| CAS Number | 60-18-4 |
|---|---|
| Molecular Formula | C9H11NO3 |
| Molecular Weight | 181.191 g/mol |
| InChI Key | OUYCCCASQSFEME-QMMMGPOBSA-N |
| LogP | -2.3 |
| Synonyms |
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Applications:
Complex Mixture of Acids, Bases, Amino Acids, and Neutral Compounds
Primesep 100 separates a mixture of amino acids (tyrosine, phenylalanine), organic acids (benzoic acid, mandelic acid), amines (benzylamine, pyridine), and neutrals (benzonitrile, toluene) in one HPLC run by combining reversed-phase, cation-exchange, and polar interactions. The method is tunable and peak order can be changed significantly by adjusting acetonitrile and trifluoroacetic acid concentrations. The separation method uses a mobile phase mixture of water, acetonitrile (MeCN, ACN) and trifluoracetic acid (TFA) and compatible with UV, mass spec (LC/MS) and evaporative light scattering (ELSD) detection.
| Column | Primesep 100, 4.6x250 mm, 5 µm, 100A |
| Mobile Phase | MeCN/H2O - 30/70% |
| Buffer | TFA - 0.2 |
| Flow Rate | 1.0 ml/min |
| Detection | UV, 210 nm |
| Class of Compounds | Drug, Acid, Hydrophilic, Ionizable, Vitamin, Supplements, Amino acid |
| Analyzing Compounds | Tyrosine, phenylalanine, Benzoic acid, mandelic acid, Benzylamine, Pyridine, Benzonitrile, Toluene |
Application Analytes:
Amino Acids
Benzoic Acid
Benzonitrile
Benzylamine
Mandelic Acid
Organic Acids
Phenylalanine
Pyridine
Toluene
Tyrosine
HPLC Analysis of the Catecholamine Pathway
Primesep 200 separates catecholamines in the catecholamine pathway in 10 minutes. Phenylalanine, tyrosine, DOPA, dopamine, norepinephrine, and epinephrine are baseline resolved by a combination of reversed-phase, ion-exchange, and ion-exclusion mechanisms. Excellent peak shape results with a mass spec compatible mobile phase of water, acetonitrile (MeCN, ACN) and trifluoracetic acid (TFA) with UV detection at 210 nm.
| Column | Primesep 200, 3.2x150 mm, 5 µm, 100A |
| Mobile Phase | MeCN/H2O - 10/90% |
| Buffer | TFA - 0.1% |
| Flow Rate | 0.5 ml/min |
| Detection | UV, 210 nm |
| Class of Compounds | Drug, Acid, Hydrophilic, Ionizable, Hormone |
| Analyzing Compounds | Tyrosine, DOPA, Phenylalanine, Norepinephrine, Epinephrine, Dopamine |
Application Analytes:
DOPA (3,4-dihydroxy-L-phenylalanine)
Dopamine
Epinephrine
Phenylalanine
Tyrosine
Bufferless Ion Separation (BLIS™) Chromatography of Amino Acids (1)
Amino acids are the building blocks of proteins and are widely used as supplements. Amino acids can exist in acidic, basic or zwitterionic form depending on the pH of their environment (mobile phase in this case). Primesep 200 is a reverse-phase column with embedded weak acidic ion-pairing groups which allows for retention and separation of amino acids without requiring a buffer making it ideal for use with UV detection, mass spectrometry (MS), evaporative light scattering detection (ELSD).
| Column | Primesep 200, 3.0x150 mm, 5 µm, 100A |
| Mobile Phase | MeCN/H2O - 40/60% |
| Buffer | No |
| Flow Rate | 0.5 ml/min |
| Detection | UV, 210 nm |
| Class of Compounds | Drug, Acid, Hydrophilic, Ionizable, Vitamin, Supplements, Amino acid |
| Analyzing Compounds | Dopa, Phenylalanine, Tyrosine |
Application Analytes:
DOPA (3,4-dihydroxy-L-phenylalanine)
Phenylalanine
Tyrosine
HPLC Analysis of Polar Basic and Acetic Compounds on Primesep AB Column
Mixture of polar acidic and basic compounds is separated on a Primesep AB mixed-mode HPLC column. Dopamine and tyrosine are retained by combination of reversed-phase and cation-exchange mechanisms. Maleic acid is retained by anion-exchange mechanism, and benzoic acid is retained by reversed-phase mechanism. Primesep AB is a trimodal column with a C12 hydrophobic chain and cation-exchange and anion exchange groups on the surface. Method utilizes UV detection but can be used with other detection techniques (ELSD, LC/MS, Corona).
| Column | Primesep AB, 4.6x150 mm, 5 µm, 100A |
| Mobile Phase | MeCN/H2O |
| Buffer | TFA - 0.1% |
| Flow Rate | 1.0 ml/min |
| Detection | UV, 210 nm |
| Class of Compounds | Drug, Acid, Monocarboxylic acid, Hydrophilic, Ionizable, Hormone |
| Analyzing Compounds | Tyrosine, Dopamine, Maleic Acid, Benzoic Acid |
Application Analytes:
Benzoic Acid
Dopamine
Maleic Acid
Tyrosine
HPLC Separation of Compounds of Catecholamine Pathway
The catecholamine neurotransmitters are amino-acid derivatives of tyrosine. DOPA, tyrosine, phenylalanine, norepinephrine, epinephrine, and dopamine and baseline are resolved on a
Primesep 200 column with UV-transparent phosphate buffer. This method can be used for the analysis of catecholamines and related impurities in various matrices. The peak order and retention time can be changed by changing the amount of ACN, buffer concentration and buffer pH. Various buffers can be used to accommodate the desired detection technique. Primesep 200 is a reversed-phase cation-exchange mixed-mode column that can be used for analysis of polar neutral, polar ionizable, polar zwitterionic, hydrophobic neutral, and hydrophobic ionic compounds in the same run. Column can be operated in reverse-phase, cation-exchange, anion-exclusion, HILIC and mixed-modes depending on the mobile phase selection and the nature of the analytes. The column is compatible with LC/MS and does not require the use of ion-pairing reagents.
| Column | Primesep 200, 4.6x150 mm, 5 µm, 100A |
| Mobile Phase | MeCN/H2O |
| Buffer | TFA, AmFm |
| Flow Rate | 1.0 ml/min |
| Detection | UV, 210 nm |
| Class of Compounds | Drug, Acid, Hydrophilic, Ionizable, Hormone |
| Analyzing Compounds | Tyrosine, DOPA, Phenylalanine, Norepinephrine, Epinephrine, Dopamine |
Application Analytes:
DOPA (3,4-dihydroxy-L-phenylalanine)
Dopamine
Epinephrine
Phenylalanine
Tyrosine
HPLC Separation of Catecholamines on Primesep 100 Column with Phosphate Buffers
The catecholamine neurotransmitters are amino-acid derivatives of tyrosine. DOPA, tyrosine, phenylalanine, norepinephrine, epinephrine, and dopamine and baseline are resolved on a Primesep 100 column with UV-transparent phosphate buffer. This method can be used for analysis of catecholamines and related impurities in various matrices. Peak order and retention time can be changed by changing the amount of ACN, buffer concentration and buffer pH. Various buffers can be used to accommodate desired detection technique. Primesep 100 is a reversed-phase cation-exchange mixed-mode column that can be used for analysis of polar neutral, polar ionizable, polar zwitter-ionic, hydrophobic neutral, and hydrophobic ionic compounds in the same run. Column can be operated in reverse-phase, cation-exchange, anion-exclusion, HILIC and mixed-modes depending on the mobile phase selection and nature of analytes. Column is compatible with LC/MS and does not require use of ion-pairing reagents.
| Column | Primesep 100, 4.6x150 mm, 5 µm, 100A |
| Mobile Phase | MeCN/H2O |
| Buffer | Na2HPO4 |
| Flow Rate | 1.0 ml/min |
| Detection | UV, 210 nm |
| Class of Compounds | Drug, Acid, Hydrophilic, Ionizable, Hormone |
| Analyzing Compounds | Tyrosine, DOPA, Phenylalanine, Norepinephrine, Epinephrine, Dopamine |
Application Analytes:
DOPA (3,4-dihydroxy-L-phenylalanine)
Dopamine
Epinephrine
Norepinephrine
Phenylalanine
Tyrosine
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.
| 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 |
| Class of Compounds | Drug, Acid, Hydrophilic, Ionizable, Vitamin, Supplements |
| Analyzing Compounds | Valine, Leucyne, Isoleycine, Tyrosine, Phenylalanine, Histidine, Lysine |
Application Analytes:
Histidine
Isoleucine
Leucine
Lysine
Phenylalanine
Tyrosine
Valine
HPLC Separation of Amino Acids in HILIC and Cation-Exchange Modes
Amino acids can be retained and separated on a Primesep S2 column in HILIC/cation-exchange mode.
| Column | Primesep S2, 4.6x150 mm, 5 µm, 100A |
| Mobile Phase | MeCN/H2O |
| Buffer | AmFm |
| Flow Rate | 1.0 ml/min |
| Detection | UV, 270 nm |
| Class of Compounds | Drug, Acid, Monocarboxylic acid, Hydrophilic, Ionizable, Hormone |
| Analyzing Compounds | Dopa, Tyrosine, Phenylalanine |
Application Analytes:
DOPA (3,4-dihydroxy-L-phenylalanine)
Phenylalanine
Tyrosine
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 Conditionally Essential Amino Acids on Primesep 100 Column
| 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 |
| 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 Analytes:
Amino Acids
Arginine
Cysteine
Glycine
L-Cysteine
L-Glutamine
Proline
Tyrosine