INVESTIGATING THE TOXICITY OF BETALAIN COMPOUNDS: IN SILICO ANALYSIS AND IN VIVO PREDICTIONS FOR STANDARDIZED BETA VULGARIS L. EXTRACT

Authors

  • SONY EKA NUGRAHA Department of Pharmaceutical Biology, Faculty of Pharmacy, Universitas Sumatera Utara, Medan, Indonesia https://orcid.org/0000-0001-8849-2481
  • JANE MELITA KELIAT Faculty of Vocational Study, Universitas Sumatera Utara, Medan, Indonesia
  • MARIANNE Department of Pharmacology, Faculty of Pharmacy, Department of Pharmacology, Universitas Sumatera Utara, Medan, Indonesia
  • RONY ABDI SYAHPUTRA Department of Pharmacology, Faculty of Pharmacy, Department of Pharmacology, Universitas Sumatera Utara, Medan, Indonesia

DOI:

https://doi.org/10.22159/ijap.2024v16i1.49189

Keywords:

Beta vulgaris L., In silico, In vivo, Toxicity

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.

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References

Singh S, Selvakumar R, Mangal M, Kalia P. P. Breeding and genomic investigations for quality and nutraceutical traits in vegetable crops-a review. Indian J Hortic. 2020;77(1):1-40. doi: 10.5958/0974-0112.2020.00001.8.

Kaushik A. Development of product through supplementation using beet greens and its sensory evaluation. J Pharmacogn Phytochem. 2020;9(6S):83-5.

Clements WT, Lee SR, Bloomer RJ. Nitrate ingestion: a review of the health and physical performance effects. Nutrients. 2014;6(11):5224-64. doi: 10.3390/nu6115224, PMID 25412154.

Prado J, Rostagno M. Natural product extraction: principles and applications. Cambridge: Royal Society of Chemistry Publishing; 2022.

Yuandani, Nugraha SE, Laila L, Satria D. Immunomodulatory effects of standardized extract of curcuma mangga val. on cytokines, antibody and delayed-type hypersensitivity response in Wistar rats. Res Pharm Sci. 2021;16(1):16-25. doi: 10.4103/1735-5362.305185, PMID 33953771.

Nugraha SE, Yuandani Y, Syahputra RA. Protective activity of beetroot extract on doxorubicin-induced hepatic and renal toxicity in rat model. Open Access Maced J Med Sci. 2021 Sep 28;9(A):1037-42. doi: 10.3889/oamjms.2021.7114.

Nugraha SE, Yuandani NES, Nasution ES, Syahputra RA. Investigation of phytochemical constituents and cardioprotective activity of ethanol extract of beetroot (Beta vulgaris. L) on doxorubicin induced toxicity in rat. Rasayan J Chem. 2020;13(2):973-8. doi: 10.31788/RJC.2020.1325601.

Strack D, Vogt T, Schliemann W. Recent advances in betalain research. Phytochemistry. 2003;62(3):247-69. doi: 10.1016/s0031-9422(02)00564-2, PMID 12620337.

Khare P, Kishore K, Sharma DK. Acute oral toxicity of bauhinia variegata and madhuca longifolia in mice. Int J Curr Pharm Sci 2022;14(2):69-71. doi: 10.22159/ijcpr.2022v14i2.1966.

Merdekawati F. In silico study of pyrazolylaminoquinazoline toxicity by lazar, protox, and admet predictor. J App Pharm Sci. 2018;8(9):119-29. doi: 10.7324/JAPS.2018.8918.

RI DP. Parameter standar umum ekstrak tumbuhan obat. Jakarta: Depkes; 2000.

Flores Mancha MA, Ruiz Gutierrez MG, Sanchez Vega R, Santellano Estrada E, Chavez Martinez A. Characterization of beet root extract (Beta vulgaris) encapsulated with maltodextrin and Inulin. Molecules. 2020;25(23):5498. doi: 10.3390/molecules25235498, PMID 33255296.

Ayipo YO, Ahmad I, Najib YS, Sheu SK, Patel H, Mordi MN. Molecular modelling and structure-activity relationship of a natural derivative of o-hydroxybenzoate as a potent inhibitor of dual NSP3 and NSP12 of SARS-CoV-2: in silico study. J Biomol Struct Dyn. 2023;41(5):1959-77. doi: 10.1080/07391102.2022.2026818, PMID 35037841.

Mallikarjunayya Mathapati, Akash More, Ujwal Gajbe, Deepti Shrivastava. Comparative study of effect of GnRH protocols on the quality and the quantity of oocytes retrieved and embryos form. Journal of Pharmaceutical Negative Results 2022;13(3):1081-4. doi: 10.47750/pnr.2022.13.03.175.

Seibel J, Bodie K, Weber S, Bury D, Kron M, Blaich G. Comparison of haematology, coagulation and clinical chemistry parameters in blood samples from the sublingual vein and vena cava in sprague dawley rats. Lab Anim. 2010;44(4):344-51. doi: 10.1258/la.2010.009049, PMID 20679324.

Argmann CA, Auwerx J. Collection of blood and plasma from the mouse. Curr Protoc Mol Biol. 2006;Chapter(29):Unit 29A.3. doi: 10.1002/0471142727.mb29a03s75, PMID 18265383.

Baum N, Dichoso CC, Carlton Jr CE. Blood urea nitrogen and serum creatinine. Physiology and interpretations. Urology. 1975;5(5):583-8. doi: 10.1016/0090-4295(75)90105-3, PMID 1093306.

Wang L, Wang L, Ding W, Zhang F. Acute toxicity of ferric oxide and zinc oxide nanoparticles in rats. J Nanosci Nanotechnol. 2010;10(12):8617-24. doi: 10.1166/jnn.2010.2483, PMID 21121374.

Yadav NP, Dixit VK. Recent approaches in herbal drug standardization. Int J Integr Biol. 2008;2(3):195-203.

Fernando GSN, Wood K, Papaioannou EH, Marshall LJ, Sergeeva NN, Boesch C. Application of an ultrasound-assisted extraction method to recover betalains and polyphenols from red beetroot waste. ACS Sustainable Chem Eng. 2021;9(26):8736-47. doi: 10.1021/acssuschemeng.1c01203.

Halim B, Syahputra RA, Adenin I, Lubis HP, Mendrofa F, Lie S. Determination of phytochemical constituent, antioxidant activity, total phenol and total flavonoid of extract ethanol phyllanthus emblica fruit. Pharmacogn J. 2022;14(1):63-7. doi: 10.5530/pj.2022.14.9.

Xu KY, Xia GH, Lu JQ, Chen MX, Zhen X, Wang S. Impaired renal function and dysbiosis of gut microbiota contribute to increased trimethylamine-N-oxide in chronic kidney disease patients. Sci Rep. 2017;7(1):1445. doi: 10.1038/s41598-017-01387-y, PMID 28469156.

Tsutsumi V, Nakamura T, Ueno T, Torimura T, Aguirre Garcia J. Structure and ultrastructure of the normal and diseased liver. Inliver Pathophysiology; 2017. p. 23-44. https://doi.org/10.1016/B978-0-12-804274-8.00002-3

Asra R, Yetti RD, Ratnasari D, Nessa N. Studi fisikokimia betasianin dan aktivitas antioksidan dari umbi bit merah (Beta vulgaris L.). J Pharm Sci. 2020;3(1):14-21. doi: 10.36490/journal-jps.com.v3i1.35.

Raies AB, Bajic VB. In silico toxicology: computational methods for the prediction of chemical toxicity. Wiley Interdiscip Rev Comput Mol Sci. 2016;6(2):147-72. doi: 10.1002/wcms.1240, PMID 27066112.

Madden JC, Thompson CV. In silico methods for predicting drug toxicity. New York. Springer; 2022.

He Q, Han C, Li G, Guo H, Wang Y, Hu Y. In silico design novel (5-imidazol-2-yl-4-phenylpyrimidin-2-yl)[2-(2-pyridylamino)ethyl]amine derivatives as inhibitors for glycogen synthase kinase 3 based on 3D-QSAR, molecular docking and molecular dynamics simulation. Comput Biol Chem. 2020;88:107328. doi: 10.1016/j.compbiolchem.2020.107328, PMID 32688011.

Saha J, Choudhuri S, Choudhuri D. Effect of sub-chronic exposure to chromium on haematological and biochemical parameters of male albino rat. Asian J Pharm Clin Res. 2017;10(5):345-8. doi: 10.22159/ajpcr.2017.v10i5.17468.

Published

07-01-2024

How to Cite

NUGRAHA, S. E., KELIAT, J. M., MARIANNE, & SYAHPUTRA, R. A. (2024). INVESTIGATING THE TOXICITY OF BETALAIN COMPOUNDS: IN SILICO ANALYSIS AND IN VIVO PREDICTIONS FOR STANDARDIZED BETA VULGARIS L. EXTRACT. International Journal of Applied Pharmaceutics, 16(1), 118–123. https://doi.org/10.22159/ijap.2024v16i1.49189

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