Integration of Butterfly Pea (Clitoria ternatea L.) Flower Extract and ZnO into a Mouthwash: Physicochemical Characteristics and Antibacterial Activity

Authors

DOI:

https://doi.org/10.35451/jfm.v9i1.3955

Keywords:

Clitoria ternatea, ZnO, mouthwash, flavonoids, physicochemical characteristics, antibacterial

Abstract

Butterfly pea (Clitoria ternatea L.) is a source of flavonoids with potential biological activity, while ZnO is known for its antibacterial properties, making their integration into a mouthwash formulation of particular interest. This study aimed to evaluate the physicochemical characteristics and antibacterial activity of a mouthwash integrating butterfly pea flower extract and ZnO. The extract was obtained by maceration using 70% ethanol, and its total flavonoid content was determined by UV-Vis spectrophotometry using the AlCl3-quercetin method. Mouthwash formulations were prepared with extract concentrations of 0.5% (F1), 1.0% (F2), and 1.5% (F3), while ZnO was maintained at 0.05%, with ZnO without extract used as a control. Evaluations included organoleptic properties, homogeneity, pH, viscosity, physical stability, and antibacterial activity against Staphylococcus aureus. The extract exhibited a mean absorbance of 0.3562 at 373.4 nm, corresponding to a total flavonoid content of 19.61 mg QE/g. F3 showed the best characteristics, with a pH of 6.08 ± 0.05, viscosity of 2.68 ± 0.07 mPa·s, good homogeneity, and no phase separation. F3 produced an inhibition zone of 13.67 ± 0.45 mm, higher than the ZnO control (8.21 ± 0.32 mm). These findings indicate that increasing the extract concentration to 1.5% at a fixed ZnO concentration produced a mouthwash with favorable physicochemical characteristics and the highest antibacterial activity among the tested formulations.

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References

[1] Newman BA, Rosebrough CN, Tamashiro RA, Dias Ribeiro AP, Whitlock JA, Sidhu G, Aukhil I, Porral DY, Progulske-Fox A, Myntti MF, Wang GP. A randomized controlled trial to evaluate the effectiveness of a novel mouth rinse in patients with gingivitis. BMC Oral Health 2022;22:461. https://doi.org/10.1186/s12903-022-02518-2.

[2] Lin Y-C, Tsai C-F, Huang H-L. Effects of hypochlorous acid mouthwash on salivary bacteria including Staphylococcus aureus in patients with periodontal disease: a randomized controlled trial. BMC Oral Health 2023;23:698. https://doi.org/10.1186/s12903-023-03358-4.

[3] Fahim MFM, Zarnigar P. Effect of prepared herbal mouthwash in maintaining the oral health of school children: A single-blind randomised control trial. EXPLORE 2024;20:535–43. https://doi.org/10.1016/j.explore.2023.12.003.

[4] Montesani L, Montesani L, Mateo L, Daep C, Huber N, Isapour G, Zhang Y-P. Antibacterial and clinical effectiveness of a mouthwash with a novel active system of amine + zinc lactate + fluoride: a randomized controlled trial. Clin Oral Investig 2024;28:90. https://doi.org/10.1007/s00784-023-05487-0.

[5] Sabarish R, A RL, Joseph RA, Palraj KK, Thayanidhi PM, Santhanakrishnan M. Evaluation of antibacterial and antifungal activities of a commercially available herbal mouthwash in accordance with the PN-EN 1040 and EN 1650 standards. Diagn Microbiol Infect Dis 2025;111:116680. https://doi.org/10.1016/j.diagmicrobio.2025.116680.

[6] Kumar RS, Ankola A V, Sankeshwari RM, Hampiholi V, Jalihal S, Pai Khot AJ, Santhosh VN, Ragu K. Clinical and microbial evaluation of mouthwash containing Achyranthes aspera and Trachyspermum ammi: A randomized controlled non-inferiority trial. J Oral Biol Craniofacial Res 2024;14:522–9. https://doi.org/10.1016/j.jobcr.2024.06.006.

[7] Iskandar B, Lukman A, Syaputra S, Al-Abrori UNH, Surboyo MDC, Lee C-K. Formulation, characteristics and anti-bacterial effects of Euphorbia hirta L. mouthwash. J Taibah Univ Med Sci 2022;17:271–82. https://doi.org/10.1016/j.jtumed.2021.08.009.

[8] Sakthivel A, Sankaran K, Rengasamy G, Vishnu Priya V, Sathishkumar P. Formulation of Mouthwash Using Combined Herbal Extracts to Control the Predominant Oral Pathogens and Biofilm. J Herb Med 2024;46:100905. https://doi.org/10.1016/j.hermed.2024.100905.

[9] Suwandi T, Sundjojo M, Panjaitan CC, Hepziba ER, Setiabudi TO. Antibacterial effects of rosella petal extract (Hibiscus sabdariffa L.) in mouthwash formulation against Streptococcus sanguinis and Porphyromonas gingivalis Biofilms: An in vitro study. Saudi Dent J 2025;37:4. https://doi.org/10.1007/s44445-025-00007-0.

[10] Kumar RS, Ankola A V, Nagamoti MB, Sankeshwari RM, Sutar KP, Jigan SI, Pai Khot AJ. Antibacterial and cytotoxicity properties of a polyherbal mouthwash containing Achyranthes aspera and Trachyspermum ammi against selected periodontal pathogens. J Ayurveda Integr Med 2024;15:101046. https://doi.org/10.1016/j.jaim.2024.101046.

[11] Mulyanti S, Laela DS, Vasall PRN, Permadi N, Aripin D, Amaliya A, Rukayadi Y, Julaeha E. Antibacterial and Anti-Inflammatory Properties of Citrus aurantiifolia Essential Oil–Based Mouthwash: Potential and Cytotoxicity Concerns. Scientifica (Cairo) 2026;2026:5570684. https://doi.org/10.1155/sci5/5570684.

[12] Harefa K, Ritonga AH, Aritonang B, Gurusinga R, Wulan S, Irmayani I. The Impact of Butterfly Pea Flower (Clitoria ternatea L.) Extract on Atherosclerosis Biomarker Profiles in Obese White Rats (Rattus norvegicus L.). J Biomed Transl Res 2024;10:7–14. https://doi.org/10.14710/jbtr.v10i1.20281.

[13] Harefa K, Ritonga AH, Safitri R, Aritonang B. Anti-inflammatory Activity of Clitoria ternatea L. and Hibiscus sabdariffa L. Extract Combination in Reducing VCAM-1 Levels in Atherosclerosis. J Keperawatan Dan Fisioter 2024;7:128–36. https://doi.org/10.35451/jkf.v7i1.2300.

[14] Harefa K, Ritonga AH, Safitri R, Aritonang B, Gurusinga R, Irmayani I, Wulan S. Butterfly Pea and Roselle Combination Extracts Reduce V-CAM, ICAM, and IL-6 Levels in High Fat Atherogenic Diet Rats. Indones Biomed J 2024;16:527–33. https://doi.org/10.18585/inabj.v16i6.3361.

[15] Escher GB, Wen M, Zhang L, Rosso ND, Granato D. Phenolic composition by UHPLC-Q-TOF-MS/MS and stability of anthocyanins from Clitoria ternatea L. (butterfly pea) blue petals. Food Chem 2020;331:127341. https://doi.org/10.1016/j.foodchem.2020.127341.

[16] Thuy NM, Minh VQ, Ben TC, Thi Nguyen MT, Ha HT, Tai N V. Identification of Anthocyanin Compounds in Butterfly Pea Flowers (Clitoria ternatea L.) by Ultra Performance Liquid Chromatography/Ultraviolet Coupled to Mass Spectrometry. Molecules 2021;26:4539. https://doi.org/10.3390/molecules26154539.

[17] Fu X, Wu Q, Wang J, Chen Y, Zhu G, Zhu Z. Spectral Characteristic, Storage Stability and Antioxidant Properties of Anthocyanin Extracts from Flowers of Butterfly Pea (Clitoria ternatea L.). Molecules 2021;26:7000. https://doi.org/10.3390/molecules26227000.

[18] Caroline Paz Gonçalves G, Lizandra Gomes Rosas A, Carneiro de Sousa R, Regina Rodrigues Vieira T, César de Albuquerque Sousa T, Ramires T, Ferreira Ferreira da Silveira T, Barros L, Padilha da Silva W, Renato Guerra Dias Á, da Rosa Zavareze E, Dillenburg Meinhart A. A green method for anthocyanin extraction from Clitoria ternatea flowers cultivated in southern Brazil: Characterization, in vivo toxicity, and biological activity. Food Chem 2024;435:137575. https://doi.org/10.1016/j.foodchem.2023.137575.

[19] Sai-Ut S, Teksee A, Pongsetkul J, Sinthusamran S, Rawdkuen S. Optimization of ultrasonic assisted ethanolic extraction for natural pigments from butterfly pea flower applied in Thai dessert using Box-Behnken approach. Food Chem X 2024;22:101484. https://doi.org/https://doi.org/10.1016/j.fochx.2024.101484.

[20] Indrianingsih AW, Wulanjati MP, Windarsih A, Bhattacharjya DK, Suzuki T, Katayama T. In vitro studies of antioxidant, antidiabetic, and antibacterial activities of Theobroma cacao, Anonna muricata and Clitoria ternatea. Biocatal Agric Biotechnol 2021;33:101995. https://doi.org/10.1016/j.bcab.2021.101995.

[21] Satria D, Sofyanti E, Wulandari P, Fajarini, Pakpahan SD, Limbong SA. Antibacterial activity of Medan Butterfly pea (Clitoria ternatea L.) corolla extract against Streptococcus mutans ATCC®25175TM and Staphylococcus aureus ATCC®6538TM. Pharmacia 4AD;69:195–202. https://doi.org/10.3897/pharmacia.69.e77076.

[22] Sathiaseelan A, Song KP, Tan HS, Choo WS. Antibiofilm activity of Clitoria ternatea flowers anthocyanin fraction against biofilm-forming oral bacteria. FEMS Microbiol Lett 2025;372:fnaf035. https://doi.org/10.1093/femsle/fnaf035.

[23] Ray RR, Pattnaik S. Evaluating the antibacterial and antibiofilm activities of the extract of Clitorea ternatea petals, present in blue tea, against Alcaligenes faecalis. Pharmacol Res - Nat Prod 2026;10:100572. https://doi.org/10.1016/j.prenap.2026.100572.

[24] Isfa B, Jamarun N, Emriadi E, Arief S, Ritonga AH, Tanjung DA, Sisca V. Characterization and Application of TiO2/ZnO Nanoparticles as Pigments in Matt-Type Water-Based Paint. Int J Technol 2023;14:291–319. https://doi.org/10.14716/ijtech.v14i2.5501.

[25] Ritonga AH, Santika I, Harahap HY, Aritonang B. Antibacterial Activity of Taro Leaves Extract Combined with ZnO and Organo-ZnO Against Staphylococcus aureus. Forte J 2025;5:137–46. https://doi.org/10.51771/fj.v5i1.1197.

[26] Ritonga AH, Sisca V, Aritonang B, Meilani D, Putri GE, Siahaan EWB. Integrated PLA/LLDPE nanocomposites with compatibilizer and hydroxyapatite-zinc oxide: Mechanical, physical, thermal, and morphological properties. South African J Chem Eng 2025;52:1–7. https://doi.org/10.1016/j.sajce.2025.01.005.

[27] Ritonga AH, Aritonang B, Meilani D, Herlina, Sisca V, Putri GE, Siregar J, Siahaan EWB. Oleic acid-modified zinc oxide incorporated compatibilized PLA/LLDPE nanocomposites: Physical, mechanical, thermal, and morphological properties. South African J Chem Eng 2026;56:100869. https://doi.org/10.1016/j.sajce.2026.100869.

[28] Mendes CR, Dilarri G, Forsan CF, Sapata V de MR, Lopes PRM, de Moraes PB, Montagnolli RN, Ferreira H, Bidoia ED. Antibacterial action and target mechanisms of zinc oxide nanoparticles against bacterial pathogens. Sci Rep 2022;12:2658. https://doi.org/10.1038/s41598-022-06657-y.

[29] Abdelghafar A, Yousef N, Askoura M. Zinc oxide nanoparticles reduce biofilm formation, synergize antibiotics action and attenuate Staphylococcus aureus virulence in host; an important message to clinicians. BMC Microbiol 2022;22:244. https://doi.org/10.1186/s12866-022-02658-z.

[30] Barma MD, Muthupandiyan I, Samuel SR, Amaechi BT. Inhibition of Streptococcus mutans, antioxidant property and cytotoxicity of novel nano-zinc oxide varnish. Arch Oral Biol 2021;126:105132. https://doi.org/10.1016/j.archoralbio.2021.105132.

[31] Anita P, Sathyanarayana HP, Kumar K, Ramanathan K, Kailasam V. Antimicrobial efficacy of zinc oxide nanoparticle-coated aligners on Streptococcus mutans and Candida albicans. Am J Orthod Dentofac Orthop 2023;163:338–46. https://doi.org/10.1016/j.ajodo.2021.11.020.

[32] Sevinc-Sasmaz C, Torlak E. Synthesis-dependent surface attachment and antimicrobial mechanisms of ZnO nanoparticles against oral pathogens. Appl Surf Sci 2026;749:167901. https://doi.org/10.1016/j.apsusc.2026.167901.

[33] Pascual J, Mira Otal J, Torrent-Silla D, Porcar M, Vilanova C, Vivancos Cuadras F. A mouthwash formulated with o-cymen-5-ol and zinc chloride specifically targets potential pathogens without impairing the native oral microbiome in healthy individuals. J Oral Microbiol 2023;15:2185962. https://doi.org/10.1080/20002297.2023.2185962.

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Published

2026-10-30

How to Cite

Integration of Butterfly Pea (Clitoria ternatea L.) Flower Extract and ZnO into a Mouthwash: Physicochemical Characteristics and Antibacterial Activity. (2026). JURNAL FARMASIMED (JFM), 9(1), 640-650. https://doi.org/10.35451/jfm.v9i1.3955