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CAROT-03 (FEBRUARY 2017) - MEF FORM
FOR ERP USE ONLY
DO NOT DISTRIBUTE
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
All analytes defined in the
applicability statement are
measured.
Method Measures cis and trans lutein, cis and trans beta-
carotene, and cis and trans lycopene.
1
Lutein
1-1300 mcg/100g RTF
1-1300 mcg/100g RTF
beta-Carotene
1-1300 mcg/100g RTF
1-1300 mcg/100g RTF
Lycopene
1-1300 mcg/100g RTF
1-1300 mcg/100g RTF
Lutein
0.2 mcg/100g RTF
NA
beta-Carotene
0.04 mcg/100g RTF
NA
Lycopene
0.1 mcg/100g RTF
NA
Lutein
0.4 mcg/100g RTF
1 mcg/100g RTF
beta-Carotene
0.1 mcg/100g RTF
1 mcg/100g RTF
Lycopene
0.3 mcg/100g RTF
1 mcg/100g RTF
Spike recovery (%)
89.3-108.8% See Table I
90-100%
2
Bias vs SRM
NA
NA
3
Lutein
Average of matrices above 1 mcg/100g , 4.13%
1-100 mcg/100g RTF
8%
beta-Carotene
Average of matrices above 1 mcg/100g , 4.10%
1-100 mcg/100g RTF
8%
Lycopene
Average of matrices above 1 mcg/100g , 4.60%
1-100 mcg/100g RTF
8%
See Table III
Lutein
Average of matrices above 1 mcg/100g, 5.63%
NA
beta-Carotene
Average of matrices above 1 mcg/100g , 6.92%
NA
Lycopene
Average of matrices above 1 mcg/100g , 6.89%
NA
See Table III
b
Units
Bias against established method
Analytical equipment
1
Adequate proof of performance via system suitability
Is there a bias Yes/No ?
Analytical equipment is commonly available in most labs.
move to Final Action/repeal/remove/expand 2 year term
0
No unique proprietary equipment/accessories are required.
Method does not require any special safety precautions e.g. personal protection from highly toxic solvents.
3
1
2
AOAC SMPR: …………..
Limit of detection (LOD)
1
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
Notes:
Overall Score
Recommendation of ERP
2 years after First Action Status
Laboratory safety
Other Considerations
Evaluation of Method Performance vs. SMPR requirements.
Feedback from Users of the Method since being awarded
First Action
Official Methods
Status
Repeatability (RSDr)
Reproducibility (RSDR)
Accuracy/Recovery
ADDITIONAL EVALUATION PARAMETERS
Intermediate Reproducibility
(RSDiR)
Limit of quantification (LOQ)
Method Reference #
Analytical Range.
1
Method title:
Principle of the method:
Determination of beta-Carotene, Lutein and Lycopene in Infant and Adult/Pediatric Nutritional Formula Using High performance Liquid
Chromatography with UV Detection.
This HPLC method uses C30 chromatography and ultraviolet-visible spectroscopy to separate and detect cis and trans isomers of lutein, beta-carotene, and lycopene in infant, pediatric, and adult nutritionals. Carotenoids
are extracted from samples with 75:25 hexane:methyl tertiary butyl ether (MtBE) after saponification with a mixture of potassium hydroxide, tetrahydrofuran (THF), and methanol. After extraction, a portion of the organic
layer is evaporated to dryness and the residue is dissolved in 75:25 10% butylated hydroxytoluene (BHT):MtBE. Prepared samples are injected onto a C30 HPLC column where cis and trans isomers of lutein, beta-carotene,
and lycopene are separated with a methanol-MtBE gradient and detected by ultraviolet-visible spectroscopy at 445 nm.
SMPR Requirement
SPIFAN ERP Checklist v 1.0