AOAC SPIFAN Review Team Meeting Book-July 22, 2015

BVIT-02 FOR ERP/WORKING GROUP USE ONLY DO NOT DISTRIBUTE

Abbott Nutrition Division Simultaneous Determination of Total Vitamin B 6, B 2, B 3 in Infant Formula Products by LC-MS/MS Using Enzymatic Digestion and B 1

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:

[ ] PN

mg

mL g

11.15

/ 31. 124 10 1 8227 .0 m = ××

S = b) Thiamine Hydrochloride: [ ] = • • HCl Cl THI

×

000 ,1 1 = ××× × P M W THI HCl Cl THI • •

10 ×× × PM W

VSSM

mL

100

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:

100

)6.4(

4.95

−

= M

954 .0 = =

100

100

[ ] THI

mg

mL g / 51.69 10 998 .0 954 .0

65.10

m = × × ×

=

S

c) Riboflavin

100 mL P W RIB RIB

000 ,1 11

××××

[ ]

P W

10 ×× =

=

VSSM

RIB

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)

(mL) Volume IWSS Volume(µL) Standard Stoc

ng

mL

1000 ] [

1000 1

Vit

Vit

IWSS ] [

=

× × ×

S

g

L

m

m

mL µL Volume StockStd 10

) (

Vit

S ] [

=

×

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

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