ACUTE ORAL TOXICITY ASSESSMENT OF THE ETHANOL EXTRACT OF HOLOTHURIA ATRA IN MICE

PANDU SALIM HANAFI; AJI SUTRISNO; TUTIK MURNIASIH; HARIJONO; MASTERIA YUNOVILSA PUTRA; FEBRIANA UNTARI

doi:10.22159/ajpcr.2020.v13i2.36316

Authors

PANDU SALIM HANAFI Department of Agricultural Product Technology, Faculty of Agricultural Technology, Brawijaya University, Malang 65145, Indonesia.
AJI SUTRISNO Department of Agricultural Product Technology, Faculty of Agricultural Technology, Brawijaya University, Malang 65145, Indonesia. https://orcid.org/0000-0001-7971-2884
TUTIK MURNIASIH Research Center for Oceanography, Indonesian Institute of Sciences, Jakarta 14430 , Indonesia
HARIJONO Department of Agricultural Product Technology, Faculty of Agricultural Technology, Brawijaya University, Malang 65145, Indonesia.
MASTERIA YUNOVILSA PUTRA Research Center for Oceanography, Indonesian Institute of Sciences, Jakarta 14430 , Indonesia
FEBRIANA UNTARI Research Center for Oceanography, Indonesian Institute of Sciences, Jakarta 14430 , IndonesiaResearch Center for Oceanography, Indonesian Institute of Sciences, Jakarta 14430 , Indonesia

DOI:

https://doi.org/10.22159/ajpcr.2020.v13i2.36316

Keywords:

Holothuria atra, ethanol extract, safety dose, acute toxicity, LD50

Abstract

Objective: This study aimed to evaluate the toxicological potential of the ethanol extract of Holothuria atra through the acute oral toxicity – acute toxic class method.

Methods: The sample was immersed in ethanol for 72 h at room temperature and repeated 3 times. The extracts were evaporated using a vacuum rotary evaporator. The identification of compounds in the ethanol extract of H. atra was carried out using liquid chromatography–mass spectrometry (LCMS) analysis. The acute toxicity test was examined the effects of treating male mice with the ethanol extract of H. atra at 300 and 2000 mg/kg by oral administration for 14 days. On the past day of the toxicity test, liver of all experimental animals was taken for histopathological testing.

Results: LCMS analysis showed that the ethanol extract of H. atra is contained polar compounds (chlorogenic acid, coumaric acid, a glycosaminoglycan, and holothurin) and non-polar compounds (fatty acids). Acute toxicity study was performed at a dose of 300 and 2000 mg/kg for 14 consecutive days. No deaths or behavioral changes were observed during the administration of both doses. Histopathological test results on the liver showed a few changes at doses of 2000 mg/kg.

Conclusions: The LD50 is equal to 5000 mg/kg and the ethanol extracts of H. atra can be classified as practically nontoxic. However, further studies are required to proceed to clinical studies in humans.

Downloads

Download data is not yet available.

Author Biography

AJI SUTRISNO, Department of Agricultural Product Technology, Faculty of Agricultural Technology, Brawijaya University, Malang 65145, Indonesia.

Research Center for Oceanography, Indonesian Institute of Sciences, Indonesia

References

Saad DY, Baiomy AA, Mansour AA. Antiseptic effect of sea cucumber (Holothuria atra) against multi-organ failure induced by sepsis: Molecular and histopathological study. Exp Ther Med 2016;12:222-30.

Van Dyck S, Caulier G, Todesco M, Gerbaux P, Fournier I, Wisztorski M, et al. The triterpenoid glycosides of Holothuria forskali, usefulness and efficiency as a chemical defense mechanism against predatory fish. J Exp Biol 2011;214:1347-56.

Bordbar S, Anwar F, Nzamid S. High value components and bioactives from sea cucumbers for functional foods. A review. J Mar Drugs 2011;9:1761-805.

Murniasih T, Putra MY, Pangestuti R. Antioxidant capacities of Holothuria sea cucumbers. Ann Bogorienses 2015;19:21-6.

Dakrory AI, Fahmy SR, Soliman AM, Mohamed AS, Amer SA. Protective and curative effects of the sea cucumber (Holothuria atra) extract against DMBA induced hepatorenal diseases in rats. BioMed Res Int 2015;2015;563-652.

Pangestuti R, Murniasih T, Putra MY, Rasyid A, Wibowo JT, Ardiansyah A, et al. Free radical scavenging activity of selected sea cucumber species from Mataram-Lombok, Indonesia. J Teknol Sci Eng 2016;78:179-85.

Dhinakaran DI, Lipton AP. Bioactive compounds from Holothuria atra of Indian Ocean. Springerplus 2014a;3:673.

Dhinakaran DI, Lipton AP. Pharmacological potentials of sea cucumber Holothuria atra extracts from the Indian Ocean. Asian J Biomed Pharm Sci 2014b;4:36-43.

Yamanouchi T. On the poisonous substance contained in holothurians. Publ Seto Mar Biol Lab 1995;4:183-203.

Dyck SV, Gerbaux P, Patrick F. Qualitative and quantitative saponin contents in five sea cucumbers from the Indian Ocean. Mar Drugs 2010;8:173-89.

Kalinin VI, Avilov SA, Silchenko AS, Stonik VA. Triterpene glycosides of sea cucumbers (Holothuroidea, Echinodermata) as taxonomic markers. Nat Prod Commun 2015;10:21-6.

Salman M, El-Sayed S, Hameed A, Bazaid SA, Al-Shamranib MG, Mohamed HF. Liquid Chromatography-Mass Spectrometry (LC-MS) method for the determination of sugars in fresh pomegranate fruit juices. Pharm Chem 2014;6:320-33.

Belhekar SN, Chaudhari PD. Acute and sub-acute oral toxicity assessment of the polyherbal formulation in albino Wister rats. Int J Pharm Pharm Sci 2016;8:311-16.

Organization for Economic Cooperation and Development. Test Guideline 423 : Acute Oral Toxicity Acute Toxic Class Method. Paris, France: Organization for Economic Cooperation and Development; 2001. p. 1-14.

Ajput MA, Zehra T, Ali F, Kumar G. Assessment of analgesic activity of Nelumbo nucifera fruit ethanol extract. Int J Pharm Pharm Sci 2019;11:1-5.

Xiang F, Peng L, Yin Z, Jia R, Hu Z, Li Z, et al. Acute and subchronic toxicity as well as evaluation of safety pharmacology of Galla chinensis solution. J Ethnopharmacol 2015;162:181-90.

Hashim R, Azizan NA, Zamli Z, Zulkipli FH, Mazlan N, Althunibat OY. Toxicity effects of water extracts of Holothuria atra jaeger in mice. Asian Pac J Trop Biomed 2014;4:614-7.

Traesel GK, de Souza JC, de Barros AL, Souza MA, Schmitz WO, Muzzi RM, et al. Acute and subacute (28 Days) oral toxicity assessment of the oil extracted from Acrocomia aculeata Pulp in rats. Food Chem Toxicol 2014;74:320-5.

Duan WL, Liang XM. Technical Guidelines Assembly of Veterinary Medicine Research. Beijing: Chemical Industry Press; 2011.

Huberman A. Shrimp endocrinology: A review. Aquaculture 2000;191:191-208.

Chandrasoma P, Taylor CR. Concise Pathology. 3rd ed. Singapore: McGraw-Hill; 1997.

Jones TC, Hunt RD, King NW. Veterinary Pathology. 6th ed. USA: Lippincott Williams and Wilkins; 1997.

Price SA, Wilson LM. Patofisiologi: Konsep Klinis Proses-Proses Penyakit. 4th ed., Vol. 4. Jakarta: EGC; 1995.

Bahrami Y, Franco CM. Acetylated triterpene glycosides and their biological activity from Holothuroidea reported in the past six decades. Mar Drugs 2016;14:147.

Zaki MA. Effects of the crude toxin of sea cucumbers Holothuria atra on some hematological and biochemical parameters in rats. Egypt J Nat Toxins 2005;2:71-86.

Organization for Economic Cooperation and Development. Guidelines for Testing of Chemical. Acute Oral Toxicity up and Down Procedure. Paris: Organization for Economic Cooperation and Development; 2008.

de Lima FF, Traesel GK, Menegati SE, Santos AC, Souza RI, de Oliveira VS, et al. Acute and subacute oral toxicity assessment of the oil extracted from Attalea phalerata Mart Ex Spreng. pulp fruit in rats. Food Res Int 2017;91:11-7.