High Performance Liquid Chromatography (HPLC) Method for Analysis of Methionine on Primesep 100 by SIELC Technologies

Separation type: Liquid Chromatography Mixed-mode SIELC Technologies

 High Performance Liquid Chromatography (HPLC) Method for Analysis of Methionine

Methionine is an essential amino acid, meaning the body cannot synthesize it, so it must be obtained through the diet. It’s crucial for many bodily functions, including:

Protein Synthesis: Methionine is a building block for proteins.

Methylation: It acts as a precursor to S-adenosylmethionine (SAMe), which is involved in methylation processes that are vital for DNA regulation, detoxification, and neurotransmitter production.

Antioxidant Function: Methionine contributes to the synthesis of cysteine, which is a precursor to glutathione, one of the body’s most important antioxidants.

Liver Health: It’s also important for liver function and fat metabolism.

Dietary sources of methionine include meat, fish, dairy products, and some nuts and seeds. Methionine levels can be particularly important in contexts like growth, tissue repair, and maintaining healthy skin and nails.

Methionine can be retained, separated and analyzed using a Primesep 100 mixed-mode stationary phase column. The analysis employs a gradient method with a simple mobile phase comprising water, acetonitrile (MeCN), and sulfuric acid as a buffer. This method allows for detection using UV 200 nm.

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

For optimal results in HPLC analysis, it is recommended to measure absorbance at a wavelength that matches the absorption maximum of the compound(s) being analyzed. The UV spectrum shown can assist in selecting an appropriate wavelength for your analysis. Please note that certain mobile phases and buffers may block wavelengths below 230 nm, rendering absorbance measurement at these wavelengths ineffective. If detection below 230 nm is required, it is recommended to use acetonitrile and water as low UV-transparent mobile phases, with phosphoric acid and its salts, sulfuric acid, and TFA as buffers.

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 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.

Four underivatized amino acids (serine, methylserine, GABA and methionine) were separated on a Primesep 100 reversed-phase cation-exchange mixed-mode HPLC column. Two methods, one for UV and one for ELSD/LC/MS show good separation and peak shape for underivatized amino acids. This column and general HPLC approach can be used for analysis of underivatized amino acids. Primesep 100 is designed to replace reversed-phase HPLC column in combination with ion-pairing reagents.

Amino acids are building blocks for peptides and proteins. Serine is not essential to human diet and it is synthesized in human body. Methionine is not synthesized in human body and needs to be ingested. GABA is used to enhance growth of specified plants, prevent development of powdery mildew on grapes, and suppress certain other plant diseases. In humans GABA helps to maintain normal brain function. Serine, methylserine, GABA and methionine are separated on Primesep 100 column by mixed-mode mechanism. Amino acids are retained by combination of reverse phase and cation-exchange mechanisms. At lower pH carboxylic acid fragment of amino acid is suppressed and not ionized, making amino acids more basic and slightly more hydrophobic. Amount of acetonitrile, buffer concentration and buffer pH can be used to adjust retention time. Underivatized amino acids are well retained and separated with perfect peak shape and symmetry. Fast method can be used for UV, ESLD and LC/MS quantitation of serine, methylserine, GABA and methionine.

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).