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 H2O/CH3CN=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