Investigating the Toxicity of Betalain Compounds: In Silico Analysis and in Vivo Predictions for Standardized Beta Vulgaris L. Extract

Abstract

Objective: Extensive research has been conducted on beetroot's antioxidant, hematoprotective, and cardioprotective properties. However, there currently needs to be more available evidence pertaining to the toxicity assessment of the extract. The toxicity assessment was conducted using both in silico and in vivo methods. Prior to testing, the extracts were standardized in accordance with the guidelines set by the Indonesian Food Drug Authority (BPOM), which is the regulatory authority for food and drugs in Indonesia. Methods: The experimental subjects consisted of 25 male Wistar rats in good health, weighing between 150 and 170 grams. These rats were separated into five groups, each including five rats. Group 1 will serve as the control group, while groups 2 through 5 will be designated as the treatment groups. The analysis of chemical toxicity was conducted using pK-CSM, SwissADME, and Pro-Tox II methodologies. Results: The results indicated that the standardized ethanol extract contained 4.341% water, 3.67 % total ash, and 1.53 % acid-insoluble ash. Lead (Pb) and cadmium (Cd) were absent at a concentration of 0 parts per million (ppm). Subsequently, the total plate count and yeast mould count were 0.47 5 x 10-4 (CFU/g) and a of 0.382 x 10-4 (CFU/g) respectively. This finding implies that the extract meets BPOM requirement. This study also measured the betalain content of red beetroot, yielding a total concentration of 11.34 0.37 mg/100 gram of sample. Haematological experiments showed that beetroot extract affected rat blood haematology. Compared to the control group, rats given the extract had higher red blood cell and platelet counts. Additionally, the Insilico toxicity test conducted on the active component derived from beetroot revealed LD50 of the compounds ranged from 305 mg/kg so that were categorized into classes IV and presence of hepatotoxic potential. During the in vivo experiment, there has been a notable rise in hepatic and renal parameters. Furthermore, one mortality event occurred in the test subject at a 5,000 mg/kg body weight dosage. Conclusion: Single oral administration of the extract at a dose larger than 5,000 mg per kilogram of body weight does not result in lethal effects, however showed potential toxicity to the liver.