Histidine

HPLC Method for Separation of Histidine, Histidine Methyl Ester, and Histidine Lauryl Ester on BIST B+ by SIELC Technologies

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

Histidine is a naturally occurring essential amino acid that the body uses to repair damaged tissue and generate new blood cells. Histidine Methyl Ester is a histidine decarboxylase inhibitor and can be used to synthesize other biological compounds. Histidine Lauryl Ester is another derivative of Histidine. Using SIELC’s newly introduced BIST™ method, Histidine and its derivatives can be retained and separated 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 dipeptide 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, Histidine and its derivatives can be UV detected at 220 nm.

High Performance Liquid Chromatography (HPLC) Method for Analysis of Histidine, Histidine Methyl Ester and Histidine lauryl Ester

Condition

Description

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 Histidine and Histidine Methyl Ester Dihydrochloride

Histidine and Histidine Methyl Ester Dyhydrochloride are related compounds with interesting uses. Histidine is used in biosynthesis to create proteins, while histidine methyl ester dihydrochloride (a derivative of histidine) is used to create various other compounds.
This amino acid and its derivative can be detected in the low UV regime. Using a Primesep 100 mixed-mode column and a mobile phase consisting of water and acetonitrile (MeCN) with a sulfuric acid (H2SO4) buffer, Histidine and Histidine Methyl Ester Dyhydrochloride can be retained, separated, and analyzed. This analysis method can be UV detected at 200 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.

Essential and non-essential amino acids can be retained and separated in zero-organic mode on Primesep mixed-mode HPLC columns. Zero-organic mode is required to monitor isotopes of carbon. Amino acids are retained by combination of reversed-phase and cation-exchange mechanisms. At lower pH, some of the amino acids are more hydrophobic. Buffer pH will affect ionization state of amino acids, and at higher pH (above 2.5), the amino acids will be less hydrophobic and retentive in zero-organic mode. Amino acids can be monitored by low UV. Method can be used in archeological research for analysis of various molecules where presence of organic component of the mobile phase interferes with analysis.

Polysorbates are a class of emulsifiers used in some pharmaceuticals and food preparation. They are often used in cosmetics to solubilize essential oils into water-based products. Polysorbates are oily liquids derived from PEG-ylated sorbitan (a derivative of sorbitol) esterified with fatty acids. Surfactants that are esters of plain (non-PEG-ylated) sorbitan with fatty acids are usually referred to by the name Span. It is often required to quantitate Polysorbate (Polysorbate/Tween 20, Polysorbate/Tween 40, Polysorbate 60/Tween 60 and Polysorbate 80) by HPLC in various formulations. Polysorbates exist in form of oligomers.

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).

Creatine, creatinine, and histidine are all important amino acids in the human body. All of these can be separated by liquid chromatography. The mobile phase contains acetonitrile (MeCN), water, and phormic acid. Smaller 3 µm particle columns are available for fast UPLC applications. This liquid chromatography method is scalable and can be used for isolation of impurities in preparative separation. It also suitable for pharmacokinetics.

Creatine is produced by the body and plays a major role in recycling ATP. It also stabilizes pH in certain tissues. Creatine can be analyzed by this reverse phase (RP) HPLC method with simple conditions. The mobile phase contains acetonitrile (MeCN), water, and TFA. For Mass-Spec (MS) compatible applications the phosphoric acid needs to be replaced with formic acid. Smaller 3 µm particle columns are available for fast UPLC applications. This liquid chromatography method is scalable and can be used for isolation of impurities in preparative separation. It also suitable for pharmacokinetics.