Advertisement

Method to Quantify Plant Secondary Metabolites: Quantification of Neem Metabolites from Leaf, Bark, and Seed Extracts as an Example

  • Kannan RangiahEmail author
  • Malali Gowda
Chapter
Part of the Compendium of Plant Genomes book series (CPG)

Abstract

Among the medicinal plants, neem has its own value in terms of treating many known and unknown diseases. Neem plant is known to contain several thousands of secondary metabolites, which are crucial for multifunctional properties like anti-oxidation, anti-inflammation, antimalarial, and anticarcinogenic activities. Now it is important to understand the molecular details like exact quantity of the neem metabolites in different parts of the plants. Here we have showed a UHPLC-MS/SRM method to quantify five neem metabolites (Azadirachtin, Nimbin, Salanin, Azadiradione, Epoxy/Hydroxy-azadiradione) from different parts of neem plants (leaf, bark, and seed). Among the five metabolites analyzed, E/H-Azadi is present in very high concentration in neem plant (leaf: 124,239 pg/µg, bark: 906.97 pg/µg, seed: 7309.48 pg/µg) as compared to other metabolites. Interestingly, E/H-Azadi seems to be the most abundant metabolite in the neem leaf and bark extracts and azadi is the highest in the seed extract. In the leaf extract, E/H-Azadi is ~136 fold higher compared to bark and ~17 fold higher compared to seed extract. The amount of E/H-Azadi in leaf is 124,239 pg/µg of leaf extract, which constitutes ~10% in the leaf extract. The excess amount of E/H-Azadi in the neem leaf might be one of the reasons for its multifunctional properties in nature.

Keywords

Neem Neem metabolites Quantification Mass spectrometry UHPLC-MS/SRM method 

Abbreviations

Aza

Azadirachitin

Azadi

Azadiradione

CID

Collision-induced dissociation

E/H-Azadi

Epoxy or hydroxyazadiradione

SRM

Selected reaction monitoring

UHPLC-MS/SRM

Ultrahigh performance liquid chromatography/mass spectrometry/selected reaction monitoring

Notes

Acknowledgements

We would like to thank Department of Biotechnology, India for the financial assistance to setup the Metabolomics Facility at C-CAMP, NCBS Campus.

References

  1. Alexandr YY, Boris VN, Emilie C, Yakov IY (2015) Determination of the chemical composition of tea by chromatographic methods: a review. J Food Res 4(3):56–88CrossRefGoogle Scholar
  2. Ali A (1993) Textbook of pharmacognosy. Publication and Information Directorate, New Delhi, India, pp 381–384Google Scholar
  3. Athar A, Saikat H, Hirekodathakallu VT, Rahul K, Manish G, Mohd SI, Chinmay P, Sumanta D, Samik B, Souvik S, Uttam P, Nakul CM, Uday B (2012) Novel anti-inflammatory activity of epoxyazadiradione against macrophage migration inhibitory factor. J Biol Chem 287(29):24844–24861CrossRefGoogle Scholar
  4. Avinash P, Devdutta SD, Saikat H, Vairagkar U, Shinde GV, Fayaj AM, Thiagarayaselvam A, Hirekodathakallu VT (2015) Triterpenoid profiling and functional characterization of the initial genes involved in isoprenoid biosynthesis in neem (Azadirachta indica). BMC Plant Biol 15(214):1–14Google Scholar
  5. Champagne DE, Koul O, Isman MB, Scudder GGE, Towers GHN (1992) Biological activity of limonoids from the rutales. Phytochemistry 31(2):377–394.CrossRefGoogle Scholar
  6. Debas HT, Laxminarayan R, Straus SE (2006) Complementary and alternative medicine, chap 69. In: Jamison DT, Breman JG, Measham AR et al (eds) Disease control priorities in developing countries, 2nd edn. The International Bank for Reconstruction and Development, The World Bank, Washington DC. Available from https://www.ncbi.nlm.nih.gov/books/NBK11796/. Co-published by Oxford University Press, New York
  7. Estefânia VRC, Jhones LDO, Mônica P, Renata DL, Leonardo FF (2016) Neem oil and crop protection: from now to the future. Front Plant Sci 7(1494):1–8Google Scholar
  8. FAO (1997) Medicinal plants for forest conservation and health care. In: Non-wood forest products 11. FAO—Food and Agriculture Organization of the United Nations, Rome, ItalyGoogle Scholar
  9. Fauziah O, Gholamreza M, Sally LTP, Asmah R, Rusliza B, Chong PP (2012) Effect of neem leaf extract (Azadirachta indica) on c-Myc oncogene expression in 4T1 breast cancer cells of BALB/c mice. Cell J (Yakhteh) 14(1):53–60Google Scholar
  10. Gordon MC, David JN (2013) Natural products: a continuing source of novel drug leads. Biochim Biophys Acta 1830(6):3670–3695CrossRefGoogle Scholar
  11. Hossain MA, Shah MD, Sakari M (2011) Gas chromatography–mass spectrometry analysis of various organic extracts of Merremia borneensis from Sabah. Asian Pac J Trop Med 4(8):637–641CrossRefGoogle Scholar
  12. Isman MB (2006) Botanical insecticides, deterrents, and repellents in modern agriculture and an increasingly regulated world. Annu Rev Entomol 51:45–66CrossRefGoogle Scholar
  13. Kannan R (2014) Sensitive UHPLC-MS/SRM method for quantifying olanzapine metabolites and degradation products from sera. Anal Methods 6:5250–5257CrossRefGoogle Scholar
  14. Kannan R, Varalaxmi BA, Malali G (2016) UHPLC-MS/SRM method for quantification of neem metabolites from leaf extracts of Meliaceae family plants. Anal Methods 8:2020–2031CrossRefGoogle Scholar
  15. Kaushik SH, Lakshmi M, Rajendra H, Anil K (2014) NeeMDB: convenient database for neem secondary metabolites. Bioinformation 10(5):314–315CrossRefGoogle Scholar
  16. Kokate C, Purohit AP, Gokhale SB (2010) Pharmacognosy. Nirali Prakashan, Maharashtra, IndiaGoogle Scholar
  17. Mamta S, Jyoti S, Rajeev N, Dharmendra S, Abhishek G (2013) Phytochemistry of medicinal plants. J Pharmacogn Phytochem 1(6):168–182Google Scholar
  18. Mohammad AA (2016) Therapeutics role of Azadirachta indica (Neem) and their active constituents in diseases prevention and treatment. Evid Based Complement Altern Med 2016(Article ID 7382506):11. http://dx.doi.org/10.1155/2016/7382506
  19. Nagesh AK, Vijay Y, Kannan R, Mahesh HB, Anantharamanan R, Meghana S, Heikham R, Ramya ML, Chandana SL, Shilpa S, Aishwarya R, Sathyanarayana BN, Malali G (2015) Comprehensive analyses of genomes, transcriptomes and metabolites of neem tree. Peer J 3:e1066CrossRefGoogle Scholar
  20. National Research Council (1992) Neem: a tree for solving global problems. The National Academies Press, Washington DC.  https://doi.org/10.17226/1924CrossRefGoogle Scholar
  21. National Research Council (US) Panel on Neem (1992) Effects on insects, chap 5. In: Neem: a tree for solving global problems. National Academies Press (US), Washington DC. Available from https://www.ncbi.nlm.nih.gov/books/NBK234642/
  22. Nivedita N, Padma R, Vairavan L, Dasaradhi P, Kannan R (2015) A quantitative metabolomics peek into planarian regeneration. Analyst 140:3445–3464CrossRefGoogle Scholar
  23. Priyadarshini K, Keerthi AU (2012) Paclitaxel against cancer: a short review. Med Chem 2(7):139–141Google Scholar
  24. Ruchi T, Amit KV, Sandip C, Kuldeep D, Shoor VS (2013) Neem (Azadirachta indica) and its potential for safeguarding health of animals and humans: a review. J Biol Sci.  https://doi.org/10.3923/jbs.2013
  25. Saikat H, Fayaj AM, Thiagarayaselvam A, Devdutta SD, Hirekodathakallu VT (2014) Expedient preparative isolation and tandem mass spectrometric characterization of C-seco triterpenoids from neem oil. J Chromatogr A 1366:1–14CrossRefGoogle Scholar
  26. Schmutterer H (1990) Properties and potential of natural pesticides from the neem tree, Azadirachta indica. Annu Rev Entomol 35:271–297CrossRefGoogle Scholar
  27. Selma B, Eltahir AK, Sami AK, Mohammad AA, Abdlmarouf M, Yousef HA, Paul FSE, Mohammed F (2013) Azadirachta indica ethanolic extract protects neurons from apoptosis and mitigates brain swelling in experimental cerebral malaria. Malaria J 12(298):1–9Google Scholar
  28. Simon JH, Benjamin PYL, Rainer S, Rudolf K (2014) Liquid chromatography-mass spectrometry for the determination of chemical contaminants in food. Trends Anal Chem 59:59–72CrossRefGoogle Scholar
  29. Tan QG, Luo XD (2011) Meliaceous limonoids: chemistry and biological activitiesCrossRefGoogle Scholar
  30. Vincent PKT, Denis Z, Moses NN (2008) The antimalarial potential of medicinal plants used for the treatment of malaria in cameroonian folk medicine. Afr J Tradit CAM 5(3):302–321Google Scholar
  31. Wungsem R, Sreya D, Debajyoti D, Jayram H (2013) A brief review on the botanical aspects and therapeutic potentials of important Indian medicinal plants. Int J Herb Med 1(3):38–45Google Scholar

Copyright information

© Springer Nature Switzerland AG 2019

Authors and Affiliations

  1. 1.Food Safety & Analytical Quality Control LaboratoryCSIR-Central Food Technological Research Institute (CFTRI)MysoreIndia
  2. 2.Centre for Functional Genomics and Bioinformatics, The University of TransDisciplinary Health Sciences and Technology, Foundation for Revitalization of Local Health TraditionsBengaluruIndia

Personalised recommendations