INTRODUCTION:

The skin, the body's most extensive organ, has important roles in protection, regulation of body temperature, and sensation. It acts as the first line of defence against external threats, including microorganisms, harmful chemicals, and ultraviolet (UV) radiation. Skin ageing is a complex process determined by internal factors like genes and natural ageing phenomenon, and external factors, for example, environmental influences. These external factors include UV radiation, infections, tobacco use, and air pollutants¹.

Among these, exposure to UV radiation is a significant contributor, accounting for up to 80% of visible signs of skin ageing². Cosmetic products are typically designed not only to enhance the appearance of the skin but also to moisturize, protect, and sometimes treat it. A fundamental requirement for marketing these products is ensuring their safety and minimizing the risk of harmful side effects, such as allergic reactions or long-term health impacts. Cosmetics generally contain a combination of lipid-based ingredients like oils, fats, and waxes, as well as emulsifiers, water, various chemicals, and fragrances. Given their extensive use and direct application to the skin, the safety of these products is paramount. Although the skin serves as a protective barrier, certain components of cosmetics can permeate the skin and enter the systemic circulation, potentially leading to broader health concerns³.

Women have adorned themselves for beauty since antiquity, a tradition that has evolved over time but continues today, especially in rural areas where natural remedies, such as plant extracts, are still favoured for cosmetic purposes. While synthetic products dominate in urban settings, many women in rural regions continue to rely on herbal cosmetics for their beauty routines. These products, designed to cleanse and enhance the skin, often claim additional benefits, though it is essential to distinguish between cosmetic and medicinal claims. The primary advantage of herbal cosmetics lies in their plant-based composition, which is often perceived as safer and more nourishing for the skin⁴. These natural ingredients, rich in vitamins and minerals, provide essential nutrients while minimizing harmful side effects. Plants are a treasure trove of beneficial compounds that continue to fuel the cosmetics industry, especially those with a long history of use in traditional medicine⁵. Therefore, it is crucial to continuously explore and integrate these natural ingredients into modern cosmetic formulations. High-energy UV radiation generates reactive oxygen species (ROS) that, in turn, activate dermal proteases such as collagenase and elastase, which are enzymes responsible for breaking down collagen and elastin fibres in the skin. Botanical extracts rich in phytochemicals (e.g. anthocyanins and flavonoids) often inhibit these proteases through metal-chelation and active-site interactions⁶, thereby providing an anti-ageing effect beyond simple radical scavenging. Likewise, in vitro SPF testing quantifies an extract’s ability to absorb UV and prevent ROS formation⁷. This study assessed collagenase/elastase inhibition and photoprotective SPF, thus offering mechanistic insight into skin-protective effects.

A significant disadvantage of synthetic face cream is its potential to cause skin irritation or allergic reactions, often due to the presence of certain synthetic chemicals and artificial fragrances. These adverse effects range from mild redness and itching to more severe dermatological conditions. As a result, there has been a growing demand for cosmetics made with natural ingredients, particularly as consumers become more aware of the benefits of plant-based products. This trend is not limited to rural regions, where traditional cosmetics incorporating plant extracts have long been favoured, but is also expanding into urban markets. Scientific research increasingly supports the use of plant-derived ingredients, revealing that they contain a diverse array of potent phytochemicals capable of soothing and enhancing the skin, as well as aiding in its restoration, healing, and protection⁸. Consequently, the cosmetics industry is seeing a continuous influx of new products formulated with biological ingredients. Among these, face cream is one of the most important products, and a key quality factor is its colour. However, the synthetic colours traditionally used in face creams may pose risks when applied over extended periods. This has led to a preference among consumers for cosmetics with natural ingredients, including natural colourants.

The research gap identified by us is that so far, no face cream is available in the market that contains natural ingredients such as refined mango butter, virgin coconut oil, aloe vera gel and flower extracts. Given these considerations, this research project aimed to develop four different types of face creams with natural ingredients enriched with flower extracts. Specifically, we utilised the flowers of Ixora coccinea L., Clitoria ternatea L., Quisqualis indica L., and Hibiscus rosa-sinensis L. to create products that meet the growing demand for safe, effective, and naturally formulated skincare solutions.

I. coccinea (Rubiaceae) is an evergreen shrub widely distributed in India. In the Indian traditional system of medicine, Ayurveda, as well as in various folk medicine practices, different parts of the plant, such as the flowers, leaves, roots, and stems, are utilised to treat a range of ailments depending on the specific medical condition⁹. The flowers of I. coccinea are reported to contain several bioactive compounds, including rutin, leucocyanidin glycoside, cyanidin-3-rutinoside, and delphinidin monoglycoside, which may contribute to their therapeutic properties¹⁰. C. ternatea (Fabaceae), commonly known as blue pea or butterfly pea flower, is a traditional Ayurvedic medicine that has been used for centuries for its wide range of therapeutic effects, including as a memory enhancer, nootropic, antistress, anxiolytic, antidepressant, anticonvulsant, tranquillizing, and sedative agent¹¹. The flower is not only edible but also widely utilised in culinary practices and as a natural food colourant, owing to its vibrant blue hue. A distinctive feature of C. ternatea is its high content of polyacylated anthocyanins, specifically ternatins, which are responsible for its striking colour and are noted for their stability and potential health benefits¹². The presence of these unique anthocyanins underscores the plant's value both in traditional medicine and modern applications. Q. indica (Combretaceae family), commonly known as Rangoon Creeper, is native to Africa and parts of the Indo-Malaysian region and is extensively cultivated throughout India. This climbing shrub blooms year-round, producing a profusion of flowers that start as white and gradually transition to a vibrant red hue¹³. The flowers of Q. indica are noted for their antioxidative properties, which have been documented in several studies¹⁴. These antioxidative properties suggest that Q. indica flowers could provide significant antioxidant benefits for the skin when incorporated into cosmetic formulations. H. rosa-sinensis (Malvaceae), commonly known as hibiscus, is a lavishly flowering shrub prevalent in tropical regions. Studies have shown that aqueous extracts of hibiscus are rich in tannins and anthocyanins and exhibit significant ferric reducing antioxidant power. Both aqueous and ethanolic extracts of hibiscus have demonstrated the ability to inhibit the growth of food-borne pathogens such as Salmonella typhimurium and Staphylococcus aureus¹⁵. These findings highlight the potential of hibiscus extracts for use in antioxidant and antimicrobial applications.

The objectives of this research work were (i) preparation of ethanolic flower extracts of various plants such as I. coccinea, C. ternatea, Q. indica and H. rosa-sinensis, (ii) formulation of face creams using the flower extracts, (iii) evaluation of the quality of the face creams through various tests, including stability, safety, sensory characteristics, antiradical potential, anti-ageing effects (through inhibition of collagenase and elastase), and (iv) investigation of in vitro SPF.

MATERIALS AND METHODS:

Flower collection:

Full-bloomed flowers of I. coccinea, C. ternatea, Q. indica, and H. rosa-sinensis were collected from the Herbal Garden of the authors’ institution during Jan 2024. The flowers included red-coloured I. coccinea, blue-coloured C. ternatea, red-coloured Q. indica (collected three days after anthesis), and red-coloured H. rosa-sinensis (Fig. 1). Plant identification was performed by the corresponding author of this manuscript, who also submitted the specimens to the institution affiliated with him (Voucher number for I. coccinea, C. ternatea, Q. indica, and H. rosa-sinensis is 2024-01-12-001, 2024-01-12-002, 2024-01-12-003, and 2024-01-12-004, respectively).

Fig. 1. Flowers used in this work. (A) Ixora coccinea; (B) Clitoria ternatea; (C) Quisqualis indica; (D) Hibiscus rosa-sinensis

Flower extract preparation:

To prepare the flower extracts, 100g of fresh flower petals were macerated in 500mL of 75% ethanol in a cold room maintained at 4°C for 72hours. The mixture was then filtered through Whatman No. 1 filter paper, and the ethanol was removed using a rotary evaporator at 27°C. The remaining aqueous solution was freeze-dried using a lyophilizer to yield the dried extract powder, which was stored at 4°C for use in subsequent experiments. The extraction yield (%) was calculated using the following formula:

Weight of freeze – Dried extract

Percent Yield = -------------------------------------- x 100

Fresh Weight of flower petals

The yield of the flower extracts of I. coccinea, C. ternatea, Q. indica and H. rosa-sinensis was 8.8 %, 7.3 %, 8.5 % and 7.1 %, respectively.

Cream recipe:

Face creams incorporating various flower extracts were prepared following the method described earlier², with specific modifications to suit the ingredients used. First, the oil phase ingredients (refined mango butter, 10g; virgin coconut oil, 8g; Olivem 1000, 3g) were weighed into beaker A, while the aqueous phase ingredients (aloe vera gel, 24g; vegetable glycerine, 5g; distilled water, q.s. to 100g) were weighed into beaker B. The temperature of the contents in each beaker was raised to 75°C in a water bath. The aqueous phase was gradually mixed with the oil phase with continuous stirring to ensure proper emulsification. Once the mixture cooled to 35°C, D-panthenol (3g), hyaluronic acid (HA; 2g) and other heat-sensitive excipients (Optiphen BD, 0.3g; rose essential oil, 0.1g) were incorporated with continuous stirring to preserve their efficacy and volatility. Finally, the ethanolic flower extract (3g) was added and mixed thoroughly to obtain a uniform cream. The resulting cream was transferred to an amber vial and stored at ambient temperature to protect it from light and ensure its stability for further analysis.

Analysis of cream:

a) Type of cream test:

The type of emulsion [oil-in-water (O/W) or water-in-oil (W/O)] was determined using methods described earlier¹⁶. Scarlet red dye was mixed into the cream, and a drop was placed on a microscope slide. Under microscopic examination, red globules against a colourless background indicated an O/W type, while colourless globules on a red background indicated a W/O type. A dilution test with water was also conducted, with O/W creams exhibiting good miscibility and W/O creams showing poor miscibility.

b) Irritation test:

A small quantity of cream was applied to a 1 cm² area on the dorsal surface of left hand. The site was observed for 24hours, with periodic checks every 4 hours to assess for signs of irritation, erythema, or edema¹⁷.

c) Antioxidant activity:

The antioxidant activity of the flower extracts and face creams was evaluated using the DPPH assay, based on protocols described earlier¹⁸ with modifications. This method assesses the sample’s capacity to reduce the DPPH radical dissolved in methanol at a concentration of 0.2mM. A reaction mixture was prepared by mixing 3 mL of the DPPH solution with 3 mL of one of the following: (i) flower extract dissolved in ethanol (10 mg/mL), (ii) ascorbic acid dissolved in ethanol (60 μg/mL) as a standard, (iii) face cream dissolved in ethanol (100mg/mL), or (iv) ethanol (99.9%) as a control. The absorbance of the mixture was recorded at 517nm after 30min incubation in the dark. The antioxidant activity was expressed as the percentage of DPPH scavenging activity, calculated using the formula:

cA - sA

Percent DPPH Scavenging Activity = ------------- x100

Where cA is absorbance of control and sA is absorbance of sample (flower extract, standard or face cream).

d) Collagenase Inhibition Assay:

Collagenase inhibitory activity was measured by a modified FALGPA (N-[3-(2-furyl)acryloyl]-Leu-Gly-Pro-Ala) spectrophotometric assay¹⁹. Briefly, Clostridium histolyticum collagenase (0.1 U) was pre-incubated with test samples (flower extract or cream) in 50mM Tricine buffer (pH 7.5, containing 400mM NaCl, 10mM CaCl₂) for 15min at 25°C. The reaction was initiated by adding 0.8 mM FALGPA substrate. Hydrolysis of FALGPA was monitored at 335nm over 20 min. Per cent inhibition was calculated relative to an enzyme-only control. EGCG (250μM) served as a positive control. Experimental conditions follow Thring et al.¹⁹

e) Elastase Inhibition Assay:

Elastase inhibition was determined using N-succinyl-Ala-Ala-Ala-p-nitroanilide (AAAPVN) as substrate. Porcine pancreatic elastase (1 U/mL) and samples were incubated in 0.2 M Tris–HCl buffer (pH 8.0) for 15min at 25°C. The reaction was started by adding 0.8mM AAAPVN, and the release of p-nitroaniline was measured at 410nm (detected between 381–402nm) for 20min. Per cent inhibition was calculated relative to the enzyme control. EGCG (250μM) was again used as positive control. This protocol is adapted from established assays¹⁹.

The following formula was used to calculate per cent inhibition of the enzymes.

ABS_control - ABS_sample

Collagenase / (%) = --------------------------------- x 100

Elastase Inhibition ABS _control

Where ABS control is the absorbance of control and ABS sample is the absorbance of sample.

f) In Vitro SPF Determination (UV Absorbance):

Photoprotective capacity was assessed by UV spectrophotometry⁷. Samples (0.1mg/mL hydroalcoholic solutions of extract or cream) were scanned from 290–320nm in a UV–Vis spectrophotometer. Sun Protection Factor (SPF) was calculated:

Where CF (correlation factor) = 10, and E(λ) and I(λ) are the erythemal effect and solar intensity factors respectively^7,20. The constants E(λ) and I(λ) were taken from the literature²¹.

g) Sensory characteristics:

The sensory characteristics of the prepared creams were assessed using methods reported in previous studies². The creams were carefully examined for colour, flow behaviour, consistency, phase separation, uniformity, and smell. A tiny quantity of cream was squeezed between the thumb and index finger to assess its consistency and to detect any rough particles. Additionally, the cream's firmness and oiliness during application were qualitatively evaluated to gauge its overall sensory appeal.

h) Washability:

100 mg of cream was spread evenly on a defined area of the skin and then rinsed gently with water at room temperature. Better washability is indicated when the cream is easily removed, leaving the skin surface free of residue and non-greasy to the touch, which enhances user comfort and product effectiveness²².

i) Spreadability:

To assess the spreadability of the cream, 500mg of the product was kept within a circle with a 10mm diameter at the center of a glass slide. Another glass slide of the same size was carefully positioned on top of the cream. A uniform weight of 100g was applied to the upper slide for two minutes at room temperature to ensure consistent spreading²³. Afterward, the spread diameter of the cream was measured three times using a caliper to calculate the average spread diameter ± standard error (SE). This process was repeated at intervals of 20, 40, and 60 days to evaluate any changes in spreadability over time.

j) pH of the cream:

The pH meter was first calibrated at pH 4 and pH 7. 0.5 g of the cream was thoroughly dissolved in 50mL of distilled water, with gentle stirring to ensure complete dissolution. The pH of the resulting solution was then measured at room temperature.

k) Accelerated shelf-life assessment of face creams:

The face creams were placed in amber vials, and sealed to protect them from light exposure. In accordance with the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH) guidelines (specifically ICH Q1A(R2)), an accelerated shelf-life assessment was conducted in a stability chamber set at 40°C and 75% relative humidity (RH) for two months, simulating long-term storage under stress conditions²⁴. The creams were initially assessed for their organoleptic properties (colour, texture, smell, and separation of phases), pH, and spreadability. These variables were then monitored every twenty days, with assessments conducted at 0, 20, 40, and 60 days.

Data analysis:

All analyses were carried out in triplicate, and the mean values were presented with their SE. The data were analysed using Analysis of Variance with a significance level of P<0.05. To separate the means and determine significant differences, Duncan's Multiple Range Test (DMRT) was used for post-hoc analysis.

RESULTS AND DISCUSSION:

The dispersed globules in all herbal face creams prepared in this study appeared red against a colourless background, confirming their classification as O/W emulsions. This was further validated by the creams' excellent miscibility with water, a typical feature of O/W emulsions. O/W creams are generally favoured in cosmetic formulations due to their superior washability and reduced greasiness compared to W/O creams¹⁷. Previous studies have demonstrated that O/W creams containing virgin coconut oil (VCO) or structured VCO enhance skin hydration, improve elasticity, and reduce transepidermal water loss²⁵. Additionally, O/W creams formulated with coconut oil and co-surfactants exhibit stable viscosity and homogenous oil globule dispersion even after temperature cycling, ensuring excellent stability²⁶. These attributes, combined with their spreadability, penetration ability, and non-greasy texture, contribute to a positive sensory experience²⁷.

The creams were confirmed to be safe for use, as no irritation, erythema, or edema was observed upon application. This aligns with the traditional use of the flower extracts, which are known for their beneficial effects on the skin. The flowers of I. coccinea have long been applied externally to treat skin ailments such as sores, chronic ulcers, and scabies⁹. In Thailand, C. ternatea flower extracts are commonly incorporated into cosmetics, with the flowers exhibiting significant antioxidant activity. The total phenolic content of C. ternatea flowers has been reported to be 1.9mg/g of extract, expressed as gallic acid equivalents²⁸. The flowers of Q. indica are rich in bioactive compounds such as alkaloids, phenols, flavonoids, saponins, and tannins²⁹. Q. indica flower extracts have demonstrated notable scavenging activity against DPPH radicals, achieving 90.2% inhibition at a concentration of 500 mg/L³⁰. Similarly, H. rosa-sinensis, a prominent herb in Indian traditional medicine, has shown potential in cosmetics, particularly due to its high solar protection factor values, which suggest its use as a natural additive in skincare formulations³¹. Additionally, the total phenolic content in H. rosa-sinensis flower extract has been reported as 235.77mg/100g³⁰.

The antioxidant activities of various flower extracts (10 mg/mL), as well as the herbal face creams (100mg/mL) prepared using these extracts, are presented in Fig. 2. Among the extracts, C. ternatea exhibited the highest antioxidant potential, followed by Q. indica, H. rosa-sinensis, and I. coccinea (Fig. 2A). These significant antioxidant properties justify their inclusion in anti-ageing herbal face creams.

Fig. 2. DPPH scavenging activity (%) of ethanolic flower extracts (A), herbal face creams made using the ethanolic flower extracts (B), and the standard, ascorbic acid; error bars indicate ± SE; different alphabets above the error bars indicate significance at 5% by DMRT.

In contrast to synthetic antioxidants like butylated hydroxytoluene (BHT), which carries potential risks such as skin irritation and pulmonary toxicity³², plant-based flower extracts provide safer, natural alternatives. Antioxidants are essential in anti-ageing formulations as they neutralise free radicals, reducing oxidative stress on the skin². The strong antioxidant activity of C. ternatea petals is primarily attributed to their high phenolic and flavonoid content, particularly anthocyanins like ternatins¹². These compounds have demonstrated effective free radical scavenging and protection against oxidative damage. Similarly, the ethanolic extract of Q. indica flowers, rich in phenolic compounds and flavonoids, plays a significant role in free radical scavenging³³. The presence of quinoline-4-carbonitrile (QCN) and other compounds further enhances its antioxidant properties³⁴. H. rosa-sinensis flowers contain various bioactive compounds, including anthocyanins, flavonols, and flavan-3-ols, all contributing to its antioxidant activity³⁵.

The herbal face creams in this study were formulated with 3% flower extract to retain antioxidant potential (Fig. 2B). The cream with C. ternatea extract exhibited significantly higher antioxidant potential (p<0.05) compared to other formulations. However, the creams prepared with Q. indica, H. rosa-sinensis, and I. coccinea extracts showed comparable DPPH scavenging activities. The antioxidant potential of the creams can also be attributed to other ingredients like HA, D-panthenol, and aloe vera gel. HA is well-known for maintaining skin moisture and elasticity and possesses antioxidant properties that protect against oxidative damage. D-panthenol improves skin hydration and supports wound healing, while aloe vera gel offers antioxidant, anti-inflammatory, and skin-soothing benefits. The natural pigmentation of the creams, derived from the anthocyanin-rich petals, enhances their visual appeal (Fig. 3). These formulations exhibited desirable attributes such as smooth texture, spreadability, uniformity, and excellent washability under running water (Table 1).

Fig. 3. Herbal face creams prepared using various flower extracts

Accelerated stability testing, conducted at 40°C and 75% relative humidity (RH), revealed that the creams remained stable for 60 days, with no phase separation or significant changes in colour, consistency, or scent. The pH values ranged from 5.03 to 5.50 (Table 2), and spreadability varied between 3.9 and 6 cm (Table 3). Although slight changes in pH and spreadability were observed during storage, the creams remained within the optimal pH range (4-6), which supports skin health and minimizes irritation risks³⁶. The stability of the creams under elevated temperatures, coupled with their favourable skin absorption and spreadability, suggests that they are well-suited for topical applications. The overall performance of these formulations remained within acceptable limits throughout the study, ensuring their potential for consumer use.

Table 2. Effect of ambient temperature and accelerated shelf-life assessment on the pH of herbal face creams

Face cream prepared using	pH ± SE
	Ambient temperature				Accelerated shelf-life assessment
	0 day	20 days	40 days	60 days	0 day	20 days	40 days	60 days
Ixora coccinea	5.03 ± 0.01 e	5.10 ± 0.02 d	5.22 ± 0.03 c	5.33 ± 0.03 ab	5.03 ± 0.01 e	5.21 ± 0.01 c	5.31 ± 0.02 b	5.38 ± 0.01 a
Clitoria ternatea	5.16 ± 0.04 f	5.24 ± 0.02 e	5.34 ± 0.02 c	5.42 ± 0.01 b	5.16 ± 0.04 f	5.29 ± 0.01 d	5.40 ± 0.01 b	5.50 ± 0.01 a
Quisqualis indica	5.06 ± 0.01 d	5.16 ± 0.01 c	5.26 ± 0.02 b	5.35 ± 0.02 a	5.06 ± 0.01 d	5.29 ± 0.01 b	5.36 ± 0.01 a	5.38 ± 0.01 a
Hibiscus rosa-sinensis	5.24 ± 0.01 e	5.33 ± 0.01 d	5.38 ± 0.01 c	5.43 ± 0.01 b	5.24 ± 0.01 e	5.37 ± 0.01 c	5.44 ± 0.01 b	5.50 ± 0.03 a

Values are mean ± SE. Different alphabets in a row indicate statistical significance at 5% by DMRT.

Table 3. Influence of ambient temperature and accelerated shelf-life assessment on the spreadability of herbal face creams

Face cream prepared using	Spreadability (cm) ± SE
	Ambient temperature				Accelerated shelf-life assessment
	0 day	20 days	40 days	60 days	0 day	20 days	40 days	60 days
Ixora coccinea	5.23 ± 0.09 a	4.67 ± 0.09 b	4.40 ± 0.06 c	4.20 ± 0.06 c	5.23 ± 0.09 a	4.77 ± 0.03 b	4.37 ± 0.07 c	4.20 ± 0.06 c
Clitoria ternatea	5.50 ± 0.06 a	4.53 ± 0.09 bc	4.37 ± 0.18 c	3.90 ± 0.06 d	5.50 ± 0.06 a	4.70 ± 0.06 b	4.63 ± 0.03 b	4.53 ± 0.03 bc
Quisqualis indica	6.00 ± 0.06 a	5.60 ± 0.06 c	5.10 ± 0.06 d	4.80 ± 0.10 e	6.00 ± 0.06 a	5.80 ± 0.06 b	5.50 ± 0.06 c	5.10 ± 0.06 d
Hibiscus rosa-sinensis	5.70 ± 0.06 a	5.20 ± 0.06 c	4.73 ± 0.07 de	4.27 ± 0.09 f	5.70 ± 0.06 a	5.40 ± 0.06 b	4.90 ± 0.06 d	4.67 ± 0.03 e

Values are mean ± SE. Different alphabets in a row indicate statistical significance at 5% by DMRT.

Table 4. Collagenase and elastase inhibition (%) at 500 µg/mL and in vitro SPF values for flower extracts and creams (mean ± SE).

Sample	Collagenase Inhibition (%)	Elastase Inhibition (%)	*In vitro* SPF
Clitoria extract	40.7 ± 2.4 a	31.8 ± 2.2 a	3.4 ± 0.2 b
Clitoria cream	25.4 ± 1.2 b	18.4 ± 1.5 b	7.2 ± 0.5 a
Quisqualis extract	35.3 ± 2.6 a	27.3 ± 2.7 a	3.5 ± 0.2 b
Quisqualis cream	20.2 ± 1.9 b	12.4 ± 1.6 b	6.8 ± 0.5 a
Hibiscus extract	30.5 ± 2.3 a	19.3 ± 2.8 a	12.5 ± 0.2 b
Hibiscus cream	15.7 ± 1.5 b	10.6 ± 1.5 b	18.3 ± 0.5 a
Ixora extract	25.1 ± 2.9 a	16.7 ± 2.2 a	5.5 ± 0.2 b
Ixora cream	12.8 ± 1.3 b	7.8 ± 1.4 b	9.0 ± 0.5 a

For a particular species, different alphabets in a column indicate statistical significance at 5% by DMRT.

The SPF results indicate notable UV-absorption by the extracts, especially Hibiscus. Hibiscus rosa-sinensis extract showed the highest SPF (12.5), reflecting its anthocyanin content. The cream formulations further increased SPF (18.3 for Hibiscus), likely due to film thickness and scattering by other cream ingredients. These SPF values observed in this study are comparable to those reported in the literature: for example, Saglam et al.³¹ found Hibiscus flower extract SPF 11.77 and leaf extract 22.1. Clitoria and Ixora extracts gave lower SPF (3.4–5.5) but still contributed measurable UV protection. These results confirm that the flower pigments impart significant UV filtering, supporting their use as natural photo-protectants.

CONCLUSION:

Herbal face creams made using flower extracts of C. ternatea, Q. indica, H. rosa-sinensis, and I. coccinea exhibited strong antioxidant and anti-ageing activities alongside favourable sensory characteristics. Their O/W emulsion structure provided excellent miscibility with water, a non-greasy texture, and easy washability, enhancing their cosmetic appeal. The creams had stability during accelerated testing at 75% RH and 40°C for 60 days, with no significant changes in colour, texture, odour, or phase separation, affirming their shelf stability. The pH values, ranging from 5.03 to 5.50, remained within the optimal acidic range, supporting skin health and minimizing irritation risks. While there was a slight decrease in spreadability during storage, the values stayed within acceptable limits, ensuring ease of application and consumer satisfaction. Among the flower extracts, C. ternatea exhibited highest antioxidant potential, collagenase and elastase inhibition (%), and in vitro SPF followed by Q. indica, H. rosa-sinensis, and I. coccinea, validating their suitability for anti-ageing skincare formulations. These bioactive ingredients, in combination with natural antioxidants, provide an effective and safer alternative to synthetic additives like BHT. To conclude, the herbal face creams developed in this study not only offer potent antioxidant and anti-ageing benefits but also meet key criteria for cosmetic performance. These formulations hold significant commercial potential in the natural skincare market, catering to consumers seeking biologically derived, safe, and effective alternatives to conventional synthetic products.

CONFLICT OF INTEREST:

The authors have no conflicts of interest regarding this investigation.

REFERENCES:

1. Giacomoni PU, Rein G. Factors of skin ageing share common mechanisms. Biogerontology. 2001; 2(4): 219–229. doi: 10.1023/A:1013222629919

2. Tan PL, Rajagopal M, Chinnappan S, Selvaraja M, Leong MY, Tan LF, et al. Formulation and Physicochemical Evaluation of Green Cosmeceutical Herbal Face Cream Containing Standardized Mangosteen Peel Extract. Cosmetics. 2022 Apr 27; 9(3): 46. doi: 10.3390/cosmetics9030046

3. Loretz LJ, Api AM, Barraj LM, Burdick J, Dressler WE, Gettings SD, et al. Exposure data for cosmetic products: lipstick, body lotion, and face cream. Food and Chemical Toxicology. 2005 Feb; 43(2): 279–291. doi: https://doi.org/10.1016/j.fct.2004.09.016

4. Deshmukh HS, Babar VB, Jagtap PS, Doshi RV, Deokate SV, Todkari AV, et al. A Comprehensive Review Article on Herbal Cosmetics. South Asian Res J Pharm Sci. 2024 May 10; 6(03): 50–68. doi: 10.36346/sarjps.2024.v06i03.003

5. Kowalczyk S, Grymel M, Bilik J, Kula W, Wawoczny A, Grymel P, et al. Selected Plants as Sources of Natural and Active Ingredients for Cosmetics of the Future. Applied Sciences. 2024 Apr 20; 14(8): 3487. doi: 10.3390/app14083487

6. Madan K, Nanda S. In-vitro evaluation of antioxidant, anti-elastase, anti-collagenase, anti-hyaluronidase activities of safranal and determination of its sun protection factor in skin photoaging. Bioorganic Chemistry. 2018 Apr; 77: 159–167. doi: 10.1016/j.bioorg.2017.12.030

7. Kaur CD, Saraf S. In vitro sun protection factor determination of herbal oils used in cosmetics. Pharmacognosy Res. 2010 Jan; 2(1): 22–25. doi: 10.4103/0974-8490.60586

8. Gediya SK, Mistry RB, Patel UK, Blessy M, Jain HN. Herbal Plants: Used as a cosmetics. J Nat Prod Plant Resour. 2011; 1(1): 24-32. https://www.scholarsresearchlibrary.com/articles/herbal-plants-used-as-a-cosmetics.pdf

9. Baliga MS, Kurian PJ. Ixora coccinea Linn.: Traditional uses, phytochemistry and pharmacology. Chin J Integr Med. 2012 Jan; 18(1): 72–79. doi: 10.1007/s11655-011-0881-3

10. Latha P, Nayar M, Singh O, George V, Panikkar K, Pushpangadan P. Isolation of antigenotoxic ursolic acid from Ixora coccinea flowers. Actual Biol. 2017 Nov 22; 23(74): 21–24. doi: 10.17533/udea.acbi.329614

11. Mukherjee PK, Kumar V, Kumar NS, Heinrich M. The Ayurvedic medicine Clitoria ternatea—From traditional use to scientific assessment. Journal of Ethnopharmacology. 2008 Dec; 120(3): 291–301. doi: 10.1016/j.jep.2008.09.009

12. Vidana Gamage GC, Lim YY, Choo WS. Anthocyanins From Clitoria ternatea Flower: Biosynthesis, Extraction, Stability, Antioxidant Activity, and Applications. Front Plant Sci. 2021 Dec 17; 12: 792303. doi: 10.3389/fpls.2021.792303

13. Khare CP, editor. Indian Medicinal Plants: An Illustrated Dictionary [Internet]. New York, NY: Springer New York; 2007 [cited 2023 Sep 28]. doi: 10.1007/978-0-387-70638-2

14. Habeeeb HM. A comparative phytochemical investigation and antioxidant activity of Quisqualis indica L. DASJ. 2024 Jun 30; 16(1): 137–144. doi: 10.52951/dasj.24160112

15. Mak YW, Chuah LO, Ahmad R, Bhat R. Antioxidant and antibacterial activities of hibiscus (Hibiscus rosa-sinensis L.) and Cassia (Senna bicapsularis L.) flower extracts. Journal of King Saud University - Science. 2013 Oct; 25(4): 275–282. doi: 10.1016/j.jksus.2012.12.003

16. Aswal A, Kalra M, Rout A. Preparation and evaluation of polyherbal cosmetic cream. Der Pharmacia Lettre. 2013; 5(1): 83–88. https://www.scholarsresearchlibrary.com/articles/preparation-and-evaluation-of-polyherbal-cosmetic-cream.pdf

17. Mishra AP, Saklani S, Milella L, Tiwari P. Formulation and evaluation of herbal antioxidant face cream of Nardostachys jatamansi collected from Indian Himalayan region. Asian Pacific Journal of Tropical Biomedicine. 2014 Jul; 4: S679–S682. doi: 10.12980/APJTB.4.2014APJTB-2014-0223

18. Gobikanila K, Jeyaramraja PR. Throwing light on the antidiabetic, anti-inflammatory and antioxidant nature of the leaves of tree tomato (Solanum betaceum Cav.). Discov Med. 2024 Sep 27; 1(1): 55. doi: 10.1007/s44337-024-00061-3

19. Thring TS, Hili P, Naughton DP. Anti-collagenase, anti-elastase and anti-oxidant activities of extracts from 21 plants. BMC Complement Altern Med. 2009 Dec; 9(1): 27. doi: 10.1186/1472-6882-9-27

20. Mansur J, Breder M, Mansur M, Azulay R. Determinação do fator de proteção solar por espectrofotometria. An Bras Dermatol, Rio de Janeiro. 1986; 61: 121–124.

21. Sayre RM, Agin PP, LeVee GJ, Marlowe E. A comparison of in vivo and in vitro testing of sunscreening formulas. Photochem and Photobiology. 1979 Mar; 29(3): 559–566. doi: 10.1111/j.1751-1097.1979.tb07090.x

22. Mahawar V, Patidar K, Joshi N. Development and evaluation of herbal antiaging cream formulation containing Annona squamosa leaf extract. Asian J Pharm Clin Res. 2019; 12(2): 210–214. doi: 10.22159/AJPCR.2019.V12I2.29026

23. Berenguer D, Sosa L, Alcover M, Sessa M, Halbaut L, Guillén C, et al. Development and Characterization of a Semi-Solid Dosage Form of Meglumine Antimoniate for Topical Treatment of Cutaneous Leishmaniasis. Pharmaceutics. 2019 Nov 15; 11(11): 613. doi: 10.3390/pharmaceutics11110613

24. Netto G, Jose J. Development, characterization, and evaluation of sunscreen cream containing solid lipid nanoparticles of silymarin. J of Cosmetic Dermatology. 2018 Dec; 17(6): 1073–1083. doi: 10.1111/jocd.12470

25. Ibrahim SI, Matmin J, Hairul NM, Abdul Majeed AB. Enhancing Emollient Properties of natural and Structured Virgin Coconut Oil Creams on Skin Conditions using Non-invasive Skin Biophysical Methods. MAB. 2022 Oct 31; 51(4): 109–117. doi: 10.55230/mabjournal.v51i4.18

26. Lwin WW, Puyathorn N, Chantadee T. Cold-Pressed Coconut Oil Cream: Preparation and Evaluation. KEM. 2022 Mar 21; 914: 23–29. doi: 10.4028/p-408j21

27. Bogdan C, Moldovan M, Man I, Crisan M. Preliminary study on the development of an antistretch marks water-in-oil cream: ultrasound assessment, texture analysis, and sensory analysis. CCID. 2016 Sep; 9: 249–255. doi: 10.2147/CCID.S107298

28. Kamkaen N, Wilkinson JM. The antioxidant activity of Clitoria ternatea flower petal extracts and eye gel. Phytotherapy Research. 2009 Nov; 23(11): 1624–1625. doi: 10.1002/ptr.2832

29. Barik B, Das S, Hussain T. Pharmacognostic properties of Quisqualis indica Linn. against human pathogenic microorganisms: An insight review. EJMP. 2020; 31(20): 87–103. https://journalejmp.com/index.php/EJMP/article/download/961/1928/1907

30. Afify AEMMR, Hassan HMM. Free radical scavenging activity of three different flowers-Hibiscus rosa-sinensis, Quisqualis indica and Senna surattensis. Asian Pacific Journal of Tropical Biomedicine. 2016 Sep; 6(9): 771–777. doi: 10.1016/j.apjtb.2016.07.006

31. Sağlam A, Aşan Özüsağlam M, Çeli̇K İ. Determination of Sun Protection Potential of Hibiscus rosa-sinensis as Natural Additive for Cosmetic Industry. Bulletin of Biotechnology. 2023 Jun 26; 4(1): 13–16. doi: 10.51539/biotech.1215927

32. Lanigan R, Yamarik T. Final Report on the Safety Assessment of BHT. Int J Toxicol. 2002; 21(2_suppl): 19–94. doi: 10.1080/10915810290096513

33. Singh S, Rai A, Maity S, Sarkar S, Maji S, Saha S. Effect of ethanolic extract of Quisqualis indica L. flower on experimental esophagitis in albino Wistar rats. Indian J Exp Biol. 2017; 55: 122–126. https://scispace.com/pdf/effect-of-ethanolic-extract-of-quisqualis-indica-l-flower-on-72q1halddp.pdf

34. Rout PK, Kumar P, Rao YR, Kumar A, Bawankule DU, Singh R, et al. A quinoline alkaloid rich Quisqualis indica floral extract enhances the bioactivity. Natural Product Research. 2021 May 19; 35(10): 1632–1638. doi: 10.1080/14786419.2019.1634709

35. Indumathi P, Abi Beaulah G. Antifungal and Cytotoxic Activities of Hibiscus rosa-sinensis Flower Extract: A Pharmacological Study. Intern J Zool Invest. 2023; 9(2): 671–685. doi: 10.33745/ijzi.2023.v09i02.077

36. Lukić M, Pantelić I, Savić SD. Towards Optimal pH of the Skin and Topical Formulations: From the Current State of the Art to Tailored Products. Cosmetics. 2021 Aug 4; 8(3): 69. doi: 10.3390/cosmetics8030069

37. Kim YJ, Uyama H, Kobayashi S. Inhibition effects of (+)-catechin–aldehyde polycondensates on proteinases causing proteolytic degradation of extracellular matrix. Biochemical and Biophysical Research Communications. 2004 Jul; 320(1): 256–261. doi: 10.1016/j.bbrc.2004.05.163

38. Oyinloye BE, Agbebi EA, Agboola OE, Ubah CS, Owolabi OV, Aruleba RT, et al. Skin Anti-Aging Potentials of Phytochemicals from Peperomia pellucida against Selected Metalloproteinase Targets: An In Silico Approach. Cosmetics. 2023 Nov 2; 10(6): 151. doi: 10.3390/cosmetics10060151

39. Nutho B, Tungmunnithum D. Exploring Major Flavonoid Phytochemicals from Nelumbo nucifera Gaertn. as Potential Skin Anti-Aging Agents: In Silico and In Vitro Evaluations. IJMS. 2023 Nov 21; 24(23): 16571. doi: 10.3390/ijms242316571

40. Nutho B, Tungmunnithum D. Anti-Aging Potential of the Two Major Flavonoids Occurring in Asian Water Lily Using In Vitro and In Silico Molecular Modeling Assessments. Antioxidants. 2024 May 14; 13(5): 601. doi: 10.3390/antiox13050601

Received on 07.06.2025 Revised on 04.10.2025

Accepted on 03.12.2025 Published on 05.06.2026

Available online from June 06, 2026

Research J. Pharmacy and Technology. 2026;19(6):2751-2759.

DOI: 10.52711/0974-360X.2026.00393

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. Creative Commons License.

Parameters	Face cream prepared using the flower extracts of
	Ixora coccinea	Clitoria ternatea	Quisqualis indica	Hibiscus rosa-sinensis
Colour	Orange	Purple	Light orange	Red
Flow behaviour	No flow when turned upside down
Texture	Silky, creamy, and dense
Phase separation	Absent
Uniformity	Uniform
Smell	Rose scent
Consistency	Excellent
Coarse particles	Absent
Firmness	Absent
Oiliness	Absent
Absorption	in a minute or two