HPLC Determination of Seleno Amino Acids on Newcrom AH Column

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Methylselenocysteine is used in chemotherapy to induce apoptosis (cell death) of the cancerous cells.  Methylselenocysteine is an amino acid with antioxidant properties and a main source of selenium in several foods.  Selenocysteine is an analog of cysteine with the selenium replacing sulfur.  It has been used as a radioactive tracer.  All three analytes are zwitterions and have similar structures which can present difficulties in reversed-phase separation in HPLC.  By using a mixed mode Newcrom AH column which has ion-exchange properties in addition to hydrophobic properties allows the separation of the seleno amino acids with an MS-compatible mobile phase of water and acetonitrile (ACN) with formic acid buffer.


Column Newcrom AH, 4.6×150 mm, 5 µm, 100A
Mobile Phase MeCN – 20%
Buffer Formic Acid – 4%
Flow Rate 1.0 ml/min
Detection CAD



Class of Compounds
Drug, Acid, Hydrophilic, Ionizable, Vitamin, Supplements, Amino acid
Analyzing Compounds Methylselenocysteine, Selenomethionine, Seleno-L-cystine


Application Column

Newcrom AH

The Newcrom columns are a family of reverse-phase-based columns. Newcrom A, AH, B, and BH are all mixed-mode columns with either positive or negative ion-pairing groups attached to either short (25 Å) or long (100 Å) ligand chains. Newcrom R1 is a special reverse-phase column with low silanol activity.

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