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Separation type: Bridge Ion Separation Technology, or BIST™ by SIELC Technologies
HPLC Method for Analysis of Arginine, Lysine and Histidine Amino Acids on a BIST™ B+ Column.
Basic amino acids, like Arginine, Lysine, and Histidine, can be difficult to separate and retain in typical reverse-phase chromatography. Arginine is a naturally-occurring, essential amino acid typically found in meat, poultry, and fish. Lysine is another essential amino acid typically found in meat, and also cereal grains. A third essential amino acid, Histidine, is also typtically found in meat, poultry, and also cereal grains (like rice, wheat, and rye). Using SIELC’s newly introduced BIST™ method, these three essential amino acids, which protonate in water, can be retained on a positively-charged anion-exchange BIST™ B column. There are two keys to this retention method: 1) a multi-charged, negative buffer, such as Sulfuric acid (H2SO4), which acts as a bridge, linking the positively-charged amino acid analytes to the positively-charged column surface and 2) a mobile phase consisting mostly of organic solvent (such as MeCN) to minimize the formation of a solvation layer around the charged analytes. Using this new and unique analysis method, Arginine, Lysine, and Histidine can be retained and UV detected at 210 nm.
High Performance Liquid Chromatography (HPLC) Method for Analysis of Arginine, Lysine, Histidine
Column | BIST™ B+, 4.6×150 mm, 5µm, 100A |
Mobile Phase | MeCN – 70% |
Buffer | H2SO4 – 0.2% |
Flow Rate | 1.0 ml/min |
Detection | UV 210 nm |
Class of Compounds | Amino acid |
Analyzing Compounds | Arginine, Lysine, Histidine |