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Abbott Nutrition Division
Simultaneous Determination of Total Vitamin B
6,
B
2,
B
3
and B
1
in Infant
Formula Products by LC-MS/MS Using Enzymatic Digestion
D.
CHEMICALS, STANDARDS AND SOLUTIONS
1.
Chemicals/Standards
a)
2
H
3
-Pyridoxal: Isosciences; Catalog # 7098.
b)
2
H
3
-Pyridoxamine: Isosciences; Catalog # D-7099.
c)
13
C
4
-Pyridoxine: Pyridoxine:HCl (4,5-BIS(hydroxymethyl)-
13
C
4
); Cambridge Isotope
Laboratory; Catalog # CLM-7563-0
d)
13
C
3
-Thiamine: Thiamine Chloride (4,5,4-methyl-
13
C
3
); Cambridge Isotope Laboratory; Catalog
# CLM 7667-0.
e)
13
C
4
,
15
N
2
-Riboflavin: IsoSciences
f)
2
H
4
-Niacin: CDN Isotopes; Catalog # D-3457.
g)
2
H
4
-Nicotinic acid: CDN Isotopes; Catalog # D-4368.
h)
Niacinamide: USP Reference Standard (U.S.P.C., Inc. Rockville, MD) Official Lot; Catalog #
1462006. Store as indicated on label.
i)
Niacin (nicotinic acid): USP Reference Standard (U.S.P.C., Inc. Rockville, MD) Official Lot;
Catalog # 1461003. Stored as indicated on label.
j)
Pyridoxine Hydrochloride, USP Reference Standard #1587001, current lot. Store as indicated on
label.
k)
Pyridoxal hydrochloride, Sigma P9130
l)
Pyridoxamine dihydrochloride >99%, Fluka 82885
m)
Riboflavin: USP Reference Standard (U.S.P.C., Inc. Rockville, MD) Official Lot; Catalog #
1603006. Store in desiccator protected from white light. Dry according to manufacturer’s
instructions prior to use.
n)
Thiamine Hydrochloride: USP Reference Standard (U.S.P.C., Inc. Rockville, MD) Official Lot;
Catalog # 1656002. Store in desiccator protected from white light. Measure the moisture content
of the powder prior to use.
o)
Formic Acid: Sigma ACS Reagent Grade, or equivalent.
p)
Ethanol (Reagent Alcohol): ACS Reagent Grade, or equivalent.
q)
Control Sample: current lot. Store frozen.
r)
Ammonium Formate ACS grade or equivalent.
s)
Laboratory Water
t)
Methanol: Fisher LC-MS/MS Optima grade or EMD Omni-Solve LC-MS grade. NOTE:
METHANOL MUST BE LC-MS GRADE. Other grades including HPLC grade are not
acceptable for use.
u)
Papain Purified Pwd, Spectrum chemical (PA125), or equivalent
31
BVIT-02
FOR ERP/WORKING GROUP USE ONLY
DO NOT DISTRIBUTE