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FOR EXPERT REVIEW PANEL USE ONLY
NOT FOR DISTRIBUTION
1
Expert Review Panel for Infant Formula and Adult Nutrition
Evaluation of Method:
Carot-01
#2
Title:
Determination of Carotenoids in Infant Formula and Adult/Pediatric Nutritional Formula using High
Performance Liquid Chromatography with Photo Diode Array Detection
Author:
Martijn Vermeulen, Ausnutria
Summary of Method:
Carotenoid esters are saponified using potassium hydroxide in ethanol/water in the presence of sodium
ascorbate, sodium sulfide and glycerol to prevent oxidation. Dimethyltocol (DMT) is added as an internal
standard and carotenoids are extracted using diisopropyl ether. The extract is subsequently washed and
concentrated. Carotenoids are determined by RPLC-UV at 450 nm and DMT is measured at 292 nm.
Method Scope/Applicability:
Infant Formula and Adult/Pediatric Formulas
General comments about the method:
Method appears to measure carotenoid forms specified in the SMPR, however, this is not explicitly stated.
Using this chromatography, it is not possible to get sufficient resolution to unambiguously integrate the cis-
isomers of lutein and a-carotene which are explicitly to be determined in the applicability statement.
Method Clarity:
Clear directions for operation of the method, instruments. No details about how to make up a-carotene,
lutein, lycopene standards, or mixed calibration standards. Clear equations describing how the standards
and samples are quantitated is required.
Method Safety Concerns:
None other than standard laboratory safety
Pros/Strengths:
Very simple straightforward analysis
Cons/Weaknesses
Supporting Data
SPIFAN kit was used in the validation of the method.
Results reported on dry weight basis, making interpretation against SMPR difficult.
Performance Characteristics:
Carot-01 Review Forms