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United States Department of Agriculture

Agricultural Research Service

Title: Development of Enzyme-Linked Immunosorbent Assays for Toxic Larkspur (Delphinium SPP.)ALKALOIDS

Authors
item Lee, Stephen
item Schoch, Thomas
item Gardner, Dale
item Stegelmeier, Bryan

Submitted to: Journal of Agricultural and Food Chemistry
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: August 4, 2000
Publication Date: September 15, 2000
Citation: LEE, S.T., SCHOCH, T.K., GARDNER, D.R., STEGELMEIER, B.L. DEVELOPMENT OF ENZYME-LINKED IMMUNOSORBENT ASSAYS FOR TOXIC LARKSPUR ALKALOIDS. JOURNAL OF AGRICULTURAL FOOD & CHEMICAL. 2000.

Interpretive Summary: Larkspur (Delphinium spp.) poisons thousands of cattle on western rangelands each year. As poisoning does not cause specific lesions and poisoned animals are rarely found before they die, identifying poisoned animals is difficult. Additionally, toxin concentrations in larkspur plants vary with environment, plant, and location. Rapid, sensitive and specific diagnostic techniques are needed to identify poisoned animals and to determine when and what plants are likely to poison livestock. In this study, three competitive inhibition enzyme-linked immunosorbent assays (CI- ELISA) for toxic larkspur alkaloids were developed. One assay is class specific. The other two assays are specific for individual alkaloids. The assay with the lowest limit of detection had a limit of detection of 30.5 pg for toxin. Studies using these assays demonstrated the toxic larkspur alkaloids could be detected in bovine blood and brain tissue. These findings suggest that with additional development these techniques are likely to be excellent tools to diagnose poisoned animals and identify highly toxic plants.

Technical Abstract: Larkspur (Delphinium spp.) poisons thousands of cattle on western rangelands each year. As poisoning does not cause specific lesions and poisoned animals are rarely found before they die, definitively identifying poisoned animals is difficult. Additionally, toxin concentrations in larkspur plants vary with environment, plant, and location. Rapid, sensitive and specific diagnostic techniques are needed to identify poisoned animals and to determine when and what plants are likely to poison livestock. In this study, three competitive inhibition enzyme-linked immunosorbent assays (CI-ELISA) for toxic larkspur alkaloids were developed. One assay is class specific toward the N- (methylsuccinimido)anthranoyllycoctonine (MSAL) alkaloids, while two assays are specific for individual alkaloids. The assay with the lowest limit of detection had an I50 of 191 pg with a limit of detection of 30.5 pg for methyllycaconitine. Spike and recovery studies using bovine blood and brain tissue ranged from 52-89%. These findings suggest that with additional development these techniques are likely to be excellent tools to diagnose poisoned animals and identify highly toxic plants.

Last Modified: 12/27/2014
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