• Manal Mortady Hamed Medicinal Chemistry Department, Theodor Bilharz Research Institute, Giza, Egypt
  • Aboelfetoh Mohamed Abdalla Horticultural Crops Technology Department, National Research Center, Dokki, Cairo, Egypt
  • Mosad Ahmed Ghareeb Department of Chemistry, College of Science and Arts, Sajir, Shaqra University, Kingdom of Saudi Arabia
  • Said Abdelhalim Saleh Horticultural Crops Technology Department, National Research Center, Dokki, Cairo, Egypt


Objective: The objective of this study was undertaken to estimate the total phenolic contents (TPCs), in vitro antioxidant of different solvent extracts of M. oleifera leaves, oral acute toxicity and LD50 determination of the 85% methanolic extract as well as the chromatographic isolation and identification of the extract constituents.

Methods: The antioxidant activity of different solvent extracts of Moringa oleifera leaves were estimated using three antioxidant assays and the total phenolic contents (TPCs) were also evaluated using Folin-Ciocalteu’s assay. The n-BuOH extract undergoes further chromatographic isolation owing to the high antioxidant activity using 2, 2'-diphenyl-1-picrylhydrazyl radical (DPPH) method, which resulted in the isolation of seven compounds.

Results: The results showed that the TPCs values of the tested extracts were varied from 309.52 to 43.28 mg gallic acid equivalent/g dry extract. The reducing power antioxidant activities (RPAA) were 0.434, 0.402, 0.395, 0.149, 0.143 and 0.124, while the total antioxidant capacity (TAC) values were 316.43, 203.35, 181.56, 86.70, 76.62 and 50.83 mg ascorbic acid equivalent/g dry extract; for n-BuOH, EtOAc, 85% MeOH, H2O, CH2Cl2, and pet. ether extracts, respectively. The oral acute toxicity study of the 85% methanol extracts of M. oleifera and M. peregrina revealed that; their LD50 values were 3458.3 and 4125 mg/kg respectively, thus the two plants could be classified as slightly toxic in the scale of Hodge and Sterner which reflected their nutrient values as edible plants. The isolated compounds were identified on the basis of their 1H and 13C-NMR spectra as; cis-p-coumaric acid 4-O-(2'-O-β-D-apiofuranosyl)-β-D-glucopyranoside (1), chlorogenic acid (2), niazirin (3), 3,4-dihydroxy-β-phenylethoxy-O-α-L-rhamnopyranosyl-(l→2)-α-L-rhamnopyranosyl-(1→3)-4-O-caffeoyl-β-D-glucopyranoside (4), gallic acid (5), taxifolin (6), and benzyl-carbamo-thioethionate (7).

Conclusion: The M. oleifera leaves showed promising antioxidant activities and slightly toxic behavior.

Keywords: Moringa oleifera (Lam.), In vitro antioxidants, TPCs, Oral acute toxicity, Phenolic compounds


Download data is not yet available.


1. Anwar F, Latif S, Ashraf M, Gilani AH. Moringa oleifera: a food plant with multiple medicinal uses. Phytother Res 2007;21:17-25.
2. Makkar HPS, Becker K. Nutritional value and antinutritional components of whole and extracted Moringa oleifera leaves. Anim Feed Sci Technol 1996;63:211-28.
3. Makkar HPS, Becker K. Nutrients and anti-quality factor in different morphological part of Moringa oleifera tree. J Agric Sci 1997;128:211-322.
4. Dhakar RC, Maurya SD, Pooniya BK, Bairwa N, Gupta M. Moringa: the herbal gold to combat malnutrition. Chron Young Sci 2011;2:119-25.
5. Faizi S, Sumbul S, Versiani MA, Saleem R, Sana A, Siddiqui H. GC/GCMS analysis of the petroleum ether and dichloromethane extracts of Moringa oleifera roots. Asia Pac J Trop Biomed 2014;4:650-4.
6. Kumar A, Pari L. Antioxidant action of Moringa oleifera Lam (drumstick) against antitubercular drugs induced lipid peroxidation in rats. J Med Food 2003;6:255-9.
7. Sharma R, Singh VJ. In vivo antioxidant activity of Moringa oleifera leaf and pod extracts against carbon tetrachloride induced liver damage in albino mice. J Chem Pharm Res 2010;2:275-83.
8. Kumar V, Pandey N, Mohan N, Singh RP. Antibacterial and antioxidant activity of a different extract of Moringa oleifera leaves an in vitro study. Int J Pharm Sci Rev Res 2012;12:89-94.
9. Guevara AP, Vargas C, Sakurai H. An antitumor promoter from Moringa oleifera Lam. Mutat Res 1999;440:181-8.
10. Nikkon F, Saud ZA, Rahman MH, Haque ME. In vitro antimicrobial activity of the compound isolated from the chloroform extract of Moringa oleifera Lam. Pakistan J Biol Sci 2003;6:1888-90.
11. Pari L, Kumar NA. Hepatoprotective activity of Moringa oleifera on antitubercular drug induced liver damage in rats. J Med Food 2002;5:171-7.
12. Ezejindu DN, Udemezue OO, Chinweife KC. Hepatoprotective effects of Moringa oleifera extract on liver of Wistar rats. Int J Res Med Health Sci 2014;3:23-7.
13. Roy SK, Chandra K, Ghosh K, Mondal S, Maiti D, Ojha AK, et al. Structural investigation of a heteropolysaccharide isolated from the pods (fruits) of Moringa oleifera (Sajina). Carbohyd Res 2007;342:2380-9.
14. Manguro LO, Lemmen P. Phenolics of Moringa oleifera leaves. Nat Prod Res 2007;21:56-68.
15. Yammuenart D, Chavasiri W, Pongrapeeporn K. Chemical constituents of Moringa oleifera Lam. Sci Forum 2008;3:80-1.
16. Bennett RN, Mellon FA, Foidl N, Pratt JH, Dupont MS, Perkins L, et al. Profiling glucosinolates and phenolics in vegetative and reproductive tissues of the multi-purpose trees Moringa oleifera L. (horseradish tree) and Moringa stenopetala L. J Agric Food Chem 2003;51:3546-53.
17. Siddhuraju P, Becker K. Antioxidant properties of various solvent extracts of total phenolic constituents from three different agroclimatic origins of drumstick tree (Moringa oleifera Lam.) leaves. J Agric Food Chem 2003;51:2144-55.
18. Ghareeb MA, Saad AM, Abdel-Aleem AH, Abdel-Aziz MS, Hamed MM, Hadad AH. Antioxidant, antimicrobial, cytotoxic activities and biosynthesis of silver and gold nanoparticles using Syzygium jambos leave growing in Egypt. Der Pharm Chem 2016;8:277-86.
19. Ghareeb MA, Ahmed WS, Refahy LA, Abdou AM, Hamed MM, Abdel-Aziz MS. Isolation and characterization of the bioactive phenolic compounds from Morus alba leaves growing in Egypt. Pharmacologyonline 2016;3:157-67.
20. Hamed MM, Refahy LA, Abdel-Aziz MS. Assessing the bioactivity and antioxidative properties of some compounds isolated from Abutilon hirtum (lam.). Asian J Pharm Clin Res 2017;10:333-40.
21. El-Sayed MM, Maher AM, Hanan AE, Sayed AE, Eman AE, Ghareeb MA. Bio-guided isolation and structure elucidation of antioxidant compounds from the leaves of Ficus sycomorus. Pharmacologyonline 2010;3:317-32.
22. Ghareeb MA, Saad AM, Abdou AM, Refahy LA, Ahmed WS. A new kaempferol glycoside with antioxidant activity from Chenopodium ambrosioides growing in Egypt. Orient J Chem 2016;32:3053-61.
23. Wilbrandt W. Behrens methods for calculation of LD50. Arzneimittelforschung 1952;2:501-3.
24. Sharma RK, Chatterji S, Rai DK, Mehta S, Rai PK, Singh RK, et al. Antioxidant activities and phenolic contents of the aqueous extracts of some Indian medicinal plants. J Med Plants Res 2009;3:944-8.
25. Kalpna R, Mital K, Sumitra C. Vegetable and fruit peels as a novel source of antioxidants. J Med Plants Res 2011;5:63-71.
26. Sreelatha S, Padma PR. Antioxidant activity and total phenolic content of Moringa oleifera leave in two stages of maturity. Plant Foods Hum Nutr 2009;64:303-11.
27. El-Awady MA, Hassan MM, Abdel-Hameed ES, Gaber A. Comparison of the antioxidant activities, phenolic and flavonoids contents of the leaves-crud extracts of Moringa peregrina and Moringa oleifera. Int J Biosci 2016;8:55-62.
28. Mohamed MA, Hamed MM, Abdou AM, Ahmed WS, Saad AM. Antioxidant and cytotoxic constituents from Wisteria sinensis. Molecules 2011;16:4020-30.
29. Hsu FH, Houng JY, Chang CL, Wu CC, Chang FR, Wu YC. Antioxidant activity, cytotoxicity, and DNA information of glossogyne tenuifolia. J Agric Food Chem 2005;53:6117-25.
30. Mohamed MA, Abdou AM, Hamed MM, Saad AM. Characterization of bioactive phytochemical from the leaves of Vitex trifolia. Int J Pharm Appl 2012;3:419-28.
31. Ghareeb MA, Hussein AS, Hassan MFM, Laila AR, Mona AM, Amal MS. Antioxidant and cytotoxic activities of Tectona grandis Linn. leaves. Int J Phytopharmacol 2014;5:143-57.
32. El-Sayed MM, Maher ME, Eman AE, Ghareeb MA. Total phenolic contents and antioxidant activities of Ficus sycomorus and Azadirachta indica. Pharmacologyonline 2009;3:590-602.
33. Sadraei H, Asghari G, Farahnaki F. Assessment of hydroalcoholic extract of seeds and leaves of Moringa peregrina on ileum spasm. Res Pharmacol Sci 2015;10:252-8.
34. Chang WC, Kim SC, Hwang SS, Choi BK, Ahn HJ, Lee MY, et al. Antioxidant activity and free radical scavenging capacity between Korean medicinal plants and flavonoids by assay-guided comparison. Plant Sci 2002;163:1161-8.
35. Soler-Rivas C, Espin JC, Wichers HJ. An easy and fast test to compare total free radical scavenger capacity of foodstuffs. Phytochem Anal 2000;11:330-8.
36. Huang DJ, Lin CD, Chen HJ, Lin YH. Antioxidant and antiproliferative activities of sweet potato (Ipomoea batatas [L.] Lam ‘Tainong 57’) constituents. Bot Bull Acad Sinica 2005;45:179-86.
37. Saad AM, Abdel-Aleem AH, Ghareeb MA, Hamed MM, Abdel-Aziz MS, Hadad AH. In vitro antioxidant, antimicrobial and cytotoxic activities and green biosynthesis of silver and gold nanoparticles using Callistemon citrinus leaf extract. J Appl Pharm Sci 2016;6:277-86.
38. Adedapo AA, Mogbojuri OM, Emikpe BO. Safety evaluations of the aqueous extract of the leaves of Moringa oleifera in rats. J Med Plants Res 2009;3:586-91.
39. Chivapat S, Sincharoenpokai P, Saktiyasuthorn N, Shuaprom A, Thongsrirak P, Sakpetch A, et al. Acute and chronic toxicity of Moringa oleifera Linn leaves extracts. Thai J Vet Med 2011;41:417-24.
40. Ojo NA, Igwenagu E, Badau SJ, Sambo N, Ngulde SI, Adawaren EO, et al. Acute toxicity and effect of aqueous extract of Moringa oleifera leaves on organs and tissues in rats. J Environ Issues Agric Dev Count 2013;5:41-50.
41. Okumu MO, Mbaria JM, Kanja LW, Gakuya DW, Kiama SG, Ochola FO, et al. Acute toxicity of the aqueous methanolic Moringa oleifera (Lam) leaf extract on female Wistar albino rats. Int J Basic Clin Pharmacol 2016;5:1856-61.
42. Kasolo JN, Bimenya GS, Ojok L, Ogwal-okeng JW. Phytochemicals and acute toxicity of Moringa oleifera roots in mice. J Pharm Phytother 2011;3:38-42.
43. Kasolo JN, Bimenya GB, Ojok L, Ogwal-Okeng JW. Sub-acute toxicity evaluation of Moringa oleifera leaves aqueous and ethanol extracts in swiss albino rats. Int J Med Plant Res 2012;1:075-81.
44. Reddy YRR, Lokanatha O, Ratnam KSVP, Reddy CS, Raju IN, Reddy CD. Acute and subacute toxicity of Moringa oleifera stem bark extract in Swiss albino mice. Int J Life Sci Biotechnol Pharm Res 2013;2:74-82.
45. Lu Y, Foo Y. Flavonoid and phenolic glycosides from Salvia officinalis. Phytochemistry 2000;55:263-7.
46. Mabry TJ, Markham KR, Thomas MB. In: The Systematic Identification of Flavonoids. Springer-Verlag, Berlin-Heidelberg-New York; 1970. p. 354.
47. Faizi S, Siddiqui BS, Saleem R, Siddiqui S, Aftab K, Gilani AH. Fully acetylated carbamate and hypotensive thiocarbamate glycosides from Moringa oleifera. Phytochemistry 1995;38:957-63.
48. Maurya A, Gupta S, Srivastava SK. Preparative isolation of bioactive nitrile glycoside “Niazirin” from the fruits of Moringa oleifera using fast centrifugal partition chromatography. Separ Sci Technol 2011;46:1195-9.
49. Delnavazi MR, Mohammadifar F, Rustaie A, Aghaahmadi M, Yassa N. Phytochemical constituents, antioxidant activity and toxicity potential of Phlomis olivieri Benth. Res J Pharmacogn 2016;3:9-15.
50. Nikkon F, Hasan S, Salam KA, Mosaddik MA, Khondkar P, Haque ME, et al. Benzylcarbamothioethionate from root bark of Moringa oleifera Lam, and its toxicological evaluation. Lat Am Caribb Bull Med Arom Plants 2009;8:130-8.
51. Rice-Evans CA, Miller NJ, Paganga G. Structure-antioxidant activity relationships of flavonoids and phenolic acids. Free Radical Biol Med 1996;20:933-56.
52. Mazzone G, Malaj N, Russo N, Toscano M. Density functional study of the antioxidant activity of some recently synthesized resveratrol analogues. Food Chem 2013;141:2017-24.
53. Milic BLj, Djilas SM, Canadanovic-Brunet JM. Antioxidative activity of phenolic compounds on the metal-ion breakdown of lipid peroxidation system. Food Chem 1998;61:443-7.
54. Galato D, Ckless K, Susin MF. Antioxidant capacity of phenolic and related compounds: correlation among electrochemical, visible spectroscopy methods and structure-antioxidant activity. Redox Rep 2001;6:243-50.
55. Lu Z, Nie G, Belton PS. Structure-activity relationship analysis of antioxidant ability and neuroprotective effect of gallic acid derivatives. Neurochem Int 2006;48:263-74.
476 Views | 910 Downloads
How to Cite
Hamed, M. M., A. M. Abdalla, M. A. Ghareeb, and S. A. Saleh. “CHEMICAL CONSTITUENTS, IN VITRO ANTIOXIDANT ACTIVITY, ORAL ACUTE TOXICITY AND LD50 DETERMINATION OF MORINGA OLEIFERA LEAVES”. International Journal of Pharmacy and Pharmaceutical Sciences, Vol. 9, no. 5, May 2017, pp. 240-7, doi:10.22159/ijpps.2017v9i5.17834.
Original Article(s)