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.

HPLC Method for Asparagine, L-Cysteine, Cysteine, Proline, Valine, D-Valine, Methionine, L-Methionine, Isoleucine, D-Isoleucine, DL-Isoleucine, D-Leucine, Phenylalanine, Tryptophan, Histidine, Arginine, Amino Acids, Leucine, L-Threonine on Primesep 100 by SIELC Technologies

High Performance Liquid Chromatography (HPLC) Method for Analysis of Asparagine, L-Cysteine, Cysteine, Proline, Valine, D-Valine, Methionine, L-Methionine, Isoleucine, D-Isoleucine, DL-Isoleucine, D-Leucine, Phenylalanine, Tryptophan, Histidine, Arginine, Amino Acids, Leucine, L-Threonine.

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 sulfuric acid (H2SO4) as a buffer. This analysis method can be detected in the UV regime at 200 nm.

HPLC Method for Valine, D-Valine, Methionine, L-Methionine, Isoleucine, D-Isoleucine, DL-Isoleucine, L-Isoleucine, D-Leucine, Phenylalanine, Histidine, L-Histidine hydrochloride monohydrate, Tryptophan, Amino Acids, L-Threonine on Primesep 100 by SIELC Technologies

High Performance Liquid Chromatography (HPLC) Method for Analysis of Valine, D-Valine, Methionine, L-Methionine, Isoleucine, D-Isoleucine, DL-Isoleucine, L-Isoleucine, D-Leucine, Phenylalanine, Histidine, L-Histidine hydrochloride monohydrate, Tryptophan, Amino Acids, L-Threonine.

L-Threonine is an essential amino acid with the chemical formula C₄H₉NO₃.  It cannot be produced within the body and must be obtained through consuming it. It’s found in many protein-rich foods, including but not limited to eggs, meat, dairy, legumes, and seeds. It is necessary in the body as a building block of protein like collagen and elastin. The two proteins are crucial for skin, hair, and connective issue.

L-Valine is an essential amino acid with the chemical formula C₅H₁₁NO₂. It cannot be produced within the body and must be obtained through consuming it. It’s found in foods including but not limited to nuts, legumes, whole grains, and seeds. It is especially beneficial for athletes.  It is important for muscle repair,  growth, and energy regulation.

DL-Methionine is an essential amino acid with the chemical formula C₅H₁₁NO₂S.  It cannot be produced within the body and must be obtained through consuming it. It is required for protein synthesis. It also helps build and repair tissue including, but not limited to, skin, hair, muscles, and nails. In a veterinary context, DL-Methionine is used to address bladder issues in dogs.

L-Isoleucine is an essential amino acid with the chemical formula C₆H₁₃NO₂.  It cannot be produced within the body and must be obtained through consuming it. It is a building block of protein that are essential for muscle growth, repair, and other bodily functions.  It also helps regulate blood sugar levels and supports the immune system. It is found in foods like meat, fish, eggs, dairy, beans, lentils, nuts, and seeds.

L-Leucine is an essential amino acid with the chemical formula C₆H₁₃NO₂. It cannot be produced within the body and must be obtained through consuming it. It stimulates production of protein that are essential for muscle building and repair. Meats are the easiest way to get L-Leucine in significant amounts.

L-Phenylalanine is an essential amino acid with the chemical formula C₆H₉NO₂. It cannot be produced within the body and must be obtained through consuming it. It is typically found in high protein foods such as meat, eggs, and fish. Outside of being important for creation of protein, it is also used in treatment for skin disorders and depression.

L-Histidine is an essential amino acid with the chemical formula C₉H₁₁N₃O₂. It cannot be produced within the body and must be obtained through consuming it. It s fundamental for repair of damaged tissue, growth of muscles, and making of blood cells. Outside of protein, it also has the unique property of being able to act as a buffer to help maintain stable pH levels in the body. Sources of it include meat, fish, dairy products, beans, and nuts.

L-Tryptophan is an essential amino acid with the chemical formula C₁₁H₁₂N₂O₂. It cannot be produced within the body and must be obtained through consuming it. Like the other essential proteins, it is a building block for protein and muscle tissue, but it is also converted in the body into serotonin, which affects mood. L-Tryptophan is also used in treatments for severe PMS symptoms, depression, and insomnia. It is naturally found in red meat, poultry eggs, and dairy.

Valine, D-Valine, Methionine, L-Methionine, Isoleucine, D-Isoleucine, DL-Isoleucine, L-Isoleucine, D-Leucine, Phenylalanine, Histidine, L-Histidine hydrochloride monohydrate, Tryptophan, Amino Acids, L-Threonine 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 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.

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.

Amino acids are essential components of numerous formulation. Health supplements can contain various amino acids and vitamins and require quantitation of each ingredients. Amino acids are very polar compounds with limited or no retention in reversed-phase chromatography. The most common approaches are reversed-phase chromatography with ion-pairing reagent and hydrophilic interaction chromatography (HILIC). Underivatized amino acids can be retained by combination of reversed-phase and cation exchange mechanism on Primesep 100 mixed-mode. Retention time is controlled by amount of acetonitrile, buffer and buffer pH. Method does not require ion-pairing reagent. This method is for UV detection. LC/MS, ELSD or Corona CAD can be employed for analysis of amino acids with trifluoroacteic acid or ammonium formate in the mobile phase. This approach can be used for HPLC analysis of all underivatized amino acids.

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.

Primesep 100 separates a mixture of amino acids (tyrosine, phenylalanine), organic acids (benzoic acid, mandelic acid), amines (benzylamine, pyridine), and neutrals (benzonitrile, toluene) in one HPLC run by combining reversed-phase, cation-exchange, and polar interactions. The method is tunable and peak order can be changed significantly by adjusting acetonitrile and trifluoroacetic acid concentrations. The separation method uses a mobile phase mixture of water, acetonitrile (MeCN, ACN) and trifluoracetic acid (TFA) and compatible with UV, mass spec (LC/MS) and evaporative light scattering (ELSD) detection.

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).

Adding on to the previous HPLC separation of amino acids using Bufferless Ion Separation (BLIS) Chromatography; here we have additional amino acids separated on Primesep 200 column using only water and acetonitrile (MeCN, ACN) in the mobile phase.  Primesep 200 is a reverse-phase (RP) column with weak acidic ion-pairing groups embedded. With no buffer present in the mobile phase, detection can be achieved with UV, mass spectrometry (MS), evaporative light scattering detection (ELSD).

Primesep 100 separates a mixture of polar and nonpolar compounds in one analytical run. The amino acid cysteine; amino acid derivatives L-cystine, 2,2-dimethylcystine, and 2-methylcysteine; the polar acid benzoic acid; and the nonpolar neutral toluene are separated by a gradient using a combination of polar and hydrophobic interactions. The separation method uses a mobile phase mixture of water, acetonitrile (MeCN, ACN) and sulfuric acid (H2SO4) with UV detection at 210 nm.