Benefits of Stevia ( Stevia rebaudiana B .) and Sirih ( Piper betle L .) Leaves Combination Againts Dental Care

Oral health is the most important part, to improve its hygiene by maintaining teeth. Cavities are a common problem. Betel leaf as an antibacterial. Stevia leaves as bacteriostatic and bactericidal also eliminate the cause of tooth decay and gingivitis. The purpose of the study was to determine the antibacterial activity of toothpaste combined with betel leaf extract with stevia leaves against the growth of Streptococcus mutans bacteria. The study used an experimental method consisting of making betel leaf and stevia leaf extracts using the maceration method. Preparation of toothpaste with a combination of betel leaf extract with stevia leaves at a concentration of 15% with the ratio of betel leaves and stevia leaves, namely formula 1 (5%: 10%), formula 2 (7.5%: 7.5%), formula 3 (10%: 5%). Testing antibacterial activity by measuring the clear zone using the disc paper method with blood agar media. Data analysis using ANOVA and Paired T test. The results of the study obtained formula (5%: 10%) clear zone of 16.42 mm, formula (7.5%: 7.5%) of 17.24mm, formula (10%: 5%) of 17.94mm, positive control of 18.66mm, negative control of 2.75mm. Conclusion 10:5 combination toothpaste has antibacterial activity.


INTRODUCTION
Oral health is a component of whole body health that cannot be handled separately through other health aspects because it will worsen overall health.Efforts to improve oral hygiene include maintaining oral hygiene.For human health and safety, the role of the oral cavity is enormous.Usually, people are described as healthy not only because of their healthy bodies, but because of healthy oral cavities and teeth.Thus, oral health addresses the overall well-being of the individual.The teeth serve as a food delivery system.Before food enters the digestive tract, it must first be chewed.To prevent difficulties from bacteria on the teeth and delivered by food, it is important to maintain dental hygiene.Cavities are one of the most common dental problems (Nurhamidah et al., 2016).
The most common problem with dental health is caries.Dental caries develops as a result of food debris undergoing a fermentation process caused by oral bacteria, which causes plaque to form on the teeth.Dental caries cause teeth to calcify, which makes them porous, prone to cavities, or even broken (Widayanti, 2014).According to (Ramayanti & Purnakarya, 2013) microorganisms, the tooth itself, food intake, and time are the four basic elements that affect dental caries.Dental caries is mostly caused by Streptococcus mutans.
One of the metabolic products required for bacterial life is lactate, which is produced by the gram-positive facultative anaerobic bacterium Streptococcus mutans.Streptococcus mutans produce glucans and water-insoluble polysaccharides that help detect bacteria on teeth, which gives it the ability to bind sugar to the tooth surface.
Oral pH is capable of being altered by Streptococcus mutans to a level that is favorable for the organism's metabolism but not for other species residing in the same environment.Streptoccocus mutans, a microorganism that can convert sugar into acid.The acid becomes bound to tooth enamel as a result, which results in demineralization of tooth tissue and cavities.(Simon, 2010).
The only component that has been used to clean teeth for a long time is toothpaste.In addition, toothpaste can smooth the tooth surface, minimize or eradicate bad breath, give the mouth a fresh taste, and maintain dental health.Herbal toothpastes are usually described as toothpastes with gentle effects and safe and healthy components.Natural substances that are beneficial, safe, and efficient to improve oral health can be added when producing toothpaste (Telrandhe et al., 2017).
One of the herbs often used in traditional medicine is betel leaf (Piper betle L.).Antibacterial properties are found in betel leaves (Piper betle L.).According to (Putri efendi et al., 2020).Flavonoid, polyphenol, steroid and tannin compounds are the content of betel leaves (Piper betle L.).Based on previous research conducted (Anas et al., 2018) proves that betel leaf extract (Piper betle L.) has antibacterial activity against Streptococcus mutans.The results of the study stated that concentrations of 0.1%, 1%, 10% and 100% were able to inhibit the growth of Streptococcus mutans bacteria .At a concentration of 0.1% the inhibition zone was 13.08 mm, at a concentration of 1% the inhibition zone was 18.46 mm, at a concentration of 10% the inhibition zone was 24.02 mm, and at a concentration of 100% the inhibition zone was 30.95 mm.\ Stevia (Stevia rebaudiana B.) promotes oral health, has antibacterial and anti-fungal characteristics, and combats the causes of gingivitis and tooth decay (Widarsih & Mahdalin, 2017).Stevia is a natural sweetener with high nutritional value that can be used to treat dental caries.Due to the presence of bioactive substances such as stevioside, flavonoids, and tannins, stevia leaves have antimicrobial properties (Wenda et al., 2017).
Based on previous research conducted (Putri et al., 2017) proved that stevia leaf extract (Stevia rebaudiana B.) has antibacterial activity against Streptococcus mutans.The results of the study stated that concentrations of 5%, 10%, 20%, 40% and 80% were able to inhibit the growth of Streptococcus mutans bacteria .The zone of inhibition obtained was 1.53 mm; 2.44 mm; 2.74 mm; 2.96 mm; the novelty of this research focuses on the activity of toothpaste with a combination of betel leaf and stevia leaf extracts that have been tested for antibacterial activity in previous studies.

Making toothpaste of betel leaf extract (Piper betle L) combined with stevia leaf (Stevia rebaudiana B):
1. Weighing all ingredients according to the calculation.2. Develop Na-CMC sorted using hot water as much as 20x the amount of CMC grind until homogeneous (mass 1). 3. Dissolving calcium carbonate with glycerin sorted until homogeneous (mass 2). 4. Adding nipagini and sodium lauryl sulfate into mass 2 slowly grind so that no foam is formed.5. Adding extracts of betel leaf (Piper betle L.) and stevia leaf (Stevia rebaudiana B.). 6.Then mass 2 is added to mass 1 and crushed until a homogeneous toothpaste is formed.
7. Put in a container, then evaluate the preparation.

Preparation of Betel Leaf Simplisia (Piper betle L.)
Betel leaves that have been collected as much as 2 kg, then carried out wet sorting, washing with running water, then knotting the betel leaves.After that, drying the betel leaves was carried out, the leaves that had been dried were sorted dry, then the powder was made by blending.The simplified powder was stored in a clean and tightly closed container.

Formulation and mode of action Preparation Extracts of Betel Leaf (Piper betle L.) and Stevia Leaf (Stevia rebaudiana B.)
Betel Leaf (Piper betle L.), prepare tools and materials, weigh 200 grams of betel ethanol solvent, let stand for 7 x 24 hours, while occasionally stirring, after 7 days do filtering using flannel cloth.The filtrate obtained is then concentrated extract using italic, the difference in weight per weight (b / b) between the results obtained and the weight of the simplisia powder used.
Then it is used to calculate the percentage of extract yield calculated by the formula: Stevia leaves (Stevia rebaudiana B.), prepare tools and materials, weigh 200 grams of stevia leaf powder, then put it in a macerator.Then add 2000 ml of 70% ethanol solvent, let it stand for 7 x 24 hours, while occasionally adding a few drops of ethanol.Stirring was carried out, after 7 days filtering using flannel cloth.The filtrate obtained then concentrates the extract using a water bath, the difference in weight per weight (b / b) between the results obtained and the weight of the simplisia powder used.
Then used to calculate the percentage yield.The extract yield was calculated by the formula: As much as 0.5 g of extract is put into a test tube, adding 0.5 mg of magnesium powder, adding 3 drops of concentrated HCL, yellow, red, or orange color indicates positive flavonoids.

Tannin Test
As much as 0.5 g of extract was put into a test tube, adding 3 drops of FeCl3, blackish green color showed catechol tannin and blackish blue showed pyrogallol tannin.

Steroid Test
As an extract of 0.5 g was put into a test tube and added with 2 mL of concentrated H2SO4, the purple color indicates the presence of steroid compounds.

Stevia leaf (Stevia rebaudiana B.) Flavonoid Test
The extract of 0.5 g is put into a test tube, adding 0.5 mg of magnesium solution, adding concentrated HCL 3 drops, yellow, red, or orange color indicates positive flavonoids.

Tannin Test
As much as 0.5 g of extract was put into a test tube, adding 3 drops of FeCl3, blackish green color showed catechol tannin and blackish blue showed pyrogallol tannin.

One-Way ANOVA Test
Tests using the anova test with a significance level = 5%.Showing the significant level of the test carried out so that this observation is stated that the extract has antibiotics.The way to read the sig value is as follows: If the value of Fcount>Ftable then H0 is rejected and H1 is accepted.If the value of Fcount <Ftable then H0 is accepted and H1 is rejected.

T -Test
One of the tests used in parametric statistical testing is the T-test.The T-test is a test that shows the extent to which one independent variable can explain the dependent variable alone.A significance level of 0.05 (= 5%) is used for the T-test statistical test (Magdalena & Angela Krisanti, 2019).
Determining the t value (t count) is by comparing with the t table.The decision making is as follows: If the significant value > 0.05, then H0 is accepted and H1 is rejected.If the significant value <0.05 then H0 is rejected and H1 is accepted.The weight of betel leaves used is 2kg.After making simplisia, the results were obtained in the form of brownish leaves with a distinctive aromatic smell of betel leaves with a dry simplisia weight of 253 grams.

Resuts of Making Sirih Leaf (Piper betle L.) and Stevia Leaf (Stevia rebaudiana B.) Extracts
The results of making betel leaf extract (Piper betle L.) are as follows: Yield Formula: Making toothpaste combination of betel leaf extract (Piper betle L.) with stevia leaves (Stevia rebaudiana B.) made at a concentration of 15% with a combination of X1 (5:10),X2 (7.5:7.5) and X3 (10:5) each 100 grams.Weighing all the ingredients according to the calculation, developing Na-CMC sorted using hot water as much as 20x the amount of CMC grind until homogeneous (mass 1), dissolving calcium carbonate with glycerin sorted grind ad homogeneous (mass 2), nipagin and sodium lauryl sulfate added slowly into mass 2 and grind slowly so that no foam is formed, then mass 2 is poured into mass 1 and grind until a homogeneous toothpaste is formed, adding betel leaf extract (Piper betle L.) and leaves (Stevia rebaudiana B.), put it in a container, then evaluate the preparation.

Evaluation Results of Toothpaste Combination of Betel Leaf Extract (Piper betle L.) with Stevia Leaf (Stevia rebaudiana B.)
The results of the evaluation of toothpaste combination of betel leaf extract (Piper betle L.) and stevia leaf (Stevia rebaudiana B.) are as follows:

Organoleptic Test Results
Formulations 1, 2 and 3 have a green color.All three formulations are pasteshaped and have a sweet taste, the aroma of formulas 1 and 2 is typical of betel leaves, while formula 3 is typical of strong betel leaves.In the negative control, the color is white, paste-shaped, tasteless and odorless.

Homogeneity Test Results
All of the formulations and the negative control were homogeneous preparations.

pH Test Results
The pH of formulas 1, 2 and the negative control were at 7, while formula 3 had an initial pH of 7 and a final pH of 6.

Foamability Test Results
Formula 1, 2, 3 and the negative control had a foam power of 1.5 cm.

ACKNOWLEDGMENTS
In this observational study, the authors have received a lot of help from related parties, maTelrandhe, R., Deshmukh, P., & Gunde, M. ( 2017).Formulation and Evalua-tion of Herbal Toothpaste: Compared With Marketed Preparation.International Journal of Pharmaceutics & Drug Analysis, 5(10), 406-410.On this occasion, I would like to thank: Rectorate of YPIB University who has provided a research grant so that the writing of this observational study is realized.
Hopefully this research will be useful for the world of health and in the field of pharmacy in particular, as well as for society in general.

Diagram 1 .
5.1 cm Observation Results of Toothpaste Activity Test Combination of Betel Leaf Extract (Piper betle L.) with Stevia Leaf (Stevia rebaudiana B.) Measurement Results of Clear Zone Diameter (mm) of Toothpaste Combination of Betel Leaf Extract (Piper betle L.) with Stevia Leaf (Stevia rebaudiana B.) on the First Day Clear zone measurement results of toothpaste extract combination of betel leaf and stevia leaf on the first day of observation Clear Zone Diameter Measurement Results (mm) Toothpaste Combination of Betel Leaf Extract (Piper betle L.) With Stevia Leaf (Stevia rebaudiana B.) on the Second Day.The following are the results of the calculation of the clear zone on the 2nd day of observation of toothpaste extracts of betel leaf and stevia leaf combinations Diagram 2. Clear zone measurement results of toothpaste extracts of betel leaf and stevia leaf combination on the 2nd day of observation.Recapitulation of Average Clear Zone (mm) of Toothpaste Combination of Betel Leaf Extract (Piper betle L.) with Stevia Leaf (Stevia rebaudiana B.) on Day One and Two From diagrams 1 and 2, a recapitulation was then carried out to find the average of how many creepage areas were obtained from the Toothpaste Combination of Betel Leaf Extract (Piper betle L.) with Stevia Leaf (Stevia rebaudiana B.).And below are the recapitulation results: Diagram 3. Recapitulation of clear zone measurements of toothpaste with a combination of betel leaf and stevia leaf extracts from day 1 and 2.extract (Piper betle L.) with stevia leaf (Stevia rebaudiana B.) combination (10:5) has antibacterial activity against Streptococcus mutans has antibacterial activity against Streptococcus mutans, toothpaste combination of betel leaf extract (Piper betle L.) with stevia leaves (Stevia rebaudiana B.) combination (10: 5) which has the greatest antibacterial activity against Streptococcus mutans, toothpaste combination of betel leaf extract (Piper bet le L.) with stevia leaves (Stevia rebaudiana B.) meets the requirements and is stable at storage temperature.

Table 1 .
Formulation of toothpaste in each sample to be tested

Table 3 .
Physical Characteristics of Preparations