BS EN 17683:2023

Animal feeding stuffs. Methods of sampling and analysis. Determination of pyrrolizidine alkaloids in animal feeding stuff by LC-MS/MS. | Status: Current

1 Scope This document specifies a method for the quantitative determination of pyrrolizidine alkaloids (PA) in the concentration ranges shown in Table 1 in complete and supplementary feed and in forages by liquid chromatography tandem mass spectrometry (LC-MS/MS) after solid phase extraction (SPE) clean-up. Table 1 — Summary of concentration ranges per PA tested in the collaborative trial Tested pyrrolizidine alkaloid (PA) Abbreviation Tested concentration range a (µg/kg) From To Echimidine Em 20 435 Echimidine-N-oxide EmN 5 30 Erucifoline Er 20 245 Erucifoline-N-oxide ErN 20 370 Europine Eu 15 330 Europine-N-oxide EuN 25 285 Heliotrine Hn 25 280 Heliotrine-N-oxide HnN 25 245 Jacobine Jb 20 230 Jacobine-N-oxide JbN 20 215 Lasiocarpine Lc 20 350 Lasiocarpine-N-oxide LcN 5 250 Intermedine Im 25 560 Intermedine-N-oxide ImN 5 395 Lycopsamine La 25 500 Lycopsamine-N-oxide LaN 20 280 Monocrotaline Mc 20 360 Monocrotaline-N-oxide McN 20 365 Retrorsine Re 250 375 Retrorsine-N-oxide ReN 5 285 Senecionineb Sc 25 205 Senecionine-N-oxide b ScN 5 300 Senecivernine b Sv 20 205 Senecivernine-N-oxide b SvN 5 165 Senkirkine Sk 20 275 Seneciphylline Sp 25 225 Seneciphylline-N-oxide SpN 5 225 Trichodesmine Td 5 250 Intermedine + Lycopsamine Im+La 50 890 Intermedine-N-oxide + Lycopsamine-N-oxide ImN+LaN 5 645 Senecivernine + Senecionine Sv+Sc 30 280 Senecivernine-N-oxide + Senecionine-N-ide SvN+ScN 10 380 a Rounded figures b Individual PA of the isomeric pairs Sv+Sc and SvN+ScN were not evaluated statistically due to insufficient chromatographic separation.

NOTE 1 A second method was part of the method validation collaborative main trial. For this method PA-N-Oxides are reduced by adding zinc powder to the extract of the feed material. The following steps correspond to the first and main method. Quantitative results for each PA except the otonecine type PA senkirkine represent the sum of the free PA base and its corresponding N-oxide.

NOTE 2 Due to insufficient numbers of data for some analyte-matrix combinations statistical evaluation was not valid for standardization. Received data indicated the methods applicability in experienced laboratories with appropriate quality assurance measures. Therefore, the method description is included as an informative annex (Annex D).