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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-02 #1
Title:
Determination of Lutein and β-Carotene in Infant Formula and Adult Nutritionals
Reversed-Phase High Performance Liquid Chromatography
Author:
Greg Hostetler, Perrigo Nutritionals, Georgia, VT USA
Summary of Method:
Samples are spiked with an internal standard and saponified with
potassium hydroxide, extracted with MTBE, THF, and hexane. The supernatants from the
liquid-liquid extraction are dried under nitrogen and reconstituted in IPA. Separation is
done by reversed-phase chromatography on a C30 column. All-trans lutein and β-
carotene are separated from their major cis isomers, as well as zeaxanthin and α-
carotene.
Method Scope/Applicability:
Beta Carotene and Lutein in Infant Formula and Adult/Pediatric
Nutritional Formula
General comments about the method: Well done work which incorporates the major learnings in the
carotenoid arena of the past 40 years.
Method Clarity: Very clearly written, and exceptionally clear calculations sections. Well Done
Method Safety Concerns:
Nothing out of ordinary laboratory safety practices.
Pros/Strengths:
Carot-02 Review Forms