INTRODUCTION:

The development of natural resources in the root canal irrigant in endodontic treatment has increased to answer biocompatibility issues¹. At the same time, caries leads to pulpitis irreversible and 36% from oral infections². Phospholipase enzymes influence the pathogenesis of periapex pulp disease, an enzyme hydrolyzed from glycerophospholipids that play a role in invading and growth of F. nucleatumin host tissue³.

The collagenolytic activity of bacteria can damage chemical bonds, so it plays a role in the propagation of cracks through the dentin substrate. Mineraldentin becomes porous due to the loss of mineral dentine and collagen matrix denaturation⁴.

Treat teeth with pulp disease, and periapical cleaning and shaping measures are carried out in endodontic treatment to eliminate microorganisms from the root canal and clean the root canal teeth from the smear layer of necrotic tissue that requires irrigation solutions. Sodium hypochlorite (NaOCl) is a recommended irrigation solution because it has antimicrobial activity and dissolves the remaining pulp tissue, but it is a toxic odor and discomfort⁵. Ethylenediaminetetraacetic acid (EDTA). The side effects of NaOCl and EDTA combination are opening the dentinal tubules, destruction of intertubular dentin, reduction of dentinal hardness, and strong erosion of dentin⁶. Chlorhexidine is also recommended but has a root canal cleaning effect⁷. Since Indonesia has thousands of developing natural ingredients, S.rarak D.C (S. rarak DC) has become an agent of cleansing and has antimicrobial effects, anti-inflammatory, analgesics, cleaning the root canals of teeth, and dissolving dental pulp tissue⁸.

Suppose S. rarak DC extract is proven to have bioreduction properties that can inhibit the growth of F. nucleatum. Using this natural material can prevent pulp and periapical diseases so that teeth can be maintained for life. Within a decade, there were dramatic advances in analytical techniques, including TLC, UV., NMR, and GC-MS, powerful tools for the separation, identification, and structure determination of phytochemicals⁹. This study aims to determine the organic compounds present in the S.rarak DC extract with the GC-MS technique, which may provide insight into its use in traditional medicine. Our study offers a platform for utilizing S.rarak DC fruit as an alternative to natural root canal irrigation in endodontic treatment.

Materials and Methods:

Plant Material:

The S.rarak DC fruitswere selected to screen theirbiopotentials based on their traditional usage. The fully mature S.rarak DCs were collected in February 2019 from Maga Village, Panyabungan District, South Tapanuli, North Sumatera Province, Indonesia (Figure 1). The collected S.rarak DC fruits were identified and authenticated by LembagaIlmuPengetahuan Indonesia, Bogor, Indonesia.

Preparation of extract:

S.rarak DC fruits 1 kg were washed under running tap water, and dust was removed from the fruits. Then, the seeds weighed and gained as much as 940g. Then cut into small pieces ± 3mm and dried in a drying cabinet at a temperature of ± 40°C for a week. It was already dried, mashed with a blender, diluted with ethanol to be macerated, and then inserted into a percolator while adding ethanol. The results of percolate evaporated with vacuum rotavapor to obtain a brown-colored viscous extract of as much as 240grams. The ethanol extract of assay material is put in a glass bottle and then stored in the refrigerator¹⁰.

GC-MS Assessment:

50mg of S rarak DC extract was decontaminated with 5 mL dichloromethane for 2 minutes at room temperature. After decontamination, The extract was incubated at 45⁰C for 2 hours in 1mL of methanol and centrifuged at 5000rpm speed for 5 minutes. The supernatant obtained is taken and evaporated using nitrogen. The residue obtained is then derivatized. A total of 10mL BSTFA with 1% TMCS was added to the residue. The tube is labeled and heated at 60⁰C for 20 minutes. After derivatization, the extract is cooled to room temperature, amount of 1mL of extracted derivative results is injected into the GCMS system. Mass spectrum GC-MS was interpreted using the National Institute Standard and Technology (NIST) database, having more than 62,000 patterns. The spectrum of the unknown component was compared with the range of the known parts stored in the NIST library. The name, molecular weight, and structure of the components of the test materials were ascertained¹¹.

Fusobacterium nucleatum Growth Assay:

The spectrophotometric growth assessment of F. nucleatum began with S.rarak DC extract preparation at 50%, 25%, 12.5percent, and 6.25percent, with 0.2 percent Chlorhexidine (CHX) as a positive control. Each well in the 96-well plate received 50L of TSB medium and was incubated for 15 minutes. Then it was washed twice with PBS pH 7.0. It was also prepared in a well of 25L of S. mutans in medium and incubated at room temperature (27⁰C) for 15 minutes before adding saliva at a predetermined concentration into a well of 100L. (1:4). The cells were then incubated in an aerobic environment for 24hours, 48 hours, and 72 hours. The stiffness of F. nucleatum was measured using a spectrophotometry-Elisa Reader (Bio-Rad, USA) and an optical density (OD) of 620nm. Mc. Farland's OD. range is 0.08-0.1nm. 0.5 (1.5x108), which is equivalent to 300 CFU¹².

Statistical Analysis:

Kruskal Wallis was used to analyze the growth quantity of F. nucleatum, with a significance limit of p0.05 and r=1. At the same time, the value of the benefits of antibacterial chemical compounds was analyzed descriptively.

Results and discussion:

Fig.1: Sapindusrarak D.C fruits

Figure 1 shows the type of fruit S.rarak D.C as the test material in this study. (GC-MS is an analysis method for smaller and volatile molecules such as benzenes, alcohols, and aromatics, and simple molecules such as steroids, fatty acids, and hormones. The advantage of this method in the compound analysis is its ability to separate complex mixtures, quantify analytes, and determine trace levels of organic contamination. The results of the GC-MS study led to the identification of several compounds from the GC fractionations of the ethanol extract of S.rarak D.C fruits. The mass spectrometry is attached to GC (Figure 2). Table 1 presented above shows the components of ethanol extract of S.rarak D.C fruit derived from GC-MS analysis and explained further below. Table 2 also reported the role S.rarak DCin inhibiting the growth of F. nucleatum.

Table 1.GC MS analysis of Lerak (Sapindusrarak DC) fruit extract.

S. No	RT	Quality (%)	Compound	Content (%)
1	7.314	93	Benzyl chloride	^6,51
2	24.049	80	1-Dodecanamine, N, N-Dimethyl-	^19,91
3	28.207	58	1-Tetradecanamine,N,N- Dimethyl	^6,57
4	28.331	64	4- (3-Dimethylaminopropoxy) Benzaldehyde	^3,28
5	28.558	43	1- (dimethylamino) –2- Butanol	^3,18
6	30.165	92	Acetamide, 2- (diethylamino)-N- (2,6-Dimethylphenyl	^1,41
7	30.420	90	9-Octadecenoic acid	^1,84
8	30.668	99	Hexadecanoic acid	^6,82
9	31.317	99	Cis- 13-Octadecenoic acid, methyl ester	^1,46
10	31.468	59	Tetrahydroquinoline-4,4,-	^4,55
11	31.827	99	6-Octadecenoic acid	^27,46
12	31.689	64	2- (Benzylmethylaminomethyl) -2- Norbornanone	^3,20
13	32.820	99	Cis-13-Eicosenoic acid	^5,54

Benzyl Chloride:

One of the components is Benzyl chloride, a reactive benzene compound with a powerful odor. It does not dissolve in cold water but decomposes slowly in warm water to form benzyl alcohol. (Figure 3.1) This compound was found at ^{6.51%
in the extract}and is extremely irritant to the eyes, mucosa, and skin; direct contact with body parts must be avoided to prevent negative effects. This compound has toxicity properties with LD50 (rat) = 1000 and LC50 (rat) = 280. Benzyl chloride is widely used as a precursor of other benzene class compounds. This compound is also used to synthesize benzyl ester compounds used as plasticizers, perfume mixers, and deodorizers in the industrial field. In addition, these compounds can be used as amphetamine-class drugs' raw materials in pharmaceuticals. Thereby, their sales in the United States are rigorous and under the supervision of the American FDA narcotics agency1.3

Fig.2: GC-MS Chromatogram peaks of ethanol extract of Sapindusrarak DC.

1-dodecanamine, N,N-dimethyl:

The second compound found is ^{19, 91% of}1-dodecanamine, N,N-dimethyl also known as Dodecylamine, N, N-dimethyl- is often abbreviated as DMDO CAS No. 112-18-5 MW.: 213.41g/mol. DMDO is a long-chain carbon compound that has one nitrogen atom. (Figure 3.2) These compounds are widely used as surfactants, resins, and natural disinfectants. Besides its free availability, this compound can be synthesized by reacting to alcohol with dimethylamine.

This compound is reported to have toxicity in animal tests. After 4 hours and 3 minutes of oral administration, it shows high irritability and corrosivity, but this compound does not cause mutagen. However, its exposure to humans has not been tested. Controlling exposure to this compound is recommended due to its corrosive nature and strong irritants. This compound decomposes quickly and is adsorbed by various aquatic organisms in the environment. Some ecotoxicity test results show as follows: Brachydaniorerio, 96 h-LC0 = 0.5ppm, LC50 = 0.71-1ppm, LC100 = 1ppm¹⁴.

Despite its reports of toxicity in animals, it also has several biological activities such as anaphylactic, antitumor, analgesic, Arylamine-N-Acetyltransferase-Inhibitor, Inhibit production of Tumor Necrosis Factor, increases NK. Cell activity, a myoneural stimulant. Besides those activities, this compound also could act as a neuroinhibitor, decrease norepinephrine production, as nauseant, NCS-depressant, NADH-Oxidise-Inhibitor, and NADH-Ubiquinone-Oxidoreduction-Inhibitor¹⁵.

1-Tetradecanamine, N,N-dimethyl:

The third component is 1-Tetradecanamine, N,N-dimethyl, or Dimethyltetradecanamine, a DMDO compound. (Figure 3.3) This compound is found in ^{6,57% of the extract.} Physical properties include a bright yellow color, a fishy odor, being insoluble in water, and floating slightly lighter than water on the water's surface. Because this compound is irritating, avoiding direct contact with the mucosal surface and membrane is best.

Furthermore, if swallowed or inhaled, it is toxic¹⁶.

4-(3-dimethylaminopropoxy) benzaldehyde:

The other compound is 4-(3-dimethylaminopropoxy) benzaldehyde which shows quite a low percentage, ^3,28%.This compound is an aldehyde derivative with an aromatic ring or benzene, also known as a benzene derivatives compound with an aldehyde functional group. (Figure 3.4) This compound is 1031 g/mL and, in 360 mmHg pressure, has a 325.4^oC boiling point. In addition to the above pathway, this compound can also be synthesized through a reaction between 3- (dimethylamino) propyl methanesulfonate and 3-Dimethylamino-1-propanol but with a yield of only 56%¹⁷.

1-(dimethylamino)butan-2-ol:

1-(dimethylamino)butan-2-ol found as much as ^3,18%.This compound is a derivative of alcohol with an amino group. (Figure 3.5) Besides its availability in nature, this compound can be synthesized by reacting Chloroacetone with ethyl bromide in an absolute ether solvent that produces 1-chloro-2-methyl butanol compound, then reacted with dimethylamine for 3 hours to produce a pure product¹⁸.

Acetamide,2-(diethylamino)-N-(2,6-dimethylphenyl):

Acetamide,2-(diethylamino)-N-(2,6-dimethyl phenyl)/Lidocainewhen placed at room temperature, takes the form of a white powder with 68.5^oC boiling point. It is slightly soluble in water, considering that this compound has an aromatic ring that makes it hydrophobic. (Figure 3.6) This compound is known as the trivial Lidocaine, xylocaine, and lidopen, which is widely used in medicine as a local anesthetic. Lidocaine is also used to treat ventricular tachycardia and nerve block.

A mixture of Lidocaine with a small amount of adrenaline (epinephrine) can be used for larger doses of anesthesia. Also, Lidocaine is used to reduce bleeding, and it gives a more prolonged numbness effect. Lidocaine typically begins working four minutes after injection and lasts for half an hour to three hours.

The lidocaine mixture can also be applied directly to the skin or mucous membranes for anesthetic purposes.This compound is found in the lowest concentration (1,41%) in the extract of S. Rarak DCBased on molinspiration prediction, the lidocaine compound has a log P of 2.13 and a TPSA of 32.34, indicating that Lidocaine meets the criteria as a drug compound. Apart from being an anesthetic, this compound is also used as a membrane stabilizing agent against ventricular arrhythmia, which is also used as an antiarrhythmic¹⁹.

Local anesthetics work to prevent the formation and conduction of nerve impulses. To be used clinically, an anesthetic must be tissue-compatible (non-irritating), and the reaction must be temporary and fully reversible. Anesthetics must also be effective at doses expected to be far below their level of toxicity, hypoallergenic, and have a rapid onset of anesthesia with a sufficient duration of action for a comfortably complete surgical procedure. Local anesthetics are divided into two functional groups based on their chemical properties: amides and esters. The ester group includes procaine, cocaine, and novocaine. However, usually topical anesthesia (applied before an injection is given) is still part of the ester compound group. There are more modern amide groups, including Lidocaine (or xylocaine), mepivacaine (orcarbocaine), prilocaine (or citanest), bupivacaine (or marcaine), and etidocaine (or duranest)²⁰.

9-octadecenoic acid:

9-octadecenoic acid,^{1,84% derived,} better known as oleic acid, has a double bond on the C9 atom of the carboxyl group, and the C9 atom came from the methyl end. Therefore these organic acids are grouped into omega-9 fatty acid compounds (MUFA). (Figure 3.7) Monounsaturated fatty acids, which are essential for the body for their antioxidant properties, are a compound that plays a role in the formation of brain cells in the fetus, so pregnant women are strongly advised to consume foods that contain lots of omega-3 fatty acids, omega-6, and omega-9 so that the fetus has brain cells that are both of which are often associated with children's intelligence levels²¹.

This compound is found to have abundant biological activities similar to Hexadecanoic acid, as an acidifier, acidulant, arachidonic acid-inhibitor, increase aromatic amino-acid decarboxylase activity, inhibit the production of uric acid, urinary acidulant, and as urine acidifier²². Through all these benefits, we could conclude that this compound has a lot of medical advantages, thus proving beneficial in medical development industries. Oleic acid is also the essential monounsaturated fatty acid for human cells. This oleic acid is vital in the formation of phospholipid cell membranes which provide appropriate cell fluidity. Cell fluidity is necessary concerning hormonal responses, pathogen infections, mineral transport, and the immune system. For cells, oleic acid is also a significant energy source wherein the metabolic system is broken down into acetyl co A to produce ATP and other essential metabolites²³.

Oleic acid is obtained by cells from endogenous biosynthesis or serum triglycerides. Biosynthesis of an organic acid such as oleic acid involves the same enzymes that play a role in elongation (increasing the length of carbon bonds) for other fatty acids, which are precursors to the production of eicosanoids (prostaglandins), autacoids lipids derived from arachidonate acids maintain homeostatic function and mediate pathogenic mechanisms including responses to inflammation. Therefore oleic acid deficiency also shows a lack of eicosanoid production, impacting the body's weak immune system and response to pathogens²⁴.

Hexadecanoic acid:

Hexadecanoic acid, also known as palmitic acid, is an organic acid most commonly found in coconut oil, palm oil, and various other fat-containing plants such as nuts and seeds. Still, palm oil is the primary source of fatty acids palmitate,containing 40-46%. These fatty acid compounds are solid at room temperature because they are saturated fatty acid compounds without double bonds. (Figure 3.8). In a form bound to triglycerides, this compound is used for various food and beverage processing purposes as an additive to food and cooking oil. Aside from its uses as food additives described above, this compound can also be anti-mitotic²⁵. Still, research also shows that excessive PA (Palmitic Acid) intake can triggerdyslipidemia and hyperglycemia. Compared to a previous study of Alstoniaboonei's root and stem-bark extraction, this compound was also found as one of its major chemical components.Hexadecanoic acid is turpentine, also located in traditional Chinese pickled peppers²⁶. This compound is the second-highest percentage derived from the GC-MS analysis, ^{6,82%, which has a lot of biological activities such as acidifier,
acidulant, arachidonic acid inhibitor, increased aromatic amino acid
decarboxylase activity, inhibit the production of uric acid, urine acidifier,
and as a urinary acidulant27.}

^{Cis-13-Octadecenoic acid:}

^{Cis-13-Octadecenoic acid, methyl ester,
1,46% found in the extract, is a}fatty acid as the primary source of fatty acid esters. (Figure 3.9) Fatty acid ester compounds are produced by esterification or transesterification reactions.Transesterification reactions generally involve alcohol compounds as starters and catalysts in acids, bases, or enzymes²⁸. The biological activities of this compound are as an acidifier, ^{arachidonic acid inhibitor, urine
acidifier, and urinary acidulant.}The reaction to form this compound is based on the exchange of group R between triglyceride compounds or fatty acids with alcohol to form an ester. Most fatty acid esters produced naturally and synthetically are used as solvents, additives in food, and alternative fuels to replace diesel or are popular with biodiesel²⁹. Since 1998, there has been much research regarding Fatty Acid Methyl Ester (FAME), and Fatty Acid Ethyl Ester (FAEE). By utilizing oil or fat from various sources, fatty acid esters as biodiesel production is currently under rapid development³⁰.

Quinoline, 1-azanaphthalene:

Quinoline, 1-azanaphthalene, or 1,2,3,4-tetrahydroquinoline-4,4- known as kuson, is an aromatic nitrogen compound with a solid ring form containing benzene, which is fused with pyridine on two corresponding carbon atoms.(Figure 3.10) (Pyridine is a compound with five cyclic carbons with one nitrogen atom). This compound was found ^{4,55% in the extract.}Quinoline is the simplest member of quinoline in oily liquid and hygroscopic (easy to bind water vapor), slightly yellowish; slightly soluble in water, alcohol, ether, carbon disulfide, and very soluble in organic solvents. This compound can be obtained by refining coal tar. In addition, Quinoline can also be obtained from the reaction between aniline and acrolein in hot sulfuric acid (Skraup synthesis). Various quinoline compounds can be synthesized using the Skraup method with various oxidizing agents³¹.

Quinoline compounds are widely used as precursors for making drugs (especially anti-malaria drugs), anti-cancer, fungicides, biocides, alkaloids, dyes, rubber chemicals, and flavoring material³². This class of compounds has antiseptic, antipyretic, and antiperiodic properties. Besides that, quinoline is also used as a catalyst, corrosion inhibitor, preservative, and solvent for resins and terpenoid class compounds. In addition, it is also used in complex catalysis of transition metals, polymerization, and antifoaming. Quinaldine, 2-methyl quinoline, is used as an anti-malaria and raw material to prepare other anti-malaria drugs. These compounds also manufacture oil colorants, food coloring, pharmaceuticals, pH indicators, and other organic compounds³³.

6-Octaenoic acid:

6-Octaenoic acid compound, also known as petroselinic acid (Figure 3.11), is the highest from the extraction of S. rarak DC, which is ^{27,46%. It}is a monoenoic fatty acid and is the main component of parsley seed oil (Parsley). Parsley is a herb with many medical benefits that helps kidney cleansing, improving digestion, healing of inflammation, and detoxification. Furthermore, parsley oil is essential for its uses in food, cosmetics, and pharmaceuticals.³⁴ Compared to other GC-MS analysis studies, this compound is also found in selected medicinal plants' leaves in Boraginaceae family Cordiadichotoma L. Octadecanoid acid is also categorized as terpentin. 6-Octadenoid has the most significant percentage in the extraction of S. rarakDC, ^{27,46
%. This compound is classified as terpenes.}

Fig.3: The chemical compound of S. rarak. These compounds have more than significant potential as antibacterial. itis a reference of the alternative root canal irrigation antibacterial

2-(Benzylmethylaminomethyl)-2-norbornanone:

2-(Benzylmethylaminomethyl)-2-norbornanone or Norborenanone (3,20%) is an organic compound that falls under the bicyclic ketone group (Figure 3.12). This compound is a derivative of camphor. The membrane was the primary target of the norbornane class of compounds, but no experiments were conducted to prove this. Furthermore, while most CAMPs are membrane-active, numerous peptides have an intracellular target. A combination of cellular targets is thought to result in increased antibacterial activity and a higher barrier to evolved resistance.³⁵

Cis-13-Eicosanoic:

Cis-13-Eicosanoic acid is a single fatty acid compound with a double bond on the seventh carbon atom, often called omega-7 fatty acid.(Figure 3.13) This compound is also known as paulinic acid because it is the main content of the seeds of Paullinia cupana var. sorbillis (Mart.), with a scope of about 7%.Besides being found in plants above, these fatty acids are metabolites in animals such as mice, rats, and herring. This compound is found at ^5,54%in ethanol extract ofS. rarak. This compound is also found in rapeseed oil and canola oil. The biological activities of this compound are acidifier, acidulant, and arachidonic acid-inhibitor.³⁶

The milog P partition coefficient (octanol-water) prediction value shows the solubility ratio in solvent water and octanol. This value indicates the level of lipophilicity of a compound or the ability to penetrate the cell's semipermeable membrane to reach the nerve center during treatment. Therefore, the log P value is significant to determine whether a drug is suitable for penetrating the nerve center.³⁷Compounds with log P above 5 tend to be challenging to adsorb by cell membranes. At the same time, TPSA (Topographic Polar Surface Area) values indicate polar molecular surfaces that affect the solubility of a drug in water. Too high a solubility level will cause low permeability.³⁸

From the results of the prediction of compounds using the online service, www.molinspiration.com obtained five compounds with a log P below 5, including Lidocaine and hydroquinone. 1,2,3,4-Tetrahydroquinoline compounds belong to the group of quinoline compounds with many derivatives and are widely used for various treatments, especially the treatment of Malaria and cancer. Lidocaine, as described above, is a local anesthetic widely produced under different patent names but is better known in the form of salt (Lidocaine HCl H2O) because of its higher water solubility, so it has better bioavailability in the body. The discussion of the Lidocaine compound is interesting because, so far, this compound is known as a synthetic compound that was first made in the laboratory by Nils Lofgren in 194.³⁹

Growth of F. nucleatumAssessments:

Table 2 shows that after a 24-hour incubation period in a 12.5 percent concentration of Sapindusrarak DC, only 0.4 percent of F. nucleatum survived in 100 percent salivary growth response. While a saliva concentration of 25% resulted in growth response of 82 percent, F. nucleatum growth was only 18 percent. The concentration of 6.25 percent was then added, followed by CHX. At 48 hours of incubation, the Sapindusrarak DC concentrations of 12.5 percent and 6.25 percent had a better growth response to F. nucleatum than other saliva concentrations. However, CHX was still higher, giving a growth response to F. nucleatum (98.42 %). All-natural antibacterial ingredients generally tend to damage the bacterial cell membrane, preventing communication and the formation of biofilms on the surface of cells or host tissues⁴⁰. Sapindusrarak DC contains antibacterial chemical compounds that may damage the bacterial cell membrane and reduce oxygen and nutrient intake by suppressing the function of the ROS and RES systems in bacteria⁴¹.

According to Kruskal Wallis analysis, there was no significant difference in the growth of F. nucleatum in the concentration of Sapindusrarak DC based on incubation time (p>0.05; 0.273) or concentration (p>0.05; 0.065). Both positively affected F. nucleatum growth in different concentrations of Sapindusrarak DC.

Conclusion:

The results of GC/MS analysis show that the ethanol extract of Sapindusrarak DC fruit has bioactive components such as benzyl chloride, 1-Dodecanamine, N, N-Dimethyl-,1-Tetradecanamine, N, N- Dimethyl, 4- (3-Dimethylaminopropoxy) benzaldehyde, 1- (Dimethylamino) –2- Butanol, Acetamide, 2-(Diethylamino)-N- (2,6-Dimethylphenyl, 9-Octadecenoic Acid, Hexadecanoic Acid, Cis- 13-Octadecenoic acid, methyl ester, Tetrahydroquinoline-4,4,-, 6-Octadecenoic acid, 2- (BenzylmethylamiNomethyl) -2-Norbornone, Cis-13-Eicosenoic acid. This study offers a platform for utilizing (Sapindusrarak DC herbal ingredients as an alternative to natural root canal irrigation in the field of Endodontics. This study shows that several antibacterial compounds found in Sapindusrarak DC can inhibit the growth of F. nucleatum, implying that this extract can be used as active material in endodontic procedures as a root canal irrigant.

Conflict of Interest:

The authors declare no conflicts of interest.

Acknowledgment:

Thank you to the Oral Biology Laboratory, Dentistry Faculty, Syiah Kuala University, Banda Aceh, Indonesia, for facilitating the assessment of Lerak extract bioactivity against Fusobacterium nucleatum.

References:

1. Sheik R, Nasim I. Newer root canal irrigants-A review. Research Journal of Pharmacy and Technology. 2016;9(12): 1451-1456.doi.org/10.5958/0974-360X.2016.00473.X

2. Subasree S, Geetha RV. Essential oils in prevention of Dental Caries-An In-Vitro study. Research Journal of Pharmacy and Technology. 2015;8(7): 909-911.doi.org/10.5958/0974-360X.2015.00148.1

3. Dennis EA, Cao J, Hsu Y-H, Magrioti V, Kokotos G. Phospholipase A2 enzymes: physical structure, biological function, disease implication, chemical inhibition, and therapeutic intervention. Chemical reviews 2011;111(10):6130-85.doi.org/10.1021/cr200085w

4. Abou Neel EA, Aljabo A, Strange A, et al. Demineralization-remineralization dynamics in teeth and bone. International journal of nanomedicine 2016;11:4743-63.doi.org/10.2147%2FIJN.S107624

5. Sabila N, Yanti N, Gani BA. Bioactivity of Lerak Fruit Extract (Sapindusrarak DC) as an Endodontic Irrigant to Inhibition the Fusobacteriumnucleatum virulence and Relate to the Fracture Resistance of Root Canal. International Journal of Innovative Science and Research Technology. 2022;7(6), 1749–1750.doi.org/10.5281/zenodo.6914344.

6. Fitria JR, Yanti N, Gani BA. In-Vitro Study of Lerak Fruit Ethanol Extract (SapindusRarak DC) on the Adhesion of Fusobacteriumnucleatum and Prevent Root Canal Wall Porosity. International Journal of Innovative Science and Research Technology. 2022;7(6), 1751–1756.doi.org/10.5281/zenodo.6914412Nandakumar M, Nasim, I. Use of antibiotics in endodontics-clinical practice guidelines. Research Journal of Pharmacy and Technology. 2019; 12(1):419-424.doi.org/10.5958/0974-360X.2019.00076.3

7. Yanti N, Dennis D, Prasetia W. The Ability of Root Canal Irrigant With Ethanol Extract of Lerak Fruit (Sapindus Rarak Dc) in Removing Root Canal Smear Layer (A Sem Study). IOSR Journal of Dental and Medical Sciences. 2017;16(01). doi.org/10.9790/0853-1601082430

8. Farges J-C, Alliot-Licht B, Renard E, et al. Dental Pulp Defence and Repair Mechanisms in Dental Caries. Mediators of inflammation 2015;2015:230251-51.doi.org/10.1155/2015/230251

9. Fajriaty I, Hariyanto I, Haryanto Y. Anti-fertility effect of ethanol extract of lerak (Sapindus rarak DC) fruits in female Sprague Dawley Rats. Nusantara Bioscience 2017;9(1):102-06.doi.org/10.13057/nusbiosci/n090118

10. Yusuf H, Husna F, Gani BA. The chemical composition of the ethanolic extract from Chromolaena odorata leaves correlates with the cytotoxicity exhibited against colorectal and breast cancer cell lines. Journal of Pharmacy and Pharmacognosy Research 2021;9(3):344-56.doi.org/10.56499/jppres20.969_9.3.344

11. Sutton S. Measurement of microbial cells by optical density. Journal of Validation technology 2011;17(1):46-49.

12. Patel, Chirag A., Sen DJ, Patel AR. Nanomedicine: Emerging Field in Medicine. Research Journal of Science and Technology. 2010; 2(3): 41-46

13. Jaishankar M, Tseten T, Anbalagan N, Mathew BB, Beeregowda KN. Toxicity, mechanism and health effects of some heavy metals. Interdisciplinary toxicology 2014;7(2):60-72.doi.org/10.2478%2Fintox-2014-0009

14. See D, Mason S, Roshan R. Increased tumor necrosis factor alpha (TNF-alpha) and natural killer cell (NK) function using an integrative approach in late stage cancers. Immunol Invest 2002;31(2):137-53.doi.org/10.1081/imm-120004804

15. Bhardwaj K, Sharma R, Abraham J, Sharma P. Pyrethroids: A Natural Product for Crop Protection. Natural Bioactive Products in Sustainable Agriculture; Springer: Singapore 2020:113-30.doi.org/10.1007/978-981-15-3024-1_8

16. Majedi S, Karamian R, Moazzami Farida SH, Asadbegy M, Alizadeh H. Synthesis and Biological Activity Evaluation of 3, 4, 7, 8-Tetrahydro-3, 3-Dimethyl-11-Aryl-2 H-Pyridazino [1, 2-a] Indazole-1, 6, 9 (11 H)-Triones by Using an Acidic Ionic Liquid 1-Methylimidazolium Trinitromethanide {[HMIM] C (NO2) 3} as a Green Catalyst. Polycyclic Aromatic Compounds 2019:1-16.doi.org/10.1080/10406638.2019.1653942

17. Saravanan G, Alagarsamy V, Prakash CR. Synthesis, characterization and in vitro antimicrobial activity of some 1-(substitutedbenzylidene)-4-(4-(2-(methyl/phenyl)-4-oxoquinazolin-3 (4H)-yl) phenyl) semicarbazide derivatives. Journal of Saudi Chemical Society 2015;19(1):3-11.doi.org/10.21276/sjams.2017.5.10.28

18. Kuchana M. In-Silico Study Of Molecular Properties, Bioactivity And Toxicity Of 2-(Substituted Benzylidene) Succinic Acids And Some Selected Anti-Inflammatory Drugs. 2020.doi.org/10.25004/IJPSDR.2020.120407

19. Becker DE, Reed KL. Local anesthetics: review of pharmacological considerations. Anesthesia progress 2012;59(2):90-103.doi.org/10.2344/0003-3006-59.2.90

20. Gowe C. Review on potential use of fruit and vegetables by-products as a valuable source of natural food additives. Food Science and Quality Management 2015;45:47-61.

21. Igwe K, Nwankudu O, Ijioma S, Madubuike A, Achi N. Screening for secondary metabolites in Huru crepitans bark ethanol extract using GCMS analysis: A preliminary study approach. Journal of Science and Technology Advances 2016;1(2):64-71.

22. Rizvi A. Heavy Metal Toxicity to certain Cereal Crops and Assessment of Bioremediation Potential of Plant Growth Promoting Rhizobacteria [Aligarh Muslim University; 2019.doi.org/10.1016/B978-0-12-821199-1.00035-3

23. Dubey SK, Batra A. Study of anti oxidant and anti-inflammatory activity from ethanol fraction of Thuja occidentalis Linn. Research Journal of Science and Technology.2009;1(1): 39-42.

24. Mortensen A. Re‐evaluation of fatty acids (E 570) as a food additive. EFSA Journal 2017;15(5):e04785.https://doi.org/10.2903/j.efsa.2017.4785

25. Dyntar D, Eppenberger-Eberhardt M, Maedler K, et al. Glucose and palmitic acid induce degeneration of myofibrils and modulate apoptosis in rat adult cardiomyocytes. Diabetes 2001;50(9):2105-13.doi.org/10.2337/diabetes.50.9.2105

26. Punasiya, Rakesh, Sujit P. In Vitro Antibacterial Activity of Leaf Extracts of Hibiscus Syriacus (L). Research Journal of Topical and Cosmetic Sciences, 2014;5(2):51-55.

27. Akacha NB, Gargouri M. Microbial and enzymatic technologies used for the production of natural aroma compounds: Synthesis, recovery modeling, and bioprocesses. Food and Bioproducts Processing 2015;94:675-706.doi.org/10.1016/J.FBP.2014.09.011

28. Hejna A, Kosmela P, Formela K, Piszczyk Ł, Haponiuk JT. Potential applications of crude glycerol in polymer technology–Current state and perspectives. Renewable and Sustainable Energy Reviews 2016;66:449-75.doi.org/10.1016/j.rser.2016.08.020

29. Huang J, Xia J, Jiang W, Li Y, Li J. Biodiesel production from microalgae oil catalyzed by a recombinant lipase. Bioresource Technology 2015;180:47-53.doi.org/10.1016/j.biortech.2014.12.072

30. Orozco D, Kouznetsov VV, Bermúdez A, et al. Recent synthetic efforts in the preparation of 2-(3, 4)-alkenyl (aryl) quinoline molecules towards anti-kinetoplastid agents. Rsc Advances 2020;10(9):4876-98.doi.org/10.1039%2Fc9ra09905k

31. Foley M, Tilley L. Quinoline antimalarials: mechanisms of action and resistance. Int J Parasitol 1997;27(2):231-40.doi.org/10.1016/s0163-7258(98)00012-6

32. AlMarzouq DS, Elnagdi NMH. Glycerol and Q-Tubes: Green Catalyst and Technique for Synthesis of Polyfunctionally Substituted Heteroaromatics and Anilines. Molecules 2019;24(9):1806.doi.org/10.3390/molecules24091806

33. Mashhadi NS, Ghiasvand R, Askari G, et al. Anti-oxidative and anti-inflammatory effects of ginger in health and physical activity: review of current evidence. International journal of preventive medicine 2013;4(Suppl 1):S36-S42.

34. Gupta I, Singh K, Varshney NK, Khan S. Delineating Crosstalk Mechanisms of the Ubiquitin Proteasome System That Regulate Apoptosis. Frontiers in cell and developmental biology 2018;6:11-11.doi.org/10.3389%2Ffcell.2018.00011

35. Soler-Velasquez MP, Brendemuhl JH, McDowell LR, et al. Effects of supplemental vitamin E and canola oil on tissue tocopherol and liver fatty acid profile of finishing swine. J Anim Sci 1998;76(1):110-7.doi.org/10.2527/1998.761110x

36. Czyrski A. Determination of the lipophilicity of ibuprofen, naproxen, ketoprofen, and flurbiprofen with thin-layer chromatography. Journal of Chemistry 2019;2019.doi.org/10.1155/2019/3407091

37. Mugumbate G, Overington JP. The relationship between target-class and the physicochemical properties of antibacterial drugs. Bioorganic and medicinal chemistry 2015;23(16):5218-24.doi.org/10.1016/j.bmc.2015.04.063

38. Bahar E, Yoon H. Lidocaine: a local anesthetic, its adverse effects and management. Medicina 2021;57(8):782.doi.org/10.3390/medicina57080782

39. Ridwan, Devijanti R, Wijayanti U. The Antibacterial activity of gingival mucoadhesive patch from Thymus vulgaris essential oil towards Aggregatibacter actinomycetemcomitans and Fusobacterium nucleatum. Research Journal of Pharmacy and Technology. 2021;14(2): 645-649. doi.org/10.5958/0974-360X.2021.00115.3

40. Cabiscol E, Tamarit J, Ros J. Oxidative stress in bacteria and protein damage by reactive oxygen species. Int Microbiol 2000;3(1):3-8.

Received on 27.01.2022 Modified on 07.07.2022

Research J. Pharm. and Tech 2023; 16(3):1231-1238.

DOI: 10.52711/0974-360X.2023.00204

*Sapindusrarak DC*	N	24 h				48 h
*Sapindusrarak DC*	N	OD.	S.Dev	Freq	Growth Response	OD	S.Dev	Freq	Growth Response
50%	3	0,362	0,272	64%	36%	0,406	0,180	34%	65,59%
25%	3	0,102	0,024	18%	82%	0,298	0,019	25%	74,80%
12,50%	3	0,002	0,006	0,4%	100%	0,222	0,068	19%	81,23%
6,25%	3	0,017	0,027	3%	97%	0,151	0,033	13%	87,19%
CHX 0,2%	3	0,016	0,007	3%	97%	0,019	0,009	2%	98,42%