Determination of dihydroergosine in sorghum ergot using an immunoassay.
Dihydroergosine (DHES) is the principal toxic alkaloid produced by sorghum ergot (Claviceps africana). It has recently been shown that DHES levels as low as 1 mg/kg in animal feed can cause significant production losses. Quantitative immunoassays for detecting the related rye ergot alkaloid, ergotamine, are described in the literature, but those assays are relatively insensitive for DHES. This paper describes competitive enzyme-linked immunosorbent assays (ELISA) for measuring the DHES concentration in grains and mixed animal feed. The assays were developed using a DHES specific mouse monoclonal antibody and rabbit polyclonal antibodies raised against DHES conjugated to bovine serum albumin. Recoveries of between 77 and 103% were obtained from spiked grain using a simple, one step extraction with 70% methanol. Both the monoclonal and the polyclonal assays are capable of detecting DHES concentrations above 0.01 mg/kg, but quantification is most reliable at concentrations of 0.1 mg/kg or higher.
Molloy
John B
JB
Agency for Food and Fiber Sciences, Department of Primary Industries, Queensland, Animal Research Institute, LMB No 4 Moorooka, Qld 4105, Australia. John.Molloy@dpi.qld.gov.au
Moore
Chris J
CJ
Bruyeres
Anthea G
AG
Murray
Sally-Ann
SA
Blaney
Barry J
BJ
D000821:Animal Feed / Q000032:analysis; D002851:Chromatography, High Pressure Liquid; D002967:Claviceps / Q000378:metabolism; D002523:Edible Grain / Q000737:chemistry / Q000382:microbiology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D004879:Ergotamines / Q000032:analysis*; D005506:Food Contamination; D012680:Sensitivity and Specificity
D004879:Ergotamines; C476487:dihydroergosine