Rapid and sensitive analysis of azadirachtin and related triterpenoids from Neem (Azadirachta indica) by high-performance liquid chromatography-atmospheric pressure chemical ionization mass spectrometry

Based on reversed-phase high-performance liquid chromatography (RP-HPLC) an d atmospheric pressure chemical ionization (APCI) mass spectrometry, a HPLC -MS method was developed to permit the rapid qualitative and quantitative a nalysis of azadirachtin and related tetranortriterpenoids from seeds and ti ssue cultures of Neem (Azadirachta indica). APCI+ standard scanning mass sp ectra of the major Neem triterpenoids were recorded and utilized to select suitable ions for selected ion monitoring (SIM). Transitions for selective reaction monitoring (SRM) were based on MS-MS experiments. Using SIM, major Neem triterpenoids were detected in callus culture material and seed kerne ls of A. indica. The Limit of detection for azadirachtin in extract samples (similar to 1 ng ml(-1) or 10 pg in SIM mode) was determined to be (with r espect to injected absolute amounts) approximately 1000-times lower than va lues quoted in the literature for existing HPLC methods (similar to 200 ng ml(-1) or 10 ng). In addition to high sensitivity, the HPLC-MS method is ab le to tolerate minimal sample preparation and purification, dramatically re ducing total analysis time. (C) 2000 Elsevier Science B.V. All rights reser ved.