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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.
Trans Vitamin K
1
Method measures all required analytes.
1-100 mcg/100g reconstituted final product
0.2-8 mcg/100 mL range of standard curve
> 1 mcg/100g ready-to-feed product
Trans vitamin K1
Not Applicable
~0.1 mcg/L in prepared sample OR
~0.03 mcg/100 g in ready-to-feed product
Trans vitamin K1
1 mcg/100g reconstituted final product
~0.4 mcg/L in prepared sample OR
~0.09 mcg/100 g in ready-to-feed product
Spike recovery
90-110%
Range: 91.9-106.1% See Table 1
Bias vs SRM
See Table 2
1-10 mcg/100g reconstituted final product :
%
Range: 1-6.8% See Table 3
>10 mcg/100g reconstituted final product :
5%
1.8% See Table 3
No SMRP requirements
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
Evaluation of Method Performance vs. SMPR requirements.
VitK-02
Determination of trans vitamin K1 in infant formula and adult nutritionals by HPLC
This normal phase HPLC method with post column reduction and fluorescence detection allows for the quantitative determination of trans vitamin K1 in infant, pediatric, and adult nutritionals. Vitamin
K1 is extracted from products with iso-octane after precipitation of proteins and liberation of lipids with methanol. Prepared samples are injected onto a silica HPLC column where cis and trans vitamin
K1 are separated with an isooctane/isopropanol mobile phase. The column eluant is mixed with a dilute ethanolic solution of zinc chloride, sodium acetate, and acetic acid and vitamin K1 is reduced to a
fluorescent derivative in a zinc reactor column. The resulting fluorescent compound is then detected by fluorescence at an excitation wavelength of 245 nm and an emission wavelength of 440 nm.
Feedback from Users of the Method since being
awarded First Action
Official Methods
Status
Repeatability (RSDr)
Reproducibility (RSDR)
Accuracy/Recovery
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: 2014.001
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 7% . 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.
Intermediate Reproducibility
(RSDiR)
SMPR Requirement
Analytical Range.
Adequate proof of performance via system suitability
Limit of detection (LOD)
Limit of detection (LOQ)
SPIFAN ERP Checklist v 1.0