Assessment of in vivo acute, subacute toxicity, in vitro anti-plasmodial activity, and UPLC-ESI-QToF-MS/MS prediction of Annickia affinis (Exell) Versteegh & Sosef (Annonaceae) roots extract

Jean Emmanuel Mbosso Teinkela; Jaures Noumedem Kenfack; Philippe Belle Ebanda Kedi; Elvira Tachie Mekjele; Thierry Fokou Nzodjou; Noella Molisa Efange; Lawrence Ayong; Else Carole Eboumbou Moukoko; Giselle Marie Marguerite Etamè Loe; and Simone Veronique Fannang. | E-mail: embosso@yahoo.fr; jauresnoume@gmail.com | Received: 2026-03-24 | Accepted: 2026-05-01 | Published: 2026-05-01

Abstract

Background: Annickia affinis is a medicinal plant widely used in African traditional medicine to treat various diseases, including malaria. This study aimed to evaluate the toxicological profile, chemical composition, and antiplasmodial activity of the ethanolic (EtOH) root extract of Annickia affinis.

Methods: Acute (14-day) and subacute (28-day) toxicity studies were conducted in accordance with guidelines 423 and 407 of the Organisation for Economic Co-operation and Development. Chemical profiling was performed using ultra-performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS) and molecular networking analysis (GNPS). Antiplasmodial activity was evaluated in vitro against the chloroquine-resistant Plasmodium falciparum Dd2 strain.

Results: In the acute toxicity study, oral administration of the extract at doses of 2000 and 5000 mg/kg body weight caused no mortality or significant changes in clinical signs, body weight, or most biochemical and hematological parameters. Similarly, subacute administration at 200, 400, and 800 mg/kg for 28 days did not induce mortality. Mild increases in alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels in male rats, gamma-glutamyl transferase (γ-GT) levels in females, and urea levels in both sexes were observed, whereas serum creatinine and most hematological parameters remained unchanged. Histopathological examination of the liver, kidneys, and heart revealed no tissue damage. Chemical analysis identified several alkaloids, mainly protoberberine alkaloids, along with sesquiterpenes. The EtOH root extract exhibited strong antiplasmodial activity with an IC₅₀ of 2.08 µg/mL against the P. falciparum Dd2 strain.

Conclusion: The EtOH root extract of A. affinis demonstrates a favorable safety profile and promising antiplasmodial activity, suggesting its potential as a source of bioactive compounds for antimalarial drug development

 

Keywords: Acute toxicity; Annickia affinis; Antiplasmodial; GNPS; Subacute toxicity; UPLC-MS/MS

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