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

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


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.


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







Collision-induced dissociation


Epoxy or hydroxyazadiradione


Selected reaction monitoring


Ultrahigh performance liquid chromatography/mass spectrometry/selected reaction monitoring



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


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© 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

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