ISO 23719:2025

Cereals and cereal products - Determination of 17 mycotoxins by ultra-high-performance liquid chromatography and tandem mass spectrometry method (UHPLC-MS/MS)

This document specifies a method for the quantitative determination of 17 mycotoxins in cereals and cereal products (e.g. wheat, maize, husked rice, rice and their products) using ultra-high-performance liquid chromatography and tandem mass spectrometry method (UHPLC-MS/MS). The 17 mycotoxins are aflatoxin B1, aflatoxin B2, aflatoxin G1, aflatoxin G2, deoxynivalenol, nivalenol, deoxynivalenol-3-glucoside, 3-acetyl-deoxynivalenol, 15-acetyl-deoxynivalenol, zearalenone, ochratoxin A, fumonisin B1, fumonisin B2, fumonisin B3, T-2 toxin, HT-2 toxin and sterigmatocystin. This document does not apply to foods for infants and young children. This document is applicable to other products (e.g. nuts) provided that the method is validated for each individual case. The calibration range of the method and the validated range during the interlaboratory study for each mycotoxin are listed in Table A.1.

Céréales et produits céréaliers — Détermination de 17 mycotoxines par chromatographie liquide à ultra haute performance et spectrométrie de masse en tandem (CLUHP-SM/SM)

General Information

• Status: Published
• Publication Date: 07-Jul-2025
• ICS: 67.060 - Cereals, pulses and derived products
• Technical Committee: ISO/TC 34/SC 4 - Cereals and pulses
• Drafting Committee: ISO/TC 34/SC 4/WG 8 - Mycotoxin
• Current Stage: 6060 - International Standard published
• Start Date: 08-Jul-2025
• Due Date: 28-Feb-2026
• Completion Date: 08-Jul-2025

Overview

ISO 23719:2025 specifies a quantitative UHPLC‑MS/MS method for the determination of 17 mycotoxins in cereals and cereal products. The standard covers common commodities such as wheat, maize, husked rice and rice products, and uses an ultra‑high‑performance liquid chromatography tandem mass spectrometry (UHPLC‑MS/MS) workflow with isotopic dilution. Target analytes include aflatoxins (B1, B2, G1, G2), deoxynivalenol (and derivatives), nivalenol, zearalenone, ochratoxin A, fumonisins (B1, B2, B3), T‑2 and HT‑2 toxins, and sterigmatocystin. ISO 23719:2025 does not apply to foods for infants and young children and can be extended to other matrices (e.g., nuts) only after matrix‑specific validation.

Key Topics

• Analytical principle: Extraction with acetonitrile–water–acetic acid (70:29:1, v/v/v), low‑temperature centrifugation, filtration, addition of isotope‑labelled internal standards, and analysis by UHPLC‑MS/MS in multiple reaction monitoring (MRM) mode.
• Isotopic dilution: Quantification via 13C‑labelled internal standards (ISTDs) for robust accuracy and compensation for matrix effects.
• Calibration and validation: Defined calibration ranges and validated ranges from an interlaboratory study (see Annex A). Calibration solutions and mixed stock/ISTD preparations are specified.
• Instrument requirements: UHPLC capable of binary gradients, column retention performance, column oven, and a tandem mass spectrometer with electrospray ionization (ESI).
• Safety and quality: Mycotoxin hazard warnings, reagent quality (LC‑grade solvents), sample preparation controls, and precision metrics (repeatability and reproducibility).
• Supporting information: Annexes provide ISTD lists, example LC‑MS/MS conditions, sample chromatograms, and interlaboratory results.

Applications

• Food testing laboratories implementing routine mycotoxin surveillance in cereals and cereal products.
• Regulatory agencies performing compliance testing and exposure assessment for aflatoxins, DON, fumonisins, ZEN, OTA and others.
• Quality assurance teams in milling, grain trading and cereal processing industries for internal control and supplier verification.
• Method developers and validation labs needing a robust UHPLC‑MS/MS protocol and interlaboratory performance benchmarks.

Using ISO 23719:2025 helps labs achieve traceable, harmonized results for multiple mycotoxins with high sensitivity and specificity, improving food safety decision‑making and trade compliance.

Related standards

• ISO 3696 - Water for analytical laboratory use (normative reference in ISO 23719:2025)
• Prepared by ISO/TC 34/SC 4 (Cereals and pulses); see full annexes in ISO 23719:2025 for calibration ranges, ISTD details and example conditions.

Standards Content (Sample)

International
Standard
ISO 23719
First edition
Cereals and cereal products —
2025-07
Determination of 17 mycotoxins
by ultra-high-performance liquid
chromatography and tandem
mass spectrometry method
(UHPLC-MS/MS)
Céréales et produits céréaliers — Détermination de 17 mycotoxines
par chromatographie liquide à ultra haute performance et
spectrométrie de masse en tandem (CLUHP-SM/SM)
Reference number
© ISO 2025
All rights reserved. Unless otherwise specified, or required in the context of its implementation, no part of this publication may
be reproduced or utilized otherwise in any form or by any means, electronic or mechanical, including photocopying, or posting on
the internet or an intranet, without prior written permission. Permission can be requested from either ISO at the address below
or ISO’s member body in the country of the requester.
ISO copyright office
CP 401 • Ch. de Blandonnet 8
CH-1214 Vernier, Geneva
Phone: [phone]
Email: [e-mail]
Website: www.iso.org
Published in Switzerland
ii
Contents Page
Foreword .iv
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 Principle . 1
5 Reagents . 2
6 Apparatus and equipment . 4
7 Procedure . 5
7.1 Sampling and preparation of the laboratory sample . .5
7.2 Test portion weighing .5
7.3 Extraction .5
7.4 Preparation of the sample test solution .6
7.5 Preparation of control samples .6
7.6 Analysis .6
7.6.1 Operating conditions .6
7.6.2 Injection sequence .6
7.6.3 Data treatment .7
8 Identification . 7
9 Calculations . 7
10 Precision . 8
10.1 General .8
10.2 Repeatability .8
10.3 Reproducibility . .8
11 Test report . 8
Annex A (informative) Calibration range of the method and the validated range during the
interlaboratory study for each mycotoxin . 10
Annex B (informative) Information on 17 mycotoxins and ISTD standards .11
Annex C (informative) Example conditions for suitable LC-MS/MS systems .13
Annex D (informative) Example chromatogram of mycotoxins and ISTDs .20
Annex E (informative) Results of the interlaboratory test .22
Bibliography .40

iii
Foreword
ISO (the International Organization for Standardization) is a worldwide federation of national standards
bodies (ISO member bodies). The work of preparing International Standards is normally carried out through
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with the International Electrotechnical Commission (IEC) on all matters of electrotechnical standardization.
The procedures used to develop this document and those intended for its further maintenance are described
in the ISO/IEC Directives, Part 1. In particular, the different approval criteria needed for the different types
of ISO document should be noted. This document was drafted in accordance with the editorial rules of the
ISO/IEC Directives, Part 2 (see www.iso.org/directives).
ISO draws attention to the possibility that the implementation of this document may involve the use of (a)
patent(s). ISO takes no position concerning the evidence, validity or applicability of any claimed patent
rights in respect thereof. As of the date of publication of this document, ISO had not received notice of (a)
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this may not represent the latest information, which may be obtained from the patent database available at
www.iso.org/patents. ISO shall not be held responsible for identifying any or all such patent rights.
Any trade name used in this document is information given for the convenience of users and does not
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For an explanation of the voluntary nature of standards, the meaning of ISO specific terms and expressions
related to conformity assessment, as well as information about ISO’s adherence to the World Trade
Organization (WTO) principles in the Technical Barriers to Trade (TBT), see www.iso.org/iso/foreword.html.
This document was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee SC 4, Cereals
and pulses.
Any feedback or questions on this document should be directed to the user’s national standards body. A
complete listing of these bodies can be found at www.iso.org/members.html.

iv
International Standard ISO 23719:2025(en)
Cereals and cereal products — Determination of
17 mycotoxins by ultra-high-performance liquid
chromatography and tandem mass spectrometry method
(UHPLC-MS/MS)
1 Scope
This document specifies a method for the quantitative determination of 17 mycotoxins in cereals and cereal
products (e.g. wheat, maize, husked rice, rice and their products) using ultra-high-performance liquid
chromatography and tandem mass spectrometry method (UHPLC-MS/MS).
The 17 mycotoxins are aflatoxin B , aflatoxin B , aflatoxin G , aflatoxin G , deoxynivalenol, nivalenol,
1 2 1 2
deoxynivalenol-3-glucoside, 3-acetyl-deoxynivalenol, 15-acetyl-deoxynivalenol, zearalenone, ochratoxin A,
fumonisin B , fumonisin B , fumonisin B , T-2 toxin, HT-2 toxin and sterigmatocystin.
1 2 3
This document does not apply to foods for infants and young children.
This document is applicable to other products (e.g. nuts) provided that the method is validated for each
individual case.
The calibration range of the method and the validated range during the interlaboratory study for each
mycotoxin are listed in Table A.1.
2 Normative references
The following documents are referred to in the text in such a way that some or all of their content constitutes
requirements of this document. For dated references, only the edition cited applies. For undated references,
the latest edition of the referenced document (including any amendments) applies.
ISO 3696, Water for analytical laboratory use — Specification and test methods
3 Terms and definitions
No terms and definitions are listed in this document.
ISO and IEC maintain terminology databases for use in standardization at the following addresses:
— ISO Online browsing platform: available at https:// www .iso .org/ obp
— IEC Electropedia: available at https:// www .electropedia .org/
4 Principle
Mycotoxins are extracted using an acetonitrile-water-acetic acid (70:29:1, v/v/v) solution. After
centrifugation, the extract is diluted with water, centrifuged in low temperature and filtered. The final
solution is mixed with isotope-labelled internal standard solution and subjected to UHPLC-MS/MS analysis.
Quantification is performed by the isotopic dilution approach using C isotopically labelled mycotoxins as
internal standards (ISTDs).
WARNING — Mycotoxins are generally considered to be carcinogenic, neurotoxic and
[1]
immunosuppressive. Observe appropriate safety precautions for handling such compounds
and avoid handling in dry form as the electrostatic nature can result in dispersion and inhalation.

Glassware can be decontaminated with 4 % sodium hypochlorite solution. Attention is drawn to the
[2][3]
statement made by the International Agency for Research on Cancer (WHO).
5 Reagents
WARNING — This document requires handling of hazardous substances. Technical, organizational
and personal safety measures shall be followed.
During the analysis, unless otherwise stated, use only reagents of recognized analytical grade and only
distilled water or water of grade 1 in accordance with ISO 3696. Solvents shall be of quality for LC analysis.
5.1 Acetonitrile, LC grade or equivalent.
5.2 Methanol, LC grade or equivalent.
5.3 Ammonium acetate, American Chemical Society (ACS) grade, > 99 %.
5.4 Acetic acid, ACS grade.
5.5 Formic acid, ACS grade.
5.6 Extraction solvent: Mix 70 volume parts of acetonitrile (5.1) and 29 volume parts of water and 1
volume part of acetic acid (5.4).
5.7 Solvent of calibration solutions: Mix 50 volume parts of extraction solvent (5.6) and 50 volume parts
of water.
5.8 Individual stock solutions, either prepared by dissolving neat (solid) certified standards in an
appropriate solvent or from individual stock solutions purchased as such.
The mycotoxins covered in this document dissolve well in acetonitrile, except for fumonisins which
are soluble in a mixture of acetonitrile and water (50:50, v/v). The information and concentration of 17
mycotoxins and the isotope-labelled internal standards is shown in Table B.1.
5.9 Mixed stock solution.
Prepare a mixed stock solution containing all individual mycotoxins at the concentration given in Table 1,
using the appropriate pipettes (6.1) and dilute with deionized water. This solution can be used for six months
if stored in the dark at −20 °C.
This mixed stock solution may be used for the preparation of positive control samples (see 7.5).
Table 1 — Concentration of mixed stock solution for 17 mycotoxins
Compounds Concentration Compounds Concentration
μg/ml μg/ml
NIV 20 FB 12,5
DON 15 FB 7,5
DON-3G 2,5 FB 9,0
3-AcDON 4 T-2 0,2
15-AcDON 2 HT-2 1
AFB 0,1 ZEN 2
TTabablele 1 1 ((ccoonnttiinnueuedd))
Compounds Concentration Compounds Concentration
μg/ml μg/ml
AFB 0,1 OTA 0,2
AFG 0,1 ST 0,1
AFG 0,1 - -
5.10 Mixed isotope-labelled internal standard (ISTD) solution.
Isotopically labelled mycotoxins are generally available as certified standard solutions. Prepare a mixed ISTD
solution in a mixture of acetonitrile and water (30:70, v/v), containing all isotopically labelled mycotoxins
at the concentration given in Table 2. This solution can be used for six months if stored in the dark at −20 °C.
Table 2 — Concentration of mixed isotope-labelled internal standard solution for 17 mycotoxins
Compounds Concentration Compounds Concentration
µg/ml µg/ml
13 13
[ C ]-NIV 2,5 [ C ]-FB 0,5
15 34 1
13 13
[ C ]-DON 2 [ C ]-FB 0,3
15 34 2
13 13
[ C ]-DON-3G 1 [ C ]-FB 0,3
21 34 3
13 13
[ C ]-3-AcDON 1 [ C ]-T-2 0,05
17 24
13 13
[ C ]-15-AcDON 1 [ C ]-HT-2 0,125
17 22
13 13
[ C ]-AFB 0,01 [ C ]-ZEN 0,2
17 1 18
13 13
[ C ]-AFB 0,01 [ C ]-OTA 0,04
17 2 20
13 13
[ C ]-AFG 0,01 [ C ]-ST 0,025
17 1 18
[ C ]-AFG 0,01 - -
17 2
5.11 Calibration solutions.
Transfer appropriate volumes of the mixed stock solutions (5.9) into six separate 1,5 ml injection vials as
described in Table 3. The concentrations for the calibration solutions are shown in Table 4. Store them at
–20 °C for no longer than two weeks before use.
Transfer a 180 µl aliquot of the calibration solutions into an injection vial with a 400 µl micro-insert (6.3),
individually. Add 20 µl mixed ISTD solution (5.10) into all the vials. The same ISTD solution as used in sample
preparation shall be used. Cap and mix.
Table 3 — Example of calibration solutions
No. Calibration Calibration Volume of Volume of acetonitrile-
standard standard standard water-acetic acid
solution no. solution solution (35:64.5:0.5, v/v/v) (5.7)
µl µl
Step 1 SS1 mixed stock solution (5.9) 50 950
Step 2 SS2 mixed stock solution (5.9) 20 980
Step 3 SS3 SS1 200 800
Step 4 SS4 SS2 200 600
Step 5 SS5 SS3 200 600
Step 6 SS6 SS4 200 600
Table 4 — Concentration of calibration solutions
Dimensions in µg/l
Mycotoxin SS6 SS5 SS4 SS3 SS2 SS1
NIV 25 50 100 200 400 1 000
DON 18,75 37,5 75 150 300 750
DON-3G 3,125 6,25 12,5 25 50 125
3-ACDON 5 10 20 40 80 200
15-ACDON 2,5 5 10 20 40 100
AFB 0,125 0,25 0,5 1 2 5
AFB 0,125 0,25 0,5 1 2 5
AFG 0,125 0,25 0,5 1 2 5
AFG 0,125 0,25 0,5 1 2 5
FB 15,625 31,25 62,5 125 250 625
FB 9,375 18,75 37,5 75 150 375
FB 11,25 22,5 45 90 180 450
T-2 0,25 0,5 1 2 4 10
HT-2 1,25 2,5 5 10 20 50
ZEN 2,5 5 10 20 40 100
OTA 0,25 0,5 1 2 4 10
ST 0,125 0,25 0,5 1 2 5
6 Apparatus and equipment
The usual laboratory equipment and, in particular, the following shall be used.
6.1 Automatic pipettes, suitable for handling volumes of 1 µl to 20 µl,10 µl to 100 µl, 200 µl to 1 000 µl
and 1 ml to 10 ml.
Instead of the latter, 10 ml graduated glass pipettes may be used as an alternative.
6.2 Centrifuge tubes, 1,5 ml and 10 ml.
6.3 Injection vials, 1,5 ml, suitable for an LC autosampler, if necessary, with 400 µl micro-inserts.
6.4 Vortex mixer.
6.5 Orbital shaker, adjustable mechanical vertical or horizontal shaker, capable to shake at 30 min
6.6 Polytetrafluoroethylene (PTFE) filters, syringe filters, 0,2 µm pore size, 13 mm internal diameter.
6.7 UHPLC-MS/MS system, with the components given in 6.7.1 to 6.7.7.
6.7.1 LC pump, capable of delivering a binary gradient at flow rates appropriate for the analytical column
in use with sufficient accuracy.
6.7.2 Degasser, optional, for degassing LC mobile phases.
6.7.3 Injection system, capable of injecting an appropriate volume of injection solution with sufficient
accuracy.
6.7.4 LC column, capable to retain the first eluting analyte at least twice the retention time corresponding
to the void volume of the column. Examples of suitable columns and gradients are given in Annex C.
6.7.5 Column oven, capable to maintain a constant temperature.
6.7.6 Tandem mass spectrometer (e.g. triple quadrupole or quadrupole linear ion trap), equipped with
an electrospray ionization (ESI) interface and operated in multiple reaction monitoring (MRM) mode. Any
ionization mode (typically negative or positive) giving sufficient yield may be employed.
6.7.7 Computer-based instrument control and data evaluation system.
6.8 Mill equipped with 0, 5 mm sieves.
6.9 Conical flask, 250 ml.
6.10 Blender, 1 jar and cover, explosion-proof.
6.11 Ceramic abrasive particles, oblique elliptical cylinder diamond, 1,5 cm (length) × 0,65 cm (outer
diameter).
6.12 Centrifuges:
a) suitable for the centrifuge tubes (10 ml and 50 ml) and capable of achieving at least 3 500g;
b) suitable for the centrifuge tubes (1,5 ml) and capable of achieving at least 7 500g at 4 °C.
6.13 Laboratory balance, accuracy of 0,01 g.
6.14 Analytical balance, accuracy of 0,1 mg.
6.15 Disposable syringe, 1,5 ml.
7 Procedure
7.1 Sampling and preparation of the laboratory sample
Sampling is not part of the method specified in this document. A recommended sampling method is given in
[4]
ISO 24333 .
A representative sample should be sent to the laboratory. It should not have been damaged or changed during
transport and storage. Grind the laboratory sample using a laboratory mill (6.8) until it passes through the
sieve (0,5 mm) and mix it thoroughly.
7.2 Test portion weighing
Weigh a test portion of 25,00 g (m) of the homogeneous laboratory sample to the nearest 0,01 g into a conical
flask (6.9) or a blender (6.10).
For samples with good homogeneity (e.g. flour), a minimum test sample amount of 5 g and an extraction
solvent volume of 20 ml may be used.
7.3 Extraction
Add 100 ml extraction solvent (5.6) (V ). Cover and shake for 30 min (6.5) or blend for 3 min (6.10). Pipette
5 ml supernatant into a 10 ml centrifuge tube (6.2) and centrifuge at 3 500g for 5 min. Add 0,5 ml supernatant

(V ) into a 1,5 ml centrifuge tube (6.2), dilute with 0,5 ml water (V ) and shake on a vortex mixer (6.4) for
2 3
1 min. Then centrifuge (7 500g, 4 °C, 10 min, 6.12 b). The supernatant shall be filtrated through a 0,2 μm
PTFE filter (6.6).
7.4 Preparation of the sample test solution
Transfer a 180 µl aliquot of the filtrate obtained in 7.3 into an injection vial with a 400 µl micro-insert
(6.3). Add 20 µl mixed ISTD solution (5.10) into each vial. The same ISTD working solution as used in the
preparation of the sample and standard curve shall be used. Cap and mix.
7.5 Preparation of control samples
With each sample batch, one reagent blank and one positive control shall be used.
To create a reagent blank, perform extraction (see 7.3) and subsequent steps without adding the sample test
portion.
The positive control should a sample free of the target mycotoxins (non-detectable) which is spiked with 10 µl
mixed stock solution (5.9) per 1 g. The spiking concentration level in samples for 17 mixed mycotoxins are
shown in Table 5. Alternatively, a reference material known to contain the target mycotoxins may be used.
Table 5 — Spiking concentration level in samples for 17 mixed mycotoxins
Compounds Concentration Compounds Concentration
μg/kg μg/kg
NIV 200 FB 125
DON 150 FB 75
DON-3G 25 FB 90
3-AcDON 40 T-2 2
15-AcDON 20 HT-2 10
AFB 1 ZEN 20
AFB 1 OTA 2
AFG 1 ST 1
AFG 1 - -
7.6 Analysis
7.6.1 Operating conditions
Optimize analytical parameters (selection of the ionization mode, selection of the masses of precursor
and product ions, optimization of source parameters and collision energies) by infusion of each individual
mycotoxin standard solution. At least two ion transition reactions should be measured for each unlabelled
mycotoxin. The ion transition reaction giving the largest signal-to-noise ratio shall be selected as the
quantifier one. Choose the MRM transition reaction of the internal standard that corresponds to it, taking
into account the degree of isotope labelling.
A combination of analytical column, mobile phase composition, gradient settings and injection volume shall
be selected so that it allows for obtaining an acceptable separation and reliable results at the required levels,
with sufficient selectivity. Annex C gives some suitable parameters.
7.6.2 Injection sequence
An example is as follows:
— Start a batch of measurements by injecting an aliquot of the diluting solution (5.7) to prove non-
contamination of the system.
— Then inject the calibration solutions from SS6 to SS1 (5.11) and carefully check that all mycotoxins and
their respective ISTDs are visible at the lowest calibration level (SS6).
Inject an aliquot of the diluting solution (5.7) to check for possible carryover.
Inject the sample test solution (see 7.4) and regularly inject an aliquot of the diluting solution (5.7) to check
for possible carryover.
For each injection sequence, a reagent blank shall be injected (7.5).
For each injection sequence, a quality control sample may be included. A reference material known to contain
the targeted mycotoxins may be used. Alternatively, an extract of a sample free of the targeted mycotoxins
(e.g. non-detectable levels) spiked with the mycotoxins within the range of the measurements may be used.
End the sequence by re-injecting the calibration solutions from SS6 to SS1 (5.11).
7.6.3 Data treatment
Process the data using an appropriate integration software. Peak areas are used for subsequent calculations.
Check peak area assignment and integration for the measured transition reactions. Annex D gives some
example chromatograms of mycotoxins and ISTDs.
8 Identification
Mycotoxins are considered as positively identified in the sample when the following criteria are fulfilled:
a) A peak with an S/N (signal/noise) ratio ≥ 3 should be observed in at least two selected ion transition
reactions.
b) The retention time of the mycotoxin should correspond to that of its labelled internal standard within a
tolerance of ±0,1 min.
c) The peak area ratio of the two product ions from sample extracts should be within ±30 % (relative) of
[5]
average of calibration solutions from same sequence.
9 Calculations
Construct the calibration curve by plotting the quantifier peak area ratio of each mycotoxin and its ISTD
(y-axis) against concentration of each mycotoxin (x-axis) using calibration solutions from SS6 to SS1. To
improve the precision on the low concentration, a weighing factor (e.g. 1/x or 1/x ) should be used for the
calibration curve. Alternatively, the regression curve can be forced to zero. Calculate the slope and the
intercept by linear regression. Check that the deviation of the back-calculated concentration of the calibrants
standards from the true concentration, using the calibration curve, shall be not more than ±20 %. If higher
deviations or nonlinearity are observed, identify the cause and, if necessary, re-run the analyses. Calculate
the slope (S) and the intercept (I) of the calibration curve.

Each mass fraction, X, of the mycotoxins, in micrograms per kilogram, shall be calculated according to
Formula (1):
()AI−×VV×+()V
12 3
X = (1)
Sm××V
where
X is the final concentration of samples, in μg/kg;
A is the peak area ratio of a given mycotoxin and its IS (quantifier transition reaction);
I is the intercept of the (weighted) regression line;
S is the slope of the (weighted) regression line;
V is the volume of the of sample extract, in ml (=100);
V is the volume of the supernatant for dilution, in ml (= 0,5);
V is the volume of the water for dilution, in ml (= 0,5);
m is the mass of the test portion, in g (=25).
10 Precision
10.1 General
[6]
Details of the interlaboratory test of the precision of the method in accordance with ISO 5725-2:2019 are
summarized in Annex E. The values derived from the interlaboratory tests are not necessarily applicable to
analyte concentration ranges and matrices other than those given in Annex E.
10.2 Repeatability
The absolute difference between two single test results found on identical test material by one operator
using the same apparatus within the shortest feasible time interval will exceed the repeatability limit r in
not more than 5 % of the cases (see Tables E.1 to E.17).
10.3 Reproducibility
The absolute difference between two single test results found on identical test material reported by two
laboratories will exceed the reproducibility limit R in not more than 5 % of the cases (see Tables E.1 to E.17).
11 Test report
The test report shall contain at least the following information:
— all information necessary for the identification of the sample;
— a reference to this document, including its year of publication, e.g. ISO 23719;
— the results and the units in which the results have been expressed;
— the date and type of sampling (if known);
— the date of receipt of the laboratory sample;
— the date of the test;
— any points observed during the test;
— any operations not specified in the method or regarded as optional which can have affected the results.

Annex A
(informative)
Calibration range of the method and the validated range during the
interlaboratory study for each mycotoxin
Table A.1 — Calibration range of the method and the validated range during the interlaboratory
study for each mycotoxin
Mycotoxin Calibration range of the method Validated range during the interlaboratory study
µg/kg µg/kg
NIV 200 to 8 000 400 to 3 200
DON 150 to 6 000 300 to 2 400
DON-3G 25 to 1 000 50 to 400
3-ACDON 40 to 1 600 80 to 640
15-ACDON 20 to 800 40 to 320
AFB 1 to 40 2 to 16
AFB 1 to 40 2 to 16
AFG 1 to 40 2 to 16
AFG 1 to 40 2 to 16
FB 125 to 5 000 250 to 2 000
FB 75 to 3 000 150 to 1 200
FB 90 to 3 600 180 to 1 800
T-2 2 to 80 4 to 32
HT-2 10 to 400 20 to 160
ZEN 20 to 800 40 to 320
OTA 2 to 80 4 to 32
ST 1 to 40 2 to 16
Annex B
(informative)
Information on 17 mycotoxins and ISTD standards
Table B.1 — Information on 17 mycotoxins and ISTD standards
Compounds Abbreviation Formula Molecular Concentration Solvent
weight
Da µg/ml
Nivalenol NIV C H O 312,32 100 Acetonitrile
15 20 7
Deoxynivalenol DON C H O 296,32 100 Acetonitrile
15 20 6
Deoxynivale-
DON-3G C H O 458,46 50 Acetonitrile
21 30 11
nol-3-glucoside
3-acetyl-
3-AcDON C H O 338,35 100 Acetonitrile
17 22 7
deoxynivalenol
15-acetyl-
15-AcDON C H O 338,35 100 Acetonitrile
17 22 7
deoxynivalenol
Aflatoxin B AFB C H O 312,27 1,0
1 1 17 12 6
Aflatoxin B AFB C H O 314,29 1,0
2 2 17 14 6
a
Acetonitrile
Aflatoxin G AFG C H O 328,27 1,0
1 1 17 12 7
Aflatoxin G AFG C H O 330,29 1,0
2 2 17 14 7
Fumonisin B FB C H NO 721,83 50 Acetonitrile/water
1 1 34 59 15
Fumonisin B FB C H NO 705,83 50 Acetonitrile/water
2 2 34 59 14
Fumonisin B FB C H NO 705,83 50 Acetonitrile/water
3 3 34 59 14
T-2 toxin T-2 C H O 466,52 100 Acetonitrile
24 34 9
HT-2 toxin HT-2 C H O 424,48 100 Acetonitrile
22 32 8
Zearalenone ZEN C H O 318,36 100 Acetonitrile
18 22 5
Ochratoxin A OTA C H ClNO 403,81 10 Acetonitrile
20 18 6
Sterigmatocystin ST C H O 324,48 50 Acetonitrile
18 12 6
13 13 13
[ C ]-nivalenol [ C ]-NIV C H O 327,17 25 Acetonitrile
15 15 15 20 7
[ C ]-
15 13 13
[ C ]-DON C H O 311,18 25 Acetonitrile
15 15 20 6
deoxynivalenol
[ C ]-deoxyniva-
15 13 13
[ C ]DON-3G C H O 479,26 10 Acetonitrile
21 21 30 11
lenol-3-glucoside
[ C ]-3-acetyl-
17 13 13
[ C ]-3-AcDON C H O 355,19 25 Acetonitrile
17 17 22 7
deoxynivalenol
[ C ]-15-acetyl-
17 13 13
[ C ]-15-AcDON C H O 355,19 10 Acetonitrile
17 17 22 7
deoxynivalenol
[ C ]-aflatoxin
17 13 13
[ C ]-AFB C H O 329,12 0,5 Acetonitrile
17 1 17 12 6
B
[ C ]-aflatoxin
17 13 13
[ C ]-AFB C H O 331,13 0,5 Acetonitrile
17 2 17 14 6
B
[ C ]-aflatoxin
17 13 13
[ C ]-AFG C H O 345,12 0,5 Acetonitrile
17 1 17 12 7
G
[ C ]-aflatoxin
17 13 13
[ C ]-AFG C H O 347,13 0,5 Acetonitrile
17 2 17 14 7
G
a
AFB , AFB , AFG and AFG were mixed solutions.
1 2 1 2
TTabablele B B.11 ((ccoonnttiinnueuedd))
Compounds Abbreviation Formula Molecular Concentration Solvent
weight
Da µg/ml
[ C ]-fumonisin
34 13 13
[ C ]-FB C H NO 755,50 25 Acetonitrile/water
34 1 34 59 15
B
[ C ]-fumonisin
13 13
[ C ]-FB C H NO 739,51 10 Acetonitrile/water
34 2 34 59 14
B
[ C ]-fumonisin
34 13 13
[ C ]-FB C H NO 739,51 10 Acetonitrile/water
34 3 34 59 14
B
13 13 13
[ C ]-T-2 toxin [ C ]-T-2 C H O 490,30 25 Acetonitrile
24 24 24 34 9
13 13 13
[ C ]-HT-2 toxin [ C ]-HT-2 C H O 446,28 25 Acetonitrile
...