Title: REPORT OF THE FOLLOW-UP COLLABORATIVE STUDY Determination of the sum of Fumonisin B1 and B2 in Compound Animal Feed and Maize by Immunoaffinity Column Clean-up and High Performance Liquid Chromatography with Fluorometric Detection
Authors: BREIDBACH AndreasBOUTEN KatrienKROEGER KatySTROKA Joerg
Publisher: OPOCE
Publication Year: 2009
JRC Publication N°: JRC52796
ISBN: 978-92-79-12849-3
ISSN: 1018-5593
Other Identifiers: EUR 23941 EN
OPOCE LA-NA-23941-EN-C
URI: http://publications.jrc.ec.europa.eu/repository/handle/JRC52796
DOI: 10.2787/12806
Type: EUR - Scientific and Technical Research Reports
Abstract: The accurate determination of mycotoxins in food and feed matrices for which EU legislative limits apply require robust and reliable analytical techniques. The robustness and reliability are best shown through validation by a collaborative study. Previous collaborative studies dealing with other mycotoxins have shown that it is possible to achieve performance characteristics which are fit-for-purpose provided suitable methodology is available. As with any interlaboratory comparison homogeneity between the test units is of utmost importance. Due to the complexity of food and feed matrices particular care has to be taken during test material preparation to achieve this. Methods for the determination of Fumonisin B1 (FB1) and Fumonisin B2 (FB2) have been subject to a collaborative study in the past and the methodology used involved immunoaffinity clean-up to purify the sample extracts. Detection was afforded by derivatisation of the Fumonisins to yield fluorescent derivatives before a chromatographic separation. The reagent used was o-phtaldialdehyde and mercaptoethanol. However, pre-column derivatisation does have disadvantages related to more demanding chromatography and the instability of the derivatives. Strict time control of all processes is required to obtain adequate repeatability which necessitates the use of programmable auto liquid samplers (ALS). This may be circumvented by using post column derivatisation instead. Here the native Fumonisins are separated and reagents are added constantly to the effluent of the chromatographic column. An additional pump, a mixing Tee, and additional tubing are needed for post column derivatisation replacing the need for a sophisticated ALS. During method development it could be shown that both methods can perform equally well with respect to the requirements by EU legislation for method performance and working range. A collaborative study to validate a method for the "Determination of Fumonisin B1 and B2 in Baby Food, Breakfast Cereals and Animal Feed by Immunoaffinity Column Clean-up with High Performance Liquid Chromatography and Fluorometric Detection" failed partially because of problems with the immunoaffinity columns (IAC) used for the study. After modifications to the method protocol regarding a check of proper performance of the IAC and the sample extract clean-up we describe below the results of a repeat of the study for compound animal feed and maize.
JRC Institute:Institute for Reference Materials and Measurements

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