Glutamic Acid

HPLC Method for Separation of Ibotenic Acid and Glutamic Acid on Primesep A by SIELC Technologies

Separation type: Liquid Chromatography Mixed-mode

Ibotenic acid is a physchoactive NDMA and metabotropic glutamate receptor agonist with neurotoxic properties. These structurally similar psychoactive drugs can be retained, separated, and analyzed on a mixed-mode Primesep A column with a mobile phase consisting of water, Acetonitrile (MeCN), and Phosphoric acid (H3PO4). This analytical method can be UV detected at 210 nm with high resolution and peak symmetry.

High Performance Liquid Chromatography (HPLC) Method for Analysis of  Ibotenic Acid and Glutamic Acid

Condition

Description

View on hplc.cloud

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.

Application Notes: Amino acids are polar ionic compounds which are not retained on reversed-phase column without ion-pairing reagent. In our application, lysine and arginine can be separated from other amino acids. Amino acids with a pH between 3 and 5 and with one basic and one acidic group become very polar. Therefore these amino acids don’t have strong ion-exchange interaction with Primesep C stationary phase. Amino acids with two amino groups still carry positive net charge and can interact with stationary phase by cations-exchange mechanism. pH variation of the mobile phase can be an effective tool to adjust selectivity of separation for zwitter-ionic, basic and acidic compounds. This method can be used for separation of mono-charged compounds from compounds having an extra charge.

Application Columns: Primesep C
Application compounds: Aspartic acid, Glutamic acid, Aspargine, Glycine, Proline, Alanine, Phenylalanine, Tyrosine, Leucine, Lysine, Arginine
Detection technique: UV, LC/MS, ELSD/CAD

Glutamic acid and GABA are neurotransmitters. Glutamic acid and GABA are non-essential amino acids. They are hydrophilic and zwitter-ionic in nature . At lower pH, carboxylic acid groups of amino acids are not ionized, making them more hydrophobic and basic. Underivatized glutamic acid and GABA were retained and separated on a Primesep 100 column using ACN/water/TFA mobile phase. Amino acids can be monitored by low UV or ELSD/CAD. Retention is provided by reversed-phase and cation-exchange mechanism. Method can be used for analysis of underivatized amino acids in various matrices including supplements, vitamin and other complex mixtures. various mobile phase can be used with corresponding detection techniques.

Polar amino acids are very often used as components of vitamin and supplement composition. Analysis of such complex composition is a challenging task. In this application, 5 amino acids (asparagine, glutamic acid, proline and arginine) and two preservatives (methyl paraben and propyl paraben) are separated on a Primesep 100 reversed-phase cation-exchange column with LC/MS compatible mobile phase. Method does not require ion-pairing reagent in the mobile phase. Compounds are monitored by ELSD and UV. Method is validated for quantitation of underivatized amino acids in complex mixtures. The method is simple and robust and can be used for analysis of various vitamin formulations.

The separation of amino acids, the building blocks of proteins, can be challenging to separate on a reverse-phase column due to their high polarity. Using a mixed-mode HPLC column, allows the separation of amino acids by cation-exchange and ion-exclusion mechanisms as well as hydrophobicity. Fine tuning of separation can be achieved with changes in organic concentration of the mobile phase as well as choice of buffer and pH.

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).

Primesep 100 separates the components of a Swiss cheese extract by HPLC using cation exchange and reversed phase as retention mechanisms. The amino acids, glutamic acid and proline, as well as glycine betaine were resolved in less than 10 minutes. A mobile phase gradient of water, acetonitrile (MeCN, ACN,) and phosphoric acid (H3PO4) and ultraviolet (UV) detection was used.