Serine

High Performance Liquid Chromatography (HPLC) Method for Analysis of Non-Essential Amino Acids on Primesep 100 by SIELC Technologies

High Performance Liquid Chromatography (HPLC) Method for Analyses of Non-Essential Amino Acids

Amino acids are the building blocks of proteins. Based on their dietary requirement, they are classified into essential and non-essential amino acids. Essential amino acids cannot be synthesized by the human body in sufficient quantities and must be obtained from the diet. Non-essential amino acids, on the other hand, can be synthesized by the body and are not dependent on dietary intake.

It’s worth noting that while these amino acids are considered “non-essential” for adults under normal circumstances because the body can synthesize them, there are situations where some may become “conditionally essential.” This means that under certain conditions like illness, stress, or trauma, the body might not produce them in sufficient quantities, and dietary intake becomes necessary. Arginine, for instance, is considered conditionally essential, especially during periods of rapid growth, illness, or trauma.

Amino acids can be retained, separeted and analyzed on a Primesep 100 mixed-mode stationary phase column using an isocratic analytical method with a simple mobile phase of water, Acetonitrile (MeCN), and a phosphoric acid (H3PO4) as a buffer. This analysis method can be detected in the UV regime at 200 nm.

Four underivatized amino acids (serine, methylserine, GABA and methionine) were separated on a Primesep 100 reversed-phase cation-exchange mixed-mode HPLC column. Two methods, one for UV and one for ELSD/LC/MS show good separation and peak shape for underivatized amino acids. This column and general HPLC approach can be used for analysis of underivatized amino acids. Primesep 100 is designed to replace reversed-phase HPLC column in combination with ion-pairing reagents.

Amino acids are building blocks for peptides and proteins. Serine is not essential to human diet and it is synthesized in human body. Methionine is not synthesized in human body and needs to be ingested. GABA is used to enhance growth of specified plants, prevent development of powdery mildew on grapes, and suppress certain other plant diseases. In humans GABA helps to maintain normal brain function. Serine, methylserine, GABA and methionine are separated on Primesep 100 column by mixed-mode mechanism. Amino acids are retained by combination of reverse phase and cation-exchange mechanisms. At lower pH carboxylic acid fragment of amino acid is suppressed and not ionized, making amino acids more basic and slightly more hydrophobic. Amount of acetonitrile, buffer concentration and buffer pH can be used to adjust retention time. Underivatized amino acids are well retained and separated with perfect peak shape and symmetry. Fast method can be used for UV, ESLD and LC/MS quantitation of serine, methylserine, GABA and methionine.