HPLC Method for L-Threonine, Valine, Methionine, Isoleucine, Leucine, Phenylalanine, Histidine, Tryptophan, Lysine, Arginine on Newcrom AH by SIELC Technologies

High Performance Liquid Chromatography (HPLC) Method for Analysis of L-Threonine, Valine, Methionine, Isoleucine, Leucine, Phenylalanine, Histidine, Tryptophan, Lysine, Arginine.

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-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.

Lysine is an essential amino acid used in the synthesis of proteins. In biological conditions, it is a basic, charged molecule.

L-Arginine is an essential amino acid used in the synthesis of proteins.

L-Threonine, Valine, Methionine, Isoleucine, Leucine, Phenylalanine, Histidine, Tryptophan, Lysine, Arginine can be retained and analyzed using the Newcrom AH stationary phase column. The analysis utilizes a gradient method with a simple mobile phase consisting of water and acetonitrile (MeCN) with a Formic Acid buffer. Detection is performed using CAD.

HPLC Method for D-Isoleucine, D-Leucine, DL-Isoleucine, L-Alloisoleucine, L-Isoleucine, allo-D-isoleucine, Isoleucine, Leucine on Primesep 100 by SIELC Technologies

High Performance Liquid Chromatography (HPLC) Method for Analysis of D-Isoleucine, D-Leucine, DL-Isoleucine, L-Alloisoleucine, L-Isoleucine, allo-D-isoleucine, Isoleucine, Leucine 

Alloisoleucine is an amino acid with C₆H₁₃NO₂ chemical formula. It is a key indicator when diagnosing Maple Syrup Urine Disease, when a body cannot metabolize amino acids in specific foods. Outside of that, it is also used in peptide synthesis and drug development.

Isoleucine is an amino acid with chemical formula C₆H₁₃NO₂. It is a building block for proteins and is used in a variety of bodily functions. It helps form hemoglobin, regulating blood sugar, plays a key role in energy production, and increases the speed of muscle healing and growth. The easiest way to prevent a deficiency of it is through eating foods rich in protein, as those typically contain necessary amino acids.

Leucine is an amino acid with chemical formula C₆H₁₃NO₂. It is key for protein synthesis, metabolism, tissue repair, and certain tasks that are also done by Isoleucine. It is known to activate mTOR cell signalng pathways, which stimulate protein synthesis and prevent breakdown of protein. It is found bin animal protein, plant-based protein, and nuts.

D-Isoleucine, D-Leucine, DL-Isoleucine, L-Alloisoleucine, L-Isoleucine, allo-D-isoleucine, Isoleucine, Leucine 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 ammonium formate buffer. Detection is performed using CAD.

HPLC Method for β‑Hydroxy-β-methylbutyric Acid (HMB), D-Leucine, Leucine, 2-Hydroxyisocaproic Acid (HICA) on Primesep 100 by SIELC Technologies

High Performance Liquid Chromatography (HPLC) Method for Analysis of β‑Hydroxy-β-methylbutyric Acid (HMB), D-Leucine, Leucine, 2-Hydroxyisocaproic Acid (HICA) 

Both β-hydroxy-β-methylbutyric acid (HMB) and 2-hydroxyisocaproic acid (HICA) are metabolites of the
essential dietary branched chain amino acid leucine.

β‑Hydroxy-β-methylbutyric Acid (HMB), D-Leucine, Leucine, 2-Hydroxyisocaproic Acid (HICA) can be retained and analyzed using the Primesep 100 stationary phase column. The analysis utilizes a gradient 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 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.

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

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.

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.

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).