Fumonisin Immunoaffinity Column


The Eagle Biosciences Fumonisin Immunoaffinity Column (IAC) instruction of fumonisins (FB1 / FB2) determination in food and feed focuses on the enrichment step of extract using immunoaffinity column (IAC) and quantification with HPLC. The Fumonisin Immunoaffinity Column (IAC) is for research use only.

SKU: BTFU314005 Categories: , ,

Fumonisin Immunoaffinity Column

3ml, 10 columns

For Research Use Only

Product Developed and Manufactured in Germany by BioTeZ Berlin-Buch GmbH

Additional Information

Assay Principle

This instruction of fumonisins (FB1 / FB2) determination in food and feed focuses on the enrichment step of extract using immunoaffinity column (IAC) and quantification with HPLC.  Accepted laboratory extraction methods could be maintained. Full performance of the IAC column is given if pronounced criteria regarding organic solvent tolerance, elution process of analyte and working range of column are followed.  Many pretreatment methods of fumonisins determination in food and feed, most of them which are based on solid phase extraction (SPE) with adsorbents or strong anion resins (SAX), show low sensitivity because of interfering substances if problematic matrices are applied.

This method of content determination of fumonisins combines the high selectivity of an immunoaffinity column (IAC) with its potential to concentrate elute and additional step of purification of derivatized fumonsins by HPLC column.  As said before, this instruction focuses on the handling with the IAC column. For the commodity extraction step a literature method is given. Please see below. The given apparatus (e.g. HPLC system) might serve as example among other possibilities. For your convenience, an example HPLC method for the analysis of fumonisins is given below.

Sample Preparation:

Samples which content of Fumonisins are to be analyzed, e.g. wheat, maize etc., are extracted by the method of Prioli et al. using methanol-water (80/20 v/v) as extraction solvent.

Enrichment Step Immunoaffinity Chromatography (IAC):
1.6ml extract (0.4g sample equivalent if above mentioned example extraction is followed) is diluted with 10mM PBS, pH=7.2 resulting a total volume of 13ml and then applied in a reservoir on top of the B-TeZ IAC Fumonisin 3ml column. If the example is followed the resulting organic solvent concentration is 10% of methanol which is tolerated by the column. The flow rate lies between 1 to 3ml/min. According to application and contents expected the applied extract volumes could vary. In case of very low contents even extract volumes of 20ml may be diluted with buffer and applied without significant loss of analyte as long as resulting pH is fairly neutral and alcohol or acetonitrile content lies under 15%. If latter is not the case the extract must further be diluted with PBS until fore mentioned maximum allowed organic solvent content during enrichment step using the B-TeZ IAC Fumonisin 3ml column is reached.

After whole sample has passed through the gel the latter is washed with 5ml of 10mM PBS. Remaining liquids in the gel are removed by applying either pressure from top of the column or under pressure from bottom.

Sample reservoir on top of the using B-TeZ IAC Fumonisin 3ml column is removed and an appropriate vial is placed below the affinity column. The bound toxins are eluted by using a total of 3ml of methanol-acetic acid (98/2 v/v)(1) as elution solvent. The elution process is performed in two steps to ensure complete release of analytes. First, a volume of 1ml elution solvent is applied. After that volume has passed through column half a minute is waited before the second portion of 2ml of elutions solvent is eluted through the column. Remaining solvent solutions should be eluted by application of slight under- or overpressure. All methanolic-acetic acid fractions are unified to give the column eluate.

The column eluate is concentrated to complete dryness (e.g. using VLM evaporator at 50°C under a permanent stream of nitrogen(2)). The residue is redissolved in e.g. 1ml of acetonitrile-water (40/60 v/v)(3) as dilution solvent and an aliquot is derivatized in the auto injector prior to injection into the system.


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