Page 51 - Application Notebook - Solution for Food Safety

Application No.C103
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n Analysis of Wheat
Fig. 3 describes the sample pretreatment procedure for value), respectively. No large contaminant peaks were
wheat. The wheat extract solution was purified using detected in the chromatograms of the pretreated
either the MultiSep #227 multi-function column (Romer samples. Furthermore, although deoxynivalenol was
Labs) or the Autoprep MF-T column (Showa Denko detected, it was at a level below that of the provisional
K.K.). The chromatograms generated using the samples reference value. The spike-and-recovery rates for the
prepared using the MultiSep #227 (unspiked samples) four components were excellent, from 101 to 107 %,
and the standard-spiked samples, respectively, are without any particular matrix effects. Even in samples
shown in Fig. 4. The standard mixture was added to pretreated using Autoprep MF-T, comparable spike-
obtain a final concentration of 25 ppb for the four and-recovery test results were obtained.
components (about 1/40 of the provisional reference

Wheat Intensity Intensity
(×10,000)
(×1,000)
1:371.05>281.20(-) 1.50 2:355.10>59.00(-)
a) Nivalenol b) Deoxynivalenol
Grind to less than 1000 µm 7.5 1.25
1.00
5.0 0.75
50.0 g
0.50
200 mL Water/Acetonitrile=15/85(v/v) 2.5
0.25
Shake for 30 min 0.0 0.00
1.5 2.0 2.5 min 3.0 3.5min
Vacuum Filtration by Whatman GF/B Spike and Recovery 101 % Spike and Recovery 107 %
Intensity Intensity
(×10,000) (×10,000)
15 mL 4:397.10 > 59.00(-) 3:397.10 > 59.00(-)
3.0 3.0 3-Acetyl
c) d)
Deoxynivalenol
Clean-up by multi function column 2.0 15-Acetyl 2.0
Deoxynivalenol
“ MultiSep#227” or “Autoprep MF-T”
Discard the first 3 mL of solution.
Extract the next 5 mL of solution. 1.0 1.0
4 mL
0.0 0.0
Evaporate by N2 gas below 45 ˚C
4.0 4.5 min 4.0 4.5 min

1 mL Water/Acetonitrile/Methanol Spike and Recovery 103 % Spike and Recovery 104 %
=90/5/5(v/v/v)
Filtration
Fig. 4 Chromatograms of Wheat
(Dotted line: Unspiked Sample, Solid line: Spiked Sample,
Sample solution Spiked at 25 ppb each)
a) Nivalenol b) Deoxynivalenol
c) 15-Acetyldeoxynivalenol d) 3-Acetyldeoxynivalenol
Fig. 3 Pretreatment

Table 2 Analytical Conditions
Column : Shim-pack XR-ODS Ⅲ (150 mm L. × 2.0 mm I.D., 2.2 µm)
Mobile Phases : A 0.5 mmol/L Ammonium Acetate - Water
: B Acetonitrile
Time Program : 5 %B (0 min) → 45 %B (5.0 min) → 95 %B (5.01-7.0 min) → 5 %B (7.01 min) → STOP (12 min)
Flowrate : 0.3 mL/min
Column Temperature : 40 ˚C
Injection Volume : 2 µL
Probe Voltage : -3.0 kV (ESI-negative mode)
DL Temperature : 100 ˚C
Block Heater Temperature : 200 ˚C
Interface Temperature : 200 ˚C
Nebulizing Gas Flow : 2 L/min
Drying Gas Flow : 10 L/min
Heating Gas Flow : 10 L/min
MRM Transition : Nivalenol 371.05 > 281.20 CE: 16.0 V
: Deoxynivalenol 355.10 > 59.00 CE: 22.0 V
: 15-Acetyldeoxynivalenol 397.10 > 59.00 CE: 22.0 V
: 3-Acetyldeoxynivalenol 397.10 > 59.00 CE: 26.0 V

First Edition: Apr. 2015

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