CAS Number | 56-84-8 |
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Molecular Formula | C4H7NO4 |
Molecular Weight | 133.103 |
InChI Key | CKLJMWTZIZZHCS-REOHCLBHSA-N |
LogP | -3.89 |
Synonyms |
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Column | Newcrom AH, 4.6×150 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) |
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.
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Column | Primesep 100, 4.6×250 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) |
The Primesep family of mixed-mode columns offer a wide variety of stationary phases with an unprecedented selectivity in the separation of a broad array of chemical compounds and in multiple applications. Corresponding Primesep guard columns are available with all stationary phases and do not require holders. SIELC offers a method development service which is available for all customers. Ask about our special custom LC-phases tailored for specific separations.
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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, 4.6×150 mm, 5 µm, 100A |
Mobile Phase | MeCN – 15% |
Buffer | AmAc pH 5.0- 15 mM |
Flow Rate | 1.0 ml/min |
Detection | ELSD |
Class of Compounds |
Drug, Acid, Hydrophilic, Ionizable, Vitamin, Supplements |
Analyzing Compounds | Aspartic acid, Glutamic acid, Aspargine, Glycine, Proline, Alanine, Phenylalanine, Tyrosine, Leucine, Lysine, Arginine |
The Primesep family of mixed-mode columns offer a wide variety of stationary phases with an unprecedented selectivity in the separation of a broad array of chemical compounds and in multiple applications. Corresponding Primesep guard columns are available with all stationary phases and do not require holders. SIELC offers a method development service which is available for all customers. Ask about our special custom LC-phases tailored for specific separations.
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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, 4.6×150 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 |
SIELC has developed the mixed-mode Obelisc™ columns to be the first commercially available columns with Liquid Separation Cell technology (LiSC™). This cost effective duo can replace multiple HPLC columns such as reversed-phase (RP), AQ-type reversed-phase, polar-embedded group RP columns, normal-phase, cation-exchange, anion- exchange, ion-exclusion, and HILIC (Hydrophilic Interaction Liquid Chromatography) columns. By controlling just three orthogonal method parameters - Buffer concentration, Buffer pH, and Organic Modifier Concentration - users can adjust the column properties with pinpoint precision to separate complex mixtures.
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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, 4.6×150 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 |
SIELC has developed the mixed-mode Obelisc™ columns to be the first commercially available columns with Liquid Separation Cell technology (LiSC™). This cost effective duo can replace multiple HPLC columns such as reversed-phase (RP), AQ-type reversed-phase, polar-embedded group RP columns, normal-phase, cation-exchange, anion- exchange, ion-exclusion, and HILIC (Hydrophilic Interaction Liquid Chromatography) columns. By controlling just three orthogonal method parameters - Buffer concentration, Buffer pH, and Organic Modifier Concentration - users can adjust the column properties with pinpoint precision to separate complex mixtures.
Select optionsThe 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, 4.6×150 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 |
The Primesep family of mixed-mode columns offer a wide variety of stationary phases with an unprecedented selectivity in the separation of a broad array of chemical compounds and in multiple applications. Corresponding Primesep guard columns are available with all stationary phases and do not require holders. SIELC offers a method development service which is available for all customers. Ask about our special custom LC-phases tailored for specific separations.
Select options11 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, 4.6×250 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 | Aspartic acid, Glutamic acid, Alanine, Valine, Methionine, Isoleucine, Cysteine, Phenylalanine, Histidine, Lysine, Arginine |
The Primesep family of mixed-mode columns offer a wide variety of stationary phases with an unprecedented selectivity in the separation of a broad array of chemical compounds and in multiple applications. Corresponding Primesep guard columns are available with all stationary phases and do not require holders. SIELC offers a method development service which is available for all customers. Ask about our special custom LC-phases tailored for specific separations.
Select optionsAdding 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, 4.6×150 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 |
The Primesep family of mixed-mode columns offer a wide variety of stationary phases with an unprecedented selectivity in the separation of a broad array of chemical compounds and in multiple applications. Corresponding Primesep guard columns are available with all stationary phases and do not require holders. SIELC offers a method development service which is available for all customers. Ask about our special custom LC-phases tailored for specific separations.
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