Stability and Antibacterial Property of Polyherbal Mouthwash Formulated Using Local Ingredients

Juman Nafea, Mohamed Edbeib, Kin Israel R. Notarte, Fahrul Huyop, Harisun Yaakub


The oral cavity is a home to more than 500 bacterial species. Although some of the oral bacteria are harmless, there are certain species that may cause oral plaques, bad breath, and mouth disease. Thus, maintaining a good oral hygiene is essential for a healthy mouth and body. The present study aimed to formulate a polyherbal mouthwash that may have antibacterial properties. Mouthwash formulations were prepared containing varying percentages of herbal extracts, with each formulation stored at 12 °C and 25 °C. Over the course of 12 weeks, the appearance and pH of the formulated mouthwash were measured. The mouthwash formulations maintained good homogeneity and color when stored at 25 °C, displaying lower pH level ranging between 3.71 and 4.85. Although the mouthwash stored at 12 °C maintained good homogeneity, a change in color in the formulation was evident and a more unstable pH readings were recorded. Antibacterial assay showed that mouthwash formulations stored at 25 °C had better inhibitory activity compared to those stored at 12 °C. Furthermore, mouthwash formulation containing (30% v/v) aleppo oak extract as the major ingredient conferred the greatest inhibition zone diameter (IZD = 10-18 mm) against salivary bacteria compared to formulations with (30% v/v) clove and (30% v/v) turmeric extracts as major ingredients. The best polyherbal mouthwash formulation in terms of inhibiting bacterial growth followed the 3:1:2 ratio for aleppo oak extract, clove extract, and turmeric, respectively. Therefore, the polyherbal mouthwash formulated in this study has the potential to be optimized and commercialized to antagonize growth of pathogenic oral bacteria.


Antibacterial; Polyherbal; Mouthwash; Aleppo Oak; Clove; Turmeric


Abdelkhalek, N. K., Risha, E., El-Adl, M. A., Salama, M. F., & Dawood, M. A. O. (2020). Antibacterial and antioxidant activity of clove oil against Streptococcus iniae infection in Nile tilapia (Oreochromis niloticus) and its effect on hepatic hepcidin expression. Fish & Shellfish Immunology, 104, 478-488.

Alali, A. Y., Al-Khabbaz, A., Michael, S., & Swain, M. V. (2018). Frictional coefficient during flossing of teeth. Dental Materials, 34(12), 1727-1734.

Alsammarraie, F. K., Wang, W., Zhou, P., Mustapha, A., & Lin, M. (2018). Green synthesis of silver nanoparticles using turmeric extracts and investigation of their antibacterial activities. Colloids and Surfaces B: Biointerfaces, 171, 398-405.

Amador-Medina, L. F., Alvarez, J. A., Macias, A. E., Muñoz, J. M., Mosqueda, J. L., Arreguin, V., & Collazo, E. M. (2019). Does chlorhexidine mouthwash reduce the rate of oral colonization bygram-negative bacteria in patients with chemotherapy? Aplacebo-controlled trial. American Journal of Infection Control, 47(5), 591-594.

Anderson, R. A., Wang, J., Plassman, B. L., Nye, K., Bunn, M., Poole, P. A., Drake, C., Xu, H., Ni, Z., & Wu, B. (2019). Working together to learn new oral hygiene techniques: Pilot of a carepartner-assisted intervention for persons with cognitive impairment. Geriatric Nursing, 40(3), 269-276.

Bandura, A. V., & Lvov, S. N. (2005). The ionization constant of water over wide ranges of temperature and density. Journal of Physical and Chemical Reference Data, 35(1), 15-30.

Bescos, R., Ashworth, A., Cutler, C., Brookes, Z. L., Belfield, L., Rodiles, A., Casas-Agustench, P., Farnham, G., Liddle, L., Burleigh, M., White, D., Easton, C., & Hickson, M. (2020). Effects of chlorhexidine mouthwash on the oral microbiome. Scientific Reports, 10(1), 5254.

Carvalho, R. S., Carollo, C. A., de Magalhães, J. C., Palumbo, J. M. C., Boaretto, A. G., Nunes e Sá, I. C., Ferraz, A. C., Lima, W. G., de Siqueira, J. M., & Ferreira, J. M. S. (2018). Antibacterial and antifungal activities of phenolic compound-enriched ethyl acetate fraction from Cochlospermum regium (mart. Et. Schr.) Pilger roots: Mechanisms of action and synergism with tannin and gallic acid. South African Journal of Botany, 114, 181-187.

Chiedozie, E. I., Ahamefule, O. F., & Ukamaka, A. A. (2016). Anti-inflammatory, antimicrobial and stability studies of poly-herbal mouthwashes against Streptococcus mutans. Journal of Pharmacognosy and Phytochemistry, 5(5), 354-361.

Chowdhury, B. R., Garai, A., Deb, M., & Bhattacharya, S. (2013). Herbal toothpaste: A possible remedy for oral cancer. Journal of Natural Products, 6, 44-55.

de Oliveira Marreiro, R., Bandeira, M. F. C. L., de Souza, T. P., de Almeida, M. C., Bendaham, K., Venâncio, G. N., Rodrigues, I. C., Coelho, C. N., Milério, P. S. L. L., & de Oliveira, G. P. (2014). Evaluation of the stability and antimicrobial activity of an ethanolic extract of Libidibia Ferrea. Clinical, Cosmetic and Investigational Dentistry, 6, 9-13.

Guerrero, J. J. G., & Notarte, K. I. R. (2020). Mga tanom na nakakabulong: Medicinal plant studies among the undergraduate researches of Bicol University - Department of Biology from 1991 to 2019. Philippine Journal of Health Research and Development, 24(3), 1-12.

Hadidi, M., Pouramin, S., Adinepour, F., Haghani, S., & Jafari, S. M. (2020). Chitosan nanoparticles loaded with clove essential oil: Characterization, antioxidant and antibacterial activities. Carbohydrate Polymers, 236, 116075.

Hayasaki, H., Saitoh, I., Nakakura-Ohshima, K., Hanasaki, M., Nogami, Y., Nakajima, T., Inada, E., Iwasaki, T., Iwase, Y., Sawami, T., Kawasaki, K., Murakami, N., Murakami, T., Kurosawa, M., Kimi, M., Kagoshima, A., Soda, M., & Yamasaki, Y. (2014). Tooth brushing for oral prophylaxis. Japanese Dental Science Review, 50(3), 69-77.

Javali, M. A., Abdul Khader, M., Alqahtani, R. M., Almufarrij, M. J., Alqahtani, T. M., & Addas, M. K. (2020). Spectrophotometric Analysis of dental enamel staining to antiseptic and dietary agents: In vitro study. International Journal of Dentistry, 2020, 1-5.

Jiao, Y., Tay, F. R., Niu, L., & Chen, J. (2019). Advancing antimicrobial strategies for managing oral biofilm infections. International Journal of Oral Science, 11(3), 28.

Kilian, M., Chapple, I. L. C., Hannig, M., Marsh, P. D., Meuric, V., Pedersen, A. M. L., Tonetti, M. S., Wade, W. G., & Zaura, E. (2016). The oral microbiome: An update for oral healthcare professionals. British Dental Journal, 221(10), 657-666.

Liang, P., Zhao, C., Lin, Y., Geng, J., Chen, Y., Chen, D., & Zhang, X. (2020). Effects of sodium benzoate on growth and physiological characteristics of wheat seedlings under compound heavy metal stress. Journal of Integrative Agriculture, 19(4), 1010-1018.

Lin, J., Dinis, M., Tseng, C. H., Agnello, M., He, X., Silva, D. R., & Tran, N. C. (2020). Effectiveness of the GumChucks flossing system compared to string floss for interdental plaque removal in children: a randomized clinical trial. Scientific Reports, 10(1), 3052.

Mahulette, A. S., Hariyadi, Yahya, S., & Wachjar, A. (2020). Physico-chemical properties of clove oil from three forest clove accession groups in Maluku. IOP Conference Series: Earth and Environmental Science, 418, 012028.

Mahyari, S., Mahyari, B., Emami, S. A., Malaekeh-Nikouei, B., Jahanbakhsh, S. P., Sahebkar, A., & Mohammadpour, A. H. (2016). Evaluation of the efficacy of a polyherbal mouthwash containing Zingiber officinale, Rosmarinus officinalis and Calendula officinalis extracts in patients with gingivitis: A randomized double-blind placebo-controlled trial. Complementary Therapies in Clinical Practice, 22, 93-98.

Mitha, S., Elnaem, M. H., Koh, M., En, C., Babar, M. G., Siddiqui, J., & Jamshed, S. (2016). Use and perceived benefits of mouthwash among Malaysian adults: an exploratory insight. Journal of Advanced Oral Research, 7(3), 7-14.

Paula, I. M. B., Moraes, F. C., Souza, O. V., & Yamamoto, C. H. (2014). Development of mouthwash with Rosmarinus officinalis extract. Brazilian Journal of Pharmaceutical Sciences, 50(4), 851-858.

Qu, T., Li, B., Huang, X., Li, X., Ding, Y., Chen, J., & Tang, X. (2020). Effect of peppermint oil on the storage quality of white button mushrooms (Agaricus bisporus). Food and Bioprocess Technology, 13(3), 404-418.

Sallih, M. A. E. B. M., & Bakar, M. Z. B. A. (2019). Randomised controlled trial of salt solution (sodium chloride) mouthwash vs thymol glycerine usage in sore throat with non bacterial pharyngitis. Journal of Complementary and Alternative Medical Research, 8(1), 1-5.

Scerri, E. R. (2019). Five ideas in chemical education that must die. Foundations of Chemistry, 21(1), 61-69.

Sha, A. M., & Garib, B. T. (2019). Antibacterial effect of curcumin against clinically isolated Porphyromonas gingivalis and connective tissue reactions to curcumin gel in the subcutaneous tissue of rats. BioMed Research International, 2019, 6810936.

Shahmohammadi, M., Javadi, M., & Nassiri-Asl, M. (2016). An overview on the effects of sodium benzoate as a preservative in food products. Biotechnology and Health Sciences, 3(3), 7-11.

Shetty, S. B., Mahin-Syed-Ismail, P., Varghese, S., Thomas-George, B., Kandathil-Thajuraj, P., Baby, D., Haleem, S., Sreedhar, S., & Devang-Divakar, D. (2016). Antimicrobial effects of Citrus sinensis peel extracts against dental caries bacteria: An in vitro study. Journal of Clinical and Experimental Dentistry, 8(1), e71-e77.

Siraj, E. S., Pushpanjali, K., & Manoranjitha, B. S. (2019). Efficacy of stevioside sweetener on pH of plaque among young adults. Dental Research Journal, 16(2), 104-109.

Tartaglia, G. M., Tadakamadla, S. K., Connelly, S. T., Sforza, C., & Martín, C. (2019). Adverse events associated with home use of mouthrinses: a systematic review. Therapeutic Advances in Drug Safety, 10, 2042098619854881.

Tayel, A. A., El-Sedfy, M. A., Ibrahim, A. I., & Moussa, S. H. (2018). Application of Quercus infectoria extract as a natural antimicrobial agent for chicken egg decontamination. Revista Argentina de Microbiología, 50(4), 391-397.

Tayel, A. A., El-Tras, W. F., Abdel-Monem, O. A., El-Sabbagh, S. M., Alsohim, A. S., & El-Refai, E. M. (2013). Production of anticandidal cotton textiles treated with oak gall extract. Revista Argentina de Microbiología, 45(4), 271-276.

Toyama, T., Furukawa, Y., Hayashi, Y., Araki, M., Nakano, T., & Fukuta, O. (2019). Dental floss use among elementary school children and environmental factors; a cross-sectional study. Pediatric Dental Journal, 29(2), 78-83.

Tribble, G. D., Angelov, N., Weltman, R., Wang, B. Y., Eswaran, S. V., Gay, I. C., Parthasarathy, K., Dao, D. H. V., Richardson, K. N., Ismail, N. M., Sharina, I. G., Hyde, E. R., Ajami, N. J., Petrosino, J. F., & Bryan, N. S. (2019). Frequency of tongue cleaning impacts the human tongue microbiome composition and enterosalivary circulation of nitrate. Frontiers in Cellular and Infection Microbiology, 9(39).

Tyagi, P., Singh, M., Kumari, H., Kumari, A., & Mukhopadhyay, K. (2015). Bactericidal activity of curcumin I is associated with damaging of bacterial membrane. PloS One, 10(3), e0121313.

Ulloa, P. C., van der Veen, M. H., & Krom, B. P. (2019). Modulation of the oral microbiome by the host to promote ecological balance. Odontology, 107(4), 437-448.

Ustrell-Borràs, M., Traboulsi-Garet, B., & Gay-Escoda, C. (2020). Alcohol-based mouthwash as a risk factor of oral cancer: A systematic review. Medicina Oral, Patologia Oral y Cirugia Bucal, 25(1), e1-e12.

Zhang, H., Peng, X., Li, X., Wu, J., & Guo, X. (2017). The application of clove extract protects chinese-style aausages against oxidation and quality deterioration. Korean Journal for Food Science of Animal Resources, 37(1), 114-122.



  • There are currently no refbacks.

Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.