Scribd
Upload
214 views4 pages

Sante Mycotoxins

The document provides guidance on the identification of mycotoxins in food and feed, emphasizing the importance of confirmatory analysis and method validation. It outlines specific requirements for chromatography, fluorescence detection, and mass spectrometric detection to ensure accurate identification of mycotoxins. The guidance is intended to supplement existing regulations and improve analytical methods within the EURL/NRL mycotoxin network.

Uploaded by

Borja Muñoz Solano
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PDF, TXT or read online on Scribd
214 views4 pages

Sante Mycotoxins

The document provides guidance on the identification of mycotoxins in food and feed, emphasizing the importance of confirmatory analysis and method validation. It outlines specific requirements for chromatography, fluorescence detection, and mass spectrometric detection to ensure accurate identification of mycotoxins. The guidance is intended to supplement existing regulations and improve analytical methods within the EURL/NRL mycotoxin network.

Uploaded by

Borja Muñoz Solano
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PDF, TXT or read online on Scribd
You are on page 1/ 4

SANTE/12089/2016

Guidance document on identification of


mycotoxins in food and feed

SANTE/12089 /2016

Implemented by 01/01/2017

1
Guidance document on identification of mycotoxins in food and feed

Preamble
Identification is an integral part of confirmatory analysis of mycotoxins in food and feed. This
document has been established by and has been discussed in the EURL/NRL mycotoxin network,
taking existing criteria from other domains and literature data into account [1-5]. It provides guidance
criteria for identification that should be taken into account during method validation and mycotoxin
analysis. This guidance supplements the "Specific requirements for confirmatory methods" from
Annex II of Commission Regulation (EC) No 401/2006 [6].

Identification requirements for mycotoxins

For identification of mycotoxins, chromatography combined with mass spectrometry is the method of
choice. Alternatively, liquid chromatography with fluorescence detection may be applied, but only
when an immunoaffinity-based cleanup specific for the targeted mycotoxin(s) has been employed
during sample preparation. The use of methods based on UV detection is discouraged, however
already established methods for which adequate selectivity has been demonstrated may continue to
be used, this typically applies to patulin and deoxynivalenol.

The criteria provided below are default guidance criteria that should be met in order to achieve proper
identification. During validation of the method, it should be verified that the criteria are met within the
concentration range of the method, using spiked samples or certified reference materials. This should
include the lowest level for which results will be reported, and the legislative maximum levels.
Furthermore, it should be verified that for blank samples no false positive identifications are obtained.

Requirements for chromatography


The minimum acceptable retention time for the analyte under examination should be twice the
retention time corresponding to the void volume of the column. The retention time of the analyte in the
sample extract should correspond to that of the average of the calibration standards measured in the
same sequence with a tolerance of ±0.2 min or ±50% of the peak width at half height (whichever is
larger), for both gas chromatography and liquid chromatography. With UHPLC and GC, deviations are
typically within ±0.1 min.
13
In case a C-isotopically labelled analogue of an analyte (internal standard) has been added to the
sample or extract, the retention time of the analyte should correspond to that of its labelled internal
standard with a tolerance of ±0.05 min.

Requirements for fluorescence detection


This applies to molecules that exhibit native fluorescence and to molecules that exhibit fluorescence
after either transformation or derivatisation. The selection of the excitation and emission wavelengths
in combination with the chromatographic conditions should be done in such a way as to minimise the
occurrence of interfering components in blank sample extracts. The nearest peak maximum in the
chromatogram should be separated from the designated analyte peak by at least one full peak width
at 10% of the maximum height of the analyte peak.

Requirements for mass spectrometric detection


Identification relies on proper selection of ions that are selective and specific for the analyte.
Molecular ions or (de)protonated molecules (or, if not available, adducts) should be included in the
measurement and identification procedure whenever possible. Low m/z fragments (<100) and
fragment ions arising from loss of water or common moieties are often less specific, and may
therefore be less useful for identification purposes.

Identification should be based on chromatographic peaks observed in the extracted ion


chromatograms of two or more (see Table 1) ions that are specific for the analyte. The peaks must
have a similar peak shape, overlap with each other, and the ion ratio (defined as the response of the

2
peak with the lower area divided by the response of the peak with the higher area) should be within
±30% (relative) to that obtained from the average of the calibration standards from the same
sequence. The peaks need to be within the linear range of the detector and have an S/N ratio of at
least 3. Where an extracted ion chromatogram shows evidence of significant interference, it must not
be relied upon for identification.

In addition to the degree of selectivity of the ions measured, different types and modes of mass
spectrometric detection provide different degrees of selectivity and specificity, which relates to the
confidence in identification. The requirements for identification are given in Table 1. They should be
regarded as guidance criteria, not as absolute criteria to prove presence or absence of an analyte.

Table 1. Identification requirements for different MS techniques1


Requirements for identification

MS detector / Typical systems minimum number


characteristics (examples) Acquisition of ions other

quadrupole,
Unit mass
full scan, limited m/z range, SIM 3 ions
resolution
ion trap, TOF

selected or multiple reaction


triple quadrupole, monitoring (SRM, MRM), mass
MS/MS ion trap, Q-trap, resolution for precursor-ion 2 product ions
Q-TOF, Q-Orbitrap isolation equal to or better than
unit mass resolution S/N ≥ 3d)

2 ionsa, b with

mass accuracy Analyte peaks in the


full scan, limited m/z range, SIM,
extracted ion
fragmentation with or without
≤ 5 ppm for m/z chromatograms must
precursor-ion selection, or
≥200 fully overlap.
combinations thereof
≤ 1 mDa for m/z Ion ratio within
High resolution
≤ 200
MS:
±30% (relative)
(Q-)TOF 2 ions:
Accurate
mass of average
(Q-)Orbitrap 1 molecular ion,
measurement (de)protonated of calibration
FT-ICR-MS
combined single stage MS and molecule or standards from same
adduct ion with sequence
sector MS
MS/MS with mass resolution for mass accuracy ≤ 5
precursor-ion isolation equal to ppm (or ≤ 1mDa
or better than unit mass for m/z ≤200)
resolution
plus

1 MS/MS product
ionc)

a) preferably including the molecular ion, (de)protonated molecule or adduct ion


b) including at least one fragment ion
c) no specific requirement for mass accuracy
d) in case noise is absent, a signal should be present in at least 5 subsequent scans

1 For definition of terms relating to mass spectrometry see Murray et al. (2013) Pure Appl. Chem., 85:1515–1609.

3
References

[1] Commission Decision 2002/657/EC implementing Council Directive 96/23/EC concerning the performance of
analytical methods and the interpretation of results, Off. J. Eur. Communities L221 (17.8.2002) 8–36.

[2] SANTE/2015/11945 Analytical quality control and method validation procedures for pesticide residue analysis
in food and feed.

[3] S.J. Lehotay, Y. Sapozhnikova, H.G.J. Mol, Current issues involving screening and identification of chemical
contaminants in foods by mass spectrometry, Trends in Analytical Chemistry 69 (2015) 62–75.

[4] H.G.J. Mol, P. Zomer, M. García López, R.J. Fussell, J. Scholten, A. de Kok, A. Wolheim, M. Anastassiades,
A. Lozano, A. Fernandez Alba. Identification in residue analysis based on liquid chromatography with tandem
mass spectrometry: Experimental evidence to update performance criteria, Analytica Chimica Acta 873 (2015) 1–
13.

[5] B.J.A. Berendsen,T. Meijer, R. Wegh, H.G.J. Mol, W.G. Smyth, S.A. Hewitt, L. van Ginkel, M.W.F. Nielen, A
critical assessment of the performance criteria in confirmatory analysis for veterinary drug residue analysis using
mass spectrometric detection in selected reaction monitoring mode, Drug Test. Analysis 8 (2016) 477–490.

[6] Commission Regulation (EC) No 401/2006 of 23 February 2006 laying down the methods of sampling and
analysis for the official control of the levels of mycotoxins in foodstuffs, OJ L 70, 9.3.2006, p. 12.

You might also like

pFad - Phonifier reborn

Pfad - The Proxy pFad of © 2024 Garber Painting. All rights reserved.

Note: This service is not intended for secure transactions such as banking, social media, email, or purchasing. Use at your own risk. We assume no liability whatsoever for broken pages.


Alternative Proxies:

Alternative Proxy

pFad Proxy

pFad v3 Proxy

pFad v4 Proxy