BS EN ISO 20418-3 pdf free download

BS EN ISO 20418-3 pdf free download. Textiles — Qualitative and quantitative proteomic analysis of some animal hair fibres.
6 Reagents The following analytical grade reagents shall be used. 6.1 Acetone, with purity greater than or equal to 99,5 %. 6.2 Water, grade 3 quality specified in ISO 3696. 6.3 Ammonium hydrogen carbonate (NH 4 HCO 3 ) solution (25 mmol/l) — 197,5 mg of NH 4 HCO 3 , with purity greater than or equal to 96,0 %. — Make up 100 ml by adding water (6.2). 6.4 Trypsin, sequencing-grade porcine trypsin modified by reductive methylation. 6.5 Acetonitrile, with purity greater than or equal to 99,8 %. 6.6 Formic acid, with purity greater than or equal to 98 %. 6.7 Trypsin solution — Trypsin (6.4) 20 μg. — 0,1 % formic acid (6.6) 200 μl. 7 Apparatus The usual laboratory apparatus and, in particular, the following. 7.1 Heating mantle, capable of operating at a temperature range of 50 °C to 150 °C. 7.2 Mill, beads mill, cryogenic grinder or an equivalent, capable of crushing materials into an extremely fine powder. 7.3 Membrane filter, for aqueous solutions, with a pore size of 0,45 µm. 7.4 Heat block, capable of heating microtubes at 37 °C. 7.5 Tube mixer, capable of vortex microtubes and LC vials for about 30 min. 7.6 Centrifugal evaporator, capable to deliver 5 000 g. 7.7 LC-MS, liquid chromatography–mass spectrometer, capable of detecting m/z (u) range from 200 m/z (u) to 1 500 m/z (u). NOTE (u) is for unified atomic mass unit, SI unit. 7.8 LC vial, shall be glass or polymethylpentane. 7.9 LC column, octadecyl (C-18)-silica reversed phase column. 7.10 Balance, with a resolution of at least 0,001 g. 7.11 Recovery flask (eggplant flask or round-bottom flask). 8 Test method 8.1 Sampling The general requirement is that the test specimen shall be representative for the lot of material from which it is taken. The method to obtain a fibre test specimen differs depending on the sample form. The terms relating to sampling for the various types of samples shall be in accordance with ISO 1833-1. 8.2 Preliminary identification The preliminary qualitative analysis of the animal hair fibre shall be carried out based on their morphology, which is determined using light microscopy, according to ISO 17751 (all parts), after removal of non-animal fibre. 8.3 Wash for degreasing 8.3.1 Reflux 1 g of the fibres in a recovery flask (7.11) on a heating mantle (7.1) with 200 ml of acetone (6.1) for 30 min. This washing step may be omitted in the case of clean samples. Quantity of the fibres can be changed. 8.3.2 Take the degreased fibres out of the recovery flask and dry them in the air. Alternatively, the sample preparation method of ISO 20418-1 [5] can be used. 8.4 Powderization of fibres Crush the dried fibre sample (8.3.2) using a mill (7.2) to get a fine powder with an average length of 100 µm or less by checking under the microscope and mix thoroughly for securing representative sampling of the fibres. 8.5 Trypsin digestion 8.5.1 Weigh about 10 mg of the crushed sample and place it into a microtube. If more than 10 mg of the sample is used, increase the volumes of the NH 4 HCO 3 solution in 8.5.2 and Trypsin solution in 8.5.3 proportionally. 8.5.2 Add 300 µl of the NH 4 HCO 3 solution (6.3) and vortex for 10 min to 30 min. 8.5.3 Add 10 µl of the Trypsin solution (6.7) to the sample and incubate at 37 °C for 20 h to 24 h. 8.5.4 Centrifuge the tryptic solution for 3 min using the centrifugal evaporator (7.6). Filter the supernatant through a membrane filter (7.3) to remove residual fibres. NOTE Centrifugal filter, syringe filter or other means of filtration can be used.BS EN ISO 20418-3  pdf download.