SPDS Meeting Book

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