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FOR EXPERT REVIEW PANEL USE ONLY
NOT FOR DISTRIBUTION
1
Expert Review Panel for Infant Formula and Adult Nutrition
Evaluation of Method
BVit-02
(Added information for Vit B3) #3
Title:
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
Author:
Abbott Nutrition
Summary of Method:
This is a method utilizing an enzymatic digestion and LC/MS analysis for the determination of the
total content of all the vitamins of this group
Powder samples were first reconstituted in water at 10% (w/w). Two grams of the reconstituted
sample was weighed into 50 mL disposable centrifuge tubes, internal standard was added, and
5mL of a three enzyme cocktail was added. The enzyme cocktail was prepared in 50 mM
ammonium formate solution adjusted to a pH between 4.0 and 4.5. The samples were incubated
in a 37° water bath and shaken for a minimum of 12 hours. Upon removal from the bath, the
samples were filled to 30 mL with 50 mM ammonium formate solution. The samples were then
mixed thoroughly and filtered through 0.20 μm PTFE syringe filters into autosampler vials.
Method Scope/Applicability:
This method is applicable to determination of vitamin B1, B2, B3 and B6 in infant formulas.
General comments about the method:
The method is reported to perform well and meets all the SMPR requirements. The method demonstrates
high specificity.
Method Clarity:
The method is clear, descriptive and easy to follow in its current format.
Supporting Data
BVit-02 Review Forms