This study was aimed to provide the reference frame for the safe dose design of
polyphenol-rich herbal mixture, which consist of lemon, apple cider, garlic, ginger and
honey (PRM) for the future efficacy study. Prior to this, the nutritional composition was
first conducted and the identification of metabolites that present in PRM was determined
using proton nuclear magnetic resonance (1H-NMR) spectroscopy. The acute toxicity of
the PRM was then evaluated in zebrafish embryo and Wistar rats following The
Organisation for Economic Co-operation and Development (OECD) guidelines. The PRM
nutritional composition and sugar profile showed it was high in carbohydrate, ash and
protein and the main sugar is fructose. It also contains metabolites such as
fructosefuranose, lactic acid, ascorbic acid, acetic acid, cycloalliin, pyruvate, 5-
hydroxymethylfurane, α- and β-glucose. From the zebrafish embryo acute toxicity result,
the lethal concentration, LC50 of PRM was at 487.50 μg/mL. Meanwhile, in Wistar rats’
model, no lethality was observed in the group treated with PRM at the end of the study (14
days). No changes were also observed from the behavioural and appetite as well as the
biochemical parameters (AST, ALT, total cholesterol, triglyceride, LDL, HDL, total
protein and creatinine) of the treated group. Therefore, the safe dose for PRM can be up to
2000 mg/kg b.w. in Wistar rats and below 487.50 μg/mL in zebrafish embryo model.