersion was made at 70? and cooled at room temperature till 40?. Polyethylene glycol 400 (PEG 400) and glycerin were added to the dispersion when it attained room temperature. Meanwhile, dissolve gelatin with water by heating followed by cooling it at 40? and then add it to the polymer dispersion along with citric acid with continuous stirring. After proper mixing of gelatin and citric acid, extract is added and stirred to get uniform mixing of extract. At the end, perfumery agents like lavender oil are added in a little quantity ^[27].

Table no. 2: Formulation Table of Butterfly-pea flower peel-off mask.

6. Physical evaluation of peel-off mask:

The physical evaluation involved were organoleptic, pH, drying and film forming, and spreadability. The organoleptic properties like colour, odour, texture and consistency was observed for the formulation. The pH was determined using pH paper available, drying time was performed by applying a sufficient amount of formulated base on a glass slide in a uniform layer and peeled off after drying. Drying time was determined by applying a Peel-off mask sample in a 4x4cm area and was dried at room temperature and time was noted. Film forming capability was checked by folding endurance of peel-off mask and it was performed by applying mask sample on skin in 3x3cm area and was rolled without breaking. It checks the resistance of the peel-off mask for breaking. Spreadability was determined using a glass slide on which 1gm of sample mask was placed and another slide was put on it with 5gm of weight for 5 minutes. Spreadability was determined using formula:

S= m x l/ t

Where,

S = Spreadability

l = length after spreading

m = weight of sample on slide

t = time [24-28].

RESULT AND DISCUSSSION:

Result:

1.Extraction of butterfly-pea flower:

 The dark blue colour  of extract of Clitorea ternetea Linn were obtained  with  extractive value of 18.5%  by using macerated extraction procedure.  

2. Phytochemical Screening of Butterfly-pea flower Extract:

Table no. 3: Phytochemical Screening of Butterfly-pea flower Extract

3.Antioxidant Activity Test of Butterfly-pea flower Extract:

The results of antioxidant testing on Butterfly-pea flower extract proved that Butterfly-pea flower extract is 3.239 μg/ml.

4.Formulation and Preparation of peel-off gel mask base:

With the help of organoleptic properties and physical testing we found out that B2 base is appropriate for the formulation of peel-off mask for further trials.

4.1 Physical evaluation of peel-off gel mask base:

Table no. 4: Evaluation parameter data of base trials

5. Formulation and Preparation of Peel-off mask:

The organoleptic of peel-off gel mask preparation was homogenous violet-blue gel with the characteristic odour of lavender.

5.1. Physical evaluation of peel-off mask:

Table no.5: Evaluation parameter data of formulation trials

DISCUSSION:

1.Extraction of butterfly-pea flower:

Extraction of butterfly-pea flower powder was done by maceration method using acidified methanol. The method is chosen to avoid damage of active substances due to heating. Extra pure methanol was used to extract polar substances like flavonoid and anthocyanin. Active substances taken from butterfly-pea flower powder are flavonoids and anthocyanin which have an antioxidant effect. The extraction process will produce a liquid extract which is then concentrated by evaporating the solvent at 40-45 °C. The viscous extract obtained has characteristic odor and violet-blue in color. Extraction of 100 grams of butterfly-pea flower powder resulted in 18.5g (18.5%) viscous extract.

2. Phytochemical Screening of Butterfly-pea flower Extract:

Phytochemical screening tests include alkaloid, flavonoid, tannin, phenol, terpenoid and saponin tests. Butterfly-pea flower extract positively contains alkaloid, tannins, flavonoids and anthocyanin which is shown in Table no. 3

3.Antioxidant Activity Test of Butterfly-pea flower Extract:

Antioxidant activity test was Butterfly-pea flower extract using DPPH radical scavenging assay method. The principle of the DPPH radical scavenging assay method is that when the DPPH solution is mixed with a sample that has antioxidant activity, it causes a reduction form with the change of purple to yellow color. The results of antioxidant testing on butterfly-pea flower extract proved that the IC50 of butterfly-pea flower extract is 3.239μg/ml. An antioxidant agent which has high antioxidant activity will have a low IC50 value. Butterfly-pea flower extract was able to inhibit 50% DPPH at concentration 3.239 μg/ml. In this study, Ascorbic acid was used as a reference standard. Since ascorbic acid is a powerful antioxidant, its IC50 value was higher. An IC50 value of ≤ 50 ppm indicated a strong antioxidant whereas an IC50 value of 50-100 indicated a moderate antioxidant property.

Table no. 6: DPPH radical scavenging assay of Butterfly-pea flower extract:

Graph no. 01: DPPH radical scavenging assay of Butterfly-pea flower extract: