Azadirachtins are natural triterpenoid compounds derived from Neem tree extracts with potential for use as systemic insecticides against invasive wood-boring insect pests. In this work, a sensitive and selective analytical method has been developed for the simultaneous determination of azadirachtin A and azadirachtin B (3-tigloylazadirachtol) in foliage and twigs of various tree species. Samples were mixed with C18 and primary-secondary amine (PSA), and extracted with acetonitrile. Then, an aliquot of the raw extract was 10-fold diluted with water and directly analyzed by liquid chromatography tandem mass spectrometry (LC-MS/MS). The method was validated in foliage and twig matrices of four different tree species (London Plane Tree, Red/Freemani Maple, Norway Maple and Sugar Maple) that are known hosts of the exotic invasive insect pest – Asian Longhorn Beetle (ALB). Analytical results for replicate (N = 5) samples, fortified at 0.01, 0.1 and 1 mg kg−1, showed good recoveries (86–119%) and precision (<20% RSD). The methodology was successfully applied to the analysis of 200 samples taken from a field experiment designed to investigate uptake, translocation and expression of azadirachtins in representative high-value urban trees followed by stem injection with TreeAzin™.
