232 / 620
Simultaneous Determination of Thiamine HCl, Riboflavin, Niacin (Nicotinic Acid and Nicotinamide),
Pantothenic acid, Vitamin B6 (Pyridoxine, Pyridoxal and Pyridoxamine) and Biotin in Infant
Formula and Adult Nutritionals
CONFIDENTIAL METHOD FOR ERP ASSESSMENT ONLY
PAGE 10 OF 13
IV. CALCULATION and REPORT
The LCMSMS software will automatically calculate the concentration of the sample in ng/mL for Thiamin
HCl, Riboflavin, Nicotinic Acid, Nicotinamide, Pantothenic acid, Pyridoxine, Pyridoxal, Pyridoxamine and
Biotin, provided the concentration of the working standards in ng/mL, sample weight / powder equivalent
(g) or sample volume (mL), dilution factor and conversion to 100g or 100 mL are entered correctly. Report
in 2 decimal places or as customer requirement.
100
1000
xDFx
W
CxV
))ml or(g 100 / mcg ( WSV .
Conc
Where:
C
=
Concentration of water-soluble vitamin from working curve (ng/mL)
V
=
Sample volume (ml)
= 100 mL
DF
=
Dilution Factor
= 1/0.5
W
=
Sample weight (2.78 g for powder or 15 g Liquid, or 1 g for SRM1849a) or sample
volume (15 mL)
1000
=
conversion factor from
ng to mcg
100
=
conversion factor from
g to 100 g or from mL to 100 mL
V. DISCUSSION
a. Niacin (mcg/100g(or mL)) = Nicotinamide(mcg/100g(or mL))+Nicotinic acid (ng/100g (or
mL))
b. Vitamin B6 (mcg/100g(or mL)) = Pyridoxine (mcg/100g(or ml)) + Pyridoxal (mcg/100g(or
ml)) + Pyridoxamine (mcg/100g(or mL))
c. Vitamin B6 (as Pyridoxine) (mcg/100g(or mL)) = Pyridoxine (mcg/100g(or ml)) + Pyridoxal
(mcg/100g(or ml))x(169.18/167.162) + Pyridoxamine (mcg/100g(or mL))x(169.18/168.19)
Where
Molecular weight of Pyridoxine = 169.18 g/mol
Molecular weight of Pyridoxal = 167.162 g/mol
Molecular weight of Pyridoxamine = 168.19 g/mol
d. If Thiamine (C
12
H
17
N
4
OS) is reported, Thiamine Hydrochloride (C
12
H
18
C
l2
N
4
OS) is
converted by multiplying with 0.787.
Thiamine (mcg/100g (or mL) = Thiamine HCl (mcg/100g (or mL)) x 0.787
Note : 0.787 = MW of Thiamine (265.4 g/mol)/MW of Thiamine HCl (337.3 g/ mol)
e. Comments:
Comment No.
1
The gradient given can be altered as required to maximize the analytical separation and
avoid interferences. This should only be done by an authorized analyst.
BVit-01
FOR ERP USE ONLY
DO NOT DISTRIBUTE