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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.
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
Total B1 defined as the sum of thiamin (CAS No. 70-16-6),
thiamin phosphate (CAS No. 532-40-1), thiamin
diphosphate (CAS No. 154-87-0), and thiamin triphosphate
(CAS No. 3475-65-8). All data should be mass corrected and
expressed as thiamin ion.
Thiamine (from Thiamine hydrochloride)
1
Total B2 defined as the sum of riboflavin (CAS 83-88-5),
riboflavin 5ʹ-phosphate (CAS 146-17-8), and flavin adenine
dinucleotide (CAS 146-14-5). All data should be mass
corrected and expressed as riboflavin.
Riboflavin
Total B3 defined as the sum of nicotinic acid (CAS No. 59-
67-6) and nicotinamide (CAS No. 99586-65-9), and reported
as niacin.
Nicotinc acid and Nicotinamide
Pantothenic acid 3-[(2,4-dihydroxy-3,3-
dimethylbutanoyl)amino]
Pantothenic acid
Total B6 defined as the sum of pyridoxine(CAS No. 65-23-
6), pyridoxal (CAS No. 66-72-8), pyridoxamine (CAS No. 85-
87-0), and the 5' phosphorylated forms, pyridoxal 5'
phosphate (CAS No. 54-47-7) and pyridoxamine 5'
phosphate (CAS No. 3475-65-8). All data should be mass
corrected and expressed as pyridoxine.
Pyridoxine, Pyridoxal and Pyridoxamine
d-Biotin.—5-[(3aS,4S,6aR)-2-oxohexahydrothieno[3,4-
d]imidazol-4-yl]pentanoic acid
Biotin
Specificity
No interfering peaks were observed for any
analysis in any matrices.
Thiamine
20-2500
20-2500
Riboflavin
30-4000
30-2500
Nicotinic acid
25-400
Nicotinamide
400-16000
Pantothenic acid
50-2300
30-9000
Pyridoxine
25-2000
Pyridoxal
3-300
Pyridoxamine
1-100
Biotin
0.1-150
3-100
Analyte 1: Thiamine
-
12
Analyte 2: Riboflavin
-
12
Analyte 3: Nicotinic acid
-
8
Analyte 4: Nicotinamide
-
200
Analyte 5: Pantothenic acid
-
25
Analyte 6: Pyridoxine
-
12
Analyte 7: Pyridoxal
-
1.5
Analyte 8: Pyridoxamine
-
0.5
Analyte 9: Biotin
-
1.5
Analyte 1: Thiamine
</= 20
20
Analyte 2: Riboflavin
</= 30
30
Analyte 3: Nicotinic acid
15
Analyte 4: Nicotinamide
400
Analyte 5: Pantothenic acid
</= 50
30
Analyte 6: Pyridoxine
25
Analyte 7: Pyridoxal
2.5
Analyte 8: Pyridoxamine
1.2
Analyte 9: Biotin
</= 0.1
3
Spike recovery (%)
2
Analyte 1: Thiamine
90 to 110%
90-110%
Analyte 2: Riboflavin
90 to 110%
90-110%
Analyte 3: Nicotinic acid
90-110%
Analyte 4: Nicotinamide
90-110%
Analyte 5: Pantothenic acid
90 to 110%
90-110%
Analyte 6: Pyridoxine
90-110%
Analyte 7: Pyridoxal
90-110%
Analyte 8: Pyridoxamine
90-110%
0.1-1 mcg/100g , 80-120%
> 1 mcg/100g, 90-110%
Bias vs SRM
Certified value
Certified value
3
Analyte 1: Thiamine
</= 5
</= 5
Analyte 2: Riboflavin
</= 5
</= 5
Analyte 3: Nicotinic acid
</= 5
Analyte 4: Nicotinamide
</= 5
Analyte 5: Pantothenic acid
</= 5
</= 5
Analyte 6: Pyridoxine
</= 5
Analyte 7: Pyridoxal
</=10
Analyte 8: Pyridoxamine
</= 5
0.1-1 mcg/100g , </= 8
> 1 mcg/100g, </= 6
Analyte 1: Thiamine
</= 10
</= 10
Analyte 2: Riboflavin
</= 10
</= 10
Analyte 3: Nicotinic acid
</= 10
Analyte 4: Nicotinamide
</= 10
Analyte 5: Pantothenic acid
</= 15
</= 15
Analyte 6: Pyridoxine
</= 10
Analyte 7: Pyridoxal
</= 13
Analyte 8: Pyridoxamine
</= 10
0.1-1 mcg/100g , </= 16
> 1 mcg/100g, </= 12
b
Units
90 to 110%
Analyte 9: Biotin
Analyte 9: Biotin
Analyte 9: Biotin
90-110%
</= 10
</= 5
All analytes defined in the
applicability statement are
measured.
200-10000
10-2000
</= 200
</= 10
</= 10
</= 5
</= 10
</= 5
90 to 110%
Limit of quantification (LOQ)
Method Reference #
Method title:
Principle of the method:
Simultaneous Determination of Thiamine HCl, Riboflavin, Niacin (Nicotinic Acid and Nicotinamide), Pantothenic acid, Vitamin B6 (Pyridoxine,
Pyridoxal and Pyridoxamine) and Biotin in Infant Formula and Adult Nutritionals
Water-soluble vitamins in infant formula or adult nutritional samples are extracted with a mixture of mild acid-Methanol. Quantitation of the extracted
solution is carried out via LC-MS/MS using ESI in the positive mode.
SMPR Requirement
Analytical Range. (mcg/100g)
1
Notes:
Overall Score
Intermediate Reproducibility
(RSDiR)
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
AOAC SMPR: …2015.005-Total B6 (Pyridoxine)………..
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
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 v1.0
BVit-01 - MEF FORM
FOR ERP USE ONLY
DO NOT DISTRIBUTE