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FOR EXPERT REVIEW PANEL USE ONLY
NOT FOR DISTRIBUTION
2
The method does not include:
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Example calculations
o
Height or area?
o
How the standard curve data is treated to create a calibration;
o
Sample calculation.
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System suitability
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What sequence of injections should be used for calibrants and unknowns;
o
Performance criteria for standard curve (r) and read-backs.
Method Safety Concerns:
None
Pros/Strengths:
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The sample preparation is straightforward and uses conventional, widely practiced techniques
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The quantification uses conventional HPLC w UV detection.
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The validation report documented similar analytical results for the majority of samples from a second
brand of immunoaffinity column; most samples produced results within ± 5%. The milk based IF
powder result was 10% lower - 3 mcg/100g powder, or c 0.4 mcg/100mL (4 mcg/L) as fed; although the
absolute difference is small, further investigation is warranted because of the commonness of this
matrix. (Table 5)
Cons/Weaknesses
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Text needs to be added to describe system suitability and calculations.
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The values in the SLV are not consistently reported in the SMPR units (mcg/100g reconstituted)
Supporting Data
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General Comment:
The validation report stated that while a reagent blank and a matrix blank prepared from infant elemental
powder were devoid of interferences, the placebo (non-fortified) products child formula powder, adult
nutritional RTF high fat, and infant formula RTF milk based showed a response near the retention time of
biotin. The report does not show these chromatograms nor clarify if the “response” was attributed to
(endogenous) biotin or was an interference.
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Method Optimization:
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Performance Characteristics:
Analytical Range:
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The method describes the processing of samples with biotin content over the entire concentration
range stated in the SMPR (0.1 – 150 mcg/100g as fed)
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In practice, the concentration encountered in the SLV using the SPIFAN sample set was ~1.5 – 65
reconstituted.
Bio-02 Review Forms