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Aflatoxins B/G in Hemp

MYCOTOXINS

Sample Preparation

For many laboratories, the challenge in everyday laboratory work is to analyze as many samples as possible in the shortest possible time, precisely and with maximum sensitivity. To make this task easier for you, LCTech has developed the AflaCLEAN immunoaffinity column as well as the AflaCLEAN SMART column. 

Both columns are designed for the clean-up of aflatoxins B1, B2, G1 and G2 in food and feed, including hemp. The columns are able to bind the aflatoxins in a highly specific way and have a very high matrix tolerance. 

By using the 3.5 cm SMART column, a particularly high sample throughput of up to 500 samples / week is achieved, since the processing time is significantly reduced due to the small size of the column. In addition, more than 80 % of solvents are saved during extraction, dilution, washing, sample introduction and elution.

Automated Mycotoxin Analysis with FREESTYLE ThermELUTE™

The robotic system FREESTYLE ThermELUTE™ in combination with SMART columns enables full automation with sensitive results that even in the lower ppt-range analysis can be performed around the clock, even at weekends. 

The unique technology realizes a processing „from raw extract to chromatogram“ without any manual intermediate steps. Samples are always processed in the same way and you can be sure to fullfill European regulations and limits. 

For the automated processing of hemp, extract, filter and dilute it according to the processing protocol on page 2. Then place the sample into the FREESTYLE, equip the racks with the necessary columns, parameterize the method in the software with a few clicks and press START - done.

Manual Processing Protocol

Homogenise 10 g of hemp and add 2 g of sodium chloride. Subsequently extract the mixture with 100 mL of methanol/water (80/20 (v/v)) and 50 mL of n-hexane to remove essential oils and fats. For a high extraction efficiency, perform the extraction for 30 minutes.

Filter the raw extract and dilute 2 mL with 12 mL PBS (contains 8 % Tween20). Load 2.8 mL of the sample (corresponding to 0.04 g matrix equivalents) to an AflaCLEAN SMART column. For comparison purposes, an AflaCLEAN column was loaded in parallel.

To allow an efficient binding of the toxins to the antibodies, the flow rate of the AflaCLEAN SMART column should not exceed 3 mL/min and the flow rate of the AflaCLEAN column should not exceed 2 mL/min. Wash the SMART column with 2 mL of deionized water and use the washing solution to rinse the sample reservoir before washing the column.

Elute the toxins with 0.4 mL of methanol. Let the methanol act in the column bed for 5 minutes to ensure complete denaturation of the antibodies.

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

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