LC/MS Analysis of Essential Amino Acids (NH2P-40 2D)
A polymer-based amino HILIC column, Asahipak
NH2P-40 2D, was used to analyze 20 essential amino acids that consists proteins. This method does not require the use of delivertization or ion-pair reagents. The analysis takes less than 35 min. The result show that acidic amino acids tend to be retained longer by the amino function group modified on NH2P-40 2D column packed gel.
Sample : Amino acid standards 5μg/mL each (in H
2O/CH
3CN=25/75), 5μL
Phenylalanine, Phe |
Tyrosine, Tyr |
Asparagine, Asn |
Leucine, Leu |
Lysine, Lys |
Serine, Ser |
Isoleucine, Ile |
Alanine, Ala |
Cysteine, Cys |
Tryptophan, Trp |
Histidine, His |
Glutamic acid, Glu |
Methionine, Met |
Threonine, Thr |
Aspartic acid, Asp |
Proline, Pro |
Glycine, Gly |
|
Valine, Val |
Glutamine, Gln |
|
Arginine, Arg |
|
|
Column : Shodex Asahipak NH2P-40 2D (2.0mmI.D. x 150mm)
Eluent : (A); 100mM HCOONH4 aq.
(B); CH3CN
Linear gradient; 0 to 10min 75%(B), 10 to 11min 75%(B) to 50%(B), 11 to 35min 50%(B)
Flow rate : 0.2 mL/min
Detector : ESI-MS SIM
Column temp. : 30°C