Lysine

Applications:

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Amino Acids Analysis in Acid Gradient Condition

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).

Condition

Column	Primesep 100, 4.6x250 mm, 5 µm, 100A
Mobile Phase	MeCN/H2O - 30/70%
Buffer	TFA , gradient  0.05-0.3 % , 25 min
Flow Rate	1.0 ml/min
Detection	ELSD

 

Description

Class of Compounds	Drug, Acid, Hydrophilic, Ionizable, Vitamin, Supplements
Analyzing Compounds	Gaspartic acid, Glutamic acid, Alanine, Valine, Methionine, Isoleucine, Cysteine, Phenylalanine, Histidine, Lysine, Arginine

 

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Alanine
Amino Acids
Arginine
Aspartic Acid
Cysteine
Glutamic Acid
Histidine
Isoleucine
Lysine
Methionine
Phenylalanine
Valine

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HPLC Analysis of Active Drug and Amino Acids in a Formulation

Polar amino acids are very often used as components of vitamin and supplement composition. Analysis of such complex composition is a challenging task. In this application, 5 amino acids (asparagine, glutamic acid, proline and arginine) and two preservatives (methyl paraben and propyl paraben) are separated on a Primesep 100 reversed-phase cation-exchange column with LC/MS compatible mobile phase. Method does not require ion-pairing reagent in the mobile phase. Compounds are monitored by ELSD and UV. Method is validated for quantitation of underivatized amino acids in complex mixtures. The method is simple and robust and can be used for analysis of various vitamin formulations.

Condition

Column	Primesep 100, 4.6x150 mm, 5 µm, 100A
Mobile Phase	MeCN/H2O
Buffer	AmFm
Flow Rate	1.0 ml/min
Detection	ELSD 50C, UV 250 nm

 

Description

Class of Compounds	Drug, Acid, Hydrophilic, Ionizable, Vitamin, Supplements
Analyzing Compounds	Glutamic acid, Aspargine,  Proline, Lysine, Arginine, Methyl paraben, Propyl paraben

 

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Arginine
Asparagine
Ethyl Paraben
Glutamic Acid
Lysine
Methylparaben
Proline

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HPLC Method for Determination of Lysine in Ibuprofen Lysine Composition

Ibuprofen lysine is ibuprofen based medication which is uses lysine as counter-ion of the ibuprofen. The use of lysine makes the drug more soluble in water. Lysine is very hydrophilic compound with low UV activity, while ibuprofen is hydrophobic compound with good UV activity. These properties are making straight analysis of Ibuprofen Lysine (ibuprofen lysinate) a challenging task. Ion-pairing reagent required for retention of lysine in reversed-phase chromatography. Both compounds can be quantitated in one run without use of IP reagent. Because of the huge difference in UV activity of ibuprofen and lysine, a separate method for quantitation of lysine in ibuprofen lysinate might be required. Separation involving switching valve and guard column allows to trap ibuprofen on the guard and wash it away during analysis of lysine. This HPLC method allows to quantitate lysine separately from ibuprofen. Detailed set up of switching valve/guard system is described in our newsletter. Method can be adapted to quantitation of single compound in complex mixtures.

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Ibuprofen
Lysine

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Separation of Small Lysine-based Peptide Oligomers

Short-chain peptides are usually very polar compounds. If the peptide contains lysine moiety it also become very basic in nature. Analysis in traditional reversed-phase of HILIC mode can produce poor peak shape. The method was developed for lysine-based peptide on Primesep AP column in HILIC/cation-exclusion mode. Column can be used for analysis of small peptides with multiple basic groups. Compounds can be monitored by UV, LC/MS or ELSD.

Condition

Column	Primesep AP, 4.6x150 mm, 5 µm, 100A
Mobile Phase	MeCN/H2O
Buffer	H3PO4
Flow Rate	1.0 ml/min
Detection	UV 215 nm

 

Description

Class of Compounds	Drug, Acid, Hydrophilic, Ionizable, Vitamin, Supplements
Analyzing Compounds	Lysine-based Peptide Oligomers

 

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Lysine

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HPLC Separation of Amino Acids in Zero Organic Mode on Primesep 200 column

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.

Condition

Column	Primesep 200, 4.6x250 mm, 5 µm, 100A
Mobile Phase	MeCN/H2O
Buffer	Na2HPO4
Flow Rate	1.0 ml/min
Detection	UV 210 nm

 

Description

Class of Compounds	Drug, Acid, Hydrophilic, Ionizable, Vitamin, Supplements
Analyzing Compounds	Valine, Leucyne, Isoleycine, Tyrosine, Phenylalanine, Histidine, Lysine

 

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Histidine
Isoleucine
Leucine
Lysine
Phenylalanine
Tyrosine
Valine

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HPLC Separation of Lysine and Arginine from Other Amino Acids

  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

Condition

Column	Primesep C, 4.6x150 mm, 5 µm, 100A
Mobile Phase	MeCN - 15%
Buffer	AmAc pH 5.0- 15 mM
Flow Rate	1.0 ml/min
Detection	ELSD

 

Description

Class of Compounds	Drug, Acid, Hydrophilic, Ionizable, Vitamin, Supplements
Analyzing Compounds	Aspartic acid, Glutamic acid, Aspargine, Glycine, Proline, Alanine, Phenylalanine, Tyrosine, Leucine, Lysine, Arginine

 

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Alanine
Arginine
Asparagine
Aspartic Acid
Glutamic Acid
Glycine
Leucine
Lysine
Phenylalanine
Proline
Tyrosine

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HPLC Method for Analysis of L-Lysine and L-Arginine

L-Lysine is an essential amino acid required for growth and tissue repair, it can also help with anxiety. The amino acid is most often found in red meats,beans, fish, and dairy products. L-Arginine is an essential amino acid that is used to treat congestive heart failure, combat fatigue and circulatory diseases. It is also used to stimulate the immune system. Obelisc R is a reverse-phase column. It contains embedded ionic groups and can retain L-Lysine and L-Arginine. The method is UV compatible and can be used as a general approach for analyzing similar compounds.

Condition

Column	Obelisc R, 4.6x150 mm, 5 µm, 100A
Mobile Phase	MeCN/H2O - 10/90%
Buffer	Na2HPO4 pH 6.0 - 25 mM
Flow Rate	1.0 ml/min
Detection	UV, 210 nm

 

Description

Class of Compounds	Drug, Acid, Hydrophilic, Ionizable, Vitamin, Supplements, Amino acid
Analyzing Compounds	Arginine, L-Lysine

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Arginine
D-Lysine
Lysine
N-Nitro-L-arginine

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HPLC Separation of Mixture of D-Glucose and L-Lysine on Primesep S Column

 

Condition

Column	Primesep S, 4.6x150 mm, 5 µm, 100A
Mobile Phase	Gradient MeCN - 85-60%, 10 min 2 min hold
Buffer	Gradient AmFm pH 3.0, 10-80 mM, 10 min 2 min hold
Flow Rate	1.0 ml/min
Detection	CAD (Corona) MS- compatible mobile phase

 

Description

Class of Compounds	Sugar, Amino Acid, Hydrophilic, Ionizable, Vitamin, Supplements
Analyzing Compounds	D-Glucose, L-Lysine

 

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Glucose
Lysine

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HPLC Determination of Lysine on Primesep 200 column in MS-compatible conditions

Lysine is an essential amino acid used in the synthesis of proteins. In biological conditions, it is a basic, charged molecule. Lysine can be retained on Primesep 200 mixed-mode column which has embedded weak acidic ion-pairing groups for retention of basic analytes in HPLC. The mobile phase consisting of acetonitrile (ACN) and water with ammonium formate (AmFm) makes this method analysis MS-compatible.

Condition

Column	Primesep 200, 4.6x150 mm, 5 µm, 100A
Mobile Phase	MeCN/H2O - 10/90%
Buffer	AmFM pH 3.0 - 10 mM
Flow Rate	1.0 ml/min
Detection	CAD (Corona), MS- compatible phase

 

Description

Class of Compounds	Acid, Hydrophilic, Ionizable, Carboxylic acid, Drug, Amino Acid
Analyzing Compounds	Lysine

 

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D-Lysine
Lysine

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