Normal phase liquid chromatographic analysis of toxic norditerpenoid alkaloids

Abstract

AbstractA normal phase, isocratic, analytical high performance liquid chromatographic (HPLC) method has been developed for the analysis of toxic methylsuccinimidoanthranoyllycoctonine (MSAL) norditerpenoid alkaloids occurring in poisonous larkspurs (Delphinium sp.). The method has good sensitivity, specificity for MSAL norditerpenoid alkaloid and rapid analysis times. The technique employs 5 μm neutral alumina as the stationary phase and utilizes a mixture of hexane: 95% isopropyl alcohol as the mobile phase. Accurate and reproducible analyses can be performed on 1 g samples of plant material. The method has been applied to the analysis of the chemical phenology of the MSAL norditerpenoid alkaloids methyllycaconitine and 14‐deacetylnudicauline in three species of tall larkspurs (Delphinium barbeyi, D. occidentale, D. glaucescens). Analytical results confirm the primary occurrence of these toxic alkaloids in early vegetative tissues of these plants, their concentration in reproductive plant tissues and their reduction in level with plant maturation. The chemical phenology results further confirm the observed difference in the toxicity of the three tall larkspurs (D. barbeyi > D. glaucescens > D. occidentale) to livestock. Intraspecies differences of toxic alkaloid occurrence associated with site were also demonstrated.