INTRODUCTION:

The safety and popularity of natural products have induced the need to produce herbal-based products. The herbs used in cosmetic product have various benefit like antioxidant, anti-inflammatory, antiseptic, and antibacterial properties. Herbal cosmetic products are believed to have no side effects commonly seen with products containing synthetic agents¹. Antioxidants are molecules contrary to free radicals. Some free radicals may damage the tissues, which causes aging and even cancers. Such conditions can be controlled by antioxidants². As plants produce many antioxidants to prevent the oxidative stress caused by sunbeams and oxygen, they can represent a source of new compounds with antioxidant activity³.

Cosmetic product can be used on the human body, e.g. (teeth, nails, face, and hair). These products are used to keep the body in good condition, change its appearance, and remove body odors via perfuming, cleansing, or protection⁴. Today's use of facial mask products has grow rapidly, making them the world's fastest-growing segment in personal care regimens, including in Asia⁵. Therefore, face masks have good potential to be developed into cosmetic products.

Damage to skin collagen/ elastin, accompanied by hyperpigmentation, inflammation, and dehydration can be caused by environmental influences, such as UV rays and micro/nano particles outdoors and indoors. Fine lines, wrinkles, and age spots, which trigger premature aging, can be corrected by the breakdown of important components of the extracellular matrix. Therefore, there is consumer demand for innovative face mask formulations that can neutralize the side effects of cigarette smoke, gases, particulate matter, heavy metals, ozone, free radicals and UV rays, especially for those living in urban areas. This demand arises because of the desire to have products that have moisturizing and anti-aging activities and are easy to use, low cost, and made from natural ingredients⁵.

Masks can be formulated as creams that deliver "active substances" to the skin, improving its appearance and quality. Different packaging and ingredients of this cosmetic category can be selected according to skin type; for instance, cream-based masks are best suited for dry skin⁵. Face masks are products that can be used easily and show an effect after several uses on the skin. Bioactive ingredients with different mechanisms are added to masks to provide moisturizing, brightening, and herbal ingredients⁶. A Cream is a preparation used for the application to the skin. Face Creams are used as cosmetics for softening and cleansing action⁷.

A face cream formulation from passion fruit seed oil of the Passiflora edulis has been marketed in Indonesia. The facial cream formulations in this research were different species than commercial products and types of cream. Facial creams that have been registered are used without rinsing, thus giving an effect like a vanishing cream, while the type of cream in this research was rinsed or washing cream.

Several researchers have researched sweet granadilla (P. ligularis), such as peel and pulp with seeds against DPPH, by showing strong antioxidant activity⁸ and leaves of sweet granadilla (P. ligularis) by optimizing the total flavonoid content⁹. In addition, it has also been investigated the presence of strong antioxidant activity of DPPH from ethanol extract of leaves and stems of P. edulis followed by fractionation using petroleum ether and chloroform¹⁰. This shows that the genus Passiflora has good potential to be used as an antioxidant.

The anti-aging potential of Passiflora genus plants has been tested using the ethanol extract of P. edulis f. edulis Sims with in vitro tests of several enzymes related to skin aging, namely collagenase, elastase, and tyrosinase enzymes. Tyrosinase inhibitory value IC₅₀ = 141.30 μg/ml¹¹ and other tested tyrosinase inhibitory effect 39.9 ±0.0% at 1mg/ml which is useful in functional cosmetic formulations¹².

A product requires an appropriate formulation and good product stability. A new cosmetic product or a modified product needs to undergo product stability testing to ensure that the product meets the intended physical, chemical and microbiological quality standards according to its function and aesthetics when stored under suitable conditions¹³.

Based on that, this research was conducted on the effects of antioxidants, tyrosinase inhibition, formulation, and stability of face mask cream preparations derived from the ethyl acetate fraction of sweet granadilla seeds as the natural ingredients for daily use.

MATERIALS AND METHODS:

Plant material:

The plant material used is sweet granadilla seeds (P.ligularis) obtained from Batu Dalam village, Danau Kembar sub-district, Solok district, West Sumatra province, Indonesia, and has been determined at Herbarium Bogoriensis, Center for Biological Research, LIPI, Indonesia.

Chemical material:

The chemicals used included the enzyme tyrosinase T3824 (Sigma Aldrich, USA), ascorbic acid (Merck, Germany), kojic acid (Sigma Aldrich, USA), 3,4-Dihydroxy-L-phenylalanine (L-DOPA) (Sigma Aldrich, USA), HCl (Merck, Germany), aqua demineralisata (Brataco Chemika, Indonesia), 1,1-Diphenyl-2-picrylhydrazyl (DPPH) (TCI, Japan), aqua pro injection, methanol pro analysis (Merck, Germany), potassium dihydrogen phosphate (Merck, Germany), sodium acetate (Merck, Germany), 2,4,6-Tris(2-Pyridyl)-S-Triazine (TPTZ) (Sigma Aldrich, USA), iron (III) chloride hexahydrate (Merck, Germany), iron (II) sulfate heptahydrate (Merck, Germany), propylene glycol (Dow Chemical Pacific, Singapore), polyethylene glycol 400 (Alkamidco, Iran), glycerin (PT. Sumi Asih , Indonesia), kaolin (Takehara Kagaku Kogyo, Japan), tri ethanolamine (Petronas, Malaysia), methylparaben (Gujarat Organics ltd, India), propylparaben (Gujarat Organics ltd, India), sodium metabisulfite (Aditya Birla Chemicals, Thailand), tutty fruity oil (Chemamore, Indonesia), stearic acid (PT. Sumi Asih, Indonesia), cetyl alcohol (Ecogreen Oleochemicals, Singapore), titanium dioxide (Daito Kasei Kogyo, Japan) and distilled water (CV. Satya Darmawan, Indonesia).

Extraction and Fractionation:

The extraction and fractionation processes have been carried out according to the previous studies by Wiliantari et al (2022)¹⁴.

Ethyl Acetate Fraction Face Mask Cream Formulation Passiflora ligularis Juss Seed:

Face mask cream was made in formulation 1(F1) with a fraction concentration of 1% (10,000μg/ml) and formulation 2 (F2) with a fraction concentration of 2% (20,000μg/ml). This concentration was chosen based on the study evaluation from previous research by Wiliantari et al (2022)¹⁴.

Preparation of Facial Mask Cream:

The facial mask cream consisted of the ethyl acetate fraction of P. ligularis seeds, and all other ingredients were prepared according to those presented in Table 1. The oil phase consisting of stearic acid and cetyl alcohol was melted first at a temperature of 60^oC using a hot plate. In the aqueous phase, methylparaben, propylparaben, and sodium metabisulfite are dissolved in propylene glycol first. Then the distilled water was heated at 60^oC in the container. After reaching the temperature, the water phase material was added (propylene glycol, PEG 400, Glycerin, and TEA) while stirring with a homogenizer (Ika Eurostar 20, Malaysia) at a speed of 1,000rpm. After mixing everything, add the oil phase until a creamy mass was formed and stir continuously for 10min. Then add kaolin and stirred until homogeneous. Then, add the titanium dioxide, stirred until homogeneous for 10min and add the tutty fruity oil. Gradually lower the temperature to 40^oC, add the ethyl acetate fraction of the seeds, and then stirred again until everything was distributed homogeneously.

Table 1. Face mask cream formulation (% w/w)

Ingredient	Function	Formula
Ingredient	Function	F1 (%)	F2 (%)
Seed ethyl acetate fraction	Active substance	1	2
Propylene glycol	Humectants	5	5
PEG 400	Humectants	8	8
Glycerin	Humectants	10	10
Kaolin	Opacifier	15	15
Triethanolamine (TEA)	Surfactant	1	1
Stearic acid	Emulsifier	6	6
Cetyl alcohol	Emollient	2	2
Titanium dioxide	Pigment	2	2
Methylparaben	Preservative	0.18	0.18
Propylparaben	Preservative	0.02	0.02
Sodium metabisulfite	Antioxidant	1	1
Tutty fruity oil	Deodorizer	0.069	0.069
Distilled water	Solvent	Ad 100	Ad 100

The face mask cream that had been formed was put into the prepared pot, and the final evaluation was carried out. The control preparation was similar to the face mask cream but without ethyl acetate fraction P.ligularis.

Preparation Evaluation:

Organoleptic:

The organoleptic test was carried out visually and directly by seeing the shape, color, and smell of the cream of the face mask.

Homogeneity:

The face mask cream was spread on an object glass and covered with another object glass, observed for the presence or absence of coarse particles that were formed or not homogeneous. Observations were made under the light.

pH stability:

The preparation was weighed ±1g and dissolved in 100 ml distilled water. Then the pH test was carried out. Prior to the test, the pH meter (Ohaus ST3100-F, USA) was calibrated with standard buffers at pH 4, 7, and 10. The pH electrode was immersed in the preparation, and the observed pH value was recorded. Measurements were carried out at room temperature (25±2^oC). The results obtained are expected to be in the pH range of 4.0 - 6.0 according to the pH of the face area¹⁵.

Globule Diameter Measurement:

A particle size analyzer (Malvern, UK) analyzed the globule diameter measurement to determine vesicle size, polydispersity index, and zeta potential. The preparation was diluted with distilled water, and the measurements were read at a scattering angle of 90^o and a temperature of 25±1^oC.

Testing of Viscosity and Flow Properties:

The viscosity and flow properties were measured by a viscometer (Cole Parmer Rotational Viscometer, USA). The preparation was put into a measuring cup. Please select the appropriate spindle (spindle L4), attach it to the tool, then dip it into the preparation until the limit of the spindle was immersed. Select the desired spindle type and rpm speed according to the table provided in the manual. Press the 'on' button to start measuring viscosity and wait for the spindle to rotate for up to 5 revolutions or until it stabilizes after that, read the viscosity (Cp or mPa.s units) printed on the screen. Ensure the percentage scale on the screen was more than 15% and below 100%¹⁶.

Temperature Stability Test:

Cycling test:

The face mask cream was tested by cycling test by storing it at 4^oC along 24hours, then transferred to the oven at 40±2^oC along 24hours, this is counted as one cycle. The test was repeated for 6 cycles, then organoleptic observations were carried out¹⁷.

Stability test at room temperature:

The face mask cream was stored at room temperature (30±2ºC) for 12 w. At weeks 0, 2, 4, 8, and 12, organoleptic observations (change in color, odor, and presence or absence of syneresis), pH, and homogeneity were carried out¹⁸.

Accelerated stability test:

The face mask cream was stored at 40±2ºC for 12 w. At weeks 0, 2, 4, 8, and 12, organoleptic observations (change in color, odor, and presence or absence of syneresis), pH, and homogeneity were carried out¹⁸.

Antioxidant Activity:

DPPH (2,2-diphenyl-1-picrylhydrazyl) method:

The antioxidant activity assay using the DPPH method followed the previous procedure by Hapsari et al. with a slight modification¹⁹.

Preparations (blanks, F1, and F2) were prepared first. Weigh ±1g of the preparation and dissolve it in 10ml of methanol. Then it was centrifuged (Hettich Zentrifugen, Germany) with speed 4500rpm along 30minutes. analysis was carried out using the supernatant section. The concentration of F1 became 1,000μg/ml and F2 2,000μg/ml. Then, a series of concentrations was made into 1.5, 3.0, 4.5, 6.0, and 7.5μg/ml.

An amount of 8ml of the series solution concentration was pipetted, then put into a dark-colored vial, and 2ml of 0.3mM DPPH solution was added. Then, it was vortexed for 20 s and incubated at room temperature for 30min in a dark room. Ascorbic acid was used as a standard or positive control and then measured with a T80+ UV/Vis Spectrophotometer at a maximum DPPH wavelength of 516nm. Each procedure was repeated three times. The IC₅₀ value was calculated based on the percentage of inhibition of the DPPH radical from each concentration of the sample solution with the equation:

% Inhibition = [(control absorption – sample absorption) x 100%]/control absorption

After obtaining the percentage of inhibition from each concentration, the equation y = a + bx was determined by calculating a linear regression curve, where x is the concentration (μg/ml) and y is the percentage of inhibition (%).

IC₅₀ = (50 – a)/b

FRAP (ferric reducing antioxidant power) method:

The FRAP method testing of antioxidant activity was done by following the procedure from Ernawati et al., and Vongsak et al. with slight modifications^20,21.

Preparations (blanks, F1 and F2) were prepared first. Weigh ±1g of the preparation and dissolve it in 10ml of methanol p.a. Then it was centrifuged (Hettich Zentrifugen, Germany) with speed 4500rpm along 30 minutes. analysis was carried out using the supernatant section. The concentration of F1 became 1,000μg/ml and F2 2,000μg/ml. Then a concentration series of 10 μg/ml was made with a concentration in the well of 2.5 μg/ml. The blank was carried out in the same dilution as the preparation. It was essential because the excipient of the preparation contained several metals that could participate in the reaction. The FRAP I Working Solution for the calibration curve was prepared by mixing acetate buffer, TPTZ (tris pyridyl triazine), and distilled water in a ratio of 10:1:1. Meanwhile, the FRAP II working solution for the sample or positive control was prepared by mixing acetate buffer solution, TPTZ, and iron (III) chloride hexahydrate in a ratio of 10:1:1.

To make a calibration curve, 150μl of FRAP I working solution was added to 50μl of standard solution or distilled water (blank I). As for testing samples or positive controls, 150μl of the FRAP II working solution was added to 50μl of the preparation solution or ascorbic acid. As a blank sample or positive control (blank II), a mixture of 50μl of methanol and 150μl of FRAP II working solution was used. Then, it was incubated (Gemmy Industrial Lab Incubator, Taiwan) with temperature 37°C along 8 min in dark condition. The mixture was shaken in a shaker (Biotek 800 TS Absorbance Reader, USA) for 30s, and absorption measurements were made at a wavelength of 590nm using a microplate reader. The test was carried out 3 times.

The calibration curve was made from the plot of concentration (x) in mM vs. net absorption (y) of standard series solution of iron (II) sulfate heptahydrate so that the linear regression equation y = bx + a was obtained. The FRAP value was calculated by the following equation:

FRAP value (μmol/g sample) = (C x V x Fp) / m

Where C is the concentration of standard solution Fe₂SO₄.7H₂O, equivalent to the sample (μmol/ml), is obtained by entering the net absorption value of the sample into the calibration curve equation. V is the sample volume (ml), Fp is the dilution factor, and m is the sample (g) weight. The antioxidant activity of the sample is expressed as grams of Fe₂SO₄ equivalent per 100 g of extract, which is calculated by the following equation:

Antioxidant activity = FRAP value x 10^-6 x Mr Fe₂SO₄.7H₂O x 100

Tyrosinase Inhibition:

The testing procedure was based on previous research with slight modifications. An initial sample was carried out in the face mask cream preparation before testing the tyrosinase enzyme inhibition. A total of ±1g of the sample was weighed and added 10ml of 50mM phosphate buffer pH 6.5 was. Then centrifuged (Hettich Zentrifugen, Germany) with speed 4500rpm along 30 minutes. analysis was carried out using the supernatant section. The concentration of F1 became 1,000μg/ml and F2 2,000μg/ml.

A total of 80μl of 50mM phosphate buffer pH 6.5, 40μl of sample solution (sample preparation of facial mask cream 50-1,000μg/ml), 40μl(4mM) of L-DOPA solution, and 40μl(75U/ml) of tyrosinase enzyme solution were pipetted into a 96-well microplate. The solution mixture was incubated (Friocell MMM, Germany) for 30 min at 25˚C. Then the mixture was shaken for 60 s, and the absorbance was measured at a wavelength of 490nm (GloMax Microplate Reader, USA). Control samples were made without the addition of the tyrosinase enzyme.

Blanks were prepared by pipetting 80μl of 50mM phosphate buffer pH 6.5, 40μl (4mM) of L-DOPA solution, 40μl of blank preparation, and 40μl (75U/ml) of tyrosinase enzyme solution into a 96-well microplate (Thermo Scientific, USA). The control blank was made without the addition of the tyrosinase enzyme. Tests were carried out in triples. The percentage of inhibition was calculated with this equation below:

[(A-B) – (C-D)]

% Tyrosinase Inhibition = -------------------------- x 100

(A- B)

Description: A = absorbance of blank solution with enzyme (blank); B = absorbance of blank solution without enzyme (blank control); C = absorbance of sample solution with enzyme (sample); D = absorbance of sample solution without enzyme (sample control).

RESULTS AND DISCUSSION:

Cycling test:

The cycling test was used to obtain information regarding the stability of preparations against extreme temperature changes that may occur every year or every day. The cycling stability test was carried out with 6 cycles on face mask cream preparations and blanks. F1 and F2 were stored for 24 hours at 4ºC and 24hours at 40ºC. Organoleptic observations at the end of the 6 cycles showed no significant difference with the organoleptic observations of usual mask cream preparations compared to the initial conditions. Figure 1 showed that the usual face mask cream was in semisolid form, pale-dark cream color, and had a characteristic aromatic smell. After the test, the appearance of the face mask cream preparation did not change in shape, color, and smell, so it can be concluded from the cycling test that the F1 and F2 face mask cream preparations had good stability.

The cycling test study involved storing the preparation at alternating high and low temperatures in closed containers. Samples were tested after fixed time intervals primarily for changes in their physical attributes (e.g., consistency and homogeneity). As part of accelerated stability testing, such thermal stress conditions provide an overview of the information regarding the stability and behavior of the preparation under various storage conditions in a few weeks ²².

Organoleptic:

The results of organoleptic observations of preparations from 0, 2, 4, 8, and 12 w showed a change in the preparation, as shown in Figure 2 (0 and 12w) and Table 2. Changes were mainly seen in the color of the 0-w preparation from light beige to dark beige, which changed color after 12w of storage. Changes were primarily seen at storage temperatures of 40 ± 2ºC at F1 and F2. This can occur due to the degradation of excipient ingredients and active substances, such as sodium metabisulfite, an antioxidant in topical preparations, and active substances in preparations with antioxidant properties. In F2, the concentration of the ethyl acetate fraction of seeds was 2% more, and more degradation occurred than in F1, indicated by a change in the color of the preparation to dark cream.

Initial formulation studies should be designed to subject the preparation to several "stress conditions" to identify the significant degradation pathways and levels of degradation. From these studies, it is possible to estimate the potential stability of a chemical or biological substance under environmental conditions that may occur during synthesis or extraction, manufacturing, transportation, and storage²³.

Homogeneity:

The face mask cream preparations showed homogeneous results in the blanks, F1, and F2, in the absence of coarse particles. The cream preparations were evenly distributed on the glass object, as shown in Figure 3.

Figure 1: The results of the cycling test: (A) after the 0 cycle and (B) after the 6 cycles

pH Stability:

Evaluation of the stability of the preparation against pH showed a decrease during storage. The pH value of facial mask cream F1 and F2 at the storage of 30±2^oC and 40±2^oC decreased from the initial conditions, as shown in Figure 4. The decrease in pH value was more visible at a storage temperature of 40±2^oC. The pH value of facial mask cream F1 and F2 still met the safety requirements after 12 w of storage. This decrease in pH can be caused by the degradation of the ingredients in the preparation, such as materials containing carboxylic acid groups, such as stearic acid.

The skin surface has a pH ranging from 4.0 to 7.0 which is mostly acidic with a pH value between 5.4 and 5.9. The physicochemical properties of the skin are influenced by various factors, one of which is pH. Changes in any of these features impact the overall physiology of the skin²⁴.

Table 2: Organoleptic observations

Formula	Organoleptic observations
Blank	Shape: semisolid White color Smell: aromatic
F1	Shape: semisolid Color: light cream Smell: aromatic
F2	Shape: semisolid Color: dark cream Smell: aromatic

0-wTemp. 30 0-w Temp. 40

12 w Temp. 30 12 w Temp. 40

Figure 3: Homogeneity observation

Figure 4: pH stability

Globule Diameter Measurement:

The results showed that the face mask cream preparations in blanks, F1 and F2 at 0 and 12 w had an average particle size and uniform polydispersity index, as shown in Table 3. The particle diameter of the dispersed phase in a semisolid emulsion system generally ranged from 0.1m – 10m (100nm -10000nm). Face mask cream preparations still meet the requirements. In Fahr (2018) mentioned that the particle size must be adjusted to the application of the desired preparation. For example, it does not cause mechanical irritation on application to the skin.

The Zeta potential of the preparation at 0 and 12 w tends to increase. This value was still acceptable and had a suitable category, as in Table 3. In Fahr (2018) mentioned that for electrostatically stabilized pharmaceutical suspensions, the zeta potential value must be at least 50mV (positive or negative) to prevent agglomeration and aggregation.

Table 3: Size distribution, polydispersity index, and zeta potential

Formula	Week	Average particle size D₉₀(d.nm)	The polydispersity index (PDI)	Zeta potential (mV)
F1 (30 ± 2 ^oC)	0	1120	0.380	-34.8
F1 (30 ± 2 ^oC)	12	4040	0.342	-31.0
F2 (30 ± 2 ^oC)	0	413	0.398	-30.5
F2 (30 ± 2 ^oC)	12	1090	0.254	-34.3
F1 (40 ± 2 ^oC)	0	1120	0.380	-34.8
F1 (40 ± 2 ^oC)	12	6110	0.230	-42.3
F2 (40 ± 2 ^oC)	0	413	0.398	-30.5
F2 (40 ± 2 ^oC)	12	1250	0.291	-41.8

Viscosity and Flow Properties:

The preparation has the property that it will not flow until a particular force is exceeded, which is called the yield value. The plastic flow curve does not pass through the point (0,0) but intersects the shearing stress at a certain point, namely the yield value. The yield value is caused by the contact between adjacent particles that must be broken before flow occurs. In contrast, a thixotropic flow can be found in substances that have plastic flow, which indicates a breakdown of the structure that does not form immediately if the stress is removed or reduced. The flow curve depends on the increasing and decreasing rate of shear and the length of time the substance experiences the rate of shear. The research results on facial mask cream preparations at F1 and F2 showed plastic flow properties, as shown in Figure 5A-5D, and the viscosity value in Table 4.

Table 4: Viscosity measurement

Week

Viscosity (Cp) at 60 rpm, spindle L4

Viscosity (Cp) at 30 rpm, spindle L4

temp. 30

temp. 40

temp. 30

temp. 40

8228.9

8675.2

8674.2

10097(50rpm)

10007 (50rpm)

8228.9

8262.8

8273.3

8145.2

8114.8

13617

14386

16164

16436

16730

13617

13409

13081

12133

12082

Face Mask Cream Antioxidant Activity:

DPPH method:

In the face mask cream preparation, the antioxidant activity using the DPPH method in Formula 1 (F1) and Formula 2 (F2) showed antioxidant activity in a strong category, as shown in Figure 6A, which shows antioxidant activity during storage at 0, 2, 4, 8 and 12 w. The antioxidant activity of the face mask cream showed a decrease in the IC₅₀ value during storage at temperature storage conditions of 30 ± 2 ^oC and 40 ± 2 ^oC. The DPPH method (2,2-diphenyl-1-picrylhydrazy) is commonly known method to evaluate the antioxidant activity^25–27. This method is based on electron transfer, which is characterized by a change in color to violet in ethanol solution. In addition, these free radicals are also stable at room temperature, and are easily applied using the spectrophotometer method^28–30. The increase and decline of the IC₅₀ value at 2 and 4 w can be caused by the distribution of the less homogeneous fraction in the preparation during the manufacturing process. In addition, there can also be degradation of the ingredients in the preparation, significantly compounds that are antioxidants at the value of DPPH activity decreases during 12 w of storage.

FRAP method:

Antioxidant activity using the FRAP method on the preparation of face mask cream ethyl acetate fraction of sweet granadilla fruit seed 0 w (temperature 30 ± 2 ^oC and 40 ± 2 ^oC) in Formula 1 (F1), which is 1.91 ± 0.029 g Fe₂SO₄ equivalent/100 g of sample and Formula 2 (F2) is 2.69 ± 0.015 g Fe₂SO₄ equivalent/100 g sample. F2 showed a higher activity value because the concentration of the fraction in the preparation was higher than F1. The increase and decrease of the FRAP value in the 2 and 4 w can be caused by the distribution of the fraction that is less homogeneous in the preparation during the manufacturing process, as shown in Figure 6 B. In addition, there can also be degradation of the ingredients in the preparation, especially compounds that are antioxidants, so the activity value FRAP decreased during 12 w of storage.

Table 5: Antioxidant activity, pH, and tyrosinase inhibition

Formula	Temperature (^oC)	Weeks	pH	DPPH (IC_50,μg/ml)	FRAP (g Fe2SO4 equivalent/100 g sample)	Tyrosinase inhibition (%)
F1	30 ± 2	0	6.28	7.79 ± 0.044	1.91 ± 0.029	45.6 ± 2.795
		2	5.89	9.11 ± 0.042	1.79 ± 0.090	59.9 ± 3.267
		4	5.83	10.14 ± 0.070	2.03 ± 0.087	15.8 ± 4.399
		8	5.89	9.44 ± 0.047	1.18 ± 0.032	22.6 ± 3.820
		12	5.64	9.26 ± 0.045	1.00 ± 0.104	11.0 ± 2.069
F2	30 ± 2	0	6.14	7.44 ± 0.029	2.69 ± 0.015	65.9 ± 3.352
		2	5.45	8.80 ± 0.025	1.83 ± 0.098	68.4 ± 0.381
		4	5.53	7.55 ± 0.023	2.19 ± 0.012	48.5 ± 2.429
		8	5.38	7.27 ± 0.022	1.87 ± 0.021	62.4 ± 4.042
		12	4.93	8.89 ± 0.109	1.99 ± 0.036	44.7 ± 1.906
F1	40 ± 2	0	6.28	7.79 ± 0.044	1.91 ± 0.029	45.6 ± 2.795
		2	5.81	11.09 ± 0.017	1.70 ± 0.040	48.8 ± 1552
		4	5.52	9.03 ± 0.057	1.98 ± 0.056	31.6 ± 2.104
		8	4.89	11.21 ± 0.059	1.54 ± 0.027	25.3 ± 1.636
		12	4.75	11.74 ± 0.186	1.20 ± 0.074	38.3 ± 1.790
F2	40 ± 2	0	6.14	7.44 ± 0.029	2.69 ± 0.015	65.9 ± 3.352
		2	5.20	8.18 ± 0.008	1.52 ± 0.082	48.4 ± 3.687
		4	4.99	6.99 ± 0.037	1.95 ± 0.027	38.8 ± 1.919
		8	4.68	8.71 ± 0.064	0.67 ± 0.078	51.4 ± 1.267
		12	4.45	9.25 ± 0.179	1.51 ± 0.038	50.6 ± 1.319

Data were expressed as mean value ± SD (n = 3)

CONCLUSION:

The face mask cream showed very strong antioxidant activity and tyrosinase inhibition potential in F1 and F2. The 12-w stability showed a decrease in antioxidant activity, tyrosinase inhibition, and physical evaluation of the preparation in F1 and F2. The best test results during 12 w of storage were seen at F2 with a temperature of 30 ± 2 ^oC.

CONFLICTS OF INTEREST:

The authors have no conflicts of interest in this work.

ACKNOWLEDGMENTS:

The authors thank Universitas Indonesia for funding this research through the PUTI Pascasarjana Grant with contract number NKB-074/UN2.RST/HKP.05.00/2022. The facilities, scientific, and technical support from Advanced Characterization Laboratories Serpong, National Research and Innovation Agency (BRIN) through E-Layanan Sains, also the National Institute of Health Research and Development, Ministry of Health, Republic of Indonesia.

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Received on 16.12.2022 Modified on 10.04.2023

Research J. Pharm. and Tech 2023; 16(11):5255-5263.

DOI: 10.52711/0974-360X.2023.00852

0 w Temp. 30	0 w Temp. 40

12 w Temp. 30	12 w Temp. 40

Ethyl Acetate Fraction Face Mask Cream Formulation Passiflora ligularis Juss Seed:

Preparation Evaluation:

Organoleptic:

Homogeneity:

pH stability:

Globule Diameter Measurement:

Testing of Viscosity and Flow Properties:

Temperature Stability Test:

Cycling test:

Stability test at room temperature:

Accelerated stability test:

FRAP value (μmol/g sample) = (C x V x Fp) / m

Antioxidant activity = FRAP value x 10-6 x Mr Fe2SO4.7H2O x 100

Cycling test:

Table 2: Organoleptic observations

Formula

Organoleptic observations

Blank

Shape: semisolid

White color

Smell: aromatic

F1

Shape: semisolid

Color: light cream

Smell: aromatic

F2

Shape: semisolid

Color: dark cream

Smell: aromatic

Table 3: Size distribution, polydispersity index, and zeta potential

Formula

Week

Average particle size D90 (d.nm)

The polydispersity index (PDI)

Zeta potential (mV)

F1

(30 ± 2 oC)

0

1120

0.380

-34.8

12

4040

0.342

-31.0

F2

(30 ± 2 oC)

0

413

0.398

-30.5

12

1090

0.254

-34.3

F1

(40 ± 2 oC)

0

1120

0.380

-34.8

12

6110

0.230

-42.3

F2

(40 ± 2 oC)

0

413

0.398

-30.5

12

1250

0.291

-41.8

Table 4: Viscosity measurement

Antioxidant activity = FRAP value x 10^-6 x Mr Fe₂SO₄.7H₂O x 100

Average particle size D₉₀(d.nm)

(30 ± 2 ^oC)

(30 ± 2 ^oC)

(40 ± 2 ^oC)

(40 ± 2 ^oC)