Determination of ergot alkaloids from grains with UPLC‐MS/MS

Abstract

A simple and rapid method for determining six ergot alkaloids and four of their respective epimers was developed for rye and wheat. The analytes were extracted from the sample matrix with ACN/ammonium carbonate solution. The extract was purified with a commercial push-through SPE column (Mycosep 150 Ergot). After concentration and filtration steps, the final separation of the analytes was achieved with ultra-performance LC-MS/MS. The chromatographic separation of the ergot alkaloids was achieved in 4.5 min. The method performance proved satisfactory in the preliminary validation. The calculated LOQs were low ranging from 0.01 to 1.0 microg/kg for wheat and from 0.01 to 10.0 microg/kg for rye. At the concentration levels of 10, 50 and 200 microg/kg, the recoveries were between 80 and 120% in most cases and the within-day repeatability (expressed as RSD) ranged between 1.3 and 13.9%. Despite the cleanup of the samples, some matrix effect was observed in the MS, highlighting the necessity of using matrix-assisted standards. This is the first article to describe the application of the push-through columns and ultra-performance LC in the analysis of ergot alkaloids.

Analysis of Ergot Alkaloids

“…EA can be extracted from dry samples (including sclerotia, cereals or grain-based foods) using relatively low polarity solvent mixtures with the addition of ammonium hydroxide to give an alkaline pH [ 18 , 19 , 20 , 21 ]. The solvents used typically contain ethyl acetate or dichloromethane (although the latter is out of favour on account of its toxicity and damaging environmental effects).…”

Development and validation of a new LC–MS/MS method for the simultaneous determination of six major ergot alkaloids and their corresponding epimers. Application to some food and feed commodities

“…A number of the methods mentioned above have been used for biomedical application (Friedrich et al, 2004) and /or for identification purposes (Lehner et al, 2005). Some include no or only few -inine isomers (Burk et al, 2006;Kokkonen & Jestoi, 2010;Mohamed et al, 2006), possibly because of the lack of available standards. reported an LC-MS/MS multi-analyte method including both the -ine as well as the -inine isomers of the six most common ergot alkaloids, which was validated for ten different cereal and food samples.…”

“…It is therefore important to consider both epimers when the ergot alkaloid contamination level of a cereal or cereal product has to be determined. Different methods have been reported for the analysis of ergot alkaloids, including thin layer chromatography (TLC) (Salvat & Godoy, 2001), capillary electrophoresis (CE) (Franch & Blaschke, 1998), enzyme linked immunosorbent assay (ELISA) (Hill et al, 2001), gas chromatography (GC) with electron capture detection (ECD) (Barrow & Quigley, 1975), liquid chromatography (LC) with ultraviolet (Veress, 1993), fluorescence (Komarova & Tolkachev, 2001;Storm et al, 2008) or mass spectrometric (MS) (Burk et al, 2006;Kokkonen & Jestoi, 2010;Krska, Stubbings, Macarthur, & Crews, 2008;Mohamed, Gremaud, Rychoz-Payot, Tabet, & Guy, 2006) detection. In recent years, LC-MS has become the method of choice for mycotoxin determination (Shephard et al, 2011;Spanjer, 2010), and has provided an unequivocal identification of ergot alkaloids in various matrices (Friedrich et al, 2004;Lehner, Craig, Fannin, Bush, & Tobin, 2005;Mohamed et al, 2006).…”

Rhodococcus erythropolis MTHt3 biotransforms ergopeptines to lysergic acid

“…For chanoclavine, in vitro studies suggest a weak affinity for D-2 receptors [ 55 ], whereas in vivo chanoclavine has no effect [ 56 ]. Ergometrine is one of the main ergot alkaloids produced by C. purpurea [ 57 ]. In drunken horse grass ( Achnatherum inebrians ) infected by the endophytic fungus N. gansuense , high levels of ergometrine were also detected [ 58 ], but in endophyte-infected tall fescue, only minor concentrations of ergometrine were found [ 59 ].…”