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Abstract
An improved high performance liquid Chromatographic method is reported to determine azadirachtin (AZ) content in neem formulations and neem oil. Aqueous methanol solutions of samples were extracted sequentially with hexane and dichloromethane. The dichloromethane layer, containing AZ, was taken to dryness and reconstituted in ethyl acetate. It was purified by Florisil® column cleanup, eluted with ethyl acetate and injected into the liquid Chromatograph. Separation was performed with a reversed‐phase Spherisorb C‐18 ODS 5 μm column with an acetonitrile/water gradient system. The eluent was monitored with UV detection at 210 nm. The method was found to be sufficiently rapid and reproducible for use in the analysis of AZ in neem formulations and neem oil. Limits of quantification and detection were respectively 6 μg/g and 3 μg/g.