Valine

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
Analytes
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 18 Amino Acids on Amaze HD Column
Conditions of Experiment
Column: Amaze HD
Separation Modes: HILIC, cation-exchange
Column Dimenstions: 3.0x100 mm, 5 um, 100A
Mobile Phase: 80% ACN with 20 mM AmFm pH 3
Detection: ELSD, 45°C
Sample: 0.5-0.8 mg/ml
Injection: 3 uL
Flow rate: 0.6 ml/min
Analytes
Class of compounds: Amino acid, Catecholamine, Drug, Neurotransmitter, Supplement
Nature of compounds: Acidic, Basic, Hydrophilic, Polar, Zwitterionic
Compounds: Tryptophan, Phenylalanine, Leucine, Methionine, Tyrosine, Isoleucine, DOPA, Valine, Cysteine, Proline, Alanine, Threonine, Glutamic acid, Glycine, Glutamine, Asparagine, Serine, Aspartic acid
Ultra-Fast HPLC Separation of Underivatized Amino Acids in Reversed-Phase and Cation-Exchange Modes
Application description

Amino acids are essential components of numerous formulation. Health supplements can contain various amino acids and vitamins and require quantitation of each ingredients. Amino acids are very polar compounds with limited or no retention in reversed-phase chromatography. The most common approaches are reversed-phase chromatography with ion-pairing reagent and hydrophilic interaction chromatography (HILIC). Underivatized amino acids can be retained by combination of reversed-phase and cation exchange mechanism. These  two mechanism were combined in Coresep 100 core-shell mixed-mode column. Method can be used for fast analysis of underivatized amino acids without ion-pairing reagent. Depending on the pH of the mobile phase they can exist in acidic, basic or zwitter-ionic form. At lower pH, amino acids are basic in nature and have the highest hydrophobicity. Four amino acids (SER, GLY, ALA, VAL and DOPA) were retained and separated within 2 minutes on core-shell mixed-mode column. This high efficiency separation does not require special UPLC set up and can benefit from core-shell and mixed-mode approaches.

Conditions of Experiment
Column: Coresep 100
Separation Modes: reversed-phase and cation-exchange
Column Dimenstions: 3.2 x 50 mm, 2.7 um, 90A
Mobile Phase: 3% ACN with 0.03% TFA
Detection: UV 210 nm, ELSD
Sample: 0.3 mg/ml
Injection: 10 uL
Flow rate: 1 ml/min
Analytes
Class of compounds: Amino acid, Catecholamine, Drug, Neurotransmitter, Supplement
Nature of compounds: Basic, Hydrophilic, Polar, Zwitterionic
Compounds: Glycine, Alanine, Serine, Valine, Dopamine