w w w . LC Tech.de - Grupo BioMaster
w w w . LC Tech.de - Grupo BioMaster
w w w . LC Tech.de - Grupo BioMaster
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Mycotoxins: Sample Preparation and Analysis<br />
Matrix of the Month<br />
March, 2013:<br />
Aflatoxins<br />
in Dark Chocolate<br />
Do you have a special matrix that we should test for mycotoxins<br />
Please let us know and write an e-mail to info@<strong>LC</strong><strong>Tech</strong>.<strong>de</strong>!<br />
Protocol<br />
www. <strong>LC</strong><strong>Tech</strong>.<strong>de</strong><br />
10 g sample are melted with 2 g sodium chlori<strong>de</strong>, 100 ml 80/20 methanol/water<br />
and 50 ml n-hexane are ad<strong>de</strong>d, stir strongly for 10 minutes.<br />
After filtration the lower phase (n-hexane free) is used, 2 ml are mixed with 12 mL<br />
PBS/Tween (8%) and ad<strong>de</strong>d onto the immunoaffinity column (AflaCLEAN TM ).<br />
The column is washed with 10 ml water and dryed. The toxin is eluted<br />
with 2 x 1 ml methanol. The first milliliter methanol should incubate<br />
on the column bed for 5 minutes.<br />
The eluate is caught, diluted to the conditions of the HP<strong>LC</strong> mobile phase<br />
and analysed via HP<strong>LC</strong>.<br />
HP<strong>LC</strong> Conditions<br />
HP<strong>LC</strong>: Dionex Ultimate 3000, isocratic<br />
Column oven: 36°C<br />
Separation column: Mycotoxin HP<strong>LC</strong> column with guard<br />
Flow rate: 1.2 mL/min (water/methanol/acetonitrile(60/30/15(v/v/v))<br />
Fluorescence <strong>de</strong>tection with post column <strong>de</strong>rivatisation (photochemical with UVE TM )<br />
Excitation wavelength: 365 nm<br />
Emission wavelength: 460 nm<br />
Recovery Rates<br />
Contents of Aflatoxins B1, B2, G1 and G2 in dark chocolate<br />
Chromatogramme =><br />
Aflatoxin B1 B2 G1 G2<br />
Standard* 100 100 100 100<br />
Recovery rate**<br />
Dark chocolate<br />
90 87 96 97<br />
* Standard is set = 100 % , ** corrected with non-spiked sample<br />
Chromatograms
Chromatograms<br />
50<br />
Standard, representing 100 %<br />
mV<br />
B1<br />
Dark chocolate, spiked with 10 ppb<br />
B1<br />
40<br />
G1<br />
40<br />
B2<br />
G1<br />
30<br />
G2<br />
30<br />
B2<br />
20<br />
20<br />
55 mV<br />
min<br />
G2<br />
10<br />
10<br />
- 5<br />
min<br />
0.0 1.3 2.5 3.8 5.0 6.3 7.5 8.8 10.0<br />
-5<br />
0.0 1.3 2.5 3.8 5.0 6.3 7.5 8.8 10.0<br />
www. <strong>LC</strong><strong>Tech</strong>.<strong>de</strong><br />
© <strong>LC</strong><strong>Tech</strong> GmbH, Germany, as of: May, 2013<br />
30<br />
20<br />
10<br />
B1<br />
G1 G2<br />
B2<br />
0<br />
35 mV<br />
min<br />
-5<br />
Dark chocolate, non-spiked<br />
0.0 1.3 2.5 3.8 5.0 6.3 7.5 8.8 10.0<br />
<strong>LC</strong><strong>Tech</strong> GmbH<br />
These <strong>LC</strong><strong>Tech</strong> products were used:<br />
AflaCLEAN TM ,<br />
Immunoaffinity column<br />
for the Aflatoxins B1, B2, G1, G2<br />
P/N 10514<br />
Bahnweg 41<br />
D-84405 Dorfen<br />
Overlay of the chromatograms<br />
mV<br />
125<br />
100<br />
75<br />
50<br />
B1<br />
G1 B2<br />
G2<br />
25<br />
150<br />
-10<br />
min<br />
0.0 1.3 2.5 3.8 5.0 6.3 7.5 8.8 10.0<br />
UVE TM ,<br />
Photochemical reactor<br />
for the Aflatoxin analysis<br />
P/N 10519<br />
Do you have further questions<br />
Please simple write an e-mail to info@<strong>LC</strong><strong>Tech</strong>.<strong>de</strong>!<br />
Tel. ++49(0)8081-9368-0<br />
Fax ++49(0)8081-9368-10<br />
HP<strong>LC</strong> column,<br />
for the Aflatoxin analysis<br />
P/N 10522<br />
www.<strong>LC</strong><strong>Tech</strong>.<strong>de</strong><br />
info@<strong>LC</strong><strong>Tech</strong>.<strong>de</strong>
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