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Safety assessment of the aqueous extract of the flowers of Nymphaea lotus Linn (Nymphaeaceae): Acute, neuro- and subchronic oral toxicity studies in albinos Wistar rats

  • Mireille Kameni Poumeni , Danielle Claude Bilanda , Paul Désiré Dzeufiet Djomeni EMAIL logo , Yolande Sandrine Mengue Ngadena , Marguerite Francine Mballa , Madeleine Chantal Ngoungoure , Agnès Carolle Ouafo , Théophile Dimo and Pierre Kamtchouing



Nymphaea lotus Linn (N. lotus) is a medicinal plant widely used in Cameroon popular medicine, to treat neuropsychiatric conditions, male sexual disorders or as food supplement. However, scientific data on the pharmacotoxic profile of this plant are not available. The safety of N. lotus was assessed in acute, neuro- and subchronic toxicity studies by following the OECD guidelines. Effectively, no data have been published until now in regard to its safety on the nervous system.


Aqueous extract of N. lotus at doses of 200, 400 and 600 mg/kg body weight (BW) was evaluated for nitrites contents and orally administered to rats daily for 28 days (5 male, 5 female per group). The control group received distilled water (10 mL/kg) and a satellite group was used to observe reversal effects. Neurotoxicity of the plant was determined using open field test for motor coordination, ataxia and gait analysis. Clinical signs and state of livelihood were recorded during the 24 h, then for 28 days of treatments. At the end of 28-day period, animals were anesthetized and decapitated. The whole brain was homogenized for neurobiochemical analysis. Blood samples were collected with or without anticoagulant for hematological examinations and serum analysis. Specimens of liver, kidney, testis, ovaries, and brain were fixed in 10 % formalin and processed for histopathological examinations.


Our findings indicate dose-dependent elevation of nitrites contents in the flowers aqueous extract of N. lotus. Acute toxicity study revealed no signs of toxicity neither at the dose 2,000 mg/kg nor at 5,000 mg/kg. Thus the LD50 value of aqueous extract of N. lotus flowers is superior to 5,000 mg/kg. The repeated administration of N. lotus during 28 days, induced no signs of neurobehavioral changes in male, but female rats exhibited dose-dependent response in the open field test, suggesting sex and dose-relative psychotropic effects of N. lotus. The evaluation of neurobiochemistry revealed consistent rise of brain cholesterol by 44.05 %; 158.10 % and 147.62 % respectively in male rats treated with the doses of 200, 400 and 600 mg/kg. In female rats, these levels were significantly increased (p<0.001) only at the dose of 600 mg/kg compared to control. This trend persisted after 14 days withdrawal. Brain potassium and calcium concentrations were increased in all rats compared to their respective control receiving distilled water, suggesting transmembrane current stabilizing properties of brain cells by our extract. Further, serum biochemical analysis demonstrated that 28-day administration of N. lotus flowers increased depending on the dose and sex, the levels of serum urea, proteins, creatinine and bilirubin and reduced γ-glutamyltransferase (GGT) and alkaline phosphatase (ALP) activities. These results suggest liver alterations that are endowed by lower liver relative weight and histology damages observed in female rats treated with the dose of 600 mg/kg of our extract. We also observed a rise in the low-density lipoprotein (LDL) fraction and AI of male rats undergoing N. lotus treatment. In female rats, the latter remains unaltered, confirming the dose- and sex-dependent response of our extract. The levels of white blood cells (WBC) and granulocytes were higher in male irrespective to their control, revealing stimulatory properties of the male hematopoietic system. Such variations (sex- and dose-dependent) are without biological relevance for the majority of the biochemical parameters evaluated, indicating a wide margin of safety for the traditional use of N. lotus. The alkaloids, nitrites and phytosterols contained in N. lotus flowers extract may probably account for its neuroprotective, anti-oxidant, and immunoboosting properties.


N. lotus do not possesses neurotoxicity but is able to induce behavioral changes in rats. Therefore, the application of this plant as either drug or supplementary food should be carefully considered.


The authors are also grateful to French association PCD (Pathology Cytologie Développement) for providing histological reagents.

  1. Author contributions: DDPD, KPM, BDC, DT and PK: were involved in design, interpretation, and analysis of the data. KPM, DDPD, MNYS, MMF, NMC and OAC: were involved in acquisition of data and writing the manuscript. All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

  2. Research funding: None declared.

  3. Employment or leadership: None declared.

  4. Honorarium: None declared.

  5. Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.


1. Voon HC, Bhat R, Rusul G. Flower extracts and their essential oils as potential antimicrobial agents for food uses and pharmaceutical applications. Compr Rev Food Sci Food Saf. 2012;11:34–55. .10.1111/j.1541-4337.2011.00169.xSearch in Google Scholar

2. John-Africa LB, Idriss-Usman MS, Bulus Adzu B, Gamaniel KS. Protective effects of the aqueous extract of Nymphaea lotus L. (Nymphaeaceae) against ethanol-induced gastric ulcers. Int J Biol Chem Sci. 2012;6:1917–1925.10.4314/ijbcs.v6i5.2Search in Google Scholar

3. Van der Veen JL, Bodinga Bwa-Bodinga S. Une société traditionnelle noire africaine et ses plantes utiles: les Eviya du Gabon Web pdf source, 2007:119–153.Search in Google Scholar

4. Dalziel JM. The useful plants of West Tropical Africa. London: The Crown Agents; 1937:612.Search in Google Scholar

5. Akinjogunla OJ, Adegoke AA, Udokang IP, Adebayo-Tayo BC. Antimicrobial potential of Nymphaea lotus (Nymphaeaceae) against wound pathogens. J Med Plants Res. 2009;3:138–141.Search in Google Scholar

6. Hassan A, Rahman S, Deeba F, Mahmud S. Antimicrobial activity of some plant extracts having hepatoprotective effects. J Med Plants Res. 2009;3:20–23.Search in Google Scholar

7. Yisa J. Phytochemical analysis and antimicrobial activity of Scoparia Dulcis and Nymphaea Lotus. Aust J Basic Applied Sci. 2009;3:3975–3979.Search in Google Scholar

8. Saleem A, Ahotupa M, Pihlaja K. Total phenolics concentration and antioxidant potential of extract of medicinal plants of Pak. Z Naturoforsch C. 2001;56:973–978.10.1515/znc-2001-11-1211Search in Google Scholar

9. Mukherjee PK, Saha K, Pal M, Saha BP. Effect of Nelumbo nucifera rhizome extract on blood sugar level in rats. J Ethnopharmacol. 1997;58:207–213.10.1016/S0378-8741(97)00107-4Search in Google Scholar

10. Sowemimo AA, Fakoya FA, Awopetu I, Omobuwajo OR, Adesanya SA. Toxicity and mutagenic activity of some selected Nigerian plants. J Ethnopharmacol. 2007;113:427–432.10.1016/j.jep.2007.06.024Search in Google Scholar PubMed

11. Oliver-Bever B. Medicinal plants in Tropical West Africa. Cambridge: Cambridge University Press; 1986.10.1017/CBO9780511753114Search in Google Scholar

12. Catarino L, Martins ES, Pinto-Basto MF, Diniz MA. Plantas Vasculares e Briófitos da Guiné-Bissau. Lisboa: IICT/IPAD; 2006.Search in Google Scholar

13. Romeiras MM, Duarte MC, Indjai B, Catarino L. Medicinal plants used to treat neurological disorders in West Africa: a case study with Guinea-Bissau Flora. Am J Plant Sci. 2012;3:1028–1036.10.4236/ajps.2012.327122Search in Google Scholar

14. Kameni PM, Dzeufiet DP, Bilanda DC, Mengue NY, Mballa M, Ngoungoure MC, et al. Protective effects of Nymphaea lotus Linn. (Nymphaeaceae) aqueous extract against chronic unpredictable mild stress induced testicular lipid peroxidation. Asian J Biomed Pharm Sci. 2016;6:01–06.Search in Google Scholar

15. Raja MK, Mohan M, Sethiya NK, Mishra SH. A comprehensive review on Nymphaea stellata: a traditionally used bitter. J Adv Pharm Technol Res. 2010;1:311–319. .10.4103/0110-5558.72424Search in Google Scholar PubMed PubMed Central

16. Vergeera LH, Vander VG. Phenolic content of daylight-exposed and shaded floating leaves of water lilies (Nymphaeaceae) in relation to infection by fungi. Oecologia. 1997;112:481–484.10.1007/s004420050335Search in Google Scholar PubMed

17. Robin D. Nymphaea odorata: white pond lily. Medical Herbalism. Materia Medica Pharm. 2001;11:231–233.Search in Google Scholar

18. Adnaik RS, Pai PT, Sapakal VD, Naikwade NS, Magdum CS. Anxiolytic activity of Vitex Negundo Linn. in experimental models of anxiety in mice. Int J Green Pharm. 2009;3:243–247.10.4103/0973-8258.56284Search in Google Scholar

19. Naghma K, Sarwat S. Anticarcinogenic effect of Nymphaea alba against oxidative damage and hyperproliferative response and renal carcinogenis in Wistar rats. Mol Cell Biochem. 2005;271:1–11.10.1007/s11010-005-2258-2Search in Google Scholar

20. Naghma K, Sarwat S. Inhibition of potassium bromate-induced renal oxidative stress and hyperproliferative response by Nymphaea alba in Wistar rats. J Enzyme Inhib Med Chem. 2005;20:275–283.10.1080/14756360400028119Search in Google Scholar

21. James AD. Duke’s handbook of medicinal plants of the bible. USA: Taylor and Francis group; 2008:302–305.Search in Google Scholar

22. Bagul M, Srinivasa H. A rapid densitometric method for simultaneous quantification of gallic acid and ellagic acid in herbal raw materials using HPTLC. J Sep Sci. 2005;28:581–584. .10.1002/jssc.200301695Search in Google Scholar

23. Emboden WA. Transcultural use of narcotic water lilies in ancient Egyptian and Maya drug ritual. J Ethnopharmacol. 1981;3:39–83.10.1016/0378-8741(81)90013-1Search in Google Scholar

24. Ossian CR. The most beautiful of flowers: review of water lilies and lotuses in ancient Egypt. KMT Mag. 1999;1:48–59.Search in Google Scholar

25. Elegami AA, Bates C, Gray AI, Mackay SP, Skellern GG, Waigh RD. Two very unusual macrocyclic flavonoids from the water lily Nymphaea lotus. Phytochem. 2003;63:727–731.10.1016/S0031-9422(03)00238-3Search in Google Scholar

26. Zocrato LB, Capettini LS, Rezende BA, Silva JF, Rodrigues-Machado MG, Cortes SF, et al. Increased expression of endothelial iNOS accounts for hyporesponsiveness of pulmonary artery to vasoconstrictors after paraquat poisoning. Toxicol In Vitro. 2010;24:1019–1025.10.1016/j.tiv.2009.12.003Search in Google Scholar

27. OECD. Harmonized integrated hazard classification system for human health and environmental effects of chemical substances Vol. 2. Paris: OECD; 2001:20–24.Search in Google Scholar

28. OECD. OECD guidelines for the testing of chemicals: guidance no. 425. Acute oral toxicity: Up and Down Procedure (UDP) 2008;Available at: in Google Scholar

29. Reitman S, Frankel SA. Colorimetric method for the determination of serum glutamic oxaloacetic and glutamic pyruvic transaminase. Am J Clin Pathol. 1957;28:56–63.10.1093/ajcp/28.1.56Search in Google Scholar

30. Bartels H, Bohmer M, Heierli C. Serum creatinine determination without protein precipitation. Clinica Chimica Acta. 1972;37:193–197.10.1016/0009-8981(72)90432-9Search in Google Scholar

31. Meyer OA, Tilson HA, Byrd WC, Riley MT. A method for the routine assessment of fore- and hind limb grip strength of rats and mice. Neurobehav Toxicol. 1979;1:233–236.Search in Google Scholar

32. D’Amour FE, Smith DL. A method for determining loss of pain sensation. J Pharmacol Exp Ther. 1941;72:74–79.Search in Google Scholar

33. Rao VS, Rao A, Karanth KS. Anticonvulsant and neurotoxicity profile of Nardostachys jatamansi in rats. J Ethnopharmacol. 2005;102:351–356.10.1016/j.jep.2005.06.031Search in Google Scholar PubMed

34. Kalueff AV, Minasyan A, Tuohima P. Behavioural characterization in rats using the elevated alley Suok test. Behav Brain Res. 2005;165:52–57.10.1016/j.bbr.2005.06.037Search in Google Scholar PubMed

35. Wattanathorn J, Uabundit N, Itarat W, Mucimapura S, Laopatarakasem P, Sripanidkulchai B. Neurotoxicity of Coscinium fenestratum stem, a medicinal plant used in traditional medicine. Food Chem Toxicol. 2006;2:012.10.1016/j.fct.2006.02.012Search in Google Scholar

36. Beutler E. Glutathione. In red cell metabolism, a manual of biochemical methods. New York, NY, USA: Grune and Stratton; 1975:112–114. .Search in Google Scholar

37. Ledwozwy A, Michalak J, Stepien A, Kadziolka A. The relationship plasma triglycerides, cholesterol, total lipids, and lipid peroxidation products during human atherosclerosis. Clin Chim Acta. 1986;155:275–284.10.1016/0009-8981(86)90247-0Search in Google Scholar

38. Misra HP, Frisovich I. The role of super oxide anion in the antioxidant of epinephrine and simple assay for SOD. J Biolchem. 1979;2457:3170–3175.Search in Google Scholar

39. Dröge W. Free radicals in the physiological control of cell function. Physiological Rev. 2002;82:47–95.10.1152/physrev.00018.2001Search in Google Scholar

40. Aebi H. Methods of enzymatic analysis Vol. 2. New York, NY: Academic Press; 1974:673–678.10.1016/B978-0-12-091302-2.50032-3Search in Google Scholar

41. Griess LC, Wagner DA, Glogowski J, Skippir PL, Wishnok JS, Tannenbaum SR. Analysis of nitrate and [15N] nitrate in biological fluids. Anal Biochem. 1982;126:131–138.10.1016/0003-2697(82)90118-XSearch in Google Scholar

42. Zhang QH, Zhang L. Research advance in chemical composition and pharmacological action of Chrysanthemum morifolium. Food Drug. 2007;9:60–62.Search in Google Scholar

43. Dung NT, Kim JM, Kang SC. Chemical composition, antimicrobial and antioxidant activities of the essential oil and the ethanol extract of Cleistocalyx operculatus (Roxb.). Merr Perry buds Food Chem Toxicol. 2008;46:3632–3639.10.1016/j.fct.2008.09.013Search in Google Scholar PubMed

44. Zhao L, Yang L, Liu Y, Li C, Kang W. Antimicrobial activity of seven species Chrysanthemum morifolium Ramat cultivated in Kaifeng. Modern Pharm Res. 2009;2:82–85.Search in Google Scholar

45. Madhusudhan N, Lakshmi T, Gowtham Kumar S, Ramakrishanan , Venu GR, Anitha R, et al. Invitro antioxidant and free radical scavenging activity of aqueous and ethanolic flower extract of Nymphaea alba. Int J Drug Dev Res. 2011;3:156–164. .Search in Google Scholar

46. Aboubakar OB, Tchuemdem LM, Dzeufiet DP, Bilanda DC, Ngo Lemba TE, Bella NM, et al. Mineral constituents and toxicological profile of Jateorhiza macrantha (Menispermaceae) aqueous extract. J Ethnopharmacol. 2013;149:117–122.10.1016/j.jep.2013.05.054Search in Google Scholar PubMed

47. Rodriguez-Yanez M, Castellanos M, Blanco M, Mosguera E, Castillo J. Vascular protection in brain ischemia. Cerebrovasc Dis. 2006;21:21–29.10.1159/000091700Search in Google Scholar PubMed

48. Loomis TA, Hayes AW. Loomis’s essentials of toxicology, 4th ed. California: Academic Press; 1996:208–245.10.1016/B978-012455625-6/50014-3Search in Google Scholar

49. Sharaibi OJ, Ogundipe OT, Magbagbeola OA, Kazeem MI, Afolayan AJ. Acute and sub-acute toxicity profile of aqueous leaf extract of Nymphaea lotus Linn (Nymphaeaceae) in wistar rats. Trop J Pharm Res. 2015;14:1231–1238.10.4314/tjpr.v14i7.16Search in Google Scholar

50. Tahraoui A, Israili ZH, Lyoussi B. Acute and sub-chronic toxicity of a lyophilized aqueous extract of Centaurium erythraea in rodents. J Ethnopharmacol. 2010;132:48–55.10.1016/j.jep.2010.07.038Search in Google Scholar

51. Massoma DL, Ngaha NM, Bend E, Domkam J, Oundoum Oundoum PC, Dongho DF, et al. Toxicological study on aqueous extract of Allanblackia froribunda (Clusiaceae) on rats. J Intercult Ethnopharmacol. 2013;2:23–28.10.5455/jice.20120724082841Search in Google Scholar

52. Trayhurn P. Brown adipose tissue and energy balance. In: Trayhurn P, Nicholls D, editors. Brown Adipose Tissue. Baltimore, MD: Edward Arnold. editors; 1986:299–338.Search in Google Scholar

53. Czech AD, Kazel MR, Harris J. A nitric oxide synthase inhibitor, NG-nitro-l-arginine methylester, attenuates lipoprivic feeding in mice. Physiol Behav. 2003;80:75–79.10.1016/S0031-9384(03)00220-8Search in Google Scholar

54. Farr SA, Banks WA, Kumar VB, Morley JE. Orexin-A-induced feeding is dependent on nitric oxide. Peptides. 2005;26:759–765.10.1016/j.peptides.2004.12.004Search in Google Scholar PubMed

55. Auletta CS. Acute, subchronic and chronic toxicology. In: Derelanko MJ, Hollinger MA, editors. CRC handbook of toxicology. USA: CRC Press, Inc.1995:51–104.Search in Google Scholar

56. Haslam E. Natural polyphenols (vegetable tannins) as drugs: Possible modes of action. J Nat Prod. 1996;59:205–215.10.1021/np960040+Search in Google Scholar PubMed

57. Sowemimo AA, Omobuwajo OR, Adesanya SA. Constituents of Nymphaea lotus Linn. Nig J Nat Prod Med. 2001b;11:1–2.10.4314/njnpm.v11i1.11890Search in Google Scholar

58. Harry J, Kulig B, Lotti M, Tilson H, Winneke G. Neurotoxicity risk assessment for human health. Environmental health criteria 223. Geneva: World Health Organization; 2001.Search in Google Scholar

59. Hodgson EA. Textbook of modern toxicology. In: Hodgson E, editors. Toxicology of the nervous system, 3rd ed. Hoboken, New Jersey: John Wiley & Sons, Inc. Blake, B.L. Chapter 16; 2004:279–297. .Search in Google Scholar

60. Singh D, Goel RK. Anticonvulsant effect of Ficus religiosa: Role of serotonergic pathways. J Ethnopharmacol. 2009;123:330–334.10.1016/j.jep.2009.02.042Search in Google Scholar PubMed

61. Ngo Bum E, Taïwe GS, Moto FC, Ngoupaye GT, Nkantchoua GN, Pelanken MM. Anticonvulsant, anxiolytic and sedative properties of the roots of Nauclea latifolia Smith in mice. Epilepsy Behav. 2009;15:434–440.10.1016/j.yebeh.2009.05.014Search in Google Scholar PubMed

62. Li X, Lu Y, Wang L, Li Y, Shi Y, Cui Y, et al. Acute and sub acute toxicity of ethanol extracts from Salvia przewalskii Maxim in rodents. J Ethnopharmacol. 2010;131:110–115.10.1016/j.jep.2010.06.012Search in Google Scholar PubMed

63. Omaye ST. Chapter 2: General Principles of Toxicology. Food and nutritional toxicology. Boca Raton, London: CRC PRESS LLC; 2004:32.10.1201/9780203485309Search in Google Scholar

64. Satinder KP. A note on the correlation between open-field and escape avoidance behavior in the rat. J Psychol. 1968;69:3–6.10.1080/00223980.1968.10543441Search in Google Scholar

65. Sridhar KR, Bhat R. Lotus – a potential nutraceutical source. J Agric Technol. 2007;3:143–155.Search in Google Scholar

66. Poirier J., Apolipoprotein E. Cholesterol transport and synthesis in sporadic Alzheimer’s disease. Neurobiol Aging. 2005;26:355–361.10.1016/j.neurobiolaging.2004.09.003Search in Google Scholar

67. Angulo-Guerrero O, Oliart RR. Effects of dietary polyunsaturated fatty acids on rat brain plasma membrane fatty acid composition. Arch Latinoam Nutr. 1998;48:287–292.Search in Google Scholar

68. Evans PH. Free radicals in brain metabolism and pathology. Br Med Bull. 1993;49:577–587.10.1093/oxfordjournals.bmb.a072632Search in Google Scholar

69. Ohkawa H, Ohishi N, Yagi K. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem. 1979;95:351–358.10.1016/0003-2697(79)90738-3Search in Google Scholar

70. Parke DV, Ioannides C. The effects of nutrition on chemical toxicity. Drug Metab Rev. 1994;26:739–765.10.3109/03602539408998325Search in Google Scholar PubMed

71. Ronis MJ, Cunny HC. Physiological factors affecting xenobiotic metabolism. Introduction to biochemical toxicology, 3rd ed. New York: John Wiley & Sons; 2001:137–162.Search in Google Scholar

72. Coultate TP. Food: The chemistry of its components, 4th ed. Cambridge, UK: Royal Society of Chemistry; 2002.Search in Google Scholar

73. Musa A, Birnin-Yauri UA, Muhammad C, Umar A. Proximate composition and mineral analysis of Nymphaea lotus seeds. Afr J Food Sci Technol. 2012;3(7).Search in Google Scholar

74. Afolayan AJ, Olubunmi JS, Mutiu IK. Phytochemical analysis and in vitro antioxidant activity of Nymphaea lotus L. Int J of Pharmacol. 2013;9:297–304.10.3923/ijp.2013.297.304Search in Google Scholar

75. Adeneye AA, Ajagbonna OP, Adeleke TI, Bello SO. Preliminary toxicity and phytochemical studies of the stem bark aqueous extract of Musanga cecropioides in rats. J Ethnopharmacol. 2006;105:374–379.10.1016/j.jep.2005.11.027Search in Google Scholar PubMed

76. Guyton AC, Hall JE. Textbook of medical physiology, 11th ed. USA: Elsevier Saunders; 2006:1152.Search in Google Scholar

Received: 2016-5-24
Accepted: 2016-11-28
Published Online: 2017-3-25
Published in Print: 2017-3-24

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