Nutritional composition, phytochemicals and acute toxicity of herbal mixture (lemon, apple cider, garlic, ginger and honey) in zebrafish embryo and Wistar rat

Abstract

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.

The Changes in Endogenous Metabolites in Hyperlipidemic Rats Treated with Herbal Mixture Containing Lemon, Apple Cider, Garlic, Ginger, and Honey

“…Based on our previous study, the proton NMR of PRM containing primary and secondary metabolites includes amino acids, sugars, 5-hydroxymethylfurfural (5-HMF) and ascorbic acid (Vitamin C). The nutritional composition of PRM reveals it contained about 43.40% carbohydrates, whereby 18% of it came from fructose [ 21 ]. The high content of sugar can mask the appearance of other secondary metabolites in spectra.…”

“…Nevertheless, the only significant increase ( p < 0.05) was noted in the AST level in the PRM-M and D groups when compared to the C group. Previously, elevated AST and ALT levels have been recorded in hypercholesterolemia and hyperlipidemia, herbal, and statin studies [ 21 , 26 , 27 ]. The consumption of high cholesterol diet and drugs can influence the liver’s rate of clearance.…”

“…Polyphenol rich mixture (PRM) preparation was conducted based on our previously published article by Abu Bakar Sajak et al [ 21 ]. In brief, a volume of 1000 mL of juice mixture consisting of lemon ( Citrus limon ), garlic ( Allium sativum ), ginger ( Zingiber officinale ) and apple cider (250 mL each) was simmered at 85 °C for 30 min until it reduced to 750 mL.…”

Phytochemical analysis, antioxidant, α-glucosidase inhibitory activity, and toxicity evaluation of Orthosiphon stamineus leaf extract

“…These findings highlight the significant impact of the type of herb and extraction solvent used on final toxicity. Notably, Sajak et al 13 compared toxicity testing using zebrafish embryos and Wistar rats, finding that a polyphenolic-rich herbal mixture (PRM) had an LC 50 of approximately 487.50 μg/mL in zebrafish embryos, despite no lethal effects being observed in rats at a dose of 2000 mg/kg body weight. Zebrafish embryos exhibited higher sensitivity in toxicity evaluations, allowing for more sensitive detection and for herbal extracts with LC 50 values above 500 μg/mL to be classified as non-toxic.…”