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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-04
Title: LC-MS/MS analysis of nicotinic acid and nicotinamide
Author:
S. Bhandari & T. Gallegos (Silliker, Inc. Chemistry Research Center)
Summary of Method:
LC-MS/MS method for the analysis of vitamin B3 or niacin (defined as the sum of nicotinic acid and
nicotinamide). The extraction is performed by action of protease and takadiastase to release bound forms
in an ammonium formate buffer (pH 5).
After addition of labelled internal standards, the extracts are analysed using liquid chromatography with
tandem mass spectrometric detection.
Method Scope/Applicability:
The method is applicable to the matrixes in the scope of SPIFAN.
General comments about the method:
The method is, in general clearly written and should be easy to apply. Both compounds are extracted and
analysed simultaneously.
The chromatography seems suitable to analyse most of B-vitamins.
Method Clarity:
There are a few points that should be clarified in the method:
a)
Harmonise naming of compounds through the documents is needed. Definition of niacin to match
SMPR as nicotinic acid and nicotinamide:
-
In point 4.9.3 vitamin B3 is defined as niacin and niacinamide, it should read nicotinic acid and
nicotinamide
-
Point 4.9.6 may also not be clear for labs not familiar with vitamins analysis, it mixes niacinamide
and nicotinamide
b)
Point 4.4.3 is a repetition of point 4.5 e). Please review
c)
4.7.2-10 Final volume of 40 mL should be as precise as possible, since internal standards are not
added but after this point, could this be reviewed? Would it be possible to add either a ± range? Or
transfer to 20 or 50 mL volumetric flask and make up to volume?