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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

Example:

The Pyridoxine standard weight is 15.11 mg is 100% pure. The Pyridoxine stock

concentration, [PN]

S

is 124.31

m

g/mL and is calculated as follows:

[ ]

mL g

mg

PN

S

/

31. 124

10 1 8227 .0

11.15

m

= ××

×

=

b)

Thiamine Hydrochloride:

[ ]

10

100

000 ,1 1

×× ×

=

××× ×

=

• •

• •

PM W

mL

P M W THI

HCl

Cl

THI

HCl

Cl

THI

VSSM

Example:

The Thiamine Hydrochloride standard weight is 10.65 mg is 99.8% pure and the

standard is found to have 4.6% moisture. The Thiamine stock concentration,

[THI]

S

,

is 69.51

m

g/mL and is calculated as follows:

954 .0

100

4.95

100

)6.4(

100

= =

=

M

[ ]

mL g

mg

THI

S

/

51.69 10 998 .0 954 .0

65.10

m

= × × ×

=

c)

Riboflavin

[ ]

10

100

000 ,1 11

×× =

××××

=

P W

mL

P

W RIB

RIB

RIB

VSSM

Example:

Riboflavin standard weight is 7.12mg and is 100% pure. The Riboflavin

concentration in the Vitamin Standard Stock Mixture, [RIB]

VSSM

is 71.2

m

g/mL

and is calculated as follows:

[ ]

mL g

mg

RIB

VSSM

/ 2.71 10 00.1

12.7

m

= × ×

=

4.

Calculation of the Intermediate Working Standard Solution (IWSS)

S

S

IWSS

Vit

mL

µL

Volume

StockStd

L

mL

g

ng

Vit

(mL)

Volume

IWSS

Volume(µL)

Standard

Stoc

Vit

] [

10

) (

1000

1

1000

] [

] [

×

=

×

× ×

=

m

m

where:

[Vit]

IWSS

: Concentration of the vitamin of interest in the Intermediate Working Standard

Solution, in ng/mL

[Vit]

S

: Vitamin Stock Standard Concentration, in

m

g/mL

39

BVit-02

FOR ERP USE ONLY

DO NOT DISTRIBUTE