133 / 696
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).
Combination of milk, soy, hydrolyzed protein, FOS/GOS, amino acids, with high
or low fat.
All SPIFAN matrices analyzed for repeatability.
Representative matrices from each category analyzed for
recovery during SLV, except for Infant RTF due to the
defficient sample amount.
All required analytes are
measured.
Biotin
Method measures all required analytes.
0.1-150 mcg/100g reconstituted final product
1.66-142 mcg/100g reconstituted final product
Biotin
Not Applicable
Biotin
d
0.1 mcg/100g reconstituted final product
~0.8 mcg/100g in reconsitituted powder product OR
~1.5 mcg/100 g in ready-to-feed product
See Table 1
0.1-1 mcg/100g reconstituted final product: 80-120%
Not Applicable
>1 mcg/100g recons tituted final product: 90-110%
Range: 95-111% See Table 2
Bias vs SRM
See Table 3
0.1-1 mcg/100g reconstituted final product :
d
%
Not Applicable
>1 mcg/100g reconstituted final product :
d
6%
Range: 0.5-3.0% See Table 4
No SMRP requirements
Range: 1.3-4.5% See Table 4
0.1-1 mcg/100g reconstituted final product :
d
%
>1 mcg/100g reconstituted final product :
d
%
Suitable methods will include blank check samples, and check standards at the
lowest point and midrange point of the analytical range.
90-110% recovery of SRM with different sample sizes and
being quantified withing the calibration curve range of
8.5-60 ng/mL. See Table 5
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
Feedback from Users of the Method since being
awarded First Action
Official Methods
Status
Adequate proof of performance via system suitability
Spike recovery
Accuracy/Recovery
Reproducibility (RSDR)
Intermediate Reproducibility
(RSDiR)
Evaluation of Method Performance vs. SMPR requirements.
Determination of biotin in infant, pediatric and adult nutritionals by HPLC and fluorescence detection
This reverse phase HPLC method with post column reduction and fluorescence detection allows for the quantitative determination of biotin in infant, pediatric, and adult nutritionals. Sample of
appropriate size is mixed with 6% meta-phosphoric acid to precipitate out the protein to produce a filtrate. The filtrate is injected onto a C18 HPLC column where biotin and riboflavin are separated
with a mobile phase (20% methanol in 0.02 M phosphate buffer at pH7.0). The biotin after eluting from the column binds with the streptavidin fluorescein to become a fluorescent conjugate. The
conjugate is then detected by fluorescence at an excitation wavelength of 495 nm and an emission wavelength of 518 nm.
Repeatability (RSDr)
SMPR Requirement
Analytical Range
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.
hod does not require any special safety precautions e.g. personal protection from highly toxic solve
SPIFAN ERP Checklist v 1.0
Bio-03 (February 2016)
FOR ERP USE ONLY
DO NOT DISTRIBUTE