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)
Tested pyrrolizidine alkaloid (PA)	Abbreviation	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
Senecionine^b	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).

BS EN 17683:2023

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

Current

Published:

31 Mar 2023