HPLC Method for Aspartic Acid, L-Glutamine, Glutamic Acid, Glycine on Newcrom AH by SIELC Technologies

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.

You can find detailed UV spectra of Aspartic Acid and information about its various lambda maxima by visiting the following link.

You can find detailed UV spectra of Glutamine and information about its various lambda maxima by visiting the following link.

You can find detailed UV spectra of Glutamine and information about its various lambda maxima by visiting the following link.

You can find detailed UV spectra of Glutamine and information about its various lambda maxima by visiting the following link.

Aspartic Acid, L-Glutamine, Glutamic Acid, Glycine can be retained and analyzed using the Newcrom AH stationary phase column. The analysis utilizes an isocratic method with a simple mobile phase consisting of water and acetonitrile (MeCN) with a Formic Acid buffer. Detection is performed using CAD.

Hydroxyproline seems to be the most promising amino acid used in carbon dating when isolated from bone collagen. Separation of amino acids is challenging, especially without the use of ions or inorganic buffers that can interfere with Mass spectrometer (MS) or contaminate the sample with modern carbon. Amino acids are also not retained in reverse-phase chromatography. The ideal solution would be using water only to separate the amino acids. This would allow a direct coupling to MS. We were able to separate hydroxyproline from proline and other simple amino acids like glycine and alanine in HPLC on Newcrom AH column using water only as a mobile phase. Using water also allowed UV detection at 205 nm which can’t be done if using a buffer based on acetic or formic acid.
See more information on radiocarbon dating here.
The same method can be modified to get symmetrical peaks and higher efficiency if a mobile phase with ionic modifier such as formic acid is used.

HPLC Method for L-Glutamine, Glycine, Cysteine, L-Cysteine, Proline, Tyrosine, Arginine, Amino Acids on Primesep 100 by SIELC Technologies

High Performance Liquid Chromatography (HPLC) Method for Analysis of L-Glutamine, Glycine, Cysteine, L-Cysteine, Proline, Tyrosine, Arginine, Amino Acids.

Glutamine is an amino acid with the chemical formula C₅H₁₀N₂O₃. It is a building block for protein, but it also supports the immune system, gut health, and detoxification. It is usually found in meat, fish, eggs, dairy, and whole grains; though some consider it the most abundant amino acid in the body. People also take glutamine as treatment for sickle cell disease, healing of burns, recovery after surgery, and injuries.

L-Glycine is an important amino acid compounds widely used in pharmaceutical, biochemical, and peptide research. It has he chemical formula C₂H₅NO₂. It is water-soluble and plays a critical role in protein synthesis, peptide modification, and metabolic studies. It can be found in meat., eggs. and bones.

L-Cysteine is an amino acid with the chemical formula C₅H₁₀N₂O₃. It is primarily a building block for protein, but it also has antioxidant effects. On occasion, ot os used to support people dealing with cancer, diabetes, and hangover, but there is yet to be substantial evidence that it works. Poultry, egg, beef, and whole grains are rich sources of the amino acid.

L-Proline is an amino acid with the chemical formula C₅H₉NO₂. It is naturally produced in the body, but it is also recommended to be consumed through the diet. It can be found in meat, fish, and dairy. It is used for skin healing and treatment of other skin conditions as it is an essential component of collagen. It is important for upkeep of joints and tendons, as well as heart muscles.

L-Tyrosine is an amino acid with the chemical formula C₉H₁₁NO₃. It is naturally used in the body to create neurotransmitters like dopamine and norepinephrine. While it is produced in the body naturally, it can also be consumed in beef, pork, fish, dairy products, and beans. As a supplement, it is also used to treat Phenylketonuria (PKU).

L-Arginine is an amino acid with the chemical formula C₉H₁₁NO₃. It is a precursor for nitric acid, making it crucial for blood flow and general cardiovascular health. It can help lower blood pressure, help with chest pain, and improve symptoms of Peripheral arterial disease (PAD).

L-Glutamine, Glycine, Cysteine, L-Cysteine, Proline, Tyrosine, Arginine, Amino Acids can be retained and analyzed using the Primesep 100 stationary phase column. The analysis utilizes an isocratic method with a simple mobile phase consisting of water and acetonitrile (MeCN) with a sulfuric acid buffer. Detection is performed using UV.

HPLC Method for Glycine methyl ester, Glycine on Primesep A by SIELC Technologies

High Performance Liquid Chromatography (HPLC) Method for Analysis of Glycine methyl ester, Glycine 

Glycine Methyl Ester Hydrochloride is a water soluble organic compound that is typically used in synthesis of pharmaceuticals and pesticides. Besides that, it is also used as a flavor enhancer and nutritional supplement.

Glycine Methyl Ester Hydrochloride can be retained and analyzed using the Primesep-A stationary phase column. The analysis utilizes an isocratic method with a simple mobile phase consisting of water and acetonitrile (MeCN) with a sulfuric acid buffer. Detection is performed using UV.

HPLC Method for Glycine methyl ester, Glycine on Primesep 100 by SIELC Technologies

High Performance Liquid Chromatography (HPLC) Method for Analysis of Glycine methyl ester, Glycine 

Glycine and glycine methyl ester hydrochloride are important amino acid compounds widely used in pharmaceutical, biochemical, and peptide research. Both compounds are water-soluble and play critical roles in protein synthesis, peptide modification, and metabolic studies. High-performance liquid chromatography (HPLC) offers a precise and reliable method to separate, identify, and quantify glycine and its methyl ester derivative with excellent sensitivity and reproducibility. This analytical approach is essential for quality control, pharmaceutical formulation analysis, and research applications involving amino acid derivatives. Using HPLC for these compounds ensures accurate, consistent results in laboratory and industrial settings.

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

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.

Dimethyl sulfoxide is important polar aprotic solvent, which is frequently used in pharmaceutical drug manufacturing, desolution, etc. DMSO is used as one of the solvents on protein chemistry due to universal ability to dissolve small molecules like amino acids. Amino acids and DMSO are very polar and have no retention on reversed-phase columns. In this HPLC application DMSO and glycine are separated on Primesep 100 mixed-mode column. DMSO is retained by weak reversed-phase mechanism and very low organic concentration is required in order to achieve any retention. Glycine, like any other underivatized amino acid, is retained by weak reversed-phase and moderate cation-exchange mechanism. Method uses acetonitrile/water/TFA gradient and UV detection.