Applications:
Amino Acids Analysis in Acid Gradient Condition

11 underivatized amino acids (aspartic acid, glutamic acid, alanine, valine, methionine, isoleucine, cysteine, phenylalanine, histidine, lysine, and arginine) are separated by a Primesep 100 HPLC column by reversed-phase and ion-exchange mechanisms with LC/MS compatible conditions without the use of ion-pair reagents. The HPLC separation uses a TFA (trifluoroacetic acid) gradient in a mobile phase of water acetonitrile (MeCN, ACN with evaporative light scattering detection (ELSD).
Condition
Column |
Primesep 100, 4.6x250 mm, 5 µm, 100A |
Mobile Phase |
MeCN/H2O - 30/70% |
Buffer |
TFA , gradient 0.05-0.3 % , 25 min |
Flow Rate |
1.0 ml/min |
Detection |
ELSD |
Description
Class of Compounds
|
Drug, Acid, Hydrophilic, Ionizable, Vitamin, Supplements |
Analyzing Compounds |
Gaspartic acid, Glutamic acid, Alanine, Valine, Methionine, Isoleucine, Cysteine, Phenylalanine, Histidine, Lysine, Arginine |
AlanineAmino AcidsArginineAspartic AcidCysteineGlutamic AcidHistidineIsoleucineLysineMethioninePhenylalanineValine
Hydrophobic and Hydrophilic Compound Separation

Primesep 100 separates a mixture of polar and nonpolar compounds in one analytical run. The amino acid cysteine; amino acid derivatives L-cystine, 2,2-dimethylcystine, and 2-methylcysteine; the polar acid benzoic acid; and the nonpolar neutral toluene are separated by a gradient using a combination of polar and hydrophobic interactions. The separation method uses a mobile phase mixture of water, acetonitrile (MeCN, ACN) and sulfuric acid (H2SO4) with UV detection at 210 nm.
Condition
Column |
Primesep 100, 4.6x150 mm, 5 µm, 100A |
Mobile Phase |
MeCN/H2O |
Buffer |
H2SO4 |
Flow Rate |
1.0 ml/min |
Detection |
UV 210 nm |
Description
Class of Compounds
|
Drug, Acid, Hydrophilic, Ionizable, Nonpolar, Polar, Supplements |
Analyzing Compounds |
Cysteine, L-cystine, 2,2-dimethylcystine, 2-methylcysteine, Benzoic Acid, Toluene |
2,2-Dimethylcysteine2-MethylcysteineAmino AcidsBenzoic AcidCysteineL-CystineToluene
HPLC Separation of Mixture of Conditionally Essential Amino Acids on Primesep 100 Column
Condition
Column |
Primesep 100, 4.6x250 mm, 5 µm, 100A |
Mobile Phase |
MeCN/H2O - 35/65% |
Buffer |
H2SO4 0.05% 12 min hold, gradient 0.05-0.20, 13 min |
Flow Rate |
1.0 ml/min |
Detection |
UV, 200 nm |
Description
Class of Compounds
|
Drug, Acid, Hydrophilic, Ionizable, Vitamin, Supplements, Amino acid |
Analyzing Compounds |
Glutamin (Gln/Q), L-Glycine (Gly/G), L-Cysteine (Cys/C), L-Proline Pro/p), L-Tyrosine (Tyr/Y), L-Arginine (Arg/R) |
Amino AcidsArginineCysteineGlycineL-CysteineL-GlutamineProlineTyrosine
HPLC Separation of Mixture of 12 Amino Acids on Primesep 100 Column
Condition
Column |
Primesep 100, 4.6x250 mm, 5 µm, 100A |
Mobile Phase |
MeCN/H2O - 35/65% |
Buffer |
H2SO4 0.05% 12 min hold, gradient 0.05-0.20, 13 min |
Flow Rate |
1.0 ml/min |
Detection |
UV, 200 nm |
Description
Class of Compounds
|
Drug, Acid, Hydrophilic, Ionizable, Vitamin, Supplements, Amino acid |
Analyzing Compounds |
L-Asparagine (Asn/N), L-Threonine (Thr/T), L-Cysteine (Cys/C), L-Proline (Pro/P), L-Valine (Val/V), DL-Methionine (Met/M), L-Isoleucine (Ile/I), L-Leucine (Leu/L), L-Phenylalanine (Phe/F), L- Tryptophan (Trp/W), L-Histidine (His/H), L-Arginine (Arg/R) |
Amino AcidsArginineAsparagineCysteineD-IsoleucineD-LeucineD-ValineDL-IsoleucineHistidineIsoleucineL-CysteineL-MethionineLeucineMethioninePhenylalanineProlineTryptophanValine