19 / 159
FOR EXPERT REVIEW PANEL USE ONLY
NOT FOR DISTRIBUTION
1
Expert Review Panel for Infant Formula and Adult Nutrition
Evaluation of Method:
Bio-02
#3
Title:
Determination of biotin by liquid chromatography coupled with immunoaffinity column clean-up extraction
Author:
George Joseph, AsureQuality
Summary of Method:
The sample is dissolved, then autoclaved. After cooling, the diluted sample is centrifuged and filtered. The
filtrate is collected for clean-up and extraction using an immunoaffinity column is mounted onto a SPE
manifold. Biotin from the column is eluted with methanol and collected in a reacti-vial. The eluent is
evaporated to dryness and the sample is re-constituted in 1mL of water. The biotin in the reconstituted
sample is quantified by HPLC using a UV detector set at 200nm
Method Scope/Applicability:
Infant Formula and Adult/Pediatric Formulas
General comments about the method:
Method Clarity:
Clear directions for operation of the method, instruments. Clear equations describing how the standards
and samples are quantitated is required.
Method Safety Concerns:
None, beyond standard laboratory safety
Pros/Strengths:
Simple straight forward analysis.
Cons/Weaknesses
Proprietary components (immunoaffinity cartridge)
Supporting Data
SPIFAN Kit 1 used.
Data not calculated on RTF basis
Performance Characteristics:
Analytical Range:
0.1
to 300 µg/100g meets SMPR limits (0.1-150 µg/100g)
Bio-02 Review Forms