Proline

Proline structural formula

CAS Number609-36-9
Molecular FormulaC5H9NO2
Molecular Weight115.133
InChI KeyONIBWKKTOPOVIA-UHFFFAOYSA-N
LogP-1.64
Synonyms
  • Proline
  • 609-36-9
  • (.+-.)-Proline
  • (RS)-Proline
  • 2-Pyrrolidinylcarboxylic acid
  • DL-Prolin
  • DL-prolina
  • DL-proline
  • NSC 97923
  • Proline, DL-
  • EINECS 210-189-3
  • UNII-DCS9E77JPQ
  • Hpro
  • Prolin
  • prolina
  • pyrrolidine-2-carboxylic acid

Applications:

New HPLC Amino Acids Separation Compatible With Carbon Dating Technique

February 11, 2020

 

 

Hydroxyproline seems to be the most promising amino acid used in carbon dating when isolated from bone collagen. Separation of amino acids is challenging, especially without the use of ions or inorganic buffers that can interfere with Mass spectrometer (MS) or contaminate the sample with modern carbon. Amino acids are also not retained in reverse-phase chromatography. The ideal solution would be using water only to separate the amino acids. This would allow a direct coupling to MS. We were able to separate hydroxyproline from proline and other simple amino acids like glycine and alanine in HPLC on Newcrom AH column using water only as a mobile phase. Using water also allowed UV detection at 205 nm which can’t be done if using a buffer based on acetic or formic acid.
See more information on radiocarbon dating here.
The same method can be modified to get symmetrical peaks and higher efficiency if a mobile phase with ionic modifier such as formic acid is used.

Condition

Column Newcrom AH, 4.6×150 mm, 5 µm, 100A
Mobile Phase MeCN/H2O
Flow Rate 1.0 ml/min
Detection UV, 205 nm, CAD

 

Description

Class of Compounds
Drug, Acid, Hydrophilic, Ionizable, Vitamin, Supplements, Amino acid
Analyzing Compounds Alanine, Glycine, Proline, Hydroxyproline

 

Application Column

Newcrom AH

The Newcrom columns are a family of reverse-phase-based columns. Newcrom A, AH, B, and BH are all mixed-mode columns with either positive or negative ion-pairing groups attached to either short (25 Å) or long (100 Å) ligand chains. Newcrom R1 is a special reverse-phase column with low silanol activity.

Select options
Application Analytes:
Alanine
D-Alanine
Glycine
Hydroxyproline
Proline
SIELC Technologies usually develops more than one method for each compound. Therefore, this particular method may not be the best available method from our portfolio for your specific application. Before you decide to implement this method in your research, please send us an email to research@sielc.com so we can ensure you get optimal results for your compound/s of interest.

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

March 11, 2019

 

Condition

Column Primesep 100, 4.6×250 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 L-Asparagine (Asn/N), L-Threonine (Thr/T), L-Cysteine (Cys/C), L-Proline (Pro/P), L-Valine (Val/V), DL-Methionine (Met/M), L-Isoleucine (Ile/I), L-Leucine (Leu/L), L-Phenylalanine (Phe/F), L- Tryptophan (Trp/W), L-Histidine (His/H), L-Arginine (Arg/R)

 

Application Column

Primesep 100

The Primesep family of mixed-mode columns offers a wide variety of stationary phases, boasting unprecedented selectivity in the separation of a broad array of chemical compounds across multiple applications. Corresponding Primesep guard columns, available with all stationary phases, do not require holders. SIELC provides a method development service available to all customers. Inquire about our specially-tailored custom LC-phases for specific separations.

Select options
Application Analytes:
Amino Acids
Arginine
Asparagine
Cysteine
D-Isoleucine
D-Leucine
D-Valine
DL-Isoleucine
Histidine
Isoleucine
L-Cysteine
L-Methionine
L-Threonine
Leucine
Methionine
Phenylalanine
Proline
Tryptophan
Valine
SIELC Technologies usually develops more than one method for each compound. Therefore, this particular method may not be the best available method from our portfolio for your specific application. Before you decide to implement this method in your research, please send us an email to research@sielc.com so we can ensure you get optimal results for your compound/s of interest.

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

March 11, 2019

 

Condition

Column Primesep 100, 4.6×250 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 Glutamin (Gln/Q), L-Glycine (Gly/G), L-Cysteine (Cys/C), L-Proline Pro/p), L-Tyrosine (Tyr/Y), L-Arginine (Arg/R)

 

Application Column

Primesep 100

The Primesep family of mixed-mode columns offers a wide variety of stationary phases, boasting unprecedented selectivity in the separation of a broad array of chemical compounds across multiple applications. Corresponding Primesep guard columns, available with all stationary phases, do not require holders. SIELC provides a method development service available to all customers. Inquire about our specially-tailored custom LC-phases for specific separations.

Select options
Application Analytes:
Amino Acids
Arginine
Cysteine
Glycine
L-Cysteine
L-Glutamine
Proline
Tyrosine
SIELC Technologies usually develops more than one method for each compound. Therefore, this particular method may not be the best available method from our portfolio for your specific application. Before you decide to implement this method in your research, please send us an email to research@sielc.com so we can ensure you get optimal results for your compound/s of interest.

HPLC Separation of Lysine and Arginine from Other Amino Acids

July 10, 2012

 

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, 4.6×150 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

 

Application Column

Primesep C

The Primesep family of mixed-mode columns offers a wide variety of stationary phases, boasting unprecedented selectivity in the separation of a broad array of chemical compounds across multiple applications. Corresponding Primesep guard columns, available with all stationary phases, do not require holders. SIELC provides a method development service available to all customers. Inquire about our specially-tailored custom LC-phases for specific separations.

Select options
Application Analytes:
Alanine
Arginine
Asparagine
Aspartic Acid
Glutamic Acid
Glycine
Leucine
Lysine
Phenylalanine
Proline
Tyrosine

Application Detection:
ELSD Detection
UV Detection
SIELC Technologies usually develops more than one method for each compound. Therefore, this particular method may not be the best available method from our portfolio for your specific application. Before you decide to implement this method in your research, please send us an email to research@sielc.com so we can ensure you get optimal results for your compound/s of interest.

HPLC Analysis of Active Drug and Amino Acids in a Formulation

October 14, 2010


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, 4.6×150 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

 

Application Column

Primesep 100

The Primesep family of mixed-mode columns offers a wide variety of stationary phases, boasting unprecedented selectivity in the separation of a broad array of chemical compounds across multiple applications. Corresponding Primesep guard columns, available with all stationary phases, do not require holders. SIELC provides a method development service available to all customers. Inquire about our specially-tailored custom LC-phases for specific separations.

Select options
Application Analytes:
Arginine
Asparagine
Ethylparaben
Glutamic Acid
Lysine
Methylparaben
Proline

Application Detection:
ELSD Detection
UV Detection
SIELC Technologies usually develops more than one method for each compound. Therefore, this particular method may not be the best available method from our portfolio for your specific application. Before you decide to implement this method in your research, please send us an email to research@sielc.com so we can ensure you get optimal results for your compound/s of interest.

HPLC Analysis of Cheese Extract using Mixed-Mode Chromatography

May 6, 2005

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.

Condition

Column Primesep 100, 4.6×150 mm, 5 µm, 100A
Mobile Phase MeCN/H2O
Buffer H3PO4
Flow Rate 1.0 ml/min
Detection UV 210 nm

 

Description

Class of Compounds
Drug, Acid, Hydrophilic, Ionizable, Vitamin, Supplements
Analyzing Compounds Glutamic acid,  Proline, Glycine betaine

 

Application Column

Primesep 100

The Primesep family of mixed-mode columns offers a wide variety of stationary phases, boasting unprecedented selectivity in the separation of a broad array of chemical compounds across multiple applications. Corresponding Primesep guard columns, available with all stationary phases, do not require holders. SIELC provides a method development service available to all customers. Inquire about our specially-tailored custom LC-phases for specific separations.

Select options
Application Analytes:
Cheese Extract
Glutamic Acid
Glycine Betaine
Proline

Application Detection:
UV Detection
SIELC Technologies usually develops more than one method for each compound. Therefore, this particular method may not be the best available method from our portfolio for your specific application. Before you decide to implement this method in your research, please send us an email to research@sielc.com so we can ensure you get optimal results for your compound/s of interest.