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BS ISO 23305:2020 pdf download

BS ISO 23305:2020 pdf download.Fortified milk powders, infant formula and adult nutritionals – Determination of total biotin by liquid chromatography coupled with immunoaffinity column clean-up extraction.
7 Procedure
7.1 Sample preparation
7.1.1 For mass and loading volumes for the different ranges of product, see Table 1. A slurry may be used wherever product homogeneity is suspected or unknown.
For the slurry, reconstitute 25 g of powder (ml) with warm water (—50 °C) to a total mass of 200 g (m2). Mix thoroughly on a horizontal shaker for 20 mm and then sonicate at 50 °C for 10 mm. Cool to room temperature. For liquid samples, mix well to ensure homogeneity of the sample portion and weigh the specified quantity.
7.1.2 Weigh the sample/slurry (m3) into a 100 ml amber glass screw-cap bottle. See Table 1.
7.1.3 Add 0,15 mol/l sodium phosphate buffer to an approximate volume of 50 ml.
7.1.4 Swirl gently to mix.
7.1.5 Autoclave the sample preparation at 121 °C for 25 mm.
7.1.6 Cool the sample to room temperature. Quantitatively transfer the extract into a 100 ml volumetric flask and make up to the mark with 0,15 mol/l sodium phosphate buffer, mix well.
7.1,7 Transfer the extract into a centrifuge tube and centrifuge the sample at 4 000 rpm for 15 mm.
7.1.8 Filter the sample using the glass microfibre iuiter paper (3) and collect the filtrate.
7.1.9 Set up the SPE manifold (6.5). Attach the immunoaffinmty column (IAC) connected to a 10 ml reservoir. Drain off buffer Just above the gel.
7.1.10 Load the sample filtrate onto the column in accordance with Table 1 and initialize the flow with the help of a vacuum pump.
7.1.11 Turn off the vacuum and let the solution pass through the column by gravity at a rate of one drop per second.
7.1.12 Wash the column by passing 10 ml of PBS (5J.) through the column, followed by 10 ml of water.
Initialize the flow with the help of vacuum at every step and then leave it to flow by gravity.
7.1.13 Remove any residual liquid from the column by introducing a gentle vacuum.
7.1.14 Introduce a reaction vial (6JA) and elute the analyte under gravity with 2 ml methanol. Elute further with an additional 1 ml of methanol. Backflush at least three times when eluting; this can be achieved by a gentle up and down motion of the syringe plunger to maximize the elution.
7.1.15 Evaporate the eluate to dryness using a heating block (&1.5) set at 85 °C ± S °C under a gentle nitrogen blow down.
7.1.16 Remove from the heating block and cool down to room temperature (about 15 mm).
7.1.17 Re-dissolve with 1 ml water and then cap the reaction vial (d4) and vortex for 30 s. Filter by using a syringe filter into a clean glass insert in a HPLC vial for the HPLC analysis.BS ISO 23305 pdf download.

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