Untargeted Strategy in the Investigation of in Vivo Developmental and Acute Toxicity, and Anti-Inflammatory Activity Using Hptlc and Nmr-Based Metabolomic Analysis of Vitex Negundo L. Leaf Extracts

Date of Award

12-1-2022

Document Type

Thesis

Degree Name

Master of Science in Chemistry

First Advisor

Fabian M. Dayrit, PhDOlivia Erin M. Buenafe, PhD

Abstract

Herbal medicine has experienced a boom in popularity over the past decades and the direction for its development has moved towards optimization through application of metabolic profiling technology. This study employed in vivo toxicological and therapeutic analysis of Vitex negundo, commercially used as an antitussive and anti-inflammatory using zebrafish assays, in combination with metabolomic analysis through high performance thin layer chromatography (HPTLC) and 13C nuclear magnetic resonance (NMR) spectroscopy.

No significant relationship between toxicity and anti-inflammatory activity was observed and defatting of the ethanolic V. negundo leaf extract with hexane was found to substantially remove acute lethal toxicity, cardiotoxicity, and teratogenicity risk while maintaining therapeutic activity. The crude ethanolic extract of V. negundo had moderate acute toxicity risk and teratogenic risk with a 96 hours post fertilization (hpf) LC50 of 68.2 ± 0.4 ppm and teratogenic index (TI) of 2.1 ± 0.3 which were significantly lowered to LC50 of 107 ± 3 ppm (P = 0.0001) and TI of 1.4 ± 0.1 (P = 0.02) after defatting. These observed differences suggested the potential for further minimization of teratogenic and cardiotoxicity risks for a larger margin of safety for high-risk patients. Metabolomic analysis and acute tolerability tests also showed relative toxicity of V. negundo extracts was primarily associated with volatile oil terpenoids. Further analysis with PLS-R produced HPTLC and 13C NMR fingerprints for potential studies on relative toxicity minimization.

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