Glycine
Column: | Amaze SC |
Separation Modes: | reversed-phase, cation-exchange and anion-exclusion |
Column Dimenstions: | 4.6x50 mm, 3 μm, 100A |
Mobile Phase: | 10% ACN, TFA from 0.05% to 0.25% in 3 min, 5 min hold |
Detection: | Corona CAD |
Sample: | 0.5 mg/ml |
Injection: | 2 μL |
Flow rate: | 1 ml/min |
Class of compounds: | Amino acid, Inorganic anion, Inorganic cation, Organic acid, Supplement |
Nature of compounds: | Acidic, Basic, Hydrophilic, Polar, Zwitterionic |
Compounds: | Bromide ion, Glycine, Zinc ion |
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 |
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 |
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 |
Mixed-mode stationary phases were designed to retain and separate compounds with drastic difference in properties using one column and method. Carboxylic acids, amino acids and sugars are poorly retained in reversed-phase chromatography. Derivatization procedures, ion-pairing reagents and HILIC columns can be used tp analyze these complex mixtures. The use of ion-pairing reagent makes MS detection problematic due to volatility issues of IP and signal suppression in MS. HILIC mixed-mode columns, like Amaze TH, have polar ionizable groups on the surface which help significantly increase retention time of polar ionizable analytes like amines, amino acids, carboxylic acids and sugars. We have developed a method for separation and retention of succinic acid, phenylalanine, sucrose, glycine, aspartic acid and raffinose. Isocratic conditions provide separation within 15 minutes with good peak shape and retention control. Retention time on Amaze TH column is controlled by amount of ACN, buffer concentration, buffer ph and buffer nature. Method can be used for other amines, amino acids, carboxylic acids and sugars where fast MS-compatible separation is required.
Column: | Amaze TH |
Separation Modes: | HILIC, cation-exchange, anion-exchange |
Column Dimenstions: | 4.6 x 100 mm 5 um, 100A |
Mobile Phase: | 80% ACN with 10 mM AmAc pH 4.8 |
Detection: | ELSD |
Sample: | 0.3 mg/ml |
Injection: | 3 uL |
Flow rate: | 1 ml/min |
Class of compounds: | Amino acid, Organic acid, Sugar, Supplement |
Nature of compounds: | Acidic, Hydrophilic, Neutral, Polar, Zwitterionic |
Compounds: | Succinic acid, Phenylalanine, Sucrose, Glycine, Aspartic acid, Raffinose |
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.
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 |
Class of compounds: | Amino acid, Catecholamine, Drug, Neurotransmitter, Supplement |
Nature of compounds: | Basic, Hydrophilic, Polar, Zwitterionic |
Compounds: | Glycine, Alanine, Serine, Valine, Dopamine |