Method Reference #
Method title:
Principle of the method:
Parameter
Method Performance
Please report
in units as stated in SMPR!
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.
All SPIFAN matrices analyzed during SLV.
All required analytes are
measured.
cyanocobalamin, aquocobalamin, hydroxycobalamin, methylcobalamin, and
adenosylcobalamin
Method measures all required analytes.
0.01-5.0 mcg/100g reconstituted final product
0.25-2.5 mcg/100 mL range of standard curve
> 0.025 mcg/100g reconstituted final product
vitamin B12
0.004 mcg/100g reconstituted final product
~0.025 mcg/100g reconstitued final product
vitamin B12
0.01 mcg/100g reconstituted final product
~0.025 mcg/100g reconstitued final product
Spike recovery
90-110%
See Table 1
Bias vs SRM
See Table 2
0.01 mcg/100g reconstituted final product :
15%
See Table 3
0.2, 0.5, 5.0 mcg/100g reconstituted final product :
15%
No SMRP requirements
0.3,0.6,1.0,2.5,5.0 mcg/100g in reconstituted final product :
11% See Table 4
Analytical equipment
Proprietary equipment
Laboratory safety
Other Considerations
Overall Score
0
Decision by ERP
Recommendation
ERP 2 years after
First Action Status
move to Final Action/repeal/remove/expand 2 year term
Notes
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.
b
Units
Intermediate Reproducibility
(RSDiR)
SMPR Requirement
Analytical Range.
Adequate proof of performance via system suitability
Limit of detection (LOD)
Limit of detection (LOQ)
Did Method Author Consider ERP’s Method Specific
Recommendations
(See web link to specific method
comments):
Analytical equipment is commonly available in most labs.
No unique proprietary equipment/accessories are required.
Method does not require any special safety precautions e.g. personal protection from highly toxic solvents.
AOAC SMPR: 2011.005
Peak responses of precision injections of the high working standard must have an RSD of ≤ 2.0%. At each working standard concentration,
the peak areas of standards injected at the beginning and end of a set of samples should not increase or decrease by more than 10% . A
standard curve must have a correlation of at least 0.999 to be considered acceptable for sample calculations. For each set of samples, the
control result must be within 3 standard deviations of the control means.
See comments tab
Evaluation of Method Performance vs. SMPR requirements.
2011.10
Determination of vitamin B12 in infant formula and adult nutritionals by HPLC
Vitamin B12 is extracted from the sample using sodium acetate
buffer (pH 4.5) and potassium cyanide at 105°C. Extracts are
purifi ed and concentrated with C8 or C18 solid-phase extraction
(SPE) cartridges and analyzed with size-exclusion and reversedphase
chromatography. Determination of B12 is made by liquid
chromatography with visible detection at 550 nm.
Feedback from Users of the Method since being
awarded First Action
Official Methods
Status
Repeatability (RSDr)
Reproducibility (RSDR)
Accuracy/Recovery
SPIFAN ERP Checklist v 1.0
