BS ISO 21470:2020 Infant formula and adult nutritionals — Simultaneous determination of total vitamins B 1 , B 2 , B 3 and B 6 — Enzymatic digestion and LC-MS/MS

BS ISO 21470:2020 Infant formula and adult nutritionals — Simultaneous determination of total vitamins B 1 , B 2 , B 3 and B 6 — Enzymatic digestion and LC-MS/MS.
5 Reagents and materials
During the analysis, unless otherwise stated, use only reagents of recognized analytical grade and distilled or demineralized water or water of equivalent purity.
5.1 Niacinamide (nicotinamide) (MW = 122,12), primary reference standard, e.g. USP Reference Standard, catalogue #14620061). Follow the manufacturer’s storage and handling directions.
5.2 Niacin (nicotinic acid) (MW = 123,11), primary reference standard, e.g. LiSP Reference Standard, catalogue # 14610031). Follow the manufacturer’s storage and handling directions.
5.3 Pyridoxine hydrochloride (MW = 205,64), primary reference standard, e.g. USP Reference Standard, catalogue 4 15870011). Follow the manufacturer’s storage and handling directions.
5.4 Riboflavin (MW = 376,36), primary reference standard, e.g. USP Reference Standard, catalogue 4
16030061). Follow the manufacturer’s storage and handling directions.
5.5 Thiamine hydrochloride (MW = 337,27), primary reference standard, e.g. USP Reference Standard, catalogue #1656002 1) Follow the manufacturer’s storage and handling directions. Measure the moisture content of the powder prior to use or use the supplier certificate of analysis (COA) moisture value.
5.6 Pyridoxamine dihydrochioride, Fluka Analytical Standard, catalogue #P93801).
5.7 Pyridoxal hydrochloride, Sigma, catalogue #P9130’).
5.8 2H4-Niacinamide, CDN Isotopes, catalogue #D-34571).
5.9 2H4-Nicotinic acid, CDN Isotopes, catalogue #D-43681).
5.10 ‘3C4-Pyridoxine: pyridoxine:HCI (4,5-bis(hydroxymethyl)-’3C4), Cambridge Isotope Laboratory, catalogue #CLM -7563’).
5.11 2H3-Pyridoxal, lsoSciences, catalogue #7098’).
5.12 2H3-Pyridoxamine, IsoSciences, catalogue #70991).
5.13 13C4-Thiamine chloride, IsoSciences, catalogue #9209 ).
5.14 13C4,15N2-Riboflavin, IsoSciences, catalogue #70721).
5.15 Acid phosphatase, type II from potato, 0,5 U/mg to 3,0 U/mg, Sigma, catalogue #P3752’).
5.16 Papain from Carica papaya,  3 U/mg, Sigma, catalogue #762201).
5.17 a-amylase from asperqillus oryzae, 150 U/mg, Sigma, catalogue #A9857fl.
5.18 Hydrochloric acid concentrated (substance concentration c = 12 mol/l), ACS grade, or equivalent.
5.19 Ammonium formate, for mass spectrometry (purity 99,0 %), Fluka 70221 or equivalent’).BS ISO 21470 pdf download.