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Separation of Diacid: Ion Exclusion mode |
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Application Notes:
Primesep 100 separates a mixture of dicarboxylic acids in ion-exclusion mode with a mobile phase of water, acetonitrile (MeCN, ACN), and sulfuric acid (H2SO4) with UV detection at 210 nm. Baseline resolution of fumaric, maleic, malic, and succinic acids is obtained in less than 8 minutes. The separation combines ion-exclusion and reversed-phase mechanisms in one method.
Application Columns:
- Primesep® 100 is a mixed-mode HPLC column with general hydrophobic and ion-exchange properties. The Primesep 100 stationary phase combines a hydrophobic chain and an acidic functional group as a cation exchanger. Acids, bases, zwitterions and neutrals can be retained and separated by Primesep 100 columns. Primesep 100 retains hydrophobic compounds by reversed-phase retention alone or a combination of reversed-phase and ion-exchange retention. Primesep 100 separates underivatized amino acids and amines by combining reversed-phase and ion-exchange mechanisms. In this case the acidic functional group acts as an embedded ion-pairing reagent. Organic acids are separated by reversed phase and ion exclusion. Inorganic cations can be separated by cation exchange. Methods can be developed on Primesep columns that are fully compatible with detectors such as UV, RI, conductivity, fluorescence, evaporative light scattering (ELSD) as well as mass spectrometers (LC/MS) and preparative chromatography (prep HPLC) systems.
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Separation of Diacid Hydrophobic and Ion Exclusion Modes |
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Application Notes:
Primesep 200 retains and separates the organic diacids (malic, succinic, fumaric, and maleic) by a combination hydrophobic, reversed-phase interactions and ion exclusion. The separation uses a mobile phase of water, acetonitrile (MeCN, ACN) and trifluoracetic acid (TFA) with UV detection at 210 nm.
Application Columns:
- Primesep® 200 is a mixed-mode HPLC column with general hydrophobic and ion-exchange properties. The Primesep 200 stationary phase combines a hydrophobic chain and a weakly acidic functional group as a cation exchanger. Acids, bases, zwitterions and neutrals can be retained and separated by Primesep 200 columns. Primesep 200 retains hydrophobic compounds by reversed-phase retention alone or a combination of reversed-phase and ion-exchange retention. Primesep 200 separates underivatized amino acids, amines, isomers and structurally related compounds by combining reversed phase and ion exchange. In this case the acidic functional group acts as an embedded ion-pairing reagent. Organic acids and diacids are separated by reversed phase and ion exclusion. Methods can be developed on Primesep columns that are fully compatible with detectors such as UV, RI, conductivity, fluorescence, evaporative light scattering (ELSD) as well as mass spectrometers (LC/MS) and preparative chromatography (prep HPLC) systems.
Application Analytes:
Application Detection:
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Separation of Diacid Hydrophobic and Ion Exchange Modes |
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Application Notes:
Primesep B combines a hydrophobic, reversed-phase mechanism with ion exchange to separate the diacids, fumaric, benzoic, phthalic, naphthoic, and maleic acids. Changing the acetonitrile content of the mobile phase reverses the peak order for naphthoic and maleic acids. Primesep B combines reversed-phase and anion-exchange mechanism with a mobile phase of water, acetonitrile (MeCN, ACN) and trifluoracetic acid (TFA) and UV detection at 250 nm.
Application Columns:
- Primesep® B is a mixed-mode HPLC column with general hydrophobic and ion-exchange properties. The Primesep B stationary phase combines a hydrophobic chain and a strongly basic functional group as an anion exchanger recommended for mobile phase pH's of 1.5 to 4. Primesep B separates bases by an ion-exclusion mechanism. The basic functional group acts as an embedded ion-pairing reagent. Recommended buffers for Primesep columns are trifluoroacetic acid (TFA), phosphoric acid (H3PO4), perchloric acid (HClO4), and formic acid. Methods can be developed on Primesep columns that are fully compatible with detectors such as UV, RI, conductivity, fluorescence, evaporative light scattering (ELSD) as well as mass spectrometers (LC/MS) and preparative chromatography (prep HPLC) systems.
Application Analytes:
Application Detection:
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HPLC Separation of Organics Acids |
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Application Notes:
Primesep D separates organic acids such as fumaric, benzoic, phthalic, naphthoic, and maleic acids by a mixture of anion exchange and reversed phase. Retention times and elution order can be changed by adjusting the percentage of acetonitrile in the mobile. This can not be done by traditional ion-exchange and ion-exclusion chromatography. The HPLC separation uses a mobile phase of water, acetonitrile (MeCN, ACN) and trifluoroacetic acid (TFA) and UV detection at 250 nm.
Application Columns:
- Primesep® D is a reversed-phase HPLC column with general hydrophobic and ion-exchange properties for direct injection of plasma samples. The Primesep D stationary phase combines a hydrophobic chain and a basic functional group as an anion exchanger. Primesep D excludes high molecular weight plasma compounds while retaining hydrophobic and acidic small molecules. Methods can be developed on Primesep columns that are fully compatible with detectors such as UV, RI, conductivity, fluorescence, evaporative light scattering (ELSD) as well as mass spectrometers (LC/MS) and preparative chromatography (prep HPLC) systems.
Application Analytes:
Application Detection:
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HILIC Separation of Carboxylic Acids |
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Application Notes:
Application Columns:
- Obelisc N columns are the first commercially available columns with Liquid Separation Cell technology (LiSC). With multiple patents pending, LiSC technology is based on a new chemical modification of silica gel pores that creates a liquid separation cell with its own charge characteristics, ionic strength, and hydrophobic properties. Like living cells which exist in equilibrium with the outside environment, liquid separation cells exist in constant equilibrium with the mobile phase. Obelisc N, for normal phase, has very polar characteristics and works well for polar and charged analytes. In ion-exchange mode, charged analytes interact with oppositely charged groups on the stationary phase. In traditional HILIC mode, a charged or neutral polar analyte interacts with a water layer on the polar stationary phase surface. On Obelisc N the charges are greatly separated and independently accessible which results in different selectivity compared to traditional HILIC and silica columns. Mobile phase composition changes the conformation of the long hydrophilic chain. This affects the properties of the liquid separation cell and changes separation selectivity. Typical mobile phases used with Obelisc N columns are based on acetonitrile, water, and the mass spec compatible buffers ammonium formate (pH 3) and ammonium acetate (pH 5). If it is necessary to detect in low UV (<220 nm) then phosphate buffer is recommended. Obelisc N columns are compatible with mass spectrometers (LC/MS), evaporative light scattering detectors and preparative chromatography (prep HPLC) system.
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Application Detection:
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