Day time trend in the phytochemical constituents and antioxidant activity of Azadirachta indica leaves

Document Type: Original article

Authors

Biomedicinal Research Centre, Forestry Research Institute of Nigeria, PMB 5054 Jericho Hill, Ibadan, Nigeria.

Abstract

Background & Aim:Medicinal plants have been of immense therapeutic values to many communities, some traditional folklore places emphasis on collecting plants during the day for medicinal purpose. This study examined the effect of collection time on the phytochemical composition and antioxidant activity of Azadirachta indica leaves.
Experimental: Leaves samples were collected at six hour intervals between 6AM to 6PM, phytochemical quantification was done using standard methods; antioxidant assays were done after ethanol extraction via reducing power and nitric oxide scavenging assay (NO).
Results: Saponin concentration ranged from 1.08-1.24%, Tannin 0.22-0.31%, Steroid 0.08-0.14%, Glycoside 0.090-0.104%, Alkaloids 3.88-4.27%, Flavonoids 0.050-0.080% and 1.16-1.32% for Phenolics in the leaves of A. indica collected at different time of the day. The effect of time of sample collection on the phytochemical constituents shows that phytochemicals such as saponin, tannin, glycoside, alkaloids and phenolics are significantly (p<0.05) more concentrated in the leaves in the evening time (6PM) compared to other time period while steroid and flavonoid are also significantly more concentrated in the noon time (12PM). Reducing power assay shows that leave samples collected at 12PM exhibited the highest reducing power which is comparable with the samples collected at 6PM. The early morning samples showed the least reducing power while the standard antioxidant had higher reducing power than the three sample extracts. The highest percentage NO inhibition was observed in leave samples collected at 12PM (72.40%), but this is less than the ascorbic acid (standard antioxidant) which shows 84.47% inhibition. Analysis of the IC50 revealed that the afternoon sample (12PM) had the least value (23.29 μg/ml) thus, possessing better antioxidant capacity with respect to the NO scavenging; this is to a lower extent than the IC50 value (12.71 μg/ml) of the standard antioxidant.
Recommended applications/industries: From the foregoing, variation exists in daytime concentrations of phytochemical in A. indica leaves which may impact on its pharmacological actions.

Keywords


Article Title [Persian]

روند تغییرات روزانه ترکیبات فیتوشیمیایی و فعالیت آنتی اکسیدانی برگهای Azadirchta indica

Authors [Persian]

  • پاول اگونبامو
  • الامیلکان اوتدو
مرکز تحقیقات زیست پزشکی، پژوهشکده جنگلداری نیجریه، ایبادان، نیجریه
Abstract [Persian]

Background & Aim:Medicinal plants have been of immense therapeutic values to many communities, some traditional folklore places emphasis on collecting plants during the day for medicinal purpose. This study examined the effect of collection time on the phytochemical composition and antioxidant activity of Azadirachta indica leaves.
Experimental: Leaves samples were collected at six hour intervals between 6AM to 6PM, phytochemical quantification was done using standard methods; antioxidant assays were done after ethanol extraction via reducing power and nitric oxide scavenging assay (NO).
Results: Saponin concentration ranged from 1.08-1.24%, Tannin 0.22-0.31%, Steroid 0.08-0.14%, Glycoside 0.090-0.104%, Alkaloids 3.88-4.27%, Flavonoids 0.050-0.080% and 1.16-1.32% for Phenolics in the leaves of A. indica collected at different time of the day. The effect of time of sample collection on the phytochemical constituents shows that phytochemicals such as saponin, tannin, glycoside, alkaloids and phenolics are significantly (p<0.05) more concentrated in the leaves in the evening time (6PM) compared to other time period while steroid and flavonoid are also significantly more concentrated in the noon time (12PM). Reducing power assay shows that leave samples collected at 12PM exhibited the highest reducing power which is comparable with the samples collected at 6PM. The early morning samples showed the least reducing power while the standard antioxidant had higher reducing power than the three sample extracts. The highest percentage NO inhibition was observed in leave samples collected at 12PM (72.40%), but this is less than the ascorbic acid (standard antioxidant) which shows 84.47% inhibition. Analysis of the IC50 revealed that the afternoon sample (12PM) had the least value (23.29 μg/ml) thus, possessing better antioxidant capacity with respect to the NO scavenging; this is to a lower extent than the IC50 value (12.71 μg/ml) of the standard antioxidant.
Recommended applications/industries: From the foregoing, variation exists in daytime concentrations of phytochemical in A. indica leaves which may impact on its pharmacological actions.

Keywords [Persian]

  • Azadirchta indica
  • Day time variation
  • Nitric oxide scavenging assay
  • phytochemicals
  • Reducing power
Akinmalodun, A.C., Ibukun, E.O., Afor, E., Akirinlola, B.L., Onibon, T.R., Akinboboye, A.O., Obuotor, E.M. and Farombi, E.O. 2007. Chemical Constituents and Antioxidant Activity of Alstonia boonei. African Journal of Biotechnology, 6(10): 1197-1201.

AOAC 1999. Official Methods of Analysis, 15th edn. Association of Official Analytical Chemists, Arlington, VA.

Aqil, F., Ahmad, I. and Mehmood, Z. 2006. Antioxidant And Free Radical Scavenging Properties of Twelve Traditionally used Indian Medicinal Plants. Turkey Journal of Biology, 30(1): 177-183.

Asghari, G., Gholamali, H., Mahmoudi, Z., and Asghari, M. 2014. Diurnal Variation of Essential of the Oil Components of Pycnocycla spinosa Decne. ex Boiss. Jundishapur Jurnal of Natural Pharmaceutical Products, 9(1): 35–38.

Biswas, K., Chattopadhyay, I., Banerjee, R.K. and Bandyopadhyay, U. 2002. Biological Activities and Medicinal Properties of Neem (Azadirachta indica). Current Science, 82(11): 1336-1345.

Bodeker, G., Bhat, K.K.S., Burley, J. and Vantomme, P. 1997. Medicinal Plants for Forest Conservation and Health Care. Rome. Food and Agricultural Organization (FAO).

Castelo, A.V.M., Del Menezzi, C.H.S. and Resck, I.S. 2012. Seasonal Variation in the Yield and the Chemical Composition of Essential Oils from Two Brazilian Native Arbustive Species. Journal of Applied Sciences, 12(8): 753-760.

Cohen, M.L. 2000. Changing Patterns of Infectious Disease, Nature, 406: 762-767.

Ebi, G.C. and Ofoefule, S.I. 2000. Antimicrobial Activity of Pterocarps osun Stems, Fitoterapia, 71(4): 433-435.

Ejikeme, C.M., Ezeonu, C.S., Eboatu, A.N. 2014. Determination of Physical and Phytochemical Constituents of Some Tropical Timbers Indigenous to Niger Delta Area of Nigeria. European Scientific Journal, 10(18): 247-270.

Eleazu C.O., Okafor P.N., Amajor J., Awa, E., Ikpeama, A.I., Eleazu K.C. 2011. Chemical Composition, Antioxidant Activity, Functional Properties and Inhibitory Action of Unripe Plantain (M. Paradisiacae) flour. African Journal of Biotechnology, 10(74): 16948-6952

Eleazu, C.O., Eleazu, K.C., Awa, E. and Chukwuma, S.C. 2012. Comparative study of the phytochemical composition of the leaves of five Nigerian medicinal plants. Journal of Biotechnology and Pharmaceutical Research, 3(2): 42-46.

El-hawary, S.S., El-tantawy, M.E., Rabeh, M.A. and Badr, W.K. 2013. Chemical Composition and Biological Activities of Essential Oils of Azadirachta indica  A. Juss. International Journal of Applied Research in Natural Products, 6(4): 33-42.

Enyi-Idoh, K.H., Utsalo, S.J., Epoke, J. and Arikpo, G.E. 2012. Characterization of the Antibacterial and Phytochemical Potentials of Leaves Extracts of Azadirachta indica A. Juss. Journal of Science Engineering and Technology, 1(1): 52-57.

Ezeonu, C.S. and Ejikeme, C.M. 2016. Qualitative and Quantitative Determination of Phytochemical Contents of Indigenous Nigerian Softwoods. New Journal of Science, 2016, Article ID 5601327, 9 pages http://dx.doi.org/10.1155/2016/5601327.

Farhoosh, R., Gholam, A.G. and Mohammad, H.H.K. 2007. Antioxidant Activity of Various Extracts of Old Tea Leaves and Black Tea Wastes (Camellia sinensis L.). Food Chemistry, 100(1): 231-236.

Ghimeray, A.K., Jin, C.,  Ghimire, B.K. and Dong Ha Cho. 2009. Antioxidant Activity and Quantitative Estimation of Azadirachtin and Nimbin in Azadirachta indica  A. Juss grown in Foothills of Nepal. African Journal of Biotechnology, 8(13): 3084-3091.

Harborne, J.B. 1973. Phytochemical Methods: A Guide to Modern Techniques of Plant Analysis. Chapman and Hall, UK, London pp 113.

Harry-Asobara, L.J. and Samson, E.O. 2014. Comparative Study of the Phytochemical Properties of Jatropha curcas and Azadirachta indica  Plant Extracts. Journal of Poisonous and Medicinal Plants Research, 2(2): 20-24.

Itelima, J.U., Nwokedi, V.C., Ogbonna, A.I., Nyam, M.A. 2016. Phytochemical Screening and Antimicrobial Activity Evaluation of Aqueous and Ethanolic Extracts of the Leaf of Azadirachta indica  Juss (neem) on some microorganisms. World Journal of Microbiology, 3(1): 56-60.

Jagetia, G.C., Rao, S.K., Baliga, M.S. and Babu, K. 2004. The Evaluation of Nitric Oxide Scavenging Activity of Certain Herbal Formulations in vitro: A Preliminary Study. Phytotherapy Research, 18(7): 561-565.

Kelm, M. 1999. Nitric Oxide Metabolism and Breakdown. Biochimica et Biophysica Acta, 1411(1): 273-289.

Kim, K.T., Yoo, K.M., Lee, J.W., Eom, S.H., Hwang, I.K. and Lee, C.Y. 2007. Protective Effect of Steamed American Ginseng (Panax quinquefolius L.) on V79-4 Cells Induced by Oxidative Stress. Journal of Ethnopharmacology, 111(1): 443-445.

Koruthu, D.P., Manivarnan, N.K., Gopinath, A., and Abraham, R. 2011. Antibacterial Evaluation, Reducing Power Assay and Phytochemical Screening of Moringa oleifera Leaf Extracts: Effect of Solvent Polarity. International Journal of Pharmaceutical Sciences and Research, 2(11): 2991-2995.

Kpadonou Kpoviessi, B.G., Ladekan, E.Y., Kpovessk, D.S., Gbaguidi, F., Yehouenou, B., Quetin-Leclercq, J., Figueredo, G., Moudachirou, M and Accrombessi, G.C. 2012. Chemical Variation of Essential Oil Constituents of Ocimum gratissimum L. from Benin, and Impact on Antimicrobial Properties and Toxicity Against Artemia salina Leach. Chemistry and Biodiversity, 9(1): 139-150.

Krishnaiah, D., Devi, T., Bono, A. and Sarbatly, R. 2009. Studies on Phytochemical Constituents of Six Malaysian Medicinal Plants, Journal of Medicinal Plants Research, 3(2): 067-072.

Madaki, F.M., Kabiru, A.Y., Bakare-Odunola, M.T., Mailafiya, S.C., Hamzah, R.U., and Edward, J. 2016. Phytochemical and Proximate Analyses of Methanol Leaf Extract of Neem (Azadirachta indica). European Journal of Medicinal Plants, 15(2): 1-6.

Mohammed, H.A. and Omer, A.F.A. 2015. Antibacterial Activity of Azadirachta indica  (Neem) Leaf Extract Against Bacterial Pathogens in Sudan. American Journal of Research Communication, 3(5): 246-251.

Nahak, G. and Sahu, R.K. 2010. In vitro Antioxidative Acitivity of Azadirachta indica  and Melia azedarach Leaves by DPPH Scavenging Assay. Nature and Science, 8(4): 22-28.

Navabi, S.M., Ebrahimzadeh, M.A., Nabavi, S.F., Hamidinia, A. and Bekhradnia, A.R. 2008. Determination of Antioxidant Activity, Phenol and Flavonoids Content of parrotia persica Mey. Pharmacologyonline, 2: 560-567.

Ndukwe, O.K., Awomukwu, D. and Ukpabi, C.F. 2013. Comparative Evaluation of Phytochemical and Mineral Constituents of the Leaves of Some Medicinal Plants in Abia State Nigeria. International Journal of Academic Research in Progressive Education and Development, 2(3): 244-252.

Ogbonna, O.A., Ogbonna, P.C. and Dike, M.C. 2016. Phytochemical Screening, Quantitative Estimates of Bioactive Compounds in Spondias mombin and Azadirachta indica . Research Journal of Chemical Sciences, 6(1): 38-40.

Ogunjinmi, O.E., Olateru, C.T. and Ogunjinmi, S.O. 2014. Effect of Collection Time on Phytochemical Screening and Antibacterial Activities of Chromolaena odorata Leaf Extract. Science and Engineering Perspectives, 9: 32-36.

Ogunleye, R.F. 2010. Toxicity Bioassays of Four Different Botanicals Against the House Hold Pest: Periplaneta Americana (Linnaeus). Journal of Science and Technology, 30(2): 29-35.

Okoh, S.O., Oyewole, A.O., Ishola, R.O., Odusote, A.D., Okoh, O.O. and Igwe, C.C. and Elemo, G.N. 2015. Antioxidant and Free Radical Scavenging Capacity of Crude and Refined Oil Extracted From Azadirachta indica  A. Juss. International Journal of Biology, 7(2): 78-85.

Okwu, D.E. 2004. Phytochemicals and Vitamin Content of Indigenous Species of South Eastern. Nigeria Journal of Sustainable Agriculture and Environment, 6(1): 30-37.

Onwuka, G.I. 2005. Food Analysis and Instrumentation: Theory and Practice. Naphathali Prints. pp 140-146.

Pandey, G., Verma, K.K. and Singh, M. 2014. Evaluation of Phytochemical , Antibacterial and Free Radical Scavenging Properties of Azadirachta indica  (Neem) Leaves. International Journal of Pharmacy and Pharmaceutical Sciences, 6(2): 10-13.

Pinchuk, I., Shoval, H., Dotan, Y. and Lichtenberg, D. 2012. Evaluation of Antioxidants: Scope, Limitations and Relevance of Assays. Chemistry and Physics of Lipids, 165(1): 638-647.

Silva, E.A.J., Paula da Silva, V., Alves, C.C.F., Alves, J.M., Souchie, E.L. and Barbosa, L.C. 2016. Harvest Time on the Content and Chemical Composition of Essential Oil from Leaves of Guava. Ciencia Rural Santa Maria Online, 7-12.

Soni, U., Brar, S. and Gauttam, V.K. 2015. Effect of Seasonal Variation on Secondary Metabolites of Medicinal Plants. International Journal of Pharmaceutical Science Research, 6(9): 3654-62.

Sotannde, O.A., Yager, G.O., Zira, B.D. and Usman, A. 2011. Termicidal Effect of Neem Extracts on the Wood of Khaya senegalensis, Research Journal of Forestry,5(3): 128-138. DOI: 10.3923/rjf.2011.128.138

Sousa, E.O., Colares, A.V., Rodrigues, F.F.G., Campos, A.R., Lima, S.G. and Costa, J.G.M 2010. Effect of Collection Time on Essential Oil Composition of Lantana camara Linn (Verbenaceae) Growing in Brazil Northeastern Rec. Nat. Prod. 4(1): 31-37.

Trease, G. E. and Evans, W. C. 1989. Pharmacognosy. 13ed. BacilliereTinall Ltd, London, 5-9.

Vinoth, B., Manivasagaperumal, R. and Rajaravindran, M. 2012. Phytochemical Analysis and Antibacterial Activity of SA Juss. International Journal of Research in Plant Science, 2(3): 50-55.