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Toxicity of Biosal essential oil against Callosobruchus analis by three methods Direct Application method (DAM), Glass film method (GFM) and Filter paper impregnation method (FIM) after 24 hours of treatment. The LC50 value calculated as 1.338504 µl/cm2 by DAM, 12.174 µl/cm2 by GFM and 6.945603 µl/cm2 by FIM. HPLC analysis showed the activity of Biosal against C.analis by (DAM) represented by peak area in µm. The seed extract Azadirachtin (standard) shown by one peak 102387, peak (2) and (3) were absent and in Biosal sprayable (sample) shown by two peaks 111990, 2679, peak (3) was absent, while by (GFM) Biosal sprayable (standard) shown by four peaks showed as 6743, 62559, 6765, 1842 µm and in Biosal sprayable (sample) shown by four peaks which was 10818, 37728, 2954, 1277 µm, similarly by (FIM) Azadirachtin (standard) shown one peak 1339, peak 61568 and (3) was absent and Biosal sprayable (sample) shown as 7527, 30040, 1013 µm.