Proline

Applications:

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HPLC Analysis of Cheese Extract using Mixed-Mode Chromatography

Primesep 100 separates the components of a Swiss cheese extract by HPLC using cation exchange and reversed phase as retention mechanisms. The amino acids, glutamic acid and proline, as well as glycine betaine were resolved in less than 10 minutes. A mobile phase gradient of water, acetonitrile (MeCN, ACN,) and phosphoric acid (H3PO4) and ultraviolet (UV) detection was used.

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Cheese Extract
Glutamic Acid
Glycine Betaine
Proline

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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.

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Arginine
Asparagine
Ethyl Paraben
Glutamic Acid
Lysine
Methylparaben
Proline

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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

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

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HPLC Separation of Mixture of Conditionally Essential Amino Acids on Primesep 100 Column

 

Condition

Column	Primesep 100, 4.6x250 mm, 5 µm, 100A
Mobile Phase	MeCN/H2O - 35/75%
Buffer	H2SO4 0.05% 12 min hold, gradient 0.05-0.20, 13 min
Flow Rate	1.0 ml/min
Detection	UV, 200 nm

 

Description

Class of Compounds	Drug, Acid, Hydrophilic, Ionizable, Vitamin, Supplements, Amino acid
Analyzing Compounds	Amino acids

 

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Amino Acids
Arginine
Cysteine
Glycine
L-Cysteine
L-Glutamine
Proline
Tyrosine

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HPLC Separation of Mixture of 12 Amino Acids on Primesep 100 Column

 

Condition

Column	Primesep 100, 4.6x250 mm, 5 µm, 100A
Mobile Phase	MeCN/H2O - 35/65%
Buffer	H2SO4 0.05% 12 min hold, gradient 0.05-0.20, 13 min
Flow Rate	1.0 ml/min
Detection	UV, 200 nm

 

Description

Class of Compounds	Drug, Acid, Hydrophilic, Ionizable, Vitamin, Supplements, Amino acid
Analyzing Compounds	Amino acids

 

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Amino Acids
Arginine
Asparagine
Cysteine
D-Isoleucine
D-Leucine
D-Valine
DL-Isoleucine
Histidine
Isoleucine
L-Cysteine
L-Methionine
Leucine
Methionine
Phenylalanine
Proline
Tryptophan
Valine

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