CAS Number | 150-30-1 |
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Molecular Formula | C9H11NO2 |
Molecular Weight | 165.192 |
InChI Key | COLNVLDHVKWLRT-UHFFFAOYSA-N |
LogP | -1.38 |
Synonyms |
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Essential amino acids cannot be made by the body. As a result, they must come from food.
The 9 essential amino acids are: histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, and valine.
Column | Newcrom AH, 4.6×150 mm, 5 µm, 100A |
Mobile Phase | MeCN – 5% |
Buffer | Gradient Formic Acid – 3-9%, 20 min |
Flow Rate | 1.0 ml/min |
Detection | CAD |
Class of Compounds |
Drug, Acid, Hydrophilic, Ionizable, Vitamin, Supplements, Amino acid |
Analyzing Compounds | L-Threonine (Thr/T), L-Valine (Val/V), DL-Methionine (Met/M), L-Isoleucine (Ile/I), L-Leucine (Leu/L), L-Phenylalanine (Phe/F), L-Histidine (His/H), Lysine (Lys/K) , L- Tryptophan (Trp/W), L-Arginine (Arg/R) |
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 – 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 | 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) |
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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Column | Primesep 100, 4.6×250 mm, 5 µm, 100A |
Mobile Phase | MeCN/H2O – 25/75% |
Buffer | Gradient H2SO4 0.05-0.2% 25 min |
Flow Rate | 1.0 ml/min |
Detection | UV, 200 nm |
Class of Compounds |
Drug, Acid, Hydrophilic, Ionizable, Vitamin, Supplements, Amino acid |
Analyzing Compounds | L-Threonine (Thr/T), L-Valine (Val/V), DL-Methionine (Met/M), L-Isoleucine (Ile/I), L-Leucine (Leu/L), L-Phenylalanine (Phe/F), L-Histidine (His/H), L- Tryptophan (Trp/W) |
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.
Select optionsAmino acids can be retained and separated on a Primesep S2 column in HILIC/cation-exchange mode.
Column | Primesep S2, 4.6×150 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 |
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 optionsThe 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, 4.6×150 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 |
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 optionsEssential 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, 4.6×250 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 |
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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Primesep mixed-mode columns are designed to separate basic, acidic, neutral and zwitter-ionic compounds. Elution of these compounds is facilitated by acetonitrile and ions in the mobile phase. In all cases the retention time on mixed-mode columns is significantly longer than on traditional reverse-phase columns. Presence of cation-exchange mechanism in Primesep 100 column allows to achieve better retention and peak shape for analytes like phenylalanine and benzylamine. Various buffers can be used along with different detection techniques. This HPLC method can be adopted as a universal approach for analysis of basic, acidic and zwitter-ionic compounds in complex mixtures and formulations. Compounds can be monitored by UV, ELSD, CAD and LC/MS.
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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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 optionsClosely related compounds like amino acids can be separated on an Obelisc R column by various buffers depending on the amount of baseline separation required. By choosing different buffers, the separation between compounds can be adjusted based on application needs, especially those that require low organic concentration in the mobile phase. UV detection at 250nm.
Column | Obelisc R, 4.6×250 mm, 5 µm, 100A |
Mobile Phase | MeCN/H2O – 5/95% |
Buffer | AmFm |
Flow Rate | 1.0 ml/min |
Detection | UV, 250 nm |
Class of Compounds |
Drug, Acid, Hydrophilic, Ionizable, Vitamin, Supplements, Amino acid |
Analyzing Compounds | Amino acids |
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 Phenylalanine, Glucosamine and Tryptophan demonstrates how the small changes to the organic and/or buffer concentrations in the mobile phase affect the retention of compounds on a mixed-mode column by hydrophobic, ion-exchange and ion-exclusion mechanisms in reverse-phase HPLC chromatography. Evaporative Light Scattering Detector (ELSD) used
Column | Primesep 100, 2.1×100 mm, 5 µm, 100A |
Mobile Phase | MeCN/H2O |
Buffer | AmFm |
Flow Rate | 0.25 ml/min |
Detection | ELSD |
Class of Compounds |
Drug, Acid, Hydrophilic, Ionizable, Vitamin, Supplements, Amino acid |
Analyzing Compounds | Phenylalanine, Glucosamine, Tryptophan |
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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Separating basic, acidic and zwitterionic compounds in one run in reverse-phase HPLC can be very challenging. The methods might require the use of ion-pairing reagents and complex gradients that can make MS-compatibility difficult. Obelisc R column which has both positive and negative ion-pairs embedded in the stationary phase allows for fine tuning and separation of a wide range of compounds with different ionic properties. Acids, bases, amino acids and neutral compounds were separated isocratically in one run using a simple MS-compatible mobile phase of acetonitrile (ACN) and water with Ammonium Acetate (AmAc) buffer. Can also be UV detected at 250nm.
Column | Obelisc R, 4.6×250 mm, 5 µm, 100A |
Mobile Phase | MeCN/H2O – 35/65% |
Buffer | AmAc 10 mM pH 4.0 |
Flow Rate | 1.0 ml/min |
Detection | UV, 250 nm |
Class of Compounds |
Drug, Acid, Bases, Neutral, Hydrophilic, Ionizable, Vitamin, Supplements, Amino acid |
Analyzing Compounds | Amino acids |
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 optionsPrimesep 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, 4.6×250 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 |
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 optionsThe 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, 4.6×150 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 |
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 optionsAmino 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, 3.0×150 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 |
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 optionsPrimesep 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, 3.2×150 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 |
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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