EN ISO 20418-3:2020 pdf download.Textiles – Qualitative and quantitative proteomic analysis of some animal hair fibres – Part 3: Peptide detection using LC-MS without protein reduction (ISO 20418-3:2020)
6.1 Acetone, with purity greater than or equal to 995 %.
6.2 Water, grade 3 quality specifed in Iso 3696.
6.3 Ammonium hydrogen carbonate (NH,HCO3) solution (25 mmol/I)
一197,5 mg of NH,HCO3 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 mdified by reductive metylation.
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限.
一0,1 % frmic acid (6.6) 200 l
7 Apparatus
The usual laboratory apparatus and, in particular, the fllowing.
7.1 Heating mantle, capable of operating at a temperature range of50 C to 150 °C.
7.2 Mill, beads mill, cryogenic grinder or an equivalent, apable of crushing materials into an extremely
fine powder.
7.3 Membrane filter, for aqueous slutions, with a pore sie of 0.45 pm.
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 Centrifuga evaporator, capable to deliver 5 000 g-
7.7 LC-MS. liquid chromatography mass spectrometer, capable of detecting m/z (司) range from
200 m/z (田) to 1 500 m/z ().
NOTE
(uis for unifed atomic mass unit. SI unit
7.8 LC vial, shall be glass or polymethylpentane.
7.9 LC column, octadeeyl (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 requtrement is that the test specimen shall be representative for the lot of materlal from which it is taken. The method to obtain a fibre test specimen dffers depending on the sample form. The terms relating to sampling for the varlous types of samples shall be in accordance with IS0 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 IS0 17751 (all parts]. after removal of non-animal fibre.EN ISO 20418-3 pdf download.