In Vivo Toxicity Evaluation of Sugar Adulterated Heterotrigona itama Honey Using Zebrafish Model.

Rafieh Fakhlaei,Alfi Khatib,Rashidah Sukor,Syahida Ahmad,Arman Amani Babadi,Jinap Selamat,Ahmad Faizal Abdull Razis

doi:10.3390/molecules26206222

Abstract

Honey is prone to be adulterated through mixing with sugars, cheap and low-quality honey, and other adulterants. Consumption of adulterated honey may cause several health issues such as weight gain, diabetes, and liver and kidney dysfunction. Therefore, studying the impact of consumption of adulterated honey on consumers is critical since there is a lack of study in this field. Hence, the aims of this paper were: (1) to determine the lethal concentration (LC50) of adulterated honey using zebrafish embryo, (2) to elucidate toxicology of selected adulterated honey based on lethal dose (LD50) using adult zebrafish, (3) to determine the effects of adulterated honey on histological changes of zebrafish, and (4) to screen the metabolites profile of adulterated honey by using zebrafish blood serum. The LC50 of Heterotrigona itama honey (acacia honey) and its sugar adulterants (light corn sugar, cane sugar, inverted sugar, and palm sugar in the proportion of 1–3% (w/w) from the total volume) was determined by the toxicological assessment of honey samples on zebrafish embryos (different exposure concentrations in 24, 48, 72, and 96 h postfertilization (hpf)). Pure H. itama honey represents the LC50 of 34.40 ± 1.84 (mg/mL) at 96 hpf, while the inverted sugar represents the lowest LC50 (5.03 ± 0.92 mg/mL) among sugar adulterants. The highest concentration (3%) of sugar adulterants were used to study the toxicology of adulterated honey using adult zebrafish in terms of acute, prolong-acute, and sub-acute tests. The results of the LD50 from the sub-acute toxicity test of pure H. itama honey was 2.33 ± 0.24 (mg/mL). The histological studies of internal organs showed a lesion in the liver, kidney, and spleen of adulterated treated-honey groups compared to the control group. Furthermore, the LC-MS/MS results revealed three endogenous metabolites in both the pure and adulterated honey treated groups, as follows: (1) S-Cysteinosuccinic acid, (2) 2,3-Diphosphoglyceric acid, and (3) Cysteinyl-Tyrosine. The results of this study demonstrated that adulterated honey caused mortality, which contributes to higher toxicity, and also suggested that the zebrafish toxicity test could be a standard method for assessing the potential toxicity of other hazardous food additives. The information gained from this research will permit an evaluation of the potential risk associated with the consumption of adulterated compared to pure honey.

Highlights

IntroductionThe honeybees can collect and transform it by mixing it with specific substances of their own, deposit it, dehydrate it, and store it in the honeycomb to ripen and mature
The standards of Codex Alimentarius [1] define honey as the natural sweet substance produced by honey bees from the nectar of plants or secretions of living parts of plants.The honeybees can collect and transform it by mixing it with specific substances of their own, deposit it, dehydrate it, and store it in the honeycomb to ripen and mature
Embryo and adult—in sugar adulterated H. itama honey, our experiment proved that adulterated H. itama honey is not beneficial toward human health, while pure honey does not show any significant (p ≤ 0.05) cardiotoxicity effect

Summary

Introduction

The honeybees can collect and transform it by mixing it with specific substances of their own, deposit it, dehydrate it, and store it in the honeycomb to ripen and mature. The harvested honey from honeybees can be consumed as a sweetener and as a medicine due to its therapeutic effect on human health. Honey contains mainly sugar and more than 200 other compounds [2]. Fructose and glucose are the main sugars in H. itama honey in the proportion of 13.00 ± 0.21 (g/100 g honey) for the former and 11.69 ± 0.56 (g/100 g honey) for the latter [4]. In addition to fructose and glucose, several other disaccharides and oligosaccharides, including sucrose

Objectives

Methods

Discussion

Conclusion