Ultrasound-assisted extraction of azadirachtin from dried entire fruits of Azadirachta indica A. Juss. (Meliaceae) and its determination by a validated HPLC-PDA method
Abstract
Azadirachta indica A. Juss., also known as neem, is a Meliaceae family tree from India. It is globally known for the insecticidal properties of its limonoid tetranortriterpenoid derivatives, such as azadirachtin. This work aimed to optimize the azadirachtin ultrasound-assisted extraction (UAE) and validate the HPLC-PDA analytical method for the measurement of this marker in neem dried fruit extracts. Box-Behnken design and response surface methodology (RSM) were used to investigate the effect of process variables on the UAE. Three independent variables, including ethanol concentration (%, w/w), temperature (°C), and material-to-solvent ratio (gmL(-1)), were studied. The azadirachtin content (µgmL(-1)), i.e., dependent variable, was quantified by the HPLC-PDA analytical method. Isocratic reversed-phase chromatography was performed using acetonitrile/water (40:60), a flow of 1.0mLmin(-1), detection at 214nm, and C18 column (250×4.6mm(2), 5µm). The primary validation parameters were determined according to ICH guidelines and Brazilian legislation. The results demonstrated that the optimal UAE condition was obtained with ethanol concentration range of 75-80% (w/w), temperature of 30°C, and material-to-solvent ratio of 0.55gmL(-1). The HPLC-PDA analytical method proved to be simple, selective, linear, precise, accurate and robust. The experimental values of azadirachtin content under optimal UAE conditions were in good agreement with the RSM predicted values and were superior to the azadirachtin content of percolated extract. Such findings suggest that UAE is a more efficient extractive process in addition to being simple, fast, and inexpensive.
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“The optimal conditions for neem oil extraction using the RSM and MAE method have been determined in several studies [24 – 26], but, to our knowledge, the conditions for extracting neem seed oil in hydroethanolic media by MAE-assisted RSM with a BBD have not been reported.”
Method
- Authors
- de Paula, J. A. M., Brito, L. F., Caetano, K. L. F. N., de Morais Rodrigues, M. C., Borges, L. L., & da Conceição, E. C.
- Affiliations
- Universidade Paulista (BR)
- Universidade Federal de Goiás (BR)
- Venue
- Talanta
- Publisher
- Elsevier BV
- Published
- 2016-03
- Volume
- 149
- Pages
- 77-84
- ISSN
- 0039-9140 (ISSN-L)
- OpenAlex
- W1962742147
- Cited by
- 36
- References
- 34
- Citations by year
- 2026: 1 · 2025: 1 · 2024: 4 · 2023: 3 · 2022: 3 · 2021: 2 · 2020: 5 · 2019: 4 · 2018: 6 · 2017: 6 · 2016: 1
- FWCI
- 1.67
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Talanta (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Conselho Nacional de Ciência e Tecnologia (CNPq)
- Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
- Fundação de Amparo à Pesquisa do Estado de Goiás (FAPEG)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Phytochemical compounds biological activities
- Life Sciences › Agricultural and Biological Sciences › Plant Science › Insect Pest Control Strategies
- Health Sciences › Medicine › Complementary and alternative medicine › Phytochemicals and Medicinal Plants
- Keywords
- azadirachtin, Azadirachta indica, ultrasound-assisted extraction, extraction optimization, ethanol concentration, HPLC-PDA method validation, insecticidal natural products
- MeSH terms
- Ultrasonic Waves, Ethanol, Chromatography, High Pressure Liquid, Fruit, Temperature, Reproducibility of Results, Azadirachta, Limonins, Limit of Detection
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