Scribd Logo
Upload

Welcome to Scribd!

  • Jurna Internasional H. KARIM IN VITRO BIOACTIVITY

    Uploaded by

    akfar praktek
    8 views12 pages
    jurnal

    Copyright

    © © All Rights Reserved

    Available Formats

    PDF or read online from Scribd

    Did you find this document useful?

    Is this content inappropriate?

    Report this Document
    jurnal

    Copyright:

    © All Rights Reserved
    Download as PDF or read online from Scribd
    Flag for inappropriate content
    8 views12 pages

    Jurna Internasional H. KARIM IN VITRO BIOACTIVITY

    Uploaded by

    akfar praktek
    jurnal

    Copyright:

    © All Rights Reserved
    Download as PDF or read online from Scribd
    Flag for inappropriate content
    of 12
    §nternational Knowledge Press Ca Dated 30th Aug 2021 Subject: Acceptance letter for manuscript no. (2021/PCBMB/9689) submitted in PLANT CELL BIOTECHNOLOGY AND MOLECULAR BIOLOGY Dear Colleague, We are pleased to inform that peer review process and editorial review process have been completed for your following manuscript Manuscript no: 2021 /PCBMB/9689 Title: IN VITRO BIOACTIVITY OF GALANGAL RHIZOME EXTRACT AGAINST Streptococcus mutans AND Staphylococcus aureus GROWTH AND ITS APPLICATION AS AN ACTIVE SUBSTANCE IN TOOTHPASTE”. Author(s): H. KARIM* We are ready with the final decision, We are happy to inform you that your manuscript is officially accepted for publication in PLANT CELL BIOTECHNOLOGY AND MOLECULAR BIOLOGY Once your manuscript is moved to publishing, our production editor will keep you informed of your article's progress in the production process. You will also receive a galley proof of your manuscript for final review. We're ‘excited to move forward with your submission. Please feel free to email me with any questions. ‘Thank you for submitting your paper. ZR ‘a fe) Thanking you Ms. Laboni Mukherjee International Knowledge Press www.ikprress.org eter vadaton link: hips/ www skperess.ort/index php/PCRME acceptance eter Registered Offices: Asia Pacific: International Knowledge Press, N.S. Road, Tarakeswar, Hooghly, PIN-712410, West Bengal, India UK: International Knowledge Press, $107, 3 Hardman Square, Spinningfields, Manchester, M3 3EB, UK Plant Cell Biotehnology and Molecular Biology 22(14850) 86-95; 2021 ISSN; 0972-2025 IN vitro BIOACTIVITY OF GALANGAL RHIZOME EXTRACT AGAINST Streptococcus mutans AND Staphylococcus aureus GROWTH AND ITS APPLICATION AS AN ACTIVE SUBSTANCE IN TOOTHPASTE H. KARIM” Department of Pharmacy, Pharmacy College YAMASI, Makassar 90222, Indonesia [HK] [For Correspondence: E-mail: hamingsihkarim@gmail.com) Article Information Ettore: (1) De NebalS.Et-mougy. National Research Cente, Baypt Reviewers: (() ApilitaRins Yaoi Ef, Heath Sciences Universtas Fs Ung. adonesia (@) Carlos Tomde Quirno-omeds, Universided Aiténoma Metropoitan-Xochimico, México (Gy Bena Varma. Received: 25 June 2021 Accept I: 30 August 2021 Published: 03 September 2021 Orig ial Research Article ABSTRACT ‘The study aimed to isolate red (Alpinia purpurata K. Schum) and white galangal (Alpinia galanga Linn. Swartz) rhizome extract compound, examine its bioactivity against bacteria Streptococcus ‘mutans (ATCC 25175) and Staphylococcus aureus (ATCC 13568), and its potential as an active substance in toothpaste made at the laboratory scale. The result of the study indicated that the 1 hexane extract of red galangal possesses a higher antimicrobial activity effect against S. mutans and S aureus compared to the white one. The examination of the experimental toothpaste with a1 addition of 1 % active substance (extract of n-hexane, D and J fractions) indicated that the diameter of resistance against S mutans formed by D fraction was higher (15.50.50 mm) than the control (14.00.00 mm). Meanwhile, the addition of 1.5 % active substances from D and J fractions indicated a higher inhibition zone diameter (16.240.29 mm and 14,740.58 mm) compared to positive control. The diameter of resistance against S. aureus of the toothpaste with an addition of active substances (0.5, 1.0, and 1.5 %) was higher than the positive control. Spectroscopy FTIR and GC- MS data analysis indicated that the primary compound existing in the D fra 2methyl -1-(0- | In contrast, the most dominant compound in the J fraction is 2-ethyl-S-methyl ‘The D and J fraction can be used as an active substance in toothpaste in the future. Keywords: Galangal rhizome extract; S. mutans; S. aureus; active substance; toothpaste, INTRODUCTION traditional medicine. There is a growing trend in using natural plants as medicine along with the The advancement of technology and science has issue of back to nature, a prolonged economic not been able to replace the value of simple crisis and Covid-19 pandemic, resulting in a 86 decline of public's purchasing power towards relatively expensive modem medicines. Moreover, the escalate in the incidence of multidrug-resistant bacteria, which lead to the antimicrobial resistant, resistant to traditional sanitizers and toothpast, highlights the significancy of the search for alternative compounds [1, 2]. Therefore, one of the emerging alternative treatments is to increase the use of medicinal plants in the community. To improve the role of traditional medicine in public health services, efforts are required to introduce, conduct research, carry out test and develop the efficacy and safety of a medicinal plant Plant species belongs to the famity Zingiberaceae which has been used for generations by the Indonesian people as medicinal ingredients is ‘white galangal (Alpinia galanga Linn. Swartz) and red galangal (4lpinia purpurata K. Schum). The rhizome is the commonly used part of red galangal (Alpinia purpurata K. Schum). Galangal rhizome contains essential oils of | methyl-cinnamate, cineol, camphor, Spine, galangin, and eugenol DB]. In addition, the red galangal thizome (A. Purpuraia K." Schum) contains flavonoid compounds, _kaempferol-S-rutinoside and kaempferol-3-oliueronide [4]. The white and red galangal rhizome that contain a great mumber of active compounds are often used for treatments of eczema, pityriasis versicolor, bronchitis, ulcers, coryza, gastritis, morbili, otitis intema, cholera, ‘emaciation and to clean the mouth, stimulates the digestive power, appetite and as a purgative [5]. ‘Asa medicinal plant, red galangal is an ansibactrial that is used as a bad breath remover. Some studies indicated that the cause of bad breath is the presence of dental plague, where some bacteria breed. Bacteria usually found inthe mouth include Sireptococeus mutans, Streptococcus viridans, Staphylococcus ‘epidermidis, Staphylococcus pneumoniae, and Staphylococcus aureus [6]. Among these germs, Sireptococeus mutans and Staphylococcus aureus axe often used in testing to indicate plaque in the tooth layer [7], Based on the background of the medicinal properties believed by the people from generation "7 Karim to generation, and the content of secondary metabolites from galangal rhizomes that can be used as antibacterial, this study aimed to extract and isolate the chemical compounds contained in red galangal (Alpinia purpurata K. Schum), ‘examined its bioactivity against Streptococcus ‘mutans and Staphylococeus aureus for the most critical dental plagues in humans, and used galangal rhizome extract as an active ingredient in synthetic toothpaste MATERIALS AND METHODS: Materials ‘The ingredients used consisted of red galangal rhizome (A/pinia purpurata K. Schum), white galangal rhizome (Alpinia galanga Linn. Swartz), methanol solution (technical and pro analysis), n= hexane (pro analysis), chloroform (pro analysis), ethyl acetate (pro analysis), acetone (technical and pro analysis), coarse silica gel for impregnation (Merck, catalogue no. 7733), silica gel for ‘Vacuum Liquid Column Chromatography (VLCC) system (Merck, catalogue no. 7730), ‘TLC (Thin-Layer Chromatography) plate (Merck, catalogue no. 1.05553), qualitative test material (FSO, pro analysis, HCI pro analysis, acetic anhydrous acid, FeCl;, and Mg powder), DMSO (Merck, catalogue no 802912), Nutrient agar media (Merck, catalogue no. 105450), Muller Hinton Agar (MHA) media (Himedia catalogue no. M173-500G), Sireptococcus muians (ATCC 25175) and Staphylococeus aureus (ATCC 13565) bacteria strain. Instrumentations ‘The equipment used in this study consisted of ‘lass tools commonly used in laboratories, rotary evaporators, digital scales, separating funnels, fractionation devices (columns chromatography), TLC devices (TLC chambers, capillary pipes for bottlers, pencils, cutters, ruler, UV lamp), antibacterial test equipment (autoclave, centrifugation, shaker incubator, micropipette, ose wire, peti dish), and FTIR spectrometer (Shimadzu FT-IR), GC-MS (Shimadzu GCMS- QP), Procedures Material collection and sample preparation ‘The samples used in this study were red galangal thizome (Alpini purpuruta) obtained from Traditional Daya_market in Makassar, South Sulawesi, Indonesia, and white galangal rhizome (Alpinia galanga) obtained from Terong Market in Makassar, South Sulawesi, Indonesia. The two galangal species were biologically identified in the Taxonomy Laboratory of Biology Department, Hasanuddin University, Makassar, Indonesia Fresh red galangal rhizome (Alpinia purpuruta) and white galangal thizome (Aipinia galanga Linn, Swartz) were washed, thinly cut to + 1-2 mm size, dried by aerating, and then mashed. Extraction and partition of Active Compounds About 1.25 kg of each Red galangal and white galangal rhizome powder were macerated three times with methanol pa solution at room temperature for 3 x 24 hours. The resulting ‘macerate was 10 litres and concentrated using a rotary evaporator. The extract was then partitioned by liquid-liquid extraction from non-polar solvents to polar solvents, namely from n-hexane, chloroform, and ethyl acetate. The results of the partition were then tested for antibacterial activity {3} Fractionation of Active Compounds The partition results with the highest antibacterial activity, namely n-hexane, were then fractionated through the vacuum liquid — chromatography column (VLCC) using the appropriate eluent based on analysis with TLC. Each fractionation result was also monitored through analysis with TLC. Identification of Active Compounds The fractions of vacuum liquid chromatography column (VLCC) used as active substances, were identified as compounds with FTIR spectroscopy and GC-MS analysis. Antibacterial Bioactivity Test Antibacterial activity test of the galangal active substance was started with the preparation and 88 Karim formulation of toothpaste as previously described {8, 9]. Then the antibacterial activity test was performed for total methanol extract, extracts from partitions, fractions of vacuum liquid chromatography column (VLCC), and synthetic toothpaste in vitro against S. mutans and S. aureus by agar diffusion method using membrane discs sterile as described [9]. All antibacterial activity test were repeated three times with duplo analysis. and inhibition zone diameter values were expressed as mean + SEM (standard error of mean), RESULTS AND DISCUSSION Extraction and Partition of Compounds Components from Galangal Rhizome ‘The maceration with methanol on 1.25 kg of red and white galangal rhizome dried powder, produced a concentrated methanol macerate Furthermore, each methanol macerate was Partitioned ‘continuously using an increased polarity solvent, non-polar n-hexane solvent, semipolar chloroform, and polar ethyl acetate The process then produced the concentrated extracts of mhexane, chloroform, and ethyl acetate Fractionation of Active Compounds ‘A total of 5.5 grams of red galangal n-hexane extract, which had the highest antibacterial activity, was fractionated through vacuum liquid chromatography column (VLCC) with increased polarity eluents, including n-hexane, chloroform n-hexane (4 : 6), chloroform, EtOAc: chtoroform (6 : 4), EtOAc, acetone: ExOAc (6: 4), acetone and methanol The vacuum liquid chromatography column (VLCC) fractions were then tested for bioactivity against S. muans and S. aureus to determine the active fraction. From the antibacterial activity test, the D fraction and J fraction have higher activity compared with the other fractions (Table 2). Therefore, we used them as active antibacterial substances on synthetic toothpaste, and subsequently identified furtherly using FTIR) and = GC-MS spectrophotometers. Identification of Active Compounds Spectroscopy Method Identification result of the D fraction with FTIR and GC-MS spectroscopy The D fraction took the form of an oily yellow paste with a distinctive pungent odour and showed i ii oGH- Cl Cy Hs Karim ‘several spots in the TLC chromatogram (data not shown). Analysis with FTIR spectrophotometer on the D fraction provided a spectrum as shown in Fig. GC-MS analysis of fraction D chromatogram showed 26 peaks with dominant peaks was peak five as shown in Fig. 2 CHa-Cly Fig. 2. GC-MS chromatogram from the D fraction of red galangal rhizome extract ” Karim jarele e sht—~ 3. — Och ke eee peean ti ae . I eee exategm Cy peer a ea ner na i omin Clg CH CHa Ce -CHy cl Me He ao eae as a nEwewcn + meme try Fig. 3. The fragmentation pattern of compounds in peak 5 from the D fraction of the red gal rhizome extract The results of GC-MS spectroscopy on fraction D I= (toll) pentanol with a molecular weight of showed 26 peaks in the chromatogram, with the 192 g/mol. dominant peak was peak 5 with retention time (RT) of 23.618, and abundance of 27.81%, Based Based on the spectra of compound mass at peak 5, onthe spectrum of GC-MS database, _it can be assumed that the secondary metabolites fragmentation pattern, and FTIR spectrum, it can at peak 5 with a base peak at m/z 43 experienced bbe assumed that the compound which has fragmentation, with a pattern of fragmentation as similarity to the compound at peak 5 is 2-methyl- shown in Fig. 3. Analysis of FTIR spectroscopy 0 (KBr) data showed absorption bands at numbers of 3394 and 3371 cm’, which indicated the presence of the hydroxyl OH) sroup of an alcohol. This result was supported by the presence of C-OH absorbance at 1226, 1203 and 1095 cm’, The absorption at 3100 emi? wavenumber indicated the presence of an unsaturated group (=C-H). The absorption peaks at wave numbers 1612 and 1512 em indicated that the unsaturated group was aromatic. The Presence of sharp absorption at 725 and $40 cm” showed substituted aromatic patterns. Absorbance in the 2924 and 2854 cm regions indicated that the absorbance supported the presence of an aliphatic saturated CH group in the regions 1458 and 1390 cm", which were specific for bending, methylene (-CH2-) and methyl groups (-CH3) ig 1) Based on the IR spectroscopic data analysis results, it can be proposed that the peak 5 compound of the D fraction has a hydroxyl (-OH) Karim ‘group, aliphatic C-H, and substituted aromatic patterns. The structure is shown in Fig. 4 on CH-CH-CHy CH, - CH, cH cry Fig. 4, Structure of the compound 2-metil -1-(0- tolil) pentanol at peak § of D fraction of red ‘galangal rhizome extract ‘The result of identification of J fraction with FTIR and GC-MS spectroscopy The J fraction formed a brown paste with a distinctive stinging odour, and the TLC chromatogram showed several stains. The results of the analysis by FTIR spectrophotometer on the fraction are shown in Fig. 5 5. FTIR spectrum pattern of J fraction of n-hexane extract of red galangal rhizome Fig. 6. GC-MS chromatogram pattern of J fraction from galangal rhizome extract, Fig. 7. Fragmentation pattern of the compound at peak 5 of J fraction of red galangan rhizome extract 2 Data from GC-MS chromatogram analysis. for fraction J of nehexane extract of red galangal shizome is shown in Fi Analysis of GC-MS spectroscopic data on J fraction showed 23 peaks with peak 5 as the dominant peak, retention time (RT) of 23.393 and hhad an abundance of 43.24%, ‘The mass spectrum of peak 5 of J fraction was then matched to the mass spectrum of standard ‘compounds based on the GC-MS (Wiley 7. Lib) database, and with FTIR spectrum data, Based on the spectrum of the GC-MS database, fragmentation pattern, and FTIR spectrum, it can bbe assumed that the compound that have similarities with the compound at peak 5 of the J fraction is 2- ethyl-S-methyl benzoic acid with a ‘molecular weight of 164 g/mol. Based on the mass spectra of peak $ compound, it can be assumed that the secondary metabolites at peak 5 with a base peak at m/z 133 can experience 4 fragmentation. The fragmentation pattern of peak 5 of the J fraction is shown in Fig, 7. Based on the FTIR spectroscopy (KBr) data analysis, there was a reasonably sharp absorption band () at 3332 cm" wavenumbers, which indicated the presence of a hydroxyl (0-H) group ‘of a carboxylic acid. This result was supported by the presence of C=0 carboxylic acid absorbance at 1705 em", and C-OH absorbance for carboxylic acids at 1234 cm". The absorption at 3024 cm” ‘wavenumber indicated the presence of an unsaturated group (-C-H), The presence of absorption peaks at wavenumbers 1612 and 1512 fem" indicated that the unsaturated group was. aromatic, The presence of a sharp absorption at 833 cm" showed a substituted aromatic pattern supported by overtones in 2029 and 1890 cm". Absorbance in the area of 2924 cm" indicated the presence of an aliphatic saturated C-H group Karim orted by absorbance in regions 1450 and 1365 cm", which were specific for methylene (-CH2-) and methyl (-CH3) bending, Based on the IR spectroscopy data analysis results, it can be proposed that the peak 5 compound of fraction J has a substituted carboxylic, aliphatic, and aromatic pattern. Its structure described in Fig 8. 9 | E or C-O8 Nah Fig. 8. Structure of 2-etil-S-metilbenzoie acid at peak § from J fraction of n-hexane extract from red galangal rhizome. Bioactivity Test Results of bioactivity test of galangal rhizome extract The results of the antibacterial activity test on S. ‘mutans and S. aureus bacteria on total extracts of methanol, n-hexane extract, chloroform, and ethyl acetate from red and white galangal rhizomes are shown in Table 1 ‘As can be seen in Table 1, generally the results of the antibacterial activity of red galangal rhizome extract on S. mutans and S. anreus were higher than those of the white galangal rhizome. The n- hexane extract of red galangal rhizome had the highest bioactivity compared to other galangal Thizome extracts, as shown by the inhibition zones diameter that formed were 14.20.76 mm and 16.01.00 mm, ‘Table 1. Inhibition zone diameter for red rhizome extract on the growth of S. mutians and S. aureus Tahibiion Zane Dismeter, mam 2 SEM ‘Test Material Re hizome ‘White lzome Smeal Soares Euutans 8. ourene Toial methanol eaed Thal 1S 7S Tose 15340 58 hexane extract 42076 16081 00 1330038 18340.58, Chloroform extract 1333058 1434115 i2a0sk 1234058, Edy acetate extct 13720.38 150210 1272088 4041.00 93 Bioactivity test results for fractions resulted from column chromatography Antibacterial activity test for fractions produced by the column chromatography against S. mutans and S. aureus was performed, to obtain the active fraction used as the active substance in synthetic, toothpaste. The test results for antibacterial activity are shown in Table 2. Fraction Tay Fraction? (B) H2e108 Fraction 3 (©) 1133050 Fraction 45D) ‘158402923 24029 Fraction 64716) «340881734029 Pociond <@) 153405) 1588029 Friction 10 @) —13.08100——16.28029 Fetoa 1) S089 11 74058 Frctin 12) Gao TT ROSS Fraction 13d) 1524058 1684029 Fraction 14K) 127807515 381.04 Fraction 5 () Hoon 1138126 Table 2 shows that all n-hexane fractions produced by active vacuum liquid column chromatography (VLCC) inhibit the growth of S. muans and S. aureus bacteria. They are potentially used as active antibacterial substances. ‘The D and J fractions used as active substances in synthetic. toothpaste were analyzed using FTIR spectroscopy and GC-MS. Test results for bi substance from red g: synthetic toothpaste tivity of the active ingal rhizome extract in ‘The test results of antibacterial activity of synthetic toothpaste which were each treated with the active substance of n-hexane, fraction D, and fraction J with several concentration variations (055, 1.0, and 1.5%). The synthetic toothpaste was compared to a non-herbal toothpaste of certain brand which is usualy used by Indonesian as the positive control, named Toothpaste X, and synthetic toothpaste without active substance as the negative control, as shown in Table 3. From the antibacterial activity test results on toothpaste, it was seen that toothpaste treated with the active substance from fraction D showed the highest Karim activity compared to synthetic toothpaste treated with n-hexane extract active ingredient, J fraction, ‘and positive control. Moreover, it can be seen in Table 3 that synthetic toothpaste with the n- hexane extract, fraction D, and fiaction J active ingredient demonstrated larger inhibitions areas than positive controls. In parallel with the study conducted by Sujono, et a. [10] which comparing the antibacterial activity of essential oils of betel leaves and NaF, it was found that the inhibition of essential oils of betel leaves was up to three times greater than NaF at all test concentrations. This result was in line with the research conducted by ‘Awah [11], who conducted antibacterial tests on several kinds of herbal and non-herbal toothpaste The results showed that herbal toothpaste had ‘more remarkable antibacterial ability than non- herbal ones. The satisfactory bioactivity of n- hexane extract, fraction D, and the J fraction proved that galangal rhizome extract could be Used as an active herbal substance. Table 3. The results of the antibacterial activity test for galangal rhizome extract as the active substance in synthetic toothpaste Fraction of Test Inhibition Zone Diameter, ‘material mean + SEM (wn) Smwians —— S aurens whecne Osi aria 1128076 M4080 00 ‘osixane [% exact 1238058 15 82076 besane |% east 1402000 16-540:50 Fraction D0S% 131.0014 08,00 Fraction DI% 1540.50.15 K40.75, FracioaD 15% 16280291630 Fraction J05% 1128029 1434058 Fraction 3196 Lats? 1642029 Frachons15% WTAE 18240 29 Toothpaste X (Control +) 14030100 13.08 30 ‘Synihetic totus” 608000 6040.00 (Comte: ‘This study indicated that methanol and n-hexane extract of red A. galanga rhizome could strongly inhibit the growth of S. mutans and S. aureus in vitro. The results of the antibacterial activity test fof galangal thizome extract as an active herbal ingredient in toothpaste showed a reliable ability of its antibacterial activity. This is the first fesearch which utilizing this natural material as an alternative antcaries in preventive dentistry. The results are consistent. with previous research, where the 4. galanga aqueous extract demonstrated remarkable antibacterial activity against various bacteria including Klebsiella pneumonia, Escherichia coli, Pseudomonas deruginosa, S$. aureus and Streptococeus pyogenes. However, the extract was not efficient against Staphylococcus epidermidis [12 Moreover, Alpinia galanga flower oven-dried ethanol extract was proved to be the most effective one against S. aureus. The flower extract formed 2 relatively large inhibition zone of about 26-31 mm, with the minimum inhibitory concentration (MIC) value ranging from 0.352 to 0.547 mg/mL. Overall, the oven-dried ethanol extract samples’s antimicrobial activity showed the highest inhibition zone of 8.94 mm, and MIC of 1.457 mg/mL. In contrast, freeze-dried ethanol extracted samples displayed the _ lowest antimicrobial activity (7.05 mm and 2.470 mg/mL) [13] According to the transmission lectron microscopy test, the Alpinia galanga ‘extract induced - membrane damage and cytoplasm ‘coagulation. These disruptions were indicated by the disclose of cell materials, such as nucleic acids (13, 14), Certainly, further research with various methods are required to identify the specific antibacterial mechanism. For example, further studies are necessitated to examine the antibacterial ability of galangal extract im vivo More studies are significantly important to assess the antibacterial ability of galangal rhizome extract against other bacteria which commonly found in the oral cavity. CONCLUSIONS Bioactivity test results of red galangal and white galangal rhizome extract against S. mutans and S. aureus indicated that n-hexane extract of red galangal thizome has the most desirable antibacterial activity as indicated by the formation cof large inhibition zone diameter of 15 mm for S. ‘mutans and 16 mm for S. aureus. According to the identification results of FTIR. spectroscopy and GC-MS, it is found that the dominant active compound component from the D fraction is 2- methyl-1-(c-tolil) pentanol, whereas the dominant active compound component from J fraction is 2- ethyl-5-methyl benzoic acid. In the addition of 1% active substance, the D fraction indicated higher inhibition zone diameter compared to the positive ‘control, whereas the addition of 1.5% of n-hexane, the D fraction, and the J fraction extract of the Karim active substance can properly inhibit the growth of S. mutans and S$. aureus compared to the Toothpaste X as positive control with sodium fluoride (NaF) active substance. COMPETING INTERESTS ‘Author has declared that no competing interests exist REFERENCES 1 Hsu WY, Simonne A, Weissman A, Kim JM. Antimicrobial activity of _ greater galangal [4lpinia galanga (Linn.) Swartz} flowers. Food Sci. Biotechnol. 2010, 19(4)'873-880, 2 Rao K, Ch B, Narasu IM, ef al Antibacterial activity of Alpinia sgalanga (L) Willd Crude Extracts. Appl Biochem Biotechnol, 2010;162:871-884. 3. Rialita’ T, Rahayo WP, Alfiah D. Antimicrobial activity of the combination of red galangal (4lpinia purpurata K. Schum) and cinnamon (Cinnamonum ‘burmanit) essential cils on Escherichia colt and Staphylococcus aureus bacteria Journal of Physics: Conf Series. 2019,1217: 012132 4, Vietorio CP, Kuster RM, Lage CLS. Detection of flavonoids’ in Alpinia purpuraia (Vieill.) K. Schum. leaves using high-performance liquid chromatography, Rev. Bras, Plantas Med, 2009;11 (2):147. 5. AlSnafi AE. The — pharmacological activities of Alpinia galangal - A review. International Joumal for Pharmaceutical Research Scholars. 2014:3(1-1):607- 614. 6. Sizar_ 0, Unakal CG. Gram Positive Bacteria, Treasure Island (FL): Stat Pearls Publishing, India; 2021 7. Apriansi, M. Effect of siwak wood powder extract (Salvadora persica) on the growth of Streprococcus mutans Bacteria. Journal of Agrogua. 2017;15(2):29-34 8 Gautam D, Palkar P, Maule K. Singh, Sawant S, Kuvalekar G, Rukari CT, Jagtap V. A. preparation, "evaluation and comparison of herbal toothpaste with snarketed herbal toothpaste. Asian Journal 10 u. of Pharmacy and Technology. 2020, 10(3):165. Mangilal T, Ravikuma M. Preparation and evaluation of herbal toothpaste with commercial herbal toothpastes: An in-vitro study. IJAHM. 2016;6(3):2266-2273, Sujono H, Rizal S, Purbaya S. dan Jasmansyah. Antibacterial activity of the essential cil from betel leaf (Piper betle L.) against. Streptococcus pyogenes and Staphylococeus aureus, Jurmal Kartika Kimia. 2019,1(2)30-36 Awah NS, Agu KC, Okorie CC, Okeke CB, etal. Invitro" assessment of the antibacterial quality of some commonly used herbal and non-herbal toothpastes on Streptococcus mulans, Open Journal of 4 Karim Dentistry and Oral Medicine, 2016,4(2)21- 25, Turker A, Usta C. Biological screening of some Turkish medicinal plant extracts for ‘antimicrobial and toxicity activities. Natural Product Research. 2008;22(2):136- 146. Eram S. Mujahid, M. Bagga, P. Arif, M. Ahmad MA, et al. A review on Phytopharmacological activity of Alpinia

    You might also like