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Ochratoxin A in Pumpkin Seeds

Mycotoxins

Fully automated with FREESTYLE ThermELUTE™

Fully Automated Mycotoxin Analysis

Fully automated mycotoxin analysis from raw extract to chromatogram - quite simple with the robotic
system FREESTYLE ThermELUTE™. The samples are automatically loaded, washed, thermally
eluted and quantitatively transferred into the injection system of any LC system via partial filling.
Due to the injection of the complete eluate and the omission of the adjustment of the HPLC eluent
extreme low detection limits in the lower ppt range can be reached.
At the same time you increase your sample throughput up to 500 samples per week due to the
parallel sample processing in both the robotic system and also in the LC system.

Protocol of Manual Processing

Extract 20 g of homogenised pumpkin seed and 2 g NaCl with 100 mL of the extraction solution
(methanol/water, 80/20, v/v) and 50 mL n-hexane for at least 10 minutes for defatting. Filtrate the
raw extract. You can centrifuge it for 10 minutes at 2000 x g facilitating the phase separation. Dilute
10 mL of the filtrate with 40 mL PBS. In case of precipitation or turbidity you can efficiently remove
them by filtration.

Now you have two choices. Either load the sample directly onto the immunoaffinity column
OtaCLEAN SMART via FREESTYLE ThermELUTE™ resulting in a low range measurement.
Alternatively dilute 3 mL of the filtrate again with additional 12 mL PBS and to load 10 mL
(represents 0.08 g matrix) onto the column. Ideally the maximum matrix load is 0.4 g/injection.
The loading of the column (1.5 mL/min) is already done by the FREESTYLE system. The sample
is then washed with 2 mL of water (1.5 mL/min) and thermally eluted. The eluate is quantitatively
injected into the sample loop and analysed via HPLC-FLD. 

Find more Details, Recovery Rates, HPLC-Conditions and Chromatograms here.  

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