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Updated: May 29, 2026

Post Column Derivatization Using Reaction Flow High Performance Liquid Chromatography Columns
Published on: April 26, 2016
Validated high-performance liquid chromatographic enantioseparation of selenomethionine using isothiocyanate based
1Department of Chemistry, Indian Institute of Technology Roorkee, Roorkee 247667, India. rbushfcy@iitr.ernet.in
Abstract:
A high-performance liquid chromatographic (HPLC) method for enantioseparation of selenomethionine (SeMet) was developed using two isothiocyanate-based chiral derivatizing reagents [(R)-methyl benzyl isothiocyanate (MBIC) and (S)-1-(1-naphthyl) ethyl isothiocyanate (NEIC)] and UV detection. Diastereomers of selenomethionine were synthesized either via stirring (using MBIC) or by microwave irradiation (using NEIC). Derivatization conditions were optimized and the synthesized diastereomers were successfully resolved using triethyl ammonium phosphate buffer and acetonitrile on a reversed-phase column. The method was validated for accuracy, precision and limit of detection. The mechanism of separation is also discussed.
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