SimultaneousDetermination of SevenWater SolubleVitamins inProducts byLC-MS/MS
Page
12
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20
p)
PositiveDisplacement Pipettes: GilsonMicroman—10, 100, 250, and1000μL
q)
Pipet tips.
r)
pHmeter
s)
UV shieldsA.L.P. Protect-A-Lamp, UV cutoff at 460nm.
A.
REAGENTS
1.
Solvents
a)
Methanol.—LCMSgrade.
b)
Hydrochloric acid concentrated (12M); ACSgrade, or equivalent.
c)
L-Ascorbic acid: SigmaBioXtra grade (≥99%), or equivalent.
d)
Formic acid: SigmaACSReagent Grade, or equivalent.
e)
Heptafluorobutyric acid (HFBA): for ion chromatography (>99%), or equivalent.
f)
Glacial acetic acid:Mallinckrodt ACS grade (>99%), or equivalent.
g)
Ammoniumhydroxide (NH4OH): ACS reagent grade (28-30%), or equivalent.
h)
Ethylenediaminetetracetic acid, disodium salt dihydrate (EDTA): ACSgrade (99-101%), or
equivalent.
i)
Potassiumphosphate dibasic: ACS grade (>98%), or equivalent.
j)
meta-Phosphoric acid: ACS grade (33.5-36.5%), or equivalent.
k)
Phosphoric acid, 85%, ACS grade, or equivalent.
l)
Potassiumhydroxide, 40%,ACSgrade, or equivalent.
m)
Distilledor deionized laboratorywater.—Purifiedwater, USP; ≥18Mohm/cm.
2.
Standards
a)
Folic acid;USPReferenceStandard (U.S.P.C., Inc. Rockville,MD)
b)
Biotin: USPReferenceStandard (U.S.P.C., Inc. Rockville,MD).
c)
Calciumpantothenic acid: USPReferenceStandard (U.S.P.C., Inc. Rockville,MD)
d)
Niacinamide: USPReference Standard (U.S.P.C., Inc. Rockville,MD).
e)
Niacin (nicotinic acid): USPReferenceStandard (U.S.P.C., Inc. Rockville,MD)
f)
PyridoxineHydrochloride:USPReferenceStandard (U.S.P.C., Inc. Rockville,MD)
g)
Riboflavin: USPReferenceStandard (U.S.P.C., Inc. Rockville,MD)
h)
ThiamineHydrochloride: USPReferenceStandard (U.S.P.C., Inc. Rockville,MD)Official Lot;
Catalog#1656002. Store as per label directions.
i)
13
C
5
-Biotin: Alsachim; Catalog#C1468.
j)
13
C
5
-Folic acid: L-folic acid-
13
C5;Merck&Cie Schauffhausen.
k)
13
C
3
,
15
N-Pantothenic acid: pantothenic acid (beta-alanyl
13
C
3
15
N); Cambridge IsotopeLaboratory;
Catalog#CNLM-7694.
l)
2
H
4
-Niacin: CDN Isotopes; Catalog#D-3457.
m)
2
H
4
-Nicotinic acid: CDN Isotopes; Catalog#D-4368.
n)
13
C
4
-Pyridoxine: pyridoxine:HCl (4,5-bis(hydroxymethyl)-
13
C
4
); Cambridge IsotopeLaboratory;
Catalog#CLM-7563.
o)
13
C
4
-Thiamine chloride; IsoSciences; Catalog#9209.
p)
13
C
4
,
15
N
2
-Riboflavin: IsoSciences, Catalog# 7072.
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