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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.
All 11 SPIFAN matrices met precision and
accuracy requirements for both choline and
carnitine.
1
All analytes defined in the
applicability statement are
measured.
Determination of suppl. and endogenous L-carnitine.
Methods must be able to determine free and total
carnitine... free and bound forms of choline.
Method determines free and total choline and
free and total L-carnitine (acetyl carnitine is
specifically targeted)
1
Total Choline
2-250 mg choline ion/100g RTF
4- 51 was the range of the materials tested
Total Carnitine
0.16 - 20 mg carnitine/100g RTF
0.9-22 was the range encountered
Choline
0.7 mg choline ion/100g RTF
Proven at same limit (see Table 5)
Carnitine
0.05 mg carnitine/100g RTF (implied)
Proven at same limit (see Table 5)
Analyte 3
Analyte 4
Analyte 5
Choline
2.0 mg choline ion/100g RTF
Proven at same limit (see Table 5)
Carnitine
0.16 mg carnitine/100g RTF
Proven at same limit (see Table 5)
Analyte 3
Analyte 4
Analyte 5
Spike recovery (%)
90-110 % mean spiked recovery over the Range (both
choline and carnitine)
choline avg. = 98%, low 90.5%
carnitine avg = 98%, low 95%
acetylcarn avg = 96%, low 90%
2
Bias vs SRM
not stated; note confusion on carnitine certificate
94.2% of choline ion certified mean
110.3% of "free" carnitine certified (by MW)
3
Free Choline
< 10% (2-20); < 5% (20-250)
average 1.3%, high 2.0%
Free Carnitine
< 8%
average 1.5%, high 2.0%
Total Choline
< 10% (2-20); < 5% (20-250)
average 1.3%, high 2.3%
Total Carnitine
< 8%
average 1.2%, high 1.7%
Free Choline
not stated
average 2.2%, high 5.4%
Free Carnitine
not stated
average 2.3%, high 3.8%
Total Choline
not stated
average 1.9%, high 3.0%
Total Carnitine
not stated
average 1.8%, high 2.2%
Choline
< 15% (2-20); < 10% (20-250)
not tested
Carnitine
< 15%
not tested
b
Units
Limit of quantification (LOQ)
Method Reference #
CARN-05
Method title:
Principle of the method:
Carnitine/Choline by LC-MS/MS
Free (dilution) and/or total choline and carnitine (microwave prep) by LC-MS/MS with stable isotope IS
Notes:
Overall Score
Intermediate Reproducibility
(RSDiR)
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
Repeatability (RSDr)
Reproducibility (RSDR)
Accuracy/Recovery
ADDITIONAL EVALUATION PARAMETERS
SMPR Requirement
Analytical Range.
1
AOAC SMPR: 2012.010
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
Laboratory safety
Other Considerations
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
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.
SPIFAN ERP Checklist v 1.6
27.06.2013
Carn-05 MEF Form
FOR ERP USE ONLY
DO NOT DISTRIBUTE