407 / 620
Parameter
Method Performance
Please
report in units as stated in SMPR!
Weighting
factor for
parameter
Suitability Ranking (1-
3-5)
(select from drop-down
list, 5 = best)
SPIFAN matrices
Applicable to all forms of infant, adult, and/or pediatric
formula (powders, ready-to-feed liquids, and liquid
concentrates).
Any combination of milk, soy, rice, whey, hydrolyzed
protein, starch, and amino acids, with and without intact
protein.
1
5
All analytes defined in the
applicability statement are
measured.
Folic acid and 5-methyl-tetrahydrofolic acid
Folic acid and 5-methyl-tetrahydrofolic acid, free
and protein bound. Polyglutamate forms are
excluded.
1
5
Analytical Range.
0.5-300 mcg/100 g
0.3-300 mcg/100 g
1
5
Folic acid
0.1 mcg/100 g
5-Me-THF
0.05 mcg/100g
Folic acid
0.3 mcg/100 g
5-Me-THF
0.1 mcg/100 g
Spike recovery (%)
90-110 %
89-114 %
2
5
Bias vs SRM
No bias
No bias detected (95 % confidence level)
3
5
Repeatability (RSDr)
RSD r ≤ 7%
≤ 5.2 %
3
5
Intermediate Reproducibility
(RSDiR)
not given
≤ 5.4 %
1
5
Reproducibility (RSDR)
1
5
3
5
5
b
Units: mcg/100 g
Method Reference #
Method title:
Principle of the method:
Free folates (Vitamin B9) in Infant Formula and Adult/Pediatric Nutritional Formula by UHPLC-
MS/MS
Folic acid is and 5-methyltetrahydrofolic acid are extracted from the sample using amylase/heat/protease
treatment in the presence of dithoithreitol and ascorbic acid as antioxidants. After addition of isotope
labelled internal standards, clean-up is performed on a strong anion exchange cartridge. Folic acid and 5-
methyl tetrahydrofolic acid are then determined by liquid chromatography with MS/MS detection.
Limit of detection (LOD)
1
5
Limit of quantification (LOQ)
< 0.1 mcg/100 g
< 0.5 mcg/100 g
2
5
Notes:
Overall Score
Recommendation of ERP
2 years after First Action Status
Evaluation of Method Performance vs. SMPR requirements.
Feedback from Users of the Method since being awarded
First Action
Official Methods
Status
Accuracy/Recovery
Repeatability of injection and retention time is set as system suitability
ADDITIONAL EVALUATION PARAMETERS
SMPR Requirement
AOAC SMPR: 2012.009
Bias against established method
Analytical equipment
Adequate proof of performance via system suitability
Is there a bias Yes/No ?
Analytical equipment is commonly available in most labs.
a
Concentrations apply to (1) "ready to feed liquids" "as-is"; (2) reconstituted powders (25 g into 200 g water); and (3) liquid concentrates diluted 1:1 by weight.
Did Method Author Consider ERP’s Method Specific
Recommendations
(See web link to specific method
comments):
Decision by ERP
Proprietary equipment
Laboratory safety
Other Considerations
move to Final Action/repeal/remove/expand 2 year term
93
No unique proprietary equipment/accessories are required.
Method does not require any special safety precautions e.g. personal protection from highly toxic solvents.
SPIFAN ERP Checklist v 1.6
27.06.2013
2013.13 (Fol-21) MEF Form
FOR ERP USE ONLY
DO NOT DISTRIBUTE