| |
 |
Application Notes:
Primesep C separates B vitamins with baseline resolution by a combination of cation exchange, complex formation, and hydrophobic interactions. Vitamin B1 (thiamine), vitamin B2 (riboflavin), and vitamin B6 (pyridoxine) are separated with a mobile phase of water, acetonitrile (MeCN, ACN) and triethylamine (TEA) phosphate with UV detection at 280 nm.
Application Columns:
- Primesep® C is a mixed-mode HPLC column with reversed-phase, ion-exchange and host-guest complex formation properties. The Primesep C stationary phase combines a hydrophobic chain and an acidic functional group as a cation exchanger. Acids, bases, and neutrals can be retained and separated by Primesep 100 columns. Primesep C retains hydrophobic compounds by a reversed phase. Strongly polar and basic compounds such as quaternary amines are retained without the use of ion-pairing reagent by reversed phase and ion exchange. An unusual elution order compared to other HPLC columns is found for primary, secondary, and tertiary amines. 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:
|
Separation and Retention of Vitamins in Mixed-Mode HPLC |
 |
Application Notes:
Primesep B and B2 columns separate Vitamins C (ascorbic acid) and B2 (riboflavin) with tunable selectivity. Both columns allow peak order reversal by simply changing the acetonitrile concentration in the mobile phase. Vitamin B2 is especially sensitive to acetonitrile concentration. The HPLC separation uses a mobile phase of water, acetonitrile (MeCN, ACN) and either trifluoroacetic acid (TFA) or acetic acid (HOAc) with 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.
- Primesep® B2 is a mixed-mode HPLC column with general hydrophobic and ion-exchange properties. The Primesep B2 stationary phase combines a hydrophobic chain and a basic functional group as an anion exchanger. Acids, bases, zwitterions and neutrals can be retained and separated by Primesep B2 columns. Primesep B2 also separates bases by an ion-exclusion mechanism. Primesep B2 demonstrates significant improvement in retention and selectivity for the separation of weak acid analytes by a combination of reversed phase and anion exchange. In this case the basic functional group acts as an embedded ion-pairing reagent. Primesep B2 column is one of the best separation media for natural product extracts. 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:
|
Separation of Two Vitamins: Different Polarity - Isocratic Methods |
 |
Application Notes:
Primesep B and B2 columns separate Vitamins C (ascorbic acid) and B2 (riboflavin) with tunable selectivity. Peak order reversal is exhibited on these two columns with the same mobile phases due to their different polarity. The HPLC separation uses a mobile phase of water, acetonitrile (MeCN, ACN) and either formic or acetic acid (HOAc) with 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.
- Primesep® B2 is a mixed-mode HPLC column with general hydrophobic and ion-exchange properties. The Primesep B2 stationary phase combines a hydrophobic chain and a basic functional group as an anion exchanger. Acids, bases, zwitterions and neutrals can be retained and separated by Primesep B2 columns. Primesep B2 also separates bases by an ion-exclusion mechanism. Primesep B2 demonstrates significant improvement in retention and selectivity for the separation of weak acid analytes by a combination of reversed phase and anion exchange. In this case the basic functional group acts as an embedded ion-pairing reagent. Primesep B2 column is one of the best separation media for natural product extracts. 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:
|
HPLC Separation of Vitamin C and Vitamin B2 |
 |
Application Notes:
Vitamin C (ascorbic acid) and Vitamin B2 (riboflavin) are water-soluble vitamins that are not stored in the body and must be replaced each day. Primesep D easily resolves Vitamins C and B2 and the elution order can be reversed by adjusting acetonitrile and TFA concentrations in the mobile phase. 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:
|
Separation of Vitamins in Mixed-Mode HPLC |
 |
Application Notes:
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:
|
HPLC Separation of Vitamin C, Vitamin Group B, and Related Impurities |
 |
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.
Application Analytes:
Application Detection:
|
HILIC Separation of Vitamins Group B |
 |
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.
Application Analytes:
Application Detection:
|
|
|
