A basic tool for risk assessment: A new method for the analysis of ergot alkaloids in rye and selected rye products

Abstract

As a basis for the collection of occurrence and exposure data of ergot alkaloids in food, an HPLC method coupled with fluorimetric detection (HPLC-FLD) for the determination of 12 pharmacologically active ergot alkaloids in rye and rye products was developed. Samples were extracted with a mixture of ethyl acetate, methanol, and aqueous ammonia, followed by centrifugation and purification by solid phase filtration (SPF) with basic alumina. After solvent adjustment, the samples were analyzed by HPLC-FLD using a phenyl-hexyl-column. Recoveries for five major alkaloids were between 89.3% (ergotamine) and 99.8% (alpha-ergokryptine) with a maximum LOQ of 3.3 microg/kg (ergometrine). Precision expressed as RSD ranged from 2.8% (ergocristine) to 12.4% (alpha-ergokryptine) for repeatability, and from 6.5% (ergocornine) to 14.9% (ergotamine) for within-laboratory reproducibility, respectively. In a survey of 39 rye product samples, ergocristine and ergotamine were found to be the major alkaloids in commercially available rye products with contents of 127 microg/kg (ergocristine), and 134 microg/kg (ergotamine) in rye flour, and 152.5 and 117.8 microg/kg in coarse meal, respectively.

Degradation and epimerization of ergot alkaloids after baking and in vitro digestion

“…This illustrates the broad variability among the epimeric ratios for the different EAs and confirms results reported previously [17,28]. After baking, the content of EAs decreased in all samples.…”

“…The chromatographic conditions were adapted from the literature [28]. An Agilent 1200 series HPLC equipped with a degasser, binary pump, column oven (thermostatic at 30°C), DAD, and fluorescence detector (FLD) set at 330 nm (excitation) and 415 nm (emission) was used for analyses.…”

Development and Validation of an Analytical Method for Determination of Ergot Alkaloids in Animal Feedingstuffs with High Performance Liquid Chromatography-fluorescence Detection

“…For the separation of ergot alkaloids most authors used reverse phase C18 columns (Mohamed et al 2006, Storm et al 2008, Di Mavungu et al 2012 though, the separation was also performed on a C6 phenyl column (Muller et al 2009). As far as the mobile phase is concerned, the most popular composition was ammonium carbonate at different concentrations and acetonitrile (Storm et al 2008, Muller et al 2009, Bryła et al 2015 used in an isocratic and gradient mode as well.…”

“…As far as the mobile phase is concerned, the most popular composition was ammonium carbonate at different concentrations and acetonitrile (Storm et al 2008, Muller et al 2009, Bryła et al 2015 used in an isocratic and gradient mode as well. In the presented method the separation was also performed on a C18 column with the mobile phase consisted of ammonium carbonate and acetonitrile; however, a gradient mode was used which provided good separation of ergot alkaloids, and method selectivity.…”

Investigations into the occurrence of alkaloids in ergot and single sclerotia from the 2007 and 2008 harvests

“…Based on the method of Müller et al (2006Müller et al ( , 2009, and additional private communication) for rye flour and products thereof, we made slight alterations for the analysis of pure ergot sclerotia. About 50 mg of a ground ergot sample (∼10 g, ≤0.5 mm, Ika MF10 mill; Staufen, ; and manually by a nutmeg grater for single sclerotia, respectively) were suspended in 50 ml of an extraction mixture consisting of ethyl acetate, methanol and aqueous ammonium hydroxide solution (25%) (75:5:4 v/v/v) (Müller et al 2009).…”