Solvent-Dependent Bioactive Composition and Antifungal Potential of Neem (Azadirachta indica) Seed Extracts Against Mycotoxigenic Aspergillus Species
Ali J. Akram M. Zahoor M. Shah A.B. Alotaibi A.
February 2026John Wiley and Sons Inc
Chemistry and Biodiversity
2026#23Issue 2
This study investigated the bioactive composition and antifungal efficacy of neem seed extracts (NSE) prepared using acetone, ethyl acetate, chloroform, methanol, and ethanol against Aspergillus parasiticus and Aspergillus flavus. Gas chromatography–mass spectrometry (GC–MS) analysis revealed solvent-specific compounds profiles: ethanol extract contained 35 bioactive compounds (mainly esters and fatty acids), dominated by n-hexadecanoic acid (28.65%); methanol extract had 29 diketopiperazines, with 3,6-bis(2-methylpropyl)-2,5-piperazinedione (45.45%) as major; chloroform, ethyl acetate, and acetone extracts featured long-chain esters, hydrocarbons, and 2-heptadecanone (33.15%), respectively. All extracts inhibited fungal growth at 500 mg/mL, whereas ethanol was most effective at 31.25 mg/mL, reducing mycelial biomass by 45%. Methanol and ethanol extracts (250 mg/mL) suppressed aflatoxin (AF) (AFB1, AFB2) production by >90%. Six-month feed trials confirmed neem seed powder (NSP) as a potent biopreservative, achieving complete AF inhibition at 30% concentration. These findings highlight neems potential as a natural antifungal and anti-aflatoxigenic agent.
aflatoxin inhibition , Aspergillus flavus , Aspergillus parasiticus , gas chromatography–mass spectrometry (GC–MS) , neem seed extract
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Pakistan Council of Scientific and Industrial Research Laboratories Complex, Khyber Pakhtunkhwa, Peshawar, Pakistan
Department of Biochemistry, University of Malakand, Chakdara, Pakistan
College of Pharmacy, Korea University, Sejong, South Korea
The Research Center for Medicinal Plants, Al-Farabi Kazakh National University, Almaty, Kazakhstan
Department of Basic Sciences, College of Medicine, Princess Nourah bint Abdulrahman University, Riyadh, Saudi Arabia
Pakistan Council of Scientific and Industrial Research Laboratories Complex
Department of Biochemistry
College of Pharmacy
The Research Center for Medicinal Plants
Department of Basic Sciences
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