A new, rapid and sensitive multiresidue method is reported for the simultaneous determination of tropane alkaloids (tropine, atropine, scopolamine, homatropine, anisodamine) and glycoalkaloids (α-solanine, α-chaconine) in grains and seeds (wheat, rye, maize, soybean, linseed). Dispersive solid phase extraction (DSPE) was performed with 0.5% formic acid in acetonitrile/water and a mixture of magnesium sulphate, sodium chloride and sodium citrate. For a fast and effective clean-up procedure for oily matrices such as soybean and linseed, matrix solid phase dispersion (MSPD) C(18) material was used to remove co-extracted non-polar components. No clean-up was necessary for less oily matrices following extraction. The analytes were separated by isocratic HPLC on a Chirobiotic V column and detected using a triple quadrupole mass spectrometer with electrospray ionization (ESI). All analytes were monitored in the positive ion mode. The method performance is presented in terms of linearity in the range 5-80 ng/g (r(2)=0.998), specifity, selectivity, accuracy (recoveries from 61-111%), precision (CV<5%) and ruggedness. The limits of quantitation (LOQ) were in the range 2.2-4.9 ng/g.