BS EN 16877 pdf free download
BS EN 16877 pdf free download. Animal feeding stuffs: Methods of sampling and analysis 一 Determination of T-2 and HT-2， toxins, Deoxynivalenol and Zearalenone, in feed materials and compound feed by LC-MS.
5.11 Syringe filter:
Small internal volume, Nylon, Pore size: 0,2 urn Nylon.
5.12 LC-MS:
5.12.1 Solvent delivery system capable of delivering a binary gradient at flow rates appropriate for the analytical column in use with sufficient accuracy.
5.12.2 Auto liquid sampler (ALS) capable of injecting an appropriate volume of injection solution with sufficient accuracy, cross-contamination below 0,1 %.
5.12.3 Analytical column capable of separating the four analytes with the following performance:
Peak asymmetry factor at 10 % height: 0,9 <As < 1,4; minimum apparent retention factor for any of the four analytes: k 2; minimum plate number for any of the four analytes: N 1200; minimum resolution between two adjacent analyte peaks: Rs 4.
5.12.4 Mass spectrometer:
An instrument capable of performing selected reaction monitoring (SRM) with a sufficiently wide dynamic range. Any ionization source giving sufficient yield may be employed.
5.13 Balance with readability d = 0,00 1 g or better.
6 Procedures
6.1 Sample preparation
Laboratory samples should be taken and prepared in accordance with European legislation ([1], [2]) where applicable or, in any other case, with EN ISO 6498. The laboratory sample should be finely ground and thoroughly mixed using a mill (5.1) and a mixer (5.2) or another process for which complete homogenization has been demonstrated before a test portion is removed for analysis.
In all instances everything should be at room temperature before any kind of manipulation takes place.
6.2 Extraction
Some of the steps described below are more critical for the accuracy of the results than others. These steps are marked as such and should be carried out with the necessary attention. A scale-up of the test portion size is deemed to be acceptable if such a need is assumed. In that case the amounts of added water, ethyl acetate, and sodium sulphate need to be increased at the same rate, f.i. scale-up by factor of 2: 4 g test portion, 16 ml water, 32 ml ethyl acetate, 16 g sodium sulphate. In no way shall a scale-up be seen as replacement for proper sample preparation (6.1).
— For the test portion weigh 1,9 to 2,1 g of the homogeneous sample into a conical polypropylene screw-cap tube (5.3), round and record the weight to the second decimal (the accuracy of this weight is critical for the accuracy of the final result!).
— Add 7,2 to 8,8 ml of deionized water (4.1).
7 Measurements
7.1 General
The LC-MS system shall meet the requirements laid out in 5.12 and its sub-entries.
7.2 LC conditions
Choose an analytical column, mobile phase, gradient settings, and injection volume that let you meet the requirements in 5.12.3 (for examples see Annex B).
7.3 MS conditions
Choose an ion source with sufficient ionization yield for the four analytes and ion source settings such that a stable spray is achieved.
Choose for each analyte an appropriate precursor ion (adducts of the molecule with a Proton, Sodium, Ammonia, etc. in positive mode, or deprotonation, etc. in negative mode). If more than one precursor ion per analyte is detectable choosing the strongest is a good starting point. But one shall be aware that the choice of precursor ion will affect repeatability and, by that, LOD and LOQ.
For SRM select two product ions in the MS/MS spectrum for each selected precursor ion of each analyte. Set up SRM transitions with these precursor/ product ion combinations (for SRM examples see Annex B).
The selected MS settings shall be such that for a relevant feed material with a contamination of ca.
90 .tg/kg DON, 10 gig/kg HT-2 toxin, 10 rig/kg T-2 toxin, and 10 .ig/kg ZON, prepared according to
Clause 6, signal-to-noise ratios (peak-to-peak) of larger than 10 are obtained (see Annex C).BS EN 16877 pdf download.
