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Representative existing methods
•
USP FCC 7 monograph
•
Total carotenoids
•
HPLC‐UV
•
Normal phase (silica)
•
Mobile phases: hexane, EtOAc
•
AOAC 2009.04
•
Lycopene in dietary supplements and raw materials
•
Protease digestion and extraction with DCM:EtOH
•
HPLC‐UV (alkylamide‐bonded silica)
•
Mobile phase: IPA:NH
4
OAc: ACN:MeOH:alkylamine
A complex extract requires both normal and reversed phase
chromatography for robust separation
•
Khachik et al. (1997) and Khachik et al. (1992)
•
Carotenoids and their metabolites in human milk and serum, and
Carotinoids and their oxidation products in human plasma, respectively
•
Two complementary chromatographic methods
1. Reversed phase (C
18
), ACN:MeOH:DCM, UV/MS
2. Normal phase (Nitrile‐bonded silica), hexane:DCM:MeOH:
alkylamine, UV/MS
•
AOAC 2005.07
•
β‐Carotene in dietary supplements and raw materials
•
Protease digestion and extraction with DCM:EtOH
•
Two complementary HPLC chromatographic methods
1. Reversed phase (C
18
), IPA:NH
4
OAc: ACN:MeOH:alkylamine, UV
2. Reversed phase (C
30
), MeOH:MTBE:alkylamine, UV