| CAS Number | 56-84-8 |
|---|---|
| Molecular Formula | C4H7NO4 |
| Molecular Weight | 133.103 g/mol |
| InChI Key | CKLJMWTZIZZHCS-REOHCLBHSA-N |
| LogP | -3.89 |
| Synonyms |
|
Applications:
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).
| 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 |
| Class of Compounds | Drug, Acid, Hydrophilic, Ionizable, Vitamin, Supplements |
| Analyzing Compounds | Gaspartic acid, Glutamic acid, Alanine, Valine, Methionine, Isoleucine, Cysteine, Phenylalanine, Histidine, Lysine, Arginine |
Application Analytes:
Alanine
Amino Acids
Arginine
Aspartic Acid
Cysteine
Glutamic Acid
Histidine
Isoleucine
Lysine
Methionine
Phenylalanine
Valine
HPLC Separation of Polar Compounds
The separation of amino acids, the building blocks of proteins, can be challenging to separate on a reverse-phase column due to their high polarity. Using a mixed-mode HPLC column, allows the separation of amino acids by cation-exchange and ion-exclusion mechanisms as well as hydrophobicity. Fine tuning of separation can be achieved with changes in organic concentration of the mobile phase as well as choice of buffer and pH.
| Column | Primesep 200, 4.6x150 mm, 5 µm, 100A |
| Mobile Phase | MeCN/H2O |
| Buffer | AcOH, AmFm |
| Flow Rate | 1.0 ml/min |
| Detection | ELSD |
| Class of Compounds | Drug, Acid, Hydrophilic, Ionizable, Hormone |
| Analyzing Compounds | Aspartic acid, Glutaric acid, Glycine, Hydroxytriptophan, GABA, Norepinephrine, Dopamine |
Application Analytes:
Aspartic Acid
Dopamine
Glutamic Acid
Norepinephrine
gamma-Aminobutyric Acid (GABA)
Bufferless Ion Separation (BLIS™) Chromatography of Amino Acids (2)
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).
| Column | Primesep 200, 4.6x150 mm, 5 µm, 100A |
| Mobile Phase | MeCN |
| Buffer | No |
| Flow Rate | 1.0 ml/min |
| Detection | UV, 195 nm |
| Class of Compounds | Drug, Acid, Hydrophilic, Ionizable, Vitamin, Supplements, Amino acid |
| Analyzing Compounds | Aspartic acid, Alanine, Valine, Methionine, Leucine |
Application Analytes:
Alanine
Amino Acids
Aspartic Acid
Leucine
Methionine
Valine
Effect of both pH and Organic Content on a Separation of Sugars, Amino Acids, and Carboxylic Acids
In mixed-mode HILIC chromatography, selectivity of separation can be adjusted by amount of acetonitrile, amount of buffer and buffer pH. Buffer concentration and pH will affect retention of ionizable compounds to a different degree. Retention of neutral compounds can be adjusted by the amount of acetonitrile. Carboxylic acid, three amino acids and two sugars are separated by combination of HILIC and ion-exchange mechanisms. Compounds can be monitored by ELSD, Corona (CAD), LC/MS or low UV. UV-transparent mobile phase /buffer is required for UV monitoring of this mixed-mode separation. This HPLC method can be adopted as general approach for analysis of sugars, amino acids and carboxylic acids.
| Column | Obelisc N, 4.6x150 mm, 5 µm, 100A |
| Mobile Phase | MeCN/H2O |
| Buffer | AmAc |
| Flow Rate | 1.0 ml/min |
| Detection | ELSD |
| Class of Compounds | Drug, Acid, Hydrophilic, Ionizable, Vitamin, Supplements, Amino acid |
| Analyzing Compounds | Succinic Acid, Phenylalanine, Sucrose, Glycine, Aspartic Acid, Raffinose |
Application Analytes:
Aspartic Acid
Phenylalanine
Succinic Acid
HPLC Separation of Aspartic Acid and Asparagine on Obelisc R Column
Two amino acids, aspartic acid and asparagine, are separated on an Obelisc R column by combination of reverse-phase and ion-exchange mechanism. Method can be used for analysis of these and other underivatized amino acids by HPLC with UV, ELSD, CAD and LC/MS detection.
| Column | Obelisc R, 4.6x150 mm, 5 µm, 100A |
| Mobile Phase | MeCN/H2O |
| Buffer | AmFm, AmAc |
| Flow Rate | 1.0 ml/min |
| Detection | ELSD |
| Class of Compounds | Drug, Acid, Hydrophilic, Ionizable, Vitamin, Supplements, Amino acid |
| Analyzing Compounds | Asparagine, Aspartic Acid |
Application Analytes:
Asparagine
Aspartic Acid
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
| 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 |
| 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 Analytes:
Alanine
Arginine
Asparagine
Aspartic Acid
Glutamic Acid
Glycine
Leucine
Lysine
Phenylalanine
Proline
Tyrosine
HPLC Separation of Mixture of Non-Essential Amino Acids
| Column | Primesep 100, 4.6x250 mm, 5 µm, 100A |
| Mobile Phase | MeCN/H2O - 20/80% |
| Buffer | H3PO4 - 0.1% |
| Flow Rate | 1.0 ml/min |
| Detection | UV, 200 nm |
| Class of Compounds | Drug, Acid, Hydrophilic, Ionizable, Vitamin, Supplements, Amino acid |
| Analyzing Compounds | L-Aspartic Acid (Asp/D), L-Serine (Ser/S), L-Glutamic Acid Glu/E), L-Alanine Ala/A) |
Application Analytes:
Amino Acids
Aspartic Acid
D-Alanine
DL-Alanine
GLU (L-Glutamic acid)
Glutamic Acid
Serine
HPLC Separation of Amino Acids on Newcrom AH Column
View on hplc.cloud
| Column | Newcrom AH, 4.6x150 mm, 5 µm, 100A |
| Mobile Phase | MeCN - 50% |
| Buffer | Formic Acid - 0.1% |
| Flow Rate | 1.0 ml/min |
| Detection | CAD |
| Class of Compounds | Drug, Acid, Hydrophilic, Ionizable, Vitamin, Supplements, Amino acid |
| Analyzing Compounds | Aspartic acid (Asp/D), Glutamine (Gln/Q), Glutamic acid (Glu/E), Glycine (Gly/G) |
Application Analytes:
Aspartic Acid
Glutamic Acid
Glycine
L-Glutamine