3. FOOD ChEMISTRy & bIOTEChNOLOGy 3.1. Lectures

Chem. Listy, 102, s265–s1311 (2008) Food Chemistry & Biotechnology
Tabulka II
Procedures employed for sample processing (according to producer recommendation)
Step of sample Immunoaffinity columns SPE columns
preparation DOnREP ® Easi exctract HT-2 & T-2 ® Easi exctract ZOn ® DZT Multi MyCo MycoSep ® 226
extraction
solution
dilution crude
extract
deionised water
no
methanol : water
(90 : 10, v/v)
phosphate buffered
saline
acetonitrile : water
(75 : 25, v/v)
phosphate buffered
saline
methanol : water acetonitrile : water
(75 : 25, v/v) (84 : 16, v/v)
phosphate buffered
no
saline (PBS)
mLs of extract
passed through
comumn
2 ml extract 25 ml diluted extract
25 ml diluted
extract
25 ml diluted
exctract
8 ml of extract
washing
5 ml deionised
water
20 ml deionised water 20 ml PBS 10 ml deionised water no
elution
3 × 1.5 ml
methanol
3 × 1.5 ml methanol
3 × 1.5 ml
methanol
3 × 1.5 ml
methanol
no
samples components was carried out in a reverse phase system
using the column with polar endcapping (Synergi Hydro
RP, 150 mm × 3mm × 4 μm).
MS-detector was operated in atmospheric pressure chemical
ionization (APCI) mode, selective negative ions were
aquired for DOn, D3G and ZOn, while for HT-2 and T-2
positive ions were monitored.
Results
The recoveries of fusarium toxins obtained within validation
process employing spiked samples (200 μg kg –1 ) for
repeated analysis procedures (n = 3) characterized in Table II
were summarized in Table III. The cross reactivity value for
D3G is higher than recovery for DOn in DOnPREP ® . Similarly
D3G crossreacted in DZT Multi Myco IACs, nevertheless,
it was fairly lower than aprox. 40 %. These DZT Multi
Myco IACs provided good recoveries for HT-2 and T-2 even
higher than in EASI EXTRACT HT-2�T-2 ® IACs dedicated
Table III
Recoveries of mycotoxins for various columns
Columns Analyte Recovery [%]
DOnPREP ® DOn
D3G�
76
103
EASI EXTR. HT-2T-2 ® HT-2
T-2
90
73
EASI EXTR. ZOn ® ZOn 66
DOn 79
D3G� 40
DZT Multi MyCo HT-2 102
T-2 106
ZOn 94
MycoSep ® DOn 94
D3G� 40
226 HT-2 92
T-2 84
ZOn 98
s701
these two toxins. Similarly, higher recovery was achieved
in DZT Multi Myco IACs than in EASI EXTRACT ZOn ®
IACs.
The levels of fusarium toxins determined in analysis
of contaminated maize corrected to recoveries are shown in
Table IV. When taking the results in Table I obtained by direct
LC-MS/MS as a reference (= 100 %), some more pronounced
differences for results were seen. The most pronounced are
higher concentration determined for HT-2 and T-2 by DZT
Multi Myco IACs and ZOn by EASI EXTRACT ZOn ®
IACs. On the other hand, underestimation of D3G content
accords when it is used IACs clean-up step. We can see good
agreement of generated data for most of tested approaches.
Conclusions
The usage of immunoaffinity columns represent the
challenging approach in selective pre-concentration of target
Table IV
Data of different extraction and clean up process corrected to
reference values noted in Table I
Columns Analyte %
DOnPREP ® DOn
D3G
86
44
EASI EXTR. HT-2T-2 ® HT-2
T-2
93
125
EASI EXTR. ZOn ® ZOn 151
DZT Multi MyCo
MycoSep ® 226
DOn 97
D3G 35
HT-2 152
T-2 131
ZOn 101
DOn 98
D3G 25
HT-2 104
T-2 110
ZOn 124