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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 USE ONLY

DO NOT DISTRIBUTE