HPLC Analysis of 20 Amino Acids on Amaze SC Mixed-Mode Column in RP and Cation-exchange Modes
Conditions of Experiment
Column: Amaze SC
Separation Modes: reversed-phase cation-exchange
Column Dimenstions: 4.6x250 mm, 5 μm, 100A
Mobile Phase: A: 2% ACN with 0.1% TFA B: 25% ACN with 0.3% TFA from 100% A to 100% B in 20 min, 8 min hold
Detection: Corona CAD
Sample: 0.2 mg/ml
Injection: 5 μL
Flow rate: 1 ml/min
Class of compounds: Amino acid, Neurotransmitter, Supplement
Nature of compounds: Acidic, Hydrophilic, Polar, Zwitterionic
Compounds: Aspartic acid, Serine, Asparagine, Cysteine, Glutamic acid, Threonine, Glutamine, Glycine, Alanine, Proline, Valine, Methionine, Tyrosine, Isoleucine, Leucine, Phenylalanine, Lysine, Histidine, Tryptophan, Arginine
HPLC Analysis of Glucose and Lysine on Amaze TH Mixed-Mode Column
Application description

Retaining polar ionizable and non-ionizable compounds is a challenging task in chromatography. This is due to the lack of retention and poor peak shape occuring due to secondary interactions on reversed-phase and HILIC columns. We have developed a method for the separation of glucose and lysine on the mixed-mode Amaze TH column. Glucose is a monosaccharide which is polar (hydrophilic) and non-ionizable. Lysine is one of the essential amino acids, it is polar and zwitterionic with two basic and one carboxylic groups. Glucose is retained separated by pure HILIC mechanism and lysine is retained by combination of HILIC and cation-exchange mechanisms. The retention time is controlled by the amount of ACN, buffer pH and buffer concentrations. Buffer pH affects the ionization state of lysine and the ionization state of the stationary phase. This method is robust. The Amaze TH column can be used to retain and separate amino acids, sugars, amino sugars, organic and inorganic acids as well as polar neutral compounds like sugars. The column is compatible with major detection technique (ELSD, CAD, RI, UV and mass spec)

Conditions of Experiment
Column: Amaze TH
Separation Modes: HILIC, cation-exchnage
Column Dimenstions: 4.6x50 mm, 3 um, 100A
Mobile Phase: 65% ACN with 175 mM AmAc pH 5
Detection: ELSD 45°C, RI-compatible
Sample: 1 mg/ml
Injection: 3 uL
Flow rate: 1 ml/min
Class of compounds: Amino acid, Sugar, Supplement
Nature of compounds: Hydrophilic, Neutral, Polar, Zwitterionic
Compounds: Glucose, Lysine