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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
#1
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# , Edith de Haan* , Arjan de Vries* , and Fariha Bouraada* . #
Ausnutria Hyproca Analytics, Lelystad, The Netherlands, to whom correspondence should
be addressed. * TNO Triskelion, Department of Analytical Chemistry, Utrechtseweg 48,
Zeist, The Netherlands.
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) as internal standard. Carotenoids are extracted using
diisopropyl ether. Carotenoids are determined by HPLC-UV at 450 nm and DMT is
measured at 292 nm. Separation is performed using 2 suplex pkb-100 columns of 25 cm
with a gradient.
Method Scope/Applicability:
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 (except for the accurate quantitation of the beta carotene stock
standard solution-see below)
Method Clarity: Well Done
Carot-01 Review Forms