A Toxicological Safety Assessment of a Standardized Extract of  Eulophia macrobulbon in Rodents

Watcharaporn Preedapirom; Kanokwan Changwichit; Warisaraporn Tangchang; Kornkanok Ingkaninan; Pornnarin Taepavarapruk

doi:10.14456/nujst.2019.10

Authors

Watcharaporn Preedapirom Department of Physiology, Faculty of Medical Science, Naresuan University, Phitsanulok, 65000, Thailand
Kanokwan Changwichit Department of Pharmaceutical Chemistry and Pharmacognosy, Faculty of Pharmaceutical Sciences and Center of Excellence for Innovation in Chemistry, Naresuan University, Phitsanulok 65000, Thailand
Warisaraporn Tangchang Center for Animal Research, Naresuan University, Phitsanulok 65000, Thailand
Kornkanok Ingkaninan Department of Pharmaceutical Chemistry and Pharmacognosy, Faculty of Pharmaceutical Sciences and Center of Excellence for Innovation in Chemistry, Naresuan University, Phitsanulok 65000, Thailand
Pornnarin Taepavarapruk Department of Physiology, Faculty of Medical Science, Naresuan University, Phitsanulok, 65000, Thailand

DOI:

https://doi.org/10.14456/nujst.2019.10

Keywords:

Eulophia macrobulbon, Toxicological safety assessment, Acute and Chronic toxicity, rodents

Abstract

The tuber of Eulophia macrobulbon (EM) has been traditionally used in Thailand to relieve pain and fatigue. Recently, it has been reported that EM extract could provide benefits such as anti-inflammatory, and antioxidant that may be used as herbal remedy in the pharmaceutical or food industries. Cytotoxic properties of EM extract on cancer cell lines were also reported in vitro. However, as yet, no safety data has been investigated. Our study is the first one in which the acute and chronic toxicity of the standardized extract of EM has been tested. The OECD Guideline No. 420 (limit test) was used for the acute toxicity test in which a single oral dose of 2000 mg crude extract/kg body weight was administered to Wistar rats. Observations were conducted during the 2-week period after administration. Clinical signs and mortality were used as parameters to evaluate the acute toxicity. The results showed no toxicity in terms of general behavior change, mortality, or change in gross appearance of internal organs. Chronic toxicity was studied by following OECD Guideline No. 452. EM extract was administered orally at doses of 5, 50 and 500 mg/kg for 6 months. Mortality, clinical signs, body weights, biological and hematological parameters, organ weights, as well as macro-and microscopic observation of several organs were evaluated at the end of the test period. Our tests showed that EM extract did not induce death or adverse effects, and nor did produce any abnormality of the hematological and biochemical parameters or histopathological changes.

Overall, our test results have indicated that the oral Lethal Dose 50 (LD₅₀) of EM extract is more than 2,000 mg/kg, and oral administration of EM for long period up to 6 months is safe. Based on established literature on health benefits of EM, it is important to focus attention on its active constituents and therapeutic effects in order to further develop EM as herbal remedy for treating of inflammations associated diseases as well as erectile dysfunction.

References

Andrade, F., Albuquerque, C. A., Maraschin, M., & Silva, E. L. (2012). Safety assessment of yerba mate (llex paraguariensis) dried extract: results of acute and 90 days subchronic toxicity studies in rats and rabbits. Food and Chemical Toxicology, 50, 328-334.

Antonelli-Ushirobira, T. M., Blainski, A., Fernandes, H. G., Moura-Costa, G. F., Costa, M. A., Campos-Shimada, L. B., … Mello, J. C. (2015). Acute toxicity and long-term safety evaluation of the crude extract from rhizomes of Limonium brasiliense in mice and rats. Journal of Ethnopharmacology, 174, 293-298.

Bailey, S. A., Zidell, R. H., & Perry, R. W. (2004). Relationships between organ weight and body/brain weight in the rat: what is the best analytical endpoint?. Toxicologic pathology, 32, 448-466.

Carson, C. C., & Lue, T. F. (2005). Phosphodiesterase type 5 inhibitors for erectile dysfunction. BJU International, 3, 257-280.

Chanda, R., Mohanti, J. P., Bhuyan, N. R., Kar, P. K., & Nath, L. K. (2007). Medicinal plants used against gastrointestinal tract disorders by traditional healers of Sikkim Himalayas. Indian Journal of Traditional Knowledge, 6, 606-610.

Chuakul, W. (2010) Thai traditional herbs for relief pain and fatigue. Thai Pharmaceutical and Health Science Journal, 1, 1-13.

Corbin, J. D. (2004). Mechanisms of action of PDE5 inhibition in erectile dysfunction. International Journal of Impotence Research journal, 1, 4-7.

Geary, N. (1998). The effect of estrogen on appetite. Medscape Womens Health, 3(6), 3. Retrieved from https://www.ncbi.nlm.nih.gov/pubmed/9878925

Giknis, M. L., & Clifford, C. B. (2008). Clinical Laboratory Parameters for Crl: WI (Han). Wilmington, MA: Charles River Laboratories.

Hematological data of NLAC-MU laboratory animals (n.d.). Retrieved from http://www.nlac.mahidol.ac.th/acth/index.php/quality-control/health-control#Hemato

Jagdale, S. P., Shimpi, S., & Chachad, D. (2009). Pharmacological studies of Salep. Journal of Herbal Medicine and Toxicology, 3, 153-156.

Lameire, N., Van Biesen, W., & Vanholder, R. (2005). Acute renal failure. Lancet, 365, 417–447.

Levine, B. S. (2002). Animal Clinical Pathology (2nd ed.). USA: CRC Press.

Li, F., He, X., Niu, W., Feng, Y., Bian, J., & Xiao, H. (2015). Acute and sub-chronic toxicity study of the ethanol extract from leaves of Aralia elata in rats. Journal of ethnopharmacology, 175, 499-508.

Li, X., Luo, Y., Wang, L., Li, Y., Shi, Y., Cui, Y., & Xue, M. (2010). Acute and subacute toxicity of ethanol extracts from Salvia przewalskii Maxim in rodents. Journal of ethnopharmacology, 131, 110-115.

Medhi, R., & Chakrabarti, S. (2009). Traditional knowledge of North-East people on conservation of wild orchids. Indian Journal of Traditional Knowledge, 8, 11-16.

OECD GUIDELINE FOR THE TESTING OF CHEMICALS: 420 Acute Oral Toxicity-Fixed Dose Procedure (2001). Retrieved from https://ntp.niehs.nih.gov/iccvam/suppdocs/feddocs/oecd/oecd_gl420.pdf

OECD GUIDELINE FOR THE TESTING OF CHEMICALS: 452 Chronic Toxicity Studies (2009). Retrieved from https://www.oecd-library.org/docserver/9789264071209-en.pdf?expires=1525850043&id=id&accname=guest&checksum=C1584032F70B64DEEF0475FCBBA8F1A2

Rao, A. N. (2004). Medicinal orchid wealth of Arunachal Pradesh. ENVIS Newsletter, 1, 1-5.

Schuster, R., Zeindl, L., Holzer, W., Khumpirapang, N., Okonogi, S., Viernstein, H., & Mueller, M. (2017). Eulophia macrobulbon - an orchid with significant anti-inflammatory and antioxidant effect and anticancerogenic potential exerted by its root extract. Phytomedicine, 24, 157-165.

Temkitthawon, P., Changwichit, K., Khorana, N., Viyoch, J., Suwanborirux, K., & Ingkaninan, K. (2017). Phenanthrenes from Eulophia macrobulbon as Novel Phosphodiesterase-5 Inhibitors. Natural Product Communications, 12, 79-82.

Thomas, S. A. (1998). A Preliminary Checklist of Genus Eulophia. Lindleyana, 13, 170-202.

Tuchinda, P., Udchachon, J., Khumtaveeporn, K., Taylor, W. C., Engelhardt, L. M., & White, A. H. (1988). Phenanthrenes of Eulophia nuda. Phytochemistry, 27, 3267–3271

Wisutthathum, S., Chootipa, K., Martin, H., Ingkaninan, K., Temkitthawon, P., Totoson, P., & Demougeot, C. (2018). Vasorelaxant and Hypotensive Effects of an Ethanolic Extract of Eulophia macrobulbon and Its Main Compound 1-(4′-Hydroxybenzyl)-4,8-Dimethoxyphenanthrene-2,7-Diol, Front. Pharmacol, 9, 1-12. https://doi.org/10.3389/fphar.2018.00484

Wisutthathum, S., Demougeot, C., Totoson, P., Adthapanyawanich, K., Ingkaninan, K., Temkitthawon, P., & Chootipa, K. (2018). Eulophia macrobulbon extract relaxes rat isolated pulmonary artery and protects against monocrotaline-induced pulmonary arterial hypertension. Phytomedicine, 50(15), 157-165. https://doi.org/10.1016/j.phymed.2018.05.014