GC-MS, FTIR and Antiulcer Screening of Aqueous Seed Extract and Oil of Nigella sativa in Wistar Rats
Peptic ulcer disease (PUD) is a sore in the lining of the stomach or duodenal mucosa. This study was aimed at evaluating the antiulcer activity of the aqueous extract of Nigella sativa (black seed) and its constitutents. The pharmacognostic properties of the dry seeds were determined. The seeds were extracted using two methods digestion and hydro-distillation. The acute toxicity, phytochemical constituents and the antiulcer evaluation were performed on ethanol-induced ulcer in wistar rats using a standard method. FTIR and GC-MS analysis of the aqueous seed extract was also determined using standard methods. Nigella sativa seed shows the presence of starch grains, lignified tissues, tannins, cellulose, protein and oil globules. The aqueous extract has a high safety margin. The phytochemical studies revealed the presence of saponins, flavonoids, alkaloids, tannins, glycosides, fats and oil. The black seed aqueous and oil extract at 500 mg/kg significantly reduced the acidity, total acidity, and ulcer index, and pH of gastric content when compared with the positive control (Famotidine). The FTIR analysis identified the presence of the following functional groups chloro, ether, amine, carboxylic acid, nitriles, methylene, alcohol, while the GC-MS identified five compounds such as glycerin, n-Hexadecanoic acid, 9, 12-octadecadienoic acid-methyl ester, 9, 12-octadecadienoic acid and 9, 12-octadecadienoyl chloride. The pharmacognostic properties can act as a reliable tool for the standardization of the plant part. This study suggests that aqueous and oil extract possess antiulcer properties. Thus the aqueous and oil extract of black seed can be considered as antiulcer medication traditionally.
Keywords: Nigella sativa, Black seed, Pharmacognostic, Phytochemical, Acute toxicity (LD50), Antiulcer.
2. Sharma NK, Ahirwar D, Jhade D, Gupta S, Medicinal and Phamacological Potential of Nigella sativa: A Review. Ethnobotanical Review, 2009; 13:946-55.
3. Oliveira AP, Santin JR, Lemos M, Klein-Junior LC, Couto AC, Meyre da Silva Bittencourt C, Cechinel-Eliho V, Faloni de Andrade S, Gastroprotective activity of methanol extract and marrubin obtained from leaves of Marrabinum vulgare L. (Lamiaceae). Journal of Pharmacy and Pharmacology, 2011; 63:1230-1237. https://doi.org/10.1111/j.2042-7158.2011.01321.x
4. Ramakrishnan K, Salinas RC, Peptic ulcer diseases. American Family Physician, 2007; 76:1005-1012.
5. Gupta S, Jhade D, Ahirwar D, and Sharma NK, Medicinal and pharmacological potentials of Nigella sativa. Ethnobotanical Review, 2009; 13:946-55.
6. Bruce SO, Onyemailu VO, Orji CE, Evaluation of the antiulcer activity and GC-MS spectroscopic Analysis of the crude ethanolic extract of Peuraria Phaseoloide Leaf (Roxb) Benth. (FABACEAE). World Journal of Pharmaceutical Research, 2021; 10(7):39-59.
7. Tsukimi H, Nakai S, Itoh K, Amagase S, Okabe S, Involvement of heat shock proteins in the healing of acetic acid-induced gastric ulcers in rats. Journal of Physiology and Pharmacology, 2001; 52:391-405.
8. Konturek PC, Konturek T, Brzozowski JW, Konturek JW, Pawlik WW, From nerves and hormones to bacteria in the stomach; Nobel prize for achievements in gastrology during last century. Journal of Physiology and Pharmacology, 2005; 52:507-530.
9. Kommu S, Gowrishankar NL, Shankar M, Suvetha K, Eswaraiah MC, Evaluation of anti-ulcer activity of methanolic extract of Balanites aegyptiaca L. Bark. International Journal of Phytopharmacology, 2013; 4:308-10.
10. Saha L, Bhatia A, Chakrabarti A, Gastroprotective effect of bezafibrate, a peroxisome proliferator activated receptor α agonist and its mechanism in a rat model of aspirin-induced gastric ulcer. Advances Digestive Medicine, 2016; 3:101-10. https://doi.org/10.1016/j.aidm.2016.04.001
11. Ezekwesili CN, Ghasi S, Adindu CS, Mefor NC, Evaluation of the antiulcer properties of aqueous extract of unripe Musa paradisiaca Linn. Peel in wistar rats. African Journal of Pharmacy and Pharmacology, 2014; 8(39):1006-1011.
12. Ingle KP, Deshmukh AG, Padole DA, Dudhare MS, Moharil MP, Khelurkar VC, Phytochemicals: Extraction methods, identification, and detection of bioactive compounds from plant extracts. Journal of Pharmacognosy and Phytochemistry, 2017; 6:32-6.
13. Milojevic JSZ, Stojanovic TD, Palic R, Lazic ML, Veljkovic VB, Kinetics of distillation of essential oil from comminuted ripe juniper (Juniperus communis L.) berries. Biochemical Engineering Journal, 2008; 39:547-553. https://doi.org/10.1016/j.bej.2007.10.017
14. Khandelwal KR, Practical Pharmacognosy. Pragati Books Pvt. Ltd. 2008; PP 220
15. Evans WC, Trease and Evans Pharmacognosy. WB Saunders Ltd. London, 2002; 32, 33, 95 - 99, 512, 547.
16. Eleazu C, Eleazu K, Determination of the proximate composition, total carotenoid, reducing sugars and residual cyanide levels of flours of 6 new yellow and white cassava (Manihot esculenta Crantz) varieties: American Journal of Food Technology, 2012; 7(10):642-649. https://doi.org/10.3923/ajft.2012.642.649
17. Onyegbule FA, Okoli OG, Bruce SO, "In vivo Evaluation of the Antimalarial Activity of the Aqueous Ethanol Extract of Monodora myristica Seed in Albino Mice", International Journal of Science and Research, 2019; 8(6):1530 - 1538.
18. Lorke DA, New approach to practical acute toxicity test. Archives of Toxicology, 1983; 54:279-86. https://doi.org/10.1007/BF01234480
19. Ihekwereme CP, Bruce SO, Orji CE, Ibe CI, Iloh E, Aqueous extracts of Ocimum gratissimum and Anacardium occidentale synergises in anti-diarrhoeal property. International Journal of Modern Pharmaceutical Research, 2020; 4(4):06-11.
20. Onyegbule FA, Bruce SO, Onyekwe ON, Onyealisi OL, Okoye PC, Evaluation of the in vivo antiplasmodial activity of ethanol leaf extract and fractions of Jatropha gossypifolia in Plasmodium berghei infected mice. Journal of Medicinal Plant Research, 2019; 13(11):269-279 https://doi.org/10.5897/JMPR2019.6766
21. Sabiu S, Garuba T, Sunmonu T, Ajani E, Sulyman A, Nurain I, Balogun A, Indomethacin-induced gastric ulceration in rats: Protective roles of Spondias mombin and Ficus exasperata. Toxicology Reports, 2015; 2:261-7. https://doi.org/10.1016/j.toxrep.2015.01.002
22. Mshelia HS, Karumi Y, Dibal NI, Therapeutic effect of Momordica balsamina leaf extract on ethanol-induced gastric ulcer in Wistar rats. Annals of Research Hospitals, 2017; 1:1-5. https://doi.org/10.21037/arh.2017.04.03
23. Boligon AA, de Freitas RB, de Brum TF, Waczuk EP, Klimaczewski CV, de Ávila DS, Athayde ML, de Freitas Bauermann L, Antiulcerogenic activity of Scutia buxifolia on gastric ulcers induced by ethanol in rats. Acta Pharmaceutical Sinica B, 2014; 4:358-67. https://doi.org/10.1016/j.apsb.2014.05.001
24. Farzaei MH, Khazaei M, Abbasabadei Z, Feyzmahdavi M, Mohseni GR, Protective effect of Tragopogon graminifolius DC against ethanol induced gastric ulcer. Iranian Red Crescent Medical Journal, 2013; 15:813-6. https://doi.org/10.5812/ircmj.7845
25. Van der Weerd J, Heeren, RMA, Boon JJ, Preparation methods and accessories for the infrared spectroscopic analysis of multi-layer paint. Studies in Conservation, 2004; 49:193-210. https://doi.org/10.1179/sic.2004.49.3.193
26. Onyemailu VO, Bruce SO, Iloh ES, UV-Visible and FTIR Spectroscopic Analysis of the Crude Ethanolic Extract of Peuraria Phaseoloide Leaf (Roxb) Benth. (FABACEAE). International Journal of Modern Pharmaceutical Research. 2021; 5(3):148-153.
27. Bruce SO, Onyemailu VO, Orji CE, Evaluation of the antiulcer activity and GC-MS spectroscopic analysis of the crude ethanolic extract of Peuraria Phaseoloide Leaf (Roxb) Benth. (FABACEAE). World Journal of Pharmaceutical Research, 2021; 10(7): 39-59.
28. Gnanavel V, Mary Saral A, GC-MS analysis of petroleum ether and ethanol leaf extracts from Abrus precatorius Linn. International Journal of Pharma and Bio Sciences, 2013; 4(3):37-44.
29. Chai J, Peptic Ulcer Disease. Rijeka, Croatia: InTech. 2011; ISBN: 978-953-307-976-9
30. Paguigan ND, Castillo DH, Chichioco-Hernandez CL, Anti-ulcer Activity of Leguminosae Plants. Argentina Gastroenterology, 2014; 51(1):64-7. https://doi.org/10.1590/S0004-28032014000100013
31. Sultana S, Akram M, Asif HM, Akhtar N, Complementary and Alternative Approaches to Treat Peptic Ulcer. International Research Journal of Pharmacy, 2014; 5(5):353-59. https://doi.org/10.7897/2230-8407.050574
32. Yousaf Z, Shinwari ZK, Asghar R, Parveen A, Leaf epidermal anatomy of selected Allium species, family Alliaceae from Pakistan. Pakistan Journal of Botany, 2008; 40(1):77.
33. Naz N, Hameed M, Ashraf M, Ahmad R, Arshad M, Eco-morphic variation for salt tolerance in some grasses from Cholistan Desert, Pakistan. Pakistan Journal of Botany, 2009; 41(4):1707- 1714.
34. Hameed M, Ashraf M, Naz N, Al-Qurainy F, Anatomical adaptations of Cynodon dactylon (L.) Pers. from the Salt Range Pakistan to salinity stress. I. Root and stem anatomy. Pakistan Journal of Botany, 2010; 42(1):279- 289.
35. Riaz A, Younis A, Hameed M, Kiran S, Morphological and biochemical responses of turf grasses to water deficit conditions. Pakistan Journal of Botany, 2010; 42(5):3441- 3448.
36. Khan R, Abidin SZU, Mumtaz AS, Jamsheed S, Ullah H, Comparative leaf and pollen micromorphology on some Grasses taxa (Poaceae) distributed in Pakistan. International Journal of Nature and Life Sciences, 2017; 1(2):72- 82.
37. Ahmad K, Khan MA, Ahmad M, Shaheen N, Nazir A, Taxonomic diversity in epidermal cells of some sub-tropical plant species. International journal of Agricultural and Biology, 2010; 12(1):115- 118.
38. Adedeji O, Leaf epidermal studies of the species of Emilia Cass. (Senecioneae, Asteraceae) in Nigeria. Botanica Lithuanica, 2004; 10(2):121- 133.
39. Adedeji O, Ajuwon OY, Babawale OO, Foliar epidermal studies, organographic distribution and taxonomic importance of trichomes in the family Solanaceae. International Journal of Botany, 2007; 3(3):276- 282. https://doi.org/10.3923/ijb.2007.276.282
40. Kadam MPV, Yadav KN, Patel AN, Navsare VS, Bhilwade SK, Patil MJ, Phytopharmacopoeial specifications of Garcinia indica fruit rinds. Pharmacognosy Journal, 2012; 4(31):23- 28. https://doi.org/10.5530/pj.2012.31.5
41. Ilanchezhian R, Roshy JC, Acharya RN, Harisha CR, Shukla VJ, Pharmacognostical and physicochemical analysis of bhallataka (Semecarpus anacardium Linn.)-fruit. Pharmacognosy Journal, 2011; 3(20):9- 16. https://doi.org/10.5530/pj.2011.20.3
42. Bruce SO, Onyegbule FA, Ezugwu CO, Pharmacognostic, physicochemical and phytochemical evaluation of the leaves of Fadogia cienkowskii Schweinf (Rubiaceae). Journal of Pharmacognosy and Phytotherapy, 2019; 11(3):52-60. https://doi.org/10.5897/JPP2019.0552
43. Desai D, Chanda S, Pharmacognostic study and physicochemical analysis of leaves of Terminalia arjuna. Pharmacognosy Journal, 2014; 6(6):15-19. https://doi.org/10.5530/pj.2014.6.4
44. Zhao Z, Liang Z, Guo P, Macroscopic identification of Chinese medicinal materials: traditional experiences and modern understanding. Journal of Ethnopharmacology, 2011; 131:556-561. https://doi.org/10.1016/j.jep.2011.01.018
45. Raj RSN, Radhamany PM, Pharmacognostic and physicochemical analysis on the leaves of Brunfelsia americana L. Asian Pacific Journal of Tropical Biomedicine, 2012; 543-546. https://doi.org/10.1016/S2221-1691(12)60178-6
46. Onyegbule FA, Ezenwa CJ, Bruce SO, Umeokoli BO, Standardization, chemical composition and antipyretic evaluation of methanol leaf extract and fractions of Chrysophyllum albidum (Sapotaceae). Tropical Journal of Natural Product Research, 2020; 4(6):216-222. https://doi.org/10.26538/tjnpr/v4i6.1
47. Osborn AE, "Saponins in Cereals", WHO Traditional. Phytochemistry, 2003; 62:1-4. https://doi.org/10.1016/S0031-9422(02)00393-X
48. Bruce SO, Onyegbule FA, Biosynthesis of Natural Products, IntechOpen, 2021; DOI: 10.5772/intechopen.97660. https://www.intechopen.com/onlinefirst/biosynthesis-of-natural-products https://doi.org/10.5772/intechopen.97660
49. Bruce SO, Onyegbule FA, Ihekwereme CP, Evaluation of hepato-protective and antimicrobial activities of ethanol extracts and fractions of Picralima nitida seed and pod. Journal of Phytomedicine and Therapeutic, 2016; 1(2):1 - 21.
50. Okoye VO, Bruce SO, Onyegbule FA, Phytochemical screening and pharmacognostic properties of Peuraria phaseoloides leaves (roxb) benth. (fabaceae). International Journal of Public Health, Pharmacy and Pharmacology, 2020; 5(2):11-24.
51. Awaad AS, Reham ME, Gamal AS, Natural products in treatment of ulcerative colitis and peptic ulcer. J. Saudi Chem. Soc, 2013; 17:101-124. https://doi.org/10.1016/j.jscs.2012.03.002
52. Arawwawala M, Thabrew I, Handunnetti S, Arambewela L, Anti-inflammatory activity of Trichosanthes cucumerina Linn in rats. Journal of Ethnopharmacology, 2010; 131:538-543. https://doi.org/10.1016/j.jep.2010.07.028
53. Gazzieri D, Trevisani M, Springer J, Harrison S, Cottrell GS, Andre E, Nicoletti P, Gems D, Partridge L, Stress response and aging: "that which does not kill us makes us stronger". Cell Metabolism, 2008; 7:200-203. https://doi.org/10.1016/j.cmet.2008.01.001
54. Bae, D. K., Park, D., and Lee, S.H. Different antiulcer activities of pantoprazole in stress, alcohol and pylorus ligation-induced ulcer models. Lab. Animal Res, 2011; 27(1):47-52. https://doi.org/10.5625/lar.2011.27.1.47
55. Baena JM, Lopez C, Hidalgo A, Rame F, Jimenez S, Garcia M, Relationship between alcohol consumption and the success of Helicobacter pylori eradication therapy using Omeprazole, Clarithromycin and Amoxycillin for one week. European Journal of Gastroenterology and Hepatology, 2002; 14(3):291-296. https://doi.org/10.1097/00042737-200203000-00014
56. Janfaza S, Janfaza E, The study of pharmacologic and medicinal valuation of thymoquinone of oil of Nigella sativa in the treatment of diseases. Annals of Biological Research, 2012; 3(4):1953-7.
57. Tarnawski AS, Ahluwalia A, Molecular Mechanisms of Epithelial Regeneration and Neovascularization during Healing of Gastric and Esophageal Ulcers. Current Medicinal Chemistry, 2012; 19(1):16-27. https://doi.org/10.2174/092986712803414088
58. Syamsu Rijal, Upik Anderiani Miskad, Djumadi Achmad, Rina Masadah, Dasril Daud, Cahyono Kaelan, Halida Rahawarin, Swandari Paramita, Yadi Yasir, Evaluation of Anti-ulcerogenic Activity in Oil Extract of Jintan Hitam (Nigella sativa) Against Ethanol Induced Gastric Ulcer in Mice (Mus musculus). American Journal of Clinical and Experimental Medicine, 2016; 4(6):179-184 https://doi.org/10.11648/j.ajcem.20160406.14
59. Onyemailu VO, Bruce SO, Iloh ES, UV-Visible and FTIR Spectroscopic Analysis of The Crude Ethanolic Extract of Peuraria phaseoloide Leaf (Roxb) Benth. (FABACEAE). International Journal of Modern Pharmaceutical Research, 2021; 5(3):148-153.
60. Sforcin JM, Bankova JM, Propolis: Is there a potential for the development of new drugs? Journal of Ethnopharmacol, 2011; 133:253-260. https://doi.org/10.1016/j.jep.2010.10.032
61. Gomez-Caravaca AM, Gomez-Romero M, Arraez-Roman D, Segura-Carretero A, Fernandez-Gutierrez A, Advances in the analysis of phenolic compounds in products derived from bees. Journal of Pharmaceutical and Biomedical Analysis, 2006; 41:1220-1234. https://doi.org/10.1016/j.jpba.2006.03.002
62. Kartal M, Kaya S, Kurucu S, GC-MS analysis of propolis samples from two different regions of Turkey. Zeitschrift Naturforschung C, 2002; 57:905-909. https://doi.org/10.1515/znc-2002-9-1025
63. Christov R, Trusheva B, Popova M, Bankova V, Bertrand M, Chemical composition of propolis from Canada, its antiradical activity and plant origin. Natural Product Research, 2006; 20:531-536. https://doi.org/10.1080/14786410500056918
64. Araujo MJAM, Bufalo MC, Bruno JC, Fernandes Jr A, Trusheva B, Bankova V, Sforcin, JM, The chemical composition and pharmacological activities of geopropolis produced by Melipona fasciculata Smith in Northeast Brazil. Journal of Molecular Pathophysiology, 2015; 4:12-20. https://doi.org/10.5455/jmp.20150204115607
65. Komansilan A, Abadi AL, Yanuwiadi B, Kaligis DA, Isolation and Identification of Biolarvicide from Soursop (Annona muricata Linn) Seeds to Mosquito (Aedes aegypti) Larvae. International Journal of Engineering & Technology IJET-IJENS, 2012; 12(03): 28-32.
66. Jegadeeswari P, Nishanthini A, Muthukumaraswamy S, Mohan VR, GC-MS analysis of bioactive components of Aristolochia krysagathra (Aristolochiaceae). Journal of Current Chemical and Pharmaceutical Sciences, 2012; 2:226-236.
67. Upgade A, Anusha B, Characterization and medicinal importance of phytoconstiuents of Carica papaya from down south Indian region using gas chromatography and mass spectroscopy. Asian Journal of Pharmaceutical and Clinical Research, 2013; 6(4):101-106.
68. Wu L, Gao H, Wang X, Ye J, Lu J, Liang Y, Analysis of chemical composition of Chrysanthemum indicum flowers by GC/MS and HPTLC. Journal of Medicinal Plants Research, 2010; 4(5):421-426.
69. Vohra A, Kaur H, Chemical investigation of medicinal plant Ajuga bracteosa. Journal of Natural Product Plant Resources, 2011; 1(1):37-45.
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
Authors who publish with this journal agree to the following terms:
- Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution-NonCommercial 4.0 International (CC BY-NC 4.0). that allows others to share the work with an acknowledgment of the work's authorship and initial publication in this journal.
- Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgment of its initial publication in this journal.
- Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).