Simultaneous determination of five azadirachtins in the seed and leaf extracts of Azadirachta indica by automated online solid-phase extraction coupled with LC–Q-TOF–MS

Abstract

Neem (Azadirachta indica) extract is well-known as a natural pesticide for the control of agricultural pests. Azadirachtin A and its structural analogues are considered as active compounds. However, the amounts of azadirachtins varies in neem extracts, providing a variety of insecticidal activities. In this study, a novel method of automated online solid-phase extraction coupled with liquid chromatography/quadrupole-time-of-flight mass spectrometry (SPE-LC–Q-TOF–MS) was developed and validated for simultaneous quantification of five azadirachtins (azadirachtins A, B, D, H and I) in seed and leaf extracts of A. indica. Different experimental parameters (such as SPE cartridge, injection volume and washing step) were optimized. The optimized SPE-LC–Q-TOF–MS method showed good recovery (82.0–102.8%), linearity (r2 ≥ 0.9991) and precision (0.83–4.83%). The limit of detections (LODs) for the five analytes ranged from 0.34 to 0.76 ng mL−1. The validated method was successfully applied for determination of the analytes in the neem leaves and seeds from different locations and a neem formulation. The online SPE-LC–Q-TOF–MS method was found to be a simple, precise and accurate and can be used as a powerful tool for quality control of neem extracts or its formulations.Electronic supplementary materialThe online version of this article (10.1186/s13065-018-0453-y) contains supplementary material, which is available to authorized users.

Chemical and Antifungal Variability of Several Accessions of Azadirachta indica A. Juss. from Six Locations Across the Colombian Caribbean Coast: Identification of Antifungal Azadirone Limonoids

“…Electrospray ionization interface (ESI) was operated in a positive ion mode (scan 100-2000 m/z), desolvation line temperature at 250 • C, nitrogen as nebulizer gas at 1.5 L/min, drying gas at 8 L/min, quadrupole energy at 7.0 eV, and collision energy 14 eV. Peak annotations were performed after detailed scrutiny of the ultraviolet-visible (UV-Vis) and high-resolution mass spectral (exact mass of quasi-molecular and fragments ions) data of each signal in comparison to the information previously reported in literature to Azadirachta species [31,44,45].…”

Multi-tissue transcriptome analysis using hybrid-seq revealed potential genes and biological pathways associated with azadirachtin A biosynthesis in neem (Azadirachtin indica)

“…Among five tissues, fruit with the green hard seed was reported to contain the highest azadirachtin A in all the developing stage of fruit [14,19]. Leaf from neem tree in China contained azadirachtin A at the value of 969.9 μg g −1 [20]. The rank of azadirachtin content in different tissues was consistent with that in previous report (seed kernels, 0.03%; leaves, 0.9x10 -3 %; bark, 0.5x10 -3 %; root, 0.3x10 -3 %; stem 0.2 x10 -3 %) [21].…”

Multi-tissue transcriptome analysis using hybrid-sequencing reveals potential genes and biological pathways associated with azadirachtin A biosynthesis in neem (azadirachta indica)

“…Hence, five neem tissues (fruit, leaf, stem, flower, and root) were sampled for transcriptome sequencing. Among the five tissues, fruit with the green hard seed has been reported to contain the highest amount of azadirachtin A throughout fruit development [ 14 , 19 ] Leaf sample from a neem tree in China contained azadirachtin A at a concentration of 969.9 μg/g [ 20 ]. The percentage azadirachtin content in different tissues was consistent with that in a previous report (seed kernels, 0.03%; leaves, 0.9 × 10 − 3 %; bark, 0.5 × 10 − 3 %; root, 0.3 × 10 − 3 %; stem 0.2 × 10 − 3 %) [ 21 ].…”