BS EN 16006:2011 pdf download – Animal feeding stuffs — Determination of the Sum of Fumonisin B1 & B2 in compound animal feed with immunoaffinity clean-up and RP-HPLC with fluorescence detection after pre- or post- column derivatisation These calibration levels are recommendations and may be adjusted to the individual needs. The exactconcentrations of the calibration levels should be calculated based on the final dilution and the exactconcentration of the stock solution (4.20). NOTE The above solutions may be stored for up to 5 days at 6 °C±2°C in the dark. 4.23 IAC (lmmunoaffinity column) The immunoaffinity columns must contain a stationary phase with immobilized monoclonal antibodies specificto, at least,Fumonisin B, and B To be suitable for this method they must meet the requirements statedbelow: An aliquot of more than 5 ml of an extract of a fumonisin-free representative compound animal feed material isspiked with FB, & FBz in equal parts at either 920(high) ng/ml or 110 (low) ng/ml for the sum of both. Thendilute 5,0 ml of that spiked extract to a total volume of 50,0 ml(see 6.2). Following the procedures described in 6.3 and 6.4 this will result in expected concentrations in the injectionsolutions of either 460 ng/ml or 55 ng/ml for the sum of FB,& FB2. After measuring (Clause 7) these solutions the observed concentrations of FB,& FB2 can be calculated withEquation (1) and Equation (2) of Clause 8. Dividing the sum of the observed concentrations of FB, &FBz bythe expected concentrations will result in the yield of the immunoaffinity columns. These yields must be 99 %±18 %(U, k=2) at the high level and 118 %±18 %(U,k=2) at the low level. The above column test should be performed for each level on at least three randomly selected columns ofevery new batch of immunoaffinity columns which will be used.Should the tested batch not meet the aboverequirements either a new batch which does should be obtained or the conditions described in 6.3 need to beadjusted such that the requirements are met (the user instructions supplied with the columns are a goodstarting point). Any changes to the clean-up procedures will necessitate a revalidation of the clean-up and allsubsequent steps (chromatography). 5 Apparatus Usual laboratory equipment and, in particular, the following: 5.1 Mill 5.2 Tumble mixer Creates a folding motion of the material through, for instance, a rotating drum with internal fins and paddels or moving a closed container head-over-heels. 5.3 Vortex...
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