INTRODUCTION:

White and fair skin is the ideal standard of beauty for every woman, so some who have dark or tan skin feel insecure. Survey data ZAP Beauty Index (2020) describes the results of 73.1% of Indonesian women think the definition of beautiful is having clean white skin and glowing. This encourages the desire of many women who dream of having white skin^1,2.

The skin functions as a protector from exposure to UV rays. Exposure to UV rays for a long time can often cause skin problems, namely hyperpigmentation³. Hyperpigmentation is a condition of uneven pigmentation in the skin, and an increase in excess melanin production which causes darkening or an increase in skin color.

Prevention of overproduction of melanin, one of which is by reducing the concentration of melanin or known as whitening agent. Treatment of hyperpigmentation in general uses hydroquinone, kojic acid, arbutin, but they have harmful side effects on the skin^4,5. Many several ways to reduce side effects, compounds from natural ingredients are used.

Flavonoids and glabridin are found in licorice and have been shown to inhibit pigmentation and tyrosinase 16 times greater than hydroquinon^6,7. This licorice research has also been carried out by Rakhmini et al. (2018) but in the form of a face cream preparation, which resulted in a 20% concentration of licorice extract cream having a better effect as a skin lightener. Serum preparation will make it easier to apply to the skin. Serum preparations have the advantage that the concentration of the active ingredients is high so that the skin quickly absorbs them; therefore, when used, it is more comfortable and spreads quickly on the skin's surface because of its low viscosity⁸. Serum preparations are made to facilitate application on the skin. Serum preparations have the advantage that the concentration used of the active ingredients is high so that they are easily absorbed by the skin, therefore when used it is more comfortable and spreads easily on the surface of the skin because of its low viscosity⁹. Novelty in this study will make the formulation and evaluation of serum from licorice root extract (Glycyrrhiza glabra) a facial brightener. Glycyrrhiza glabra is one of the beneficial medicinal plants with increasing demand for health products, herbal medicines, and pharmaceuticals¹⁰.

MATERIALS AND METHODS:

Materials:

Licorice root, 96% ethanol, natrosol (Ashland), glycerin, DMDM hydantoin (Lonza), ethoxydiglycol (Making cosmetics), aquadest, HCl, Mayer reagent (Nitra Kimia), Wagner reagent (Nitra Kimia) and dragendrof reagent (Nitra Kimia), FeCl3, Magnesium (Mg).

Instrument:

Blender (Miyako), digital analytical balance (Fujitsu), incubator (Memmert), pH meter (AMT20), dropper, watch glass, object glass, beaker glass (Pyrex), measuring cup (Pyrex), rotary evaporator (Eyela), water bath (YNC-WBE-8L), Brookfield viscometer (NDJ-8S), skin tone chart, serum bottle, hot plate (maspion), stir bar, thermometer, microscope (XSZ107BN), petri dish, test tube (Pyrex), funnel glass, filter paper, measuring cup (Pyrex), rotary evaporator (Eyela), water bath (YNC-WBE-8L), Brookfield viscometer (NDJ-8S), skin tone chart, serum bottle, hot plate (maspion), stir bar, thermometer, microscope (XSZ107BN), petri dish, test tube (Pyrex), funnel glass, filter paper.

Experimental Step:

Approval Ethical:

This research was approved by the research ethics commission at Universitas Respati Yogyakarta, number 039.3/FIKES/PL/IV/2023.

Plant Identification:

Plant determination was conducted at the Ahmad Dahlan University Biology Laboratory (UAD) in Yogyakarta.

Extract Preparation:

1 kg of licorice root Simplicia that has been clean and dry, then chopped into small pieces, then pulverized using a blender to form a powder. Extraction was carried out by maceration method using 500 grams of liquorice powder. Then, soak the licorice powder using 96% ethanol solvent with a ratio of 1:5, and remacerate until the filtrate is clear. The filtrate obtained was then concentrated with rotary evaporators.

Phytochemical Screening:¹¹

a) Flavonoid Test, One gram of extract was added with magnesium and concentrated HCl, then heated for 15 minutes in a water bath to see if it formed a positive red or yellow color for flavonoids (flavones, kalkon, and auron).

b) Alkaloid Test, as much as 2 grams of the extract was dripped with 5 mL of 2 N HCl, then heated and cooled, divided into 3 test tubes, 1 mL each. Tube one, add reagent Mayer, positive if the precipitate is white or yellow. Tube two add reagent Wagner, positive if brown precipitate. Tube three adds reagent Dragendrof; an orange precipitate indicated positive.

c) Saponin Test, 1 gram of extract was added to 10 mL of hot and cold water and shaken vigorously for 10 seconds. If positive it was marked with 1-10 cm high foam for not less than 10 minutes, and when adding one drop of 2 N HCl, the foam did not disappear.

d) Tannin Test, 1 gram of extract was added to 10mL of hot water, then boiled for 5 minutes, then the filtrate was added FeCl3 3-4 drops. If it is blue-green (green-black), it is positive for catechol tannins, while if it is blue-black, it is positive for pyrogallol tannins.

Formulation of Sweet Root Serum:

This study used various concentrations of liquorice (G. glabra) 1%, 10% and 20% (Source: Anggraini. et al 2021).¹²

Table 1. Formulasi Sediaan Serum

Componen	Function	Concentration (%)			Negative Control	Positive Control
Componen	Function	F1	F2	F3	Negative Control	Positive Control
Licorice extract	Active substance	1%	10%	20%	0	Garnier Bright Complete
Natrosol	Gelling agent	0,75	0,75	0,75	0,75
Glycerin	Humectants	25	25	25	25
DMDM Hydantoin	Preservative	0,5	0,5	0,5	0,5
Ethoxydiglycol	Penetrant	2	2	2	2
Aquadest	Solvent	ad 100	ad 100	ad 100	ad 100

The serum was started with natrosol heated to 45ºC while stirring until an even suspension formed, and then heating stopped. The serum base was successively added to the natrosol mass and stirred until homogeneous. Add licorice extract with a concentration of 1%, 10%, and 20% into the basic formula that has been made, then put it in a serum bottle¹².

Evaluation of Serum Preparations

a) Organoleptic Test:

Observing changes in physical form, color, smell, and texture of serum preparations using the five senses. Record the results of these changes.

b) Homogeneity Test:

As much as 0.1gram of serum is spread evenly on the glass plate. Serum preparations must show a homogeneous composition, namely no clumping particles^13,14.

c) pH test:

Measurement of serum pH using a pH meter in triplo. Dissolve 1 g of serum in 10 ml of distilled water, dip the pH meter and record the pH ^13,14.

d) Viscosity Test:

Viscosity measurement using a viscometer brookfield. Put 100 ml of serum into a beaker, use spindle number 3 at 6 rpm^13,15.

e) Spreadability Test:

0.5 grams of serum is placed in the middle of a watch glass flanked by another watch glass, give a load of 150 grams and let it sit for 1 minute. Note its diffusing power. This test is as many as triplo and the desired spreadability is 5-7cm^12,14.

f) Stability Test:

The stability test was carried out for six cycles using the method freeze-thaw, where every cycle, which is 24hours, is placed in the refrigerator with a temperature of 4°C then transferred to the oven for 24 hours at 40°C. Test parameters include organoleptic tests and pH tests¹⁶.

g) Irritation Test:

The serum preparation was tested on 15 volunteers by applying the licorice serum preparation behind the ear. Observe the reaction that has occurred since the first basting. Positive response if there is redness, itching, or swelling of the treated skin¹⁷.

h) Hedonic Test:

The hedonic test uses a questionnaire method. The hedonic test used 30 respondents¹⁵. Each parameter is scored with a score of 5 which means very like, 4 means like, 3 means rather like, 2 means dislike and 1 means really dislike.

i) Face Brightening Activity Test:

They are testing the lightning activity using 15 volunteers. The inclusion criteria of women aged 20-25 years, facial skin type is not radiant, dull and dry, have no history of cosmetic allergies, and are willing to use the serum. Exclusion criteria for pregnant and lactating women, skin infections in the target area, using skin lighteners. Volunteers were divided into five groups, where each of the three volunteers used negative control serum preparations, formula 1, formula 2, formula 3, and positive control. Volunteers measured the initial condition of the skin using a skin tone chart to measure skin brightness. Basting is done twice a day for four weeks. Changes in skin condition were measured before and after using the serum. Measurements are taken once a week^8,18.

Statistic:

The calculated data was then performed using the Shapiro-Wilk normality test to determine whether the data in each group was normally distributed (p> 0.05). The test method uses the Kruskal Wallis and the Paired Sample Test.

RESULT:

The researcher examined simplicia both macroscopically and microscopically on liquorice and found that liquorice powder showed fiber, wood vessels, oxalic acid and cork cells. While the macroscopic results are seen as yellow, have a distinctive aromatic smell, have a very sweet but chelate taste in line with the microscopic and macroscopic test results in Swati Tomar's research in 2020¹⁹.

Licorice root extract is made using the maceration method. The maceration method was used because the compound to be used in the research is glabridin from the flavonoid group which is not resistant to high temperatures. The advantages of using the maceration method are the simple way of working and the equipment used. The extraction uses 96% ethanol because it is universal. 96% ethanol has a lower water content than 70% ethanol so the effect of the extract growing on fungus or mold is not too high. The maceration storage process is carried out in a dark place because to reduce the risk of catalysts occurring between the maceration material and sunlight and to prevent discoloration^20,21.

Table 2. Results of Licorice Extract Yield Test

Sample

Simplicity weight

Extract

weight

Result (%)

Licorice

500 g

78,1 g

15,62 %

Results of Evaluation of serum Preparations

a) Organoleptic Tests

Table 3. Observation Results of Organoleptic Test

Serum preparations	Form	Smell	Color
Formula 1	It’s kind a thick	Typical aromatic	Yellow golden
Formula 2	It’s kind a thick	Typical aromatic	Chocolate
Formula 3	It’s kind a thick	Typical aromatic	Dark chocolate
Positive control	It’s kind a thick	No smell	Clear cloudy
Negative control	It’s kind a thick	Typical lemon	Clear cloudy

b) pH test:

Table 4. pH test results

Serum preparations	pH 1	pH 2	pH 3	Average ± SD
Formula 1	5.70	6.02	6.07	5.93±0.20
Formula 2	5.96	6,10	6,17	6.07±0.11
Formula 3	6.00	6,32	6,24	6,18±0.17
Positive Control	4.94	6.02	6.07	5,67±0.64
Negative Control	5.94	5.79	6.02	5.86±0.12

c) Viscosity test

Table 5. Viscosity Test Results

Sampel	Viskosity (mPas)			Average±SD
Sampel	1	2	3	Average±SD
Formula 1	2100	2140	2160	2133±30,6
Formula 2	2040	2300	2560	2300±260,0
Formula 3	2780	2660	3100	2847±227,4
Negative control	3140	3180	3380	3233±128,6

d) Spreadability Test

Table 6. Power Spread Test results

Sample	Spreadability (cm)			Average±SD
Sample	1	2	3	Average±SD
Formula 1	6,0	6,7	6,3	6,3±0,35
Formula 2	5,5	5,4	5,5	5,5±0,06
Formula 3	4,8	5,2	5,1	5,0±0,21
Negative control	7,0	6,9	6,7	6,8±0,21
Positive control	5,7	6,2	6,4	6,1±0,36

e) Stability Test

Figure 1. Graph of pH stability

f) Hedonic Test

Hedonic testing based on texture, aroma and color parameters of serum preparations can be seen in the following figure:

Figure 2. Textured Hedonic Graph

g) Face Brightening Activity Test:

The graph of facial brightening activity testing can be seen in the picture following:

Figure 3. Face Lightening Activities

DISCUSSION:

The yield of licorice root extract was 15.62%. The yield obtained was higher than the study by Sogandi et al (2019) which obtained a yield of 6.76% and also higher than the study by Sathya et al (2017) of 12.7%^22,23. The higher the yield of the extract, the higher the content of the active substance or bioactive content that is attracted. The results of the phytochemical screening experiment were positive for containing flavonoids, alkaloids, saponins, and tannins. There are in line with research conducted by Simmy Gupta in 2019 with stated that the phytochemicals screening revealed the presence of alkaloids, saponins, triterpenoids, and phytosterols in root extract of Glycyrrhiza glabra²⁴.

Results of Evaluation of serum Preparations:

Some research on liquorice has also been made into gel preparations to treat oral ulcers²⁵, optimizations of spray drying process parameters for liquorice extract²⁶, as antibacterial against multidrug resistant bacterial pathogens of E.coli²⁷, Hand washing herbal formulas are more efficient in reducing the number of organisms with less or no side effects²⁸, liquorice is widely used for dental care such as dental caries , oral infections and other related endodontic problems²⁹

a) Organoleptic Tests:

The results of organoleptic observations of the physical properties of formulas 1, 2 and 3 have the same shape and smell, except formulas 2 and 3 have a slightly foamy texture, this is due to the use of a lot of extracts and the high content of saponins in liquorice root extract. sweet due to the many volatile content in the roots such as geraniol, pentanol, hexanol.

igure 4. Facial brightening serum

This result is in line with research conducted by Swati in 2020 with the results of a facial wash gel formula that functions to brighten, namely having a formula that meets standards and is stable³⁰.

b) Homogeneous Tests:

The results of the homogeneity test carried out on all serum preparations showed that the serum was applied to the object glass already qualified. The requirement for good homogeneity is that there are no coarse grains or clumping particles in the serum preparation.

c) pH test:

This pH test was carried out using a pH meter, the results of examining the pH of serum preparations can be seen in the table 4. Based on data in five formulations, it shows that serum preparations meet pH requirements, which are in the range of 4.5-6.5 or the same as facial pH¹². If the pH value is too acidic, it will irritate the skin and if the pH is too alkaline, it will dry the skin³¹. In formulas 1, 2 and 3, the pH is increasing, this is influenced by the extract used, the higher the pH, the higher. This can also be influenced by the concentration of licorice root extract which has a pH of 4.63-5.07 so that it will increase the pH. The content of polyphenolic and saponin compounds that contain many OH- groups in liquorice extract causes the pH of the preparation to increase³².

d) Viscosity test:

The viscosity of the serum preparation should not be too high and not too low. Serum with high viscosity or too thick will reduce the release of active substances from the preparation, whereas if the viscosity is too low, it will reduce the length of stay on the skin when used^33,34. The table 5. showed that serum preparations meet the viscosity requirements according to the Indonesian National Standard, namely the range of 2000-50000 mPas or 200-5000 dPas (SNI 16-4399-1996) ¹³.

e) Spreadability Test:

The spreading power of a preparation is inversely proportional to its viscosity. The higher the viscosity, the lower the spreading power. The wider the spreading power, the more extensive the ability of the active substance to spread on the skin³⁵. The results of the spreading power test can be seen in table 6. All formulations, it shows that the serum preparation meets the requirements for good coverage, namely 5-7cm¹³.

f) Stability Test:

Stability test was carried out to determine the quality of the preparation to survive within the specified limits during storage. The stability test is also to find out things related to serum preparations and incompatibility between the constituent ingredients³⁶. The decrease in pH in the preparation was due to storage at different temperatures¹². This can happen because of the acidic nature of the licorice root extract due to the presence of acids glycirrhizinic²³. The organoleptic test results after 6 cycles did not change color, shape and texture.

g) Irritation test:

An irritation test was carried out to determine the irritating effect of the serum preparation after being used on the skin. Examination of the irritation test found that the five serum formulation formulas gave negative results to all respondents, which means they did not experience irritation.

h) Hedonic Test:

The most preferred serum preparation by respondents between formulas 1, 2 and 3 is formula 1 with the highest average. Test results Kruskal Wallis shows 0.000 (<0.05) it can be said that from the five formulas it shows that there is a significant difference between the respondents' preference for the texture of the serum preparation.

i) Face Brightening Activity Test:

The graph above illustrates an increase in skin color, which shows a decrease in the number on skin tone chart then the brightness increases. The positive control formula and the third formula being the best formula can be seen from changes in brightness levels. The increase in the brightness of the skin color is due to the glabridin content contained in liquorice, where glabridin is included in the flavonoid group. Flavonoids can inhibit the process of melanogenesis; so that an increase in melanin does not occur³⁷. In formulas 2 and 3 in one use, you can see an increased skin tone.

Test results Paired Samples tests shows a Sig (2-tailed) value (<0.05) so that the data can be said to have differences before and after using the serum. However, the sig value between formula 3 and the positive control does not show much difference.

Serum apart from being a face lightener can also treat acne, as in the study of Foukiya Butool, et al in 2013 which stated that zinc supplementation as an additional therapy in acne patients with low serum zinc levels can increase the success rate of acne treatment because zinc deficiency is one of the causes of the acne diseases^38.

CONCLUSION:

This research concluded that Licorice root ethanol extract can be formulated as facial lightening serum preparations. The evaluation results of serum preparations in pH, homogeneity, spreadability, irritation test, and stability tests showed that the serum preparations were good and by serum preparation standards. The effect of the serum preparation with the best brightness level from variations in licorice concentrations of 1%, 10%, and 20%, namely the serum formula with a concentration of 20%. The initial condition of the skin before use shows a value on the skin tone chart of 7 to 3 after using the serum. Respondents' preferred serum preparation from variations in concentrations of licorice was 1%, 10%, and 20% with parameters of color, texture, and aroma, namely the serum formula with a concentration of 1%.

CONFLICT OF INTEREST:

The authors declare that they have no conflict of interest.

ACKNOWLEDGMENTS:

Collaborative research grant between the Pharmacy Study Program of Sekolah Tinggi Ilmu Kesehatan Salsabila Serang Banten and the Faculty of Pharmacy, University of Sumatera Utara.

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Received on 08.08.2023 Modified on 22.12.2023

Research J. Pharm. and Tech 2024; 17(9):4142-4148.

DOI: 10.52711/0974-360X.2024.00641

Effectivity and Evaluation of Licorice Root (Glycyrrhiza glabra) Extract Serum Formula as a Facial Brightening

Sofi N. Stiani¹, Yusransyah Yusransyah¹, Dede Septiana¹, Imam B. Sumantri²*

¹Department of Pharmaceutics, Pharmacy Study Program, STIKes Salsabila Serang, Indonesia.

²Department of Pharmaceutical Biology, Faculty of Pharmacy, University of Sumatera Utara, Indonesia.

Instrument:

Effectivity and Evaluation of Licorice Root (Glycyrrhiza glabra) Extract Serum Formula as a Facial Brightening

Sofi N. Stiani1, Yusransyah Yusransyah1, Dede Septiana1, Imam B. Sumantri2*

1Department of Pharmaceutics, Pharmacy Study Program, STIKes Salsabila Serang, Indonesia.

2Department of Pharmaceutical Biology, Faculty of Pharmacy, University of Sumatera Utara, Indonesia.

Instrument:

Sofi N. Stiani¹, Yusransyah Yusransyah¹, Dede Septiana¹, Imam B. Sumantri²*

¹Department of Pharmaceutics, Pharmacy Study Program, STIKes Salsabila Serang, Indonesia.

²Department of Pharmaceutical Biology, Faculty of Pharmacy, University of Sumatera Utara, Indonesia.