Arginine

HPLC Method for Analysis of Arginine, Lysine and Histidine Amino Acids on BIST B+ by SIELC Technologies.

Separation type: Bridge Ion Separation Technology, or BIST™ by SIELC Technologies

High Performance Liquid Chromatography (HPLC) Method for Analyses of Arginine, Lysine and Histidine Amino Acids

Basic amino acids, like Arginine, Lysine, and Histidine, can be difficult to separate and retain in typical reverse-phase chromatography. Arginine is a naturally-occurring, essential amino acid typically found in meat, poultry, and fish. Lysine is another essential amino acid typically found in meat, and also cereal grains. A third essential amino acid, Histidine, is also typtically found in meat, poultry, and also cereal grains (like rice, wheat, and rye). Using SIELC’s newly introduced BIST™ method, these three essential amino acids, which protonate in water, can be retained on a positively-charged anion-exchange BIST™ B column. There are two keys to this retention method: 1) a multi-charged, negative buffer, such as Sulfuric acid (H2SO4), which acts as a bridge, linking the positively-charged amino acid analytes to the positively-charged column surface and 2) a mobile phase consisting mostly of organic solvent (such as MeCN) to minimize the formation of a solvation layer around the charged analytes. Using this new and unique analysis method, Arginine, Lysine, and Histidine can be retained and UV detected at 210 nm.

Description

Separation type: Liquid Chromatography Mixed-mode

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High Performance Liquid Chromatography (HPLC) Method for Analysis of L-Carnitine and Arginine

L-carnitine is a quaternary ammonium cation (also known as a quat) that is key to the metabolic processes of most living species. Arginine is an alpha amino acid that is key to the biosynthesis of proteins.
These two amine-based compounds can be detected in the low UV regime. Using a Primesep 100 mixed-mode column and a gradient mobile phase consisting of water and acetonitrile (MeCN) with a gradient of sulfuric acid (H2SO4) as a buffer, L-carnitine and Arginine can be retained, separated, and analyzed. This analysis method can be UV detected at 205 nm.

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Essential amino acids cannot be made by the body. As a result, they must come from food.
The 9 essential amino acids are: histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, and valine.

L-Lysine is an essential amino acid required for growth and tissue repair, it can also help with anxiety. The amino acid is most often found in red meats,beans, fish, and dairy products. L-Arginine is an essential amino acid that is used to treat congestive heart failure, combat fatigue and circulatory diseases. It is also used to stimulate the immune system. Obelisc R is a reverse-phase column. It contains embedded ionic groups and can retain L-Lysine and L-Arginine. The method is UV compatible and can be used as a general approach for analyzing similar compounds.

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

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.

Two underivatized amino acids, citrulline and arginine, are separated by mixed-mode chromatography on Primesep 100 and Primesep 200 HPLC columns with LC/MS compatible mobile phase. Method can be used for retention and separation of underivatized amino acids without ion-pairing reagent in the mobile phase.

Citrulline and Arginine are α-amino acids that differ only in one functional group. They can be separated on a reverse-phase Primesep 200 HPLC column by hydrophobic and cation-exchange mechanisms in a mobile phase of water and acetonitrile (MeCN, ACN) with a gradient buffer of formic acid. Evaporative Light Scattering Detector (ELSD) used.

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).

Glutamine, valine, isoleucine, and leucine are baseline resolved in a dietary supplement with a mobile phase of acetonitrile, water, and sulfuric acid with ultraviolet (UV) detection at 210 nm on a Primesep 100 column. Increasing the sulfuric acid concentration allows elution of arginine. Primesep 100 separates underivatized amino acids and amines by combining reversed-phase and ion-exchange mechanisms. The HPLC separation uses a mobile phase of water, acetonitrile (MeCN, ACN), and sulfuric acid with ultraviolet (UV) detection at 210 nm.