Effect of Extraction Conditions on Total Anthocyanin Content from Perilla frutescens (L.) Britton and Its Application in Coloring Butter Biscuit

Document Type : Research Article-en

Authors

Pham Thi My Tram

Nguyen Ngoc Thanh Phuong

Institute of Green and Sustainable Technology, Thu Dau Mot University, Ho Chi Minh City, Vietnam

https://doi.org/10.22067/ifstrj.2026.96395.1506

Abstract

The consumer trend toward safe and healthy food has spurred research and application of natural colors in traditional biscuit products to improve quality and sensory value. Perilla frutescens (L.) Britton is a member of the Lamiaceae family and is widely found in East Asian regions, where it is used as both a spice in cooking and in traditional medicine. It is rich in anthocyanins, a class of polyphenolic compounds that act as natural colorants and possess strong antioxidant properties. This study aimed to evaluate the extraction process of perilla leaf for enriching the total anthocyanin content, which is applied in coloring butter biscuits. The investigated factors included the sample-to-water ratio for extraction (1:10, 1:15, 1:20, and 1:25 g:mL), extraction temperatures (60°C, 70°C, 80°C, and 90°C), and extraction times (20, 25, 30, and 35 minutes). The obtained extract was analyzed for anthocyanin content using the pH differential method. The findings suggested that a sample-to-water ratio of 1:20 g:mL, an extraction temperature of 90°C, and an extraction duration of 30 minutes were the most suitable extraction conditions to achieve a high total anthocyanin content (1.70 mg.g^-1 DW). The addition of perilla leaf extract (PLE) significantly affected the physical properties of the biscuits (diameter, yield, brightness, and hardness of the biscuits); the higher the amount of PLE added, the darker and crispier the biscuits became. Sensory analysis showed that biscuits supplemented with PLE were rated higher than the control in terms of texture, color, and flavor. The results showed that the biscuits that received the highest scores across all parameters were supplemented with PLE at a rate of 7.5%. Overall, the analysis suggests that it is possible to produce butter biscuits with a characteristic color and improved nutritional profile by adding PLE.

Keywords

Anthocyanin

Butter biscuit

Extraction

Lamiaceae

Perilla frutescens

Subjects

• Food Biotechnology

---

Authors retain the copyright. This is an open access article distributed under Creative Commons Attribution 4.0 International License (CC BY 4.0)

References

1. Adam, G., Robu, S., Flutur, M.M., Cioanca, O., Vasilache, I.A., Adam, A.M., Mircea, C., Nechita, A., Harabor, V., Harabor, A., & Hancianu, M. (2023). Applications of Perilla frutescens extracts in clinical practice. Antioxidants, 12(3), 727. https://doi.org/10.3390/antiox12030727
2. Ahmed, H.M. (2019). Ethnomedicinal, phytochemical and pharmacological investigations of Perilla frutescens (L.) Britt. Molecules, 24(1), 102. https://doi.org/10.3390/molecules24010102
3. Alappat, B., & Alappat, J. (2020). Anthocyanin pigments: Beyond aesthetics. Molecules, 25(23), 5500. https://doi.org/10.3390/molecules25235500
4. Apoorva, M., Pooja, S., & Vidyasagar, G.M. (2021). Phytochemical screening for secondary metabolites and nutraceutical value of Sesbania grandiflora (L.) Pers leaf extract. Indo Global Journal of Pharmaceutical Sciences, 11(1), 28-32. https://doi.org/10.35652/IGJPS.2021.111004
5. Azuan, A.A., Mohd Zin, Z.Z., Hasmadi, M., Rusli, N.D., & Zainol, M.K. (2020). Physicochemical, antioxidant and sensory characteristics of cookies supplemented with different levels of spent coffee ground extract. Food Research, 4(4), 1181-1190. https://doi.org/10.26656/fr.2017.4(4).058
6. Biswas, M., Alam, M., Akhtaruzzaman, M., Ahmed, S., Uddin, M.S., Talukder, A.H.M.R., Alam, A., & Biswas, B. (2025). Valorization of red dragon fruit peel (Hylocereus polyrhizus) as a functional ingredient for fortified biscuits: Physicochemical, nutritional, and sensory implications. Future Foods, 12, 100768. https://doi.org/10.1016/j.fufo.2025.100768
7. Choi, H.Y., Oh, S.Y., & Lee, Y.S. (2009). Antioxidant activity and quality characteristics of Perilla leaves (Perilla frutescens japonica HARA) cookies. Korean Journal of Food Cookery Science, 25(5), 521-530. https://doi.org/10.3746/jkfn.2011.40.2.267
8. Ciupei, D., Colişar, A., Leopold, L., Stănilă, A., & Diaconeasa, Z.M. (2024). Polyphenols: from classification to therapeutic potential and bioavailability. Foods, 13(24), 4131. https://doi.org/10.3390/foods13244131
9. Cuong, V.T., Tung, N.Q., Duc,T., & Quynh, P.T.H. (2021). Research to complete the process of distillation of purple essential oil in Vietnam (Perilla frutescens var. crispa) and applications in food processing. Journal of Science & Technology, 57(6), 124-128. (Vietnamese). https://vjol.info.vn/index.php/dhcnhn/article/view/65189/54963
10. Dam, S.M., & Lam, K.K. (2013). Extraction of anthocyanins from leaves of Perilla ocymoides var. bicolorlaciniata grown in Vietnam. Acta Horticulturae, 989, 293-297. https://doi.org/10.17660/ActaHortic.2013.989.38
11. Enaru, B., Drețcanu, G., Pop, T.D., Stǎnilǎ, A., & Diaconeasa, Z. (2021). Anthocyanins: factors affecting their stability and degradation. Antioxidants, 10(12), 1967. https://doi.org/10.3390/antiox10121967
12. Hoang, T.N.N., Nguyen, T.Q.T., Nguyen, M.P., Tran, T.H.L., & Le, T.H.A. (2024). Optimal extraction and spray-drying anthocyanin from Perilla frutescens. JST: Engineering And Technology For Sustainable Development, 34(5), 1-10. https://doi.org/10.51316/jst.178.etsd.2024.34.5.1
13. Hou, T., Netala, V.R., Zhang, H., Xing, Y., Li, H., & Zhang, Z. (2022). Perilla frutescens: A rich source of pharmacological active compounds. Molecules, 27(11), 3578. https://doi.org/10.3390/molecules27113578
14. Lee, (2005). Determination of total monomeric anthocyanin pigment content of fruit juices, beverages, natural colorants, and wines by the pH differential method: collaborative study. Journal of AOAC International, 88(5), 1269-1278. https://doi.org/10.1093/9780197610145.003.3423
15. Lia, F., & Baron, B. (2025). Analysis of polyphenolic composition, antioxidant power and stress-response effects of fractionated Perilla leaf extract on cells in vitro. Biologics. 5(1), 2. https://doi.org/10.3390/biologics5010002
16. Mandala, I.G., Ioannou, C.A., & Kostaropoulos, A.E. (2006). Textural attributes of commercial biscuits: Effect of relative humidity on their quality. International Journal of Food Science & Technology, 41(7), 782- https://doi.org/10.1111/j.1365-2621.2005.01092.x
17. Meng, L., Lozano, Y., Bombarda, I., Gaydou, E.M., & Li, B. (2009). Polyphenol extraction from eight Perilla frutescens Comptes Rendus. Chimie, 12(5), 602-611. https://doi.org/10.1016/j.crci.2008.04.011
18. Miller, R.A., Hoseney, R.C., & Morris, C.F. (1997). Effect of formula water content on the spread of sugar‑snap cookies. Cereal Chemistry, 74(5), 669- https://doi.org/10.1094/CCHEM.1997.74.5.669
19. Moczkowska‑Wyrwisz, M., Jastrzębska, D., & Wyrwisz, J. (2022). Application of new sources of bioactive substances (Perilla frutescens and Tagetes erecta L.) in the chosen cookies production. International Journal of Environmental Research and Public Health, 19(18), 11504. https://doi.org/10.3390/ijerph191811504
20. Najjar, Z., Alkaabi, M., Alketbi, K., Stathopoulos, C., & Ranasinghe, M. (2022). Physical chemical and textural characteristics and sensory evaluation of cookies formulated with date seed powder. Foods, 11(3), 305. https://doi.org/10.3390/foods11030305
21. Nguyen, N.T.K., Truong, H.A.V., & Nguyen, H.K.T. (2024). Optimizing conditions to improve polyphenol content and screening antioxidant capacity with DNA protection activity of Perilla frutescens. Hue University Journal of Science: Natural Science, 133(1B), 35–48. https://doi.org/10.26459/hueunijns.v133i1B.7087
22. Nhon, H.T.N., Tran, N.T.Q., Phuc, N.M., Linh, T.T.H., & Anh, L.T.H. (2025). Optimal extraction and spray-drying anthocyanin from Perilla frutescens. Journal of Science and Technology: Engineering and Technology for Sustainable Development, 34(5), 001-010. https://doi.org/10.51316/jst.178.etsd.2024.34.5.1
23. Nhut, P,.T., Phu, N.T.N., Duc, L.T., Hien, T.T., Chinh, N.D., Vo, D.V.N., Le, X.T., Do, S.T., & Giang, B.L. (2019). Effects of various solvent concentration, liquid-solid ratio, temperatures and time values on the extraction yield of anthocyanin from Vietnam Hibiscus sabdariffa L.(Roselle). IOP Conference Series: Materials Science and Engineering, Xiamen, China. https://doi.org/10.1088/1757-899x/542/1/012033
24. Patra, S., Makhal, P., Jaryal, S., More, N., & Kaki, V.R. (2022). Anthocyanins: Plant-based flavonoid pigments with diverse biological activities. International Journal of Plant Based Pharmaceuticals, 2(1), 118-127. https://doi.org/10.62313/ijpbp.2022.22
25. Pham, A.G.Q., & Pham, T.T.M. (2025). Studying the extraction conditions on rosmarinic acid content and antioxidant activity of Basil (Ocimum basilicum). Indonesian Journal of Chemistry, 25(3), 709-718. https://doi.org/10.22146/ijc.99415
26. Phuc, T.B., Ngoc,T.K., & Thu, L.T. (2022). Research and development of cocoa bean shell biscuits. Journal of Science and Technology – Nguyen Tat Thanh University, Vietnam, 18, 74-80 (Vietnamese). https://doi.org/10.55401/sv5yr725
27. Quyen, D.K. (2024). Effects of ingredients on the physical properties and sensory quality of taro biscuits. Dong Thap University Journal of Science, 13(2), 58–64 (Vietnamese). https://dthujs.vn/index.php/dthujs/article/view/840/1318?lang=en_US
28. Singh, T., Pandey, V.K., Dash, K.K., Zanwar, S., & Singh, R. (2023). Natural bio-colorant and pigments: Sources and applications in food processing. Journal of Agriculture and Food Research, 12, 100628. https://doi.org/10.1016/j.jafr.2023.100628
29. Skowyra, M., Falguera, V., Azman, N.A., Segovia, F., & Almajano, M.P. (2014). The effect of Perilla frutescens extract on the oxidative stability of model food emulsions. Antioxidants, 3(1), 38-54. https://doi.org/10.3390/antiox3010038
30. Thuy, N.M., Han, D.H.N., Minh, V.Q., & Tai, N.V. (2022). Effect of extraction methods and temperature preservation on total anthocyanins compounds of Peristrophe bivalvis Merr. leaf. Journal of Applied Biology & Biotechnology, 2, 146-153. https://doi.org/10.7324/jabb.2022.100218
31. Vega, E.N., Ciudad-Mulero, M., Fernández-Ruiz, V., Barros, L., & Morales, P. (2023). Natural sources of food colorants as potential substitutes for artificial additives. Foods, 12(22), 4102. https://doi.org/10.3390/foods12224102
32. Yu, Y., Shiau, S., Pan, W., & Yang, Y. (2024). Extraction of bioactive phenolics from various anthocyanin‑rich plant materials and comparison of their heat stability. Molecules, 29(22), 5256. https://doi.org/10.3390/molecules29225256
33. Zhang, P., & Zhu, (2023). Anthocyanins in plant food: current status, genetic modification, and future perspectives. Molecules, 28(2), 866. https://doi.org/10.3390/molecules28020866
34. Zhang, X., Zhao, Y., Li, B., Guo, M., Lv, J., & Wei, Y. (2022). Comparison of three extraction methods for anthocyanins from Perilla frutescens Sustainable Chemistry and Pharmacy, 29, 100817. https://doi.org/10.1016/j.scp.2022.100817