Evaluation of the melanin synthesis-inhibitory potential of Morus alba L. leaf extract for the development of a natural anti-hyperpigmentation formulation

Authors

  • Nguyen Minh Nam Department of Biomedical Engineering, Faculty of Medicine, University of Health Sciences, Vietnam National University Ho Chi Minh City, Ho Chi Minh City 700000, Vietnam
  • Bui Thi Phuong Research Center for Genetics and Reproductive Health – CGRH, University of Health Sciences, Vietnam National University HCM City, Linh Trung Ward, Thu Duc District, Ho Chi Minh City 700000, Vietnam
  • Nguyen Ngoc Dien Vietnam National University Ho Chi Minh City, Ho Chi Minh City 700000, Vietnam
  • Nguyen Tan Tai Vietnam National University Ho Chi Minh City, Ho Chi Minh City 700000, Vietnam
  • Tran Thi Huyen Department of Pharmacognosy, Faculty of Pharmacy, University of Health Sciences, Vietnam National University Ho Chi Minh City, Ho Chi Minh City 700000, Vietnam

DOI:

https://doi.org/10.69857/joapr.v13i6.1460

Keywords:

Morus alba, melanin synthesis inhibition, tyrosinase inhibition, natural skin-brightening formulation

Abstract

Background: Although Morus alba (white mulberry) has long been used in traditional skin care, the scientific evidence for its leaf extract in anti-hyperpigmentation applications remains limited, particularly regarding its incorporation into topical formulations. This study investigated the melanin-synthesis-inhibitory, tyrosinase-inhibitory, and antioxidant activities of M. alba leaf extract and evaluated its potential in a stable topical gel formulation. Methodology: Melanin suppression was assessed in IBMX-stimulated B16F10 melanocytes using microscopic observation, pellet analysis, and quantitative melanin measurement. Tyrosinase inhibition was examined with a mushroom tyrosinase assay, while antioxidant capacity was evaluated via DPPH radical scavenging. A topical gel containing M. alba extract was developed and assessed for physicochemical properties, phenolic retention, bioactivity, and stability. Result and Discussion: The extract showed strong tyrosinase inhibition (IC₅₀ = 5.70 ± 0.28 µg/mL) and antioxidant activity (IC₅₀ = 16.22 ± 0.6 µg/mL), and significantly reduced melanin synthesis in B16F10 cells without cytotoxicity (≤ 200 µg/mL). The formulated gel maintained phenolic content, exhibited moderate tyrosinase inhibition, and demonstrated stable appearance, pH, spreadability, and bioactivity during storage. Conclusion: Morus alba leaf extract possesses potent anti-melanogenic and antioxidant properties and can be successfully incorporated into a stable topical gel, supporting its potential as a natural ingredient for anti-hyperpigmentation products. However, these findings are based on in vitro assays and preliminary formulation studies; further in vivo and clinical evaluations are needed to confirm its efficacy and safety in humans.

Downloads

Download data is not yet available.

References

Briganti S, Camera E, Picardo M. Chemical and instrumental approaches to treat hyperpigmentation. Pigment Cell Res., 16, 101–10 (2003) https://doi.org/10.1034/j.1600-0749.2003.00029.x

Solano F. Melanins: skin pigments and much more—types, structural models, biological functions, and formation routes. New J. Sci., 2014, 498276 (2014) https://doi.org/10.1155/2014/498276

Zhu W, Gao J. The use of botanical extracts as topical skin-lightening agents for the improvement of skin pigmentation disorders. J. Investig. Dermatol. Symp. Proc., 13(1), 20–4 (2008) https://doi.org/10.1038/jidsymp.2008.8

Levitt J. The safety of hydroquinone: a dermatologist's response to the 2006 Federal Register. J. Am. Acad. Dermatol., 57, 854–72 (2007) https://doi.org/10.1016/j.jaad.2007.02.020

Kim J-K, Park K-T, Lee H-S, et al. Evaluation of the inhibition of mushroom tyrosinase and cellular tyrosinase activities of oxyresveratrol: comparison with mulberroside A. J. Enzyme Inhib. Med. Chem., 27(4), 495–503 (2012) https://doi.org/10.3109/14756366.2011.598866

Chowdhury M, De PK, Maji HS, Das D. Safety Study of Carboxymethylated Basella Alba Mucilage: a Subchronic Oral Toxicity Evaluation in Wistar Albino Rats. Journal of Applied Pharmaceutical Research, 12, 16–26 (2024) https://doi.org/10.18231/j.joapr.2024.12.2.16.26.

Li HX, Park JU, Su XD, et al. Identification of anti-melanogenesis constituents from Morus alba L. leaves. Molecules, 23, 2559 (2018) https://doi.org/10.3390/molecules23102559

Roh E, Yun C-Y, Yun JY, et al. cAMP-binding site of PKA as a molecular target of bisabolangelone against melanocyte-specific hyperpigmented disorder. J. Invest. Dermatol., 133, 1072–9 (2013) https://doi.org/10.1038/jid.2012.425

Kim Y-M, Cho S-E, Seo Y-K. The activation of melanogenesis by p-CREB and MITF signaling with extremely low-frequency electromagnetic fields on B16F10 melanoma. Life Sci., 162, 25–32 (2016) https://doi.org/10.1016/j.lfs.2016.08.015

Zheng Y, Lee E-H, Lee S-Y, et al. Morus alba L. root decreases melanin synthesis via sphingosine-1-phosphate signaling in B16F10 cells. J. Ethnopharmacol., 301, 115848 (2023) https://doi.org/10.1016/j.jep.2022.115848

Masuda T, Yamashita D, Takeda Y, et al. Screening for tyrosinase inhibitors among extracts of seashore plants and identification of potent inhibitors from Garcinia subelliptica. Biosci. Biotechnol. Biochem., 69, 197–201 (2005) https://doi.org/10.1271/bbb.69.197

Marks DC, Belov L, Davey MW, et al. The MTT cell viability assay for cytotoxicity testing in multidrug-resistant human leukemic cells. Leuk. Res., 16, 1165–73 (1992) https://doi.org/10.1016/0145-2126(92)90114-M[1]

Gerasimova E, Gazizullina E, Kolbaczkaya S, et al. The novel potentiometric approach to antioxidant capacity assay based on the reaction with stable radical 2,2′-diphenyl-1-picrylhydrazyl. Antioxidants, 11, 1974 (2022) https://doi.org/10.3390/antiox11101974

Margraf T, Karnopp AR, Rosso ND, et al. Comparison between Folin–Ciocalteu and Prussian Blue assays to estimate the total phenolic content of juices and teas using 96-well microplates. J. Food Sci., 80, C2397–C2403 (2015) https://doi.org/10.1111/1750-3841.13077

Guy RC. International conference on harmonisation. 2014 https://doi.org/10.1016/B978-0-12-386454-3.00861-7

Lin Y-S, Chen H-J, Huang J-P, et al. Kinetics of tyrosinase inhibitory activity using Vitis vinifera leaf extracts. Biomed. Res. Int., 2017, 5232680 (2017) https://doi.org/10.1155/2017/5232680

Mapunya M, Hussein A, Rodriguez B, et al. Tyrosinase activity of Greyia flanaganii (Bolus) constituents. Phytomedicine, 18, 1006–12 (2011) https://doi.org/10.1016/j.phymed.2011.03.013

Pohntadavit K, Duangmano S, Osiriphan M, et al. Tyrosinase inhibitory activity of crude procyanidin extract from green soybean seed and the stability of bioactive compounds in an anti-aging skin care formulation. Cosmetics, 11, 178 (2024) https://doi.org/10.3390/cosmetics11050178

Chaiyana W, Sirithunyalug J, Somwongin S, et al. Enhancement of the antioxidant, anti-tyrosinase, and anti-hyaluronidase activity of Morus alba L. leaf extract by pulsed electric field extraction. Molecules, 25, 2212 (2020) https://doi.org/10.3390/molecules25092212

Busca R, Ballotti R. Cyclic AMP, a key messenger in the regulation of skin pigmentation. Pigment Cell Res., 13, 60–9 (2001) https://doi.org/10.1034/j.1600-0749.2000.130203.x

Polumackanycz M, Wesolowski M, Viapiana A. Morus alba L. and Morus nigra L. leaves as a promising food source of phenolic compounds with antioxidant activity. Plant Foods Hum. Nutr., 76, 458–65 (2021) https://doi.org/10.1007/s11130-021-00922-7

Santos-Buelga C, Gonzalez-Manzano S, Dueñas M, et al. Extraction and isolation of phenolic compounds. In: Natural Products Isolation. p. 427–64 (2012) https://doi.org/10.1007/978-1-61779-624-1_17

Kumar N, Goel N. Phenolic acids: natural versatile molecules with promising therapeutic applications. Biotechnol. Rep., 24, e00370 (2019) https://doi.org/10.1016/j.btre.2019.e00370

Ercisli S, Orhan E. Chemical composition of white (Morus alba), red (Morus rubra) and black (Morus nigra) mulberry fruits. Food Chem., 103, 1380–4 (2007) https://doi.org/10.1016/j.foodchem.2006.10.054

Okatan V. Phenolic compounds and phytochemicals in fruits of black mulberry (Morus nigra L.) genotypes from the Aegean region in Turkey. Folia Hortic., 30, 93–101 (2018) https://doi.org/10.2478/fhort-2018-0010

Luque GC, Moya M, Picchio ML, et al. Polyphenol iongel patches with antimicrobial, antioxidant and anti-inflammatory properties. Polymers, 15, 1076 (2023) https://doi.org/10.3390/polym15051076

Benson HA, Watkinson AC. Topical and Transdermal Drug Delivery: Principles and Practice. John Wiley & Sons (2012) https://doi.org/10.2174/1567201816666190201143457

Draelos ZD. Skin lightening preparations and the hydroquinone controversy. Dermatol. Ther., 20, 308–13 (2007) https://doi.org/10.1111/j.1529-8019.2007.00144.x

Curto EV, Kwong C, Hermersdörfer H, et al. Inhibitors of mammalian melanocyte tyrosinase: in vitro comparisons of alkyl esters of gentisic acid with other putative inhibitors. Biochem. Pharmacol., 57, 663–72 (1999) https://doi.org/10.1016/S0006-2952(98)00340-2[1]

Feng D, Fang Z, Zhang P. The melanin inhibitory effect of plants and phytochemicals: a systematic review. Phytomedicine, 107, 154449 (2022) https://doi.org/10.1016/j.phymed.2022.154449

Jeong JY, Liu Q, Kim SB, et al. Characterization of melanogenesis inhibitory constituents of Morus alba leaves and optimization of extraction conditions using response surface methodology. Molecules, 20, 8730–41 (2015) https://doi.org/10.3390/molecules20058730

Mapoung S, Semmarath W, Arjsri P, et al. Determination of phenolic content, antioxidant activity, and tyrosinase inhibitory effects of functional cosmetic creams available on the Thailand market. Plants, 10, 1383 (2021) https://doi.org/10.3390/plants10071383

Wu L, Chen C, Cheng C, et al. Evaluation of tyrosinase inhibitory, antioxidant, antimicrobial, and antiaging activities of Magnolia officinalis extracts after Aspergillus niger fermentation. Biomed. Res. Int., 2018, 5201786 (2018) https://doi.org/10.1155/2018/5201786

He Q, Wang J, Li J, et al. Polyphenol profile and antioxidant, antityrosinase, and anti-melanogenesis activities of ethanol extract of bee pollen. Pharmaceuticals, 17, 1634 (2024) https://doi.org/10.3390/ph17121634

Published

2025-12-25

How to Cite

Nam, N. M. ., Phuong, B. T., Dien, N. N. ., Tai, N. T. ., & Huyen, T. T. (2025). Evaluation of the melanin synthesis-inhibitory potential of Morus alba L. leaf extract for the development of a natural anti-hyperpigmentation formulation. Journal of Applied Pharmaceutical Research, 13(6), 101-112. https://doi.org/10.69857/joapr.v13i6.1460

Issue

Section

Articles