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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 FOL-20 #2
Title:
Analysis of Folic acid and 5-Methyltetrahydrofolate in Infant and Adult Nutritional formula using
Ultra-Performance Liquid Chromatography-Tandem Mass Spectrometry
Author:
Maria V. Chandra‐Hioe, Martin P. Bucknall, and Jayashree Arcot
Summary of Method:
Folate extracted by heating in a boiling water bath, followed by cleanup using SPE and Amicon membrane
filters. Determination by UPLC-MS/MS using 13C5 labeled internal standards.
Method Scope/Applicability:
Applicable to all infant formula matrices. Demonstrated for SPIFAN II matrices. Note from method authors
indicates that the method is not rugged for high fat/high protein SPIFAN matrices.
General comments about the method:
Seems straightforward and logical. Removes need for triple enzyme treatment and qualifications that go
along with that approach (cleaning and/or validating cleanliness of enzymes, evaluating activity of enzymes,
etc.). Doesn’t address 5-MTHF polyglutamate, which is a grey area in the SMPR. Folic acid detected in 2 of
the placebo materials, which seems strange and may indicate an interference.
Method Clarity:
The procedure is clearly written and described.
Method Safety Concerns:
None
Pros/Strengths:
•
Does not require enzyme treatment
Cons/Weaknesses
•
Values for SRM 1849a are very high for both folic acid and 5-MTHF
•
Values do not agree with (are much higher than) tri-enzyme method
•
Performance characteristics do not meet SMPR
Supporting Data
•
General Comment: the source of the validation data is not clearly described. Only values are reported
in each SPIFAN matrix compared to the tri-enzyme treatment; nothing about recovery from each
matrix. Values are highly variable between the heat treatment and tri-enzyme treatment.
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Method Optimization: not described
Fol-20 Review Forms