Methionine

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.

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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.

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).