Aflatoxin B/G and Ochratoxin A in Chocolate

MYKOTOXINS

Chocolate

Chocolate has become an indispensable part of our daily life. Hardly any Central European can do without chocolate for a month. Whether as a spread on bread, as a biscuit filling, in cakes or simply as a small „thank you“ - everyone loves it. The sweet pleasure stimulates our body to release the happiness hormone serotonin.
This works directly in the brain and we feel balanced, satisfied and happy.

The cocoa beans needed to manufacture the chocolate are mainly imported from the Ivory Coast and other countries around the equator. As the beans have a high water content after harvesting, they must be dried in their country of origin before being imported into Germany or further processed. Mycotoxins can be produced in the cocoa bean during the drying process or under incorrect storage conditions. These can be toxic at high concentration.

Sample Clean-up Made Easy by Automation with FREESTYLE SPE

During day, night and even at weekend - the automated FREESTYLE system takes over your daily routine tasks in the field of mycotoxin analysis unattended around the clock so that you have more time for other important tasks in the laboratory. Each manual SPE method, which has already proven itself in your laboratory, can be transferred directly to the robotic system FREESTYLE SPE. Already created methods can be saved and reused but can also be modified at any time.

The fields of application range from food and feed samples to environmental samples, forensic applications and doping samples.
Follow the instructions on the following page to perform the prepared processing steps. Then position the chocolate sample in the FREESTYLE SPE, change settings or parameters in the software with a few mouse clicks and start the system – done.

Processing Protocol

Homogenise 10 g of chocolate and add 2 g of sodium chloride to the sample. Extract the mixture through 100 mL of methanol/water (80/20(v/v)) and add 50 mL n-hexane to remove fat and essential oils.
To ensure high extraction efficiencies, continue the extraction for at least 30 minutes.
Filtrate the raw extract and dilute 2 mL of it with 12 mL PBS (contains 8 % Tween). In the next step, load 14 mL of the sample onto an Afla-OtaCLEAN immunoaffinity column.
To enable efficient binding of the toxins to the antibodies, the flow rate should not exceed 2 mL / min.
Rinse the column with 10 mL deionized water and also load the rinse solution onto the column to remove matrix interferences from the column. 
Dry the column with a short air flow and elute the toxin with 2 mL of methanol. Make sure that the column bed incubates with methanol for 5 minutes in order to ensure a fully denaturation of the antibodies and release of the toxin.

Find more details, recovery rates, HPLC-conditions and chromatograms here.

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