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Purpose <p>This study evaluated <i>Azadirachta indica</i> A. Juss. (neem) leaves polysaccharides (NLP) as natural anti-hyperpigmentation and antioxidant agents. This study optimized microwave-assisted extraction (MAE) parameters to maximize multiple bioactivities in a single process and elucidated the physicochemical properties underlying NLP’s efficacy and safety for topical use.</p> Methods <p>NLP was extracted with citric acid using MAE. Initial single-factor experiments defined ranges for extraction time, buffer-to-raw material (B: R) ratio, and microwave power. A Box-Behnken design optimized five responses simultaneously: extraction yield, monophenolase inhibition, diphenolase inhibition, DPPH radical scavenging, and FRAP reducing capacity. The optimized NLP was characterized by UV-Vis spectroscopy, FTIR, HPGPC, pH measurement, chemical and monosaccharide composition, molecular weight, degree of esterification, viscosity, and cytotoxicity on HaCaT keratinocytes.</p> Results <p>Under optimized microwave-assisted extraction conditions (extraction time: 1.43&#xa0;min, B:R ratio: 22.30 mL/g, microwave power: 331.84&#xa0;W), the extraction yielded 7.62% NLP. The extracted NLP exhibited notable biological activities, including 86.16% monophenolase inhibition, 68.18% diphenolase inhibition, 45.55% DPPH scavenging activity, and 0.47 mM FRAP reducing capacity. NLP showed slightly acidic properties (pH 4.35), attributed to the presence of uronic acid content. Structural analysis revealed low molecular weight (28.15&#xa0;kDa), low degree of esterification (13.24%), and low viscosity (25.10 mPa s). Cytotoxicity evaluation on HaCaT keratinocyte cells showed high biocompatibility, with &gt; 79% cell viability across all tested concentrations.</p> Conclusion <p>Citric acid-MAE‑extracted NLP exhibit strong tyrosinase inhibition, antioxidant activity, and skin cell compatibility. Their favorable physicochemical profiles support their potential as natural multifunctional ingredients for anti-hyperpigmentation and antioxidant dermatological formulations.</p>

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Harnessing Polysaccharides from Azadirachta Indica A. Juss. (neem) Leaves Using Microwave-Assisted Extraction for Anti-Hyperpigmentation Activity

  • 期刊论文

  • 发表日期：

    2025年10月20日

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  • Abu Hurairah Darwisy Alias,
  • Muhammad Hasnun Md Yusoff,
  • Nur Qistina Abdul Razak,
  • Suhaila Sujani,
  • Zahir Husain Kamari,
  • Tin-Wui Wong,
  • Idanawati Naharudin,
  • Muhammad Ameerullah Sahudin,
  • Mohammad Afiq Aizuddin Rosdi,
  • Chee-Yuen Gan,
  • Muhammad Hakimin Shafie

摘要

Purpose

This study evaluated Azadirachta indica A. Juss. (neem) leaves polysaccharides (NLP) as natural anti-hyperpigmentation and antioxidant agents. This study optimized microwave-assisted extraction (MAE) parameters to maximize multiple bioactivities in a single process and elucidated the physicochemical properties underlying NLP’s efficacy and safety for topical use.

Methods

NLP was extracted with citric acid using MAE. Initial single-factor experiments defined ranges for extraction time, buffer-to-raw material (B: R) ratio, and microwave power. A Box-Behnken design optimized five responses simultaneously: extraction yield, monophenolase inhibition, diphenolase inhibition, DPPH radical scavenging, and FRAP reducing capacity. The optimized NLP was characterized by UV-Vis spectroscopy, FTIR, HPGPC, pH measurement, chemical and monosaccharide composition, molecular weight, degree of esterification, viscosity, and cytotoxicity on HaCaT keratinocytes.

Results

Under optimized microwave-assisted extraction conditions (extraction time: 1.43 min, B:R ratio: 22.30 mL/g, microwave power: 331.84 W), the extraction yielded 7.62% NLP. The extracted NLP exhibited notable biological activities, including 86.16% monophenolase inhibition, 68.18% diphenolase inhibition, 45.55% DPPH scavenging activity, and 0.47 mM FRAP reducing capacity. NLP showed slightly acidic properties (pH 4.35), attributed to the presence of uronic acid content. Structural analysis revealed low molecular weight (28.15 kDa), low degree of esterification (13.24%), and low viscosity (25.10 mPa s). Cytotoxicity evaluation on HaCaT keratinocyte cells showed high biocompatibility, with > 79% cell viability across all tested concentrations.

Conclusion

Citric acid-MAE‑extracted NLP exhibit strong tyrosinase inhibition, antioxidant activity, and skin cell compatibility. Their favorable physicochemical profiles support their potential as natural multifunctional ingredients for anti-hyperpigmentation and antioxidant dermatological formulations.