September 2016 SPDS Book

Existing Methods

HPLC WITH PRECOLUMN DERIVATIZATION

► Method

 Reversed phase or silica column

 o-phthaldehyde (OPA), phenylisothiocyanate (PITC), 9- fluorenylmethyl chloroformate (F-MOC)  Fluorescence or UV detection ► Advantages  Derivatization conditions (reagent and detector) can be optimized for matrix and analytes of interest – Greater sensitivity  Allows for using rapid reversed-phase chromatography

11

Existing Methods

HPLC WITH POSTCOLUMN DERIVATIZATION (AOAC METHOD)

► Method

 Cation-exchange chromatography  Ninhydrin, fluorescamine, OPA  Fluorescence or UV detection

► Advantages

 Can be automated resulting in increased performance and reproducibility  Reaction efficiency less prone to sample matrix effects  Minimal sample preparation

12

6