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Abstract
The long-term use of topical non-steroidal anti-inflammatory drugs (NSAIDs) is associated with local irritation and disruption of the skin barrier, highlighting the need for safer plant-based alternatives with adequate formulation stability. Calotropis gigantea leaves contain flavonoids with reported anti-inflammatory activity; however, their topical performance is limited by poor solubility and instability. This study aimed to evaluate the effect of emulsifier balance on the physical stability and anti-inflammatory efficacy of a Calotropis gigantea leaf extract emulgel. Three emulgel formulations were prepared using different ratios of nonionic surfactants Tween 80 and Span 80 (2:8, 5:5, and 8:2) based on hydrophilic–lipophilic balance (HLB) principles. The formulations were evaluated for physicochemical characteristics, accelerated physical stability using a cycling test, and in vivo anti-inflammatory activity using a carrageenan-induced paw edema model in rats. The formulation containing a balanced emulsifier ratio (5:5) exhibited the smallest deviations in viscosity, spreadability, and pH after cycling tests and demonstrated the highest edema inhibition (68.98 ± 1.64%). These results indicate that emulsifier balance plays a critical role in maintaining emulgel stability and directly influences its biological performance. Although chemical stability and dermal safety were not evaluated, this study provides evidence that rational emulsifier selection is essential for improving the effectiveness of herbal topical formulations.
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Copyright (c) 2026 Fauzi Rahman, Azimatul Mauna, Nur Hafifah, Ana Maria Ulfa, Luluk Latifah

This work is licensed under a Creative Commons Attribution 4.0 International License.
References
- Ahmad, W. (2020). Preliminary phytochemical, antimicrobial and photochemical study of Calotropis gigantea leaf extract. Current Chemistry Letters, 9, 105–112. https://doi.org/10.5267/j.ccl.2019.10.001.
- Alafnan, A., Sridharagatta, S., Saleem, H., Khurshid, U., Alamri, A., Ansari, S. Y., Zainal Abidin, S. A., Ansari, S. A., Alamri, A. S., Ahemad, N., & Anwar, S. (2021). Evaluation of the phytochemical, antioxidant, enzyme inhibition, and wound healing potential of Calotropis gigantea (L.) dryand: a source of a bioactive medicinal product. Frontiers in Pharmacology, 12. https://doi.org/10.3389/fphar.2021.701369.
- Al-Shammari, B., Al-Fariss, T., Al-Sewailm, F., & Elleithy, R. (2011). The effect of polymer concentration and temperature on the rheological behavior of metallocene linear low density polyethylene (mLLDPE) solutions. Journal of King Saud University - Engineering Sciences, 23(1), 9–14. https://doi.org/10.1016/j.jksues.2010.07.001.
- Chebil, L., Humeau, C., Anthoni, J., Dehez, F., Engasser, J.-M., & Ghoul, M. (2007). Solubility of flavonoids in organic solvents. Journal of Chemical & Engineering Data, 52(5), 1552–1556. https://doi.org/10.1021/je7001094.
- Chen, Z., Zhang, S., Li, Z., Ma, G., & Su, Z. (2017). Construction of a stable w/o nano-emulsion as a potential adjuvant for foot and mouth disease virus vaccine. Artificial Cells, Nanomedicine, and Biotechnology, 45(5), 897–906. https://doi.org/10.1080/21691401.2016.1188396.
- Daud, N. S., & Suryanti, E. (2017). Formulasi emulgel antijerawat minyak nilam (patchouli oil) menggunakan tween 80 dan span 80 sebagai pengemulsi dan HPMC sebagai basis gel. Jurnal Mandala Pharmacon Indonesia, 3(02), 90–95. https://doi.org/10.35311/jmpi.v3i02.3.
- Emelda, E., Nugraeni, R., & Damayanti, K. (2023). Review: exploration of Indonesian herbal plants for anti inflammatory. Inpharnmed Journal (Indonesian Pharmacy and Natural Medicine Journal), 6(2), 58. https://doi.org/10.21927/inpharnmed.v6i2.1938.
- Gupta, A. K., Parasar, D., Sagar, A., Choudhary, V., Chopra, B. S., Garg, R., Ashish, & Khatri, N. (2015). Analgesic and anti-inflammatory properties of gelsolin in acetic acid induced writhing, tail immersion and carrageenan induced paw edema in mice. PLOS One, 10(8), https://doi.org/10.1371/journal.pone.0135558.
- Houhat, N., Devaux, T., Callé, S., Saengdee, L., Boucaud Gauchet, S., & Vander Meulen, F. (2025). Monitoring of polymer viscosity by simultaneous ultrasonic and rheological measurements at high and varying temperatures. AIP Advances, 15(3). https://doi.org/10.1063/5.0251850.
- Ikhsanudin, A., & Azizah, D. N. (2019). Uji sifat fisik dan uji aktivitas repelan terhadap nyamuk aedes aegypti betina sediaan emulgel minyak atsiri biji pala (Myristica fragrans Houtt.). Jurnal Farmasi , 7(2).
- Khafifa, I. N., Shabrina, A., & Rochman, M. F. (2022). Stability and sunscreen activity of nutmeg seed oil emulgel with carbopol 940 variation as gel base. Jurnal Farmasi Sains Dan Praktis, 167–176. https://doi.org/10.31603/pharmacy.v8i2.6085.
- Kumar, P. S., E, Suresh., & Kalavathy, S. (2013). Review on a potential herb Calotropis gigantea (L.) R. Br. Scholars Academic Journal of Pharmacy, 2(2), 135–143.
- Langworthy, M., Dasa, V., & Spitzer, A. I. (2024). Knee osteoarthritis: disease burden, available treatments, and emerging options. Therapeutic Advances in Musculoskeletal Disease, 16. https://doi.org/10.1177/1759720X241273009.
- Lv, G., Wang, F., Cai, W., & Zhang, X. (2014). Characterization of the addition of lipophilic Span 80 to the hydrophilic Tween 80-stabilized emulsions. Colloids and Surfaces A: Physicochemical and Engineering Aspects, 447, 8–13. https://doi.org/10.1016/j.colsurfa.2014.01.066.
- Milutinov, J., Krstonošić, V., Ćirin, D., & Pavlović, N. (2023). Emulgels: promising carrier systems for food ingredients and drugs. Polymers, 15(10), 2302. https://doi.org/10.3390/polym15102302.
- Pawestri, S. A., & Sulaiman, T. N. S. (2019). The influence of variation of hydroxypropyl methylcellulose and tween 80 concentrations on physical characteristics and physical stabilities gel of water dry extract of temulawak. International Journal of Current Pharmaceutical Research, 11(6), 44–48.
- Proksch, E. (2018). Buffering capacity. Current Problems in Dermatology, 54, https://doi.org/10.1159/000489513.
- Purwanto, U. R. E., Cahyani, I. M., Purwaningsih, Y., Sandhi, B. G. F., & Febryana, F. (2023). Optimization of polysorbate 80 and sorbitan monooleate 80 as emulsifiers in foundation makeup containing ethyl cinnamate, Indonesian Journal of Pharmacy, 34(1), 45–53.
- Puspitasari, F., Saraswati, I., & Wulandari, F. (2023). Formulasi dan evaluasi fisik sediaan emulgel ekstrak daun kelor (Moringa oleifera Lam.) sebagai antioksidan dengan gelling agent HPMC. Generics: Journal of Research in Pharmacy, 3(1), 36–44. https://doi.org/10.14710/genres.v3i1.17256.
- Putri, A. I. E., & Ariyanto, H. D. (2022). Effect of hydrophilic- lipophilic balance (HLB) value on the stability of cosmetic lotion based on walnut oil (Canarium indicium l.) oil-in-water emulsion. Journal of Vocational Studies on Applied Research, 4(2), 53–60. https://doi.org/10.14710/jvsar.v4i2.15376.
- Rahman, F., Hendradi, E., & Purwanti, T. (2023). Physicochemical characterization, release and penetration study of nanostructured lipid carriers quercetin incorporated into membrane-type patches. Tropical Journal of Natural Product Research, 7(12). https://doi.org/10.26538/tjnpr/v7i12.30.
- Rusli, N., Setiawan, M. A., & Hikmawati, N. (2021). The effect of HPMC as a gel base and tween 80 span 80 combinations as emulgators in acetosal transdermal emulgel preparations. Jurnal Farmasi Sains dan Praktis, 7(3), 249–259.
- Semis, H. S., Gur, C., Ileriturk, M., Kaynar, O., & Kandemir, F. M. (2021). Investigation of the anti-inflammatory effects of caffeic acid phenethyl ester in a model of λ-carrageenan–induced paw edema in rats. Human & Experimental Toxicology, 40(12_suppl), S721–S738. https://doi.org/10.1177/09603271211054436.
- Touma, Z., Chen, L., & Arayssi, T. (2007). Topical nonsteroidal anti-inflammatory drugs in the treatment of osteoarthritis. Future Rheumatology, 2(2), 163–175. https://doi.org/10.2217/17460816.2.2.163.
- Wahyuni, L., Wirjosentono, B., & Tamrin, T. (2020). Effect of surfactant tween 80 (polyoxyethylene sorbitan mono oleate) addition on viscosity and activation energy on making asphalt emulsion. Journal of Chemical Natural Resources, 2(2), 113–120.
- Zawawi, N. A., Ahmad, H., Madatheri, R., Fadilah, N. I. M., Maarof, M., & Fauzi, M. B. (2025). Flavonoids as natural anti-inflammatory agents in the atopic dermatitis treatment. Pharmaceutics, 17(2), 261. https://doi.org/10.3390/pharmaceutics17020261.
References
Ahmad, W. (2020). Preliminary phytochemical, antimicrobial and photochemical study of Calotropis gigantea leaf extract. Current Chemistry Letters, 9, 105–112. https://doi.org/10.5267/j.ccl.2019.10.001.
Alafnan, A., Sridharagatta, S., Saleem, H., Khurshid, U., Alamri, A., Ansari, S. Y., Zainal Abidin, S. A., Ansari, S. A., Alamri, A. S., Ahemad, N., & Anwar, S. (2021). Evaluation of the phytochemical, antioxidant, enzyme inhibition, and wound healing potential of Calotropis gigantea (L.) dryand: a source of a bioactive medicinal product. Frontiers in Pharmacology, 12. https://doi.org/10.3389/fphar.2021.701369.
Al-Shammari, B., Al-Fariss, T., Al-Sewailm, F., & Elleithy, R. (2011). The effect of polymer concentration and temperature on the rheological behavior of metallocene linear low density polyethylene (mLLDPE) solutions. Journal of King Saud University - Engineering Sciences, 23(1), 9–14. https://doi.org/10.1016/j.jksues.2010.07.001.
Chebil, L., Humeau, C., Anthoni, J., Dehez, F., Engasser, J.-M., & Ghoul, M. (2007). Solubility of flavonoids in organic solvents. Journal of Chemical & Engineering Data, 52(5), 1552–1556. https://doi.org/10.1021/je7001094.
Chen, Z., Zhang, S., Li, Z., Ma, G., & Su, Z. (2017). Construction of a stable w/o nano-emulsion as a potential adjuvant for foot and mouth disease virus vaccine. Artificial Cells, Nanomedicine, and Biotechnology, 45(5), 897–906. https://doi.org/10.1080/21691401.2016.1188396.
Daud, N. S., & Suryanti, E. (2017). Formulasi emulgel antijerawat minyak nilam (patchouli oil) menggunakan tween 80 dan span 80 sebagai pengemulsi dan HPMC sebagai basis gel. Jurnal Mandala Pharmacon Indonesia, 3(02), 90–95. https://doi.org/10.35311/jmpi.v3i02.3.
Emelda, E., Nugraeni, R., & Damayanti, K. (2023). Review: exploration of Indonesian herbal plants for anti inflammatory. Inpharnmed Journal (Indonesian Pharmacy and Natural Medicine Journal), 6(2), 58. https://doi.org/10.21927/inpharnmed.v6i2.1938.
Gupta, A. K., Parasar, D., Sagar, A., Choudhary, V., Chopra, B. S., Garg, R., Ashish, & Khatri, N. (2015). Analgesic and anti-inflammatory properties of gelsolin in acetic acid induced writhing, tail immersion and carrageenan induced paw edema in mice. PLOS One, 10(8), https://doi.org/10.1371/journal.pone.0135558.
Houhat, N., Devaux, T., Callé, S., Saengdee, L., Boucaud Gauchet, S., & Vander Meulen, F. (2025). Monitoring of polymer viscosity by simultaneous ultrasonic and rheological measurements at high and varying temperatures. AIP Advances, 15(3). https://doi.org/10.1063/5.0251850.
Ikhsanudin, A., & Azizah, D. N. (2019). Uji sifat fisik dan uji aktivitas repelan terhadap nyamuk aedes aegypti betina sediaan emulgel minyak atsiri biji pala (Myristica fragrans Houtt.). Jurnal Farmasi , 7(2).
Khafifa, I. N., Shabrina, A., & Rochman, M. F. (2022). Stability and sunscreen activity of nutmeg seed oil emulgel with carbopol 940 variation as gel base. Jurnal Farmasi Sains Dan Praktis, 167–176. https://doi.org/10.31603/pharmacy.v8i2.6085.
Kumar, P. S., E, Suresh., & Kalavathy, S. (2013). Review on a potential herb Calotropis gigantea (L.) R. Br. Scholars Academic Journal of Pharmacy, 2(2), 135–143.
Langworthy, M., Dasa, V., & Spitzer, A. I. (2024). Knee osteoarthritis: disease burden, available treatments, and emerging options. Therapeutic Advances in Musculoskeletal Disease, 16. https://doi.org/10.1177/1759720X241273009.
Lv, G., Wang, F., Cai, W., & Zhang, X. (2014). Characterization of the addition of lipophilic Span 80 to the hydrophilic Tween 80-stabilized emulsions. Colloids and Surfaces A: Physicochemical and Engineering Aspects, 447, 8–13. https://doi.org/10.1016/j.colsurfa.2014.01.066.
Milutinov, J., Krstonošić, V., Ćirin, D., & Pavlović, N. (2023). Emulgels: promising carrier systems for food ingredients and drugs. Polymers, 15(10), 2302. https://doi.org/10.3390/polym15102302.
Pawestri, S. A., & Sulaiman, T. N. S. (2019). The influence of variation of hydroxypropyl methylcellulose and tween 80 concentrations on physical characteristics and physical stabilities gel of water dry extract of temulawak. International Journal of Current Pharmaceutical Research, 11(6), 44–48.
Proksch, E. (2018). Buffering capacity. Current Problems in Dermatology, 54, https://doi.org/10.1159/000489513.
Purwanto, U. R. E., Cahyani, I. M., Purwaningsih, Y., Sandhi, B. G. F., & Febryana, F. (2023). Optimization of polysorbate 80 and sorbitan monooleate 80 as emulsifiers in foundation makeup containing ethyl cinnamate, Indonesian Journal of Pharmacy, 34(1), 45–53.
Puspitasari, F., Saraswati, I., & Wulandari, F. (2023). Formulasi dan evaluasi fisik sediaan emulgel ekstrak daun kelor (Moringa oleifera Lam.) sebagai antioksidan dengan gelling agent HPMC. Generics: Journal of Research in Pharmacy, 3(1), 36–44. https://doi.org/10.14710/genres.v3i1.17256.
Putri, A. I. E., & Ariyanto, H. D. (2022). Effect of hydrophilic- lipophilic balance (HLB) value on the stability of cosmetic lotion based on walnut oil (Canarium indicium l.) oil-in-water emulsion. Journal of Vocational Studies on Applied Research, 4(2), 53–60. https://doi.org/10.14710/jvsar.v4i2.15376.
Rahman, F., Hendradi, E., & Purwanti, T. (2023). Physicochemical characterization, release and penetration study of nanostructured lipid carriers quercetin incorporated into membrane-type patches. Tropical Journal of Natural Product Research, 7(12). https://doi.org/10.26538/tjnpr/v7i12.30.
Rusli, N., Setiawan, M. A., & Hikmawati, N. (2021). The effect of HPMC as a gel base and tween 80 span 80 combinations as emulgators in acetosal transdermal emulgel preparations. Jurnal Farmasi Sains dan Praktis, 7(3), 249–259.
Semis, H. S., Gur, C., Ileriturk, M., Kaynar, O., & Kandemir, F. M. (2021). Investigation of the anti-inflammatory effects of caffeic acid phenethyl ester in a model of λ-carrageenan–induced paw edema in rats. Human & Experimental Toxicology, 40(12_suppl), S721–S738. https://doi.org/10.1177/09603271211054436.
Touma, Z., Chen, L., & Arayssi, T. (2007). Topical nonsteroidal anti-inflammatory drugs in the treatment of osteoarthritis. Future Rheumatology, 2(2), 163–175. https://doi.org/10.2217/17460816.2.2.163.
Wahyuni, L., Wirjosentono, B., & Tamrin, T. (2020). Effect of surfactant tween 80 (polyoxyethylene sorbitan mono oleate) addition on viscosity and activation energy on making asphalt emulsion. Journal of Chemical Natural Resources, 2(2), 113–120.
Zawawi, N. A., Ahmad, H., Madatheri, R., Fadilah, N. I. M., Maarof, M., & Fauzi, M. B. (2025). Flavonoids as natural anti-inflammatory agents in the atopic dermatitis treatment. Pharmaceutics, 17(2), 261. https://doi.org/10.3390/pharmaceutics17020261.