Tosoh TSK-GEL - affinity

TSK-GEL® kolonner Normal Phase and Hydrophilic Interaction Chromatography		MD Scientific er eneste autoriserede forhandler i Danmark for Tosoh Biobioscience

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While reversed phase is the most widely used mode of retention in HPLC, very polar compounds are often not sufficiently retained in low percent organic, or even in 100% aqueous mobile phase.

By using an amide or amino bonded phase column, polar compounds can be retained by a normal phase or hydrophilic interaction chromatography (HILIC) retention mechanism using a mobile phase mixture of acetonitrile and ammonium acetate buffer. In contrast to the retention behavior in reversed phase, in HILIC solutes will be retained longer when increasing the percent acetonitrile.

	TSKgel Amide-80	TSKgel NH2-100
Bonded phase	Amide-80 bonded phase does not react with reducing sugars	NH2-100 bonded phase is more stable than conventional amino phases due to a special endcapping prior to introduction of aminoalkyl groups.
Bonded phase	Anomer formation can be prevented by raising mobile phase temperature up to 80°C.	Amino-bonded phase can react with a reducing sugar to form a Schiff base.
Detectors	Can be used with evaporative light scattering (ELS) and mass spec (MS) detectors
Mobile Phase	Stable in 100% organic eluents
Solutes	Retains very polar drug candidates and drug metabolites
Applications	saccharides	saccharides
	polyols (polyalcohols)	polyols (polyalcohols)
	drug metabolites	methotrexate polyglutamate derivatives
	peptides	pyridylaminated oligosaccharides
	melamine & cyanuric acid	water-soluble vitamins
	oligonucleotides	nucleic acid

The 'normal' or traditional way of doing liquid chromatography, normal phase liquid chromatography (NPC) uses a polar stationary phase and a non-polar mobile phase. The polar surface can either be obtained by derivatizing the support with a ligand containing a polar functional group, or it can be the original (polar) support, as in the case of silica gel.

The popularity of bare silica gel declined after the introduction of bonded phases, since bonded phases feature shorter equilibration times and allow the use of gradient elution. Cyano, amino, and diol bonded phases are the most common normal phase column types.

The order of elution in normal phase is opposite that found in RPC for the same mixture of compounds. Although non-polar organic mobile phases and a silica stationary phase were used traditionally in normal phase LC, today most normal phase separations are performed with aqueous-organic mobile phases and a more polar-bonded stationary phase. This mode of HPLC is now commonly referred to as HILIC, hydrophilic interaction chromatography.

TSKgel Amide-80

TSKgel Amide-80 columns are packed with 3, 5, or 10µm spherical silica particles that are covalently bonded with carbamoyl groups. The amide stationary phase provides a unique selectivity under regular normal phase conditions or in the hydrophilic interaction mode of chromatography. Target applications include the analysis of saccharides, glycans, oligosaccarides, peptides and polar compounds from natural products. Unique advantages of Amide-80 phase for saccharide analysis include novel hydrogen bonding retention mechanism between hydroxyl groups of the sample and the carbamoyl group in the stationary phase. The stationary phase does not react with reducing sugars and can be used at elevated temperatures (4 to 80 °C) to prevent peak splitting of carbohydrates that can occur at lower temperatures. In addition, the new, high efficiency, 3µm columns offer shorter analysis time and enhanced sensitivity for LC-MS applications.

The pore size of the bonded phase is indicated by the number in the product description, in this case TSKgel Amide-80 has 80Å nominal pore size. The nominal pore size of the starting base silica is 100Å.

Now available: TSKgel Amide-80 HR column in 5µm, 4.6mm ID x 25cm size. These new columns are prepared from the same 5 micron packing material and thus offer the same selectivity as existing 5µm TSK-GEL Amide-80 columns. In contrast to traditional 25cm x 4.6mm TSK-GEL Amide-80 columns, the new TSK-GEL Amide-80 HR columns offer higher specifications for plate count (N = > 18,000 vs. 8,000). Because HR columns are prepared from the same bonded phase lots as standard 5 micron TSK-GEL Amide-80 columns, the only change you will notice is more efficient chromatography!

Structure:

Product Attributes:

Pore size (silica):	100Å
Particle size (mean):	3, 5 or 10µm
pH stability:	2.0 - 7.5
Functional group:	Carbamoyl
Max. temperature (°C):	80
Surface area (m2/g)	450

Literature:

Application Note - Separation of Peptides by Hydrophilic Interaction Chromatography using TSKgel Amide-80

Application Note - TSK-GEL Amide-80 Normal Phase Chromatography Columns for the Separation of Saccharides - Now Available in 3μm!

Application Note - TSK-GEL Amide-80 HILIC Columns for the Analysis of Melamine and Cyanuric Acid in Milk by LC-MS/MS

Brochure -- Reversed Phase and HILIC TSK-GEL HPLC Columns

Catalog -- Chromatography Catalog, NPC/HILIC columns

Operating Conditions and Specifications

Performance Data -- TSKgel Amide-80 Columns for the Analysis of Hydrophilic Compounds

Product Overview -- TSK-GEL Amide-80 Columns

Separation Report 55 - Separation of Saccharides Using TSK-GEL Amide-80, a Packing Material for High-Performance Normal Phase Partition Chromatography (part 1)

Technical Presentation -- Newly developed, high performance HILIC columns for use in pharmaceutical analysis by LC/MS

Applications

Separation of polyalcohols on TSKgel Amide-80

Separation of pyridylaminated N-glycans from mouse brain on TSKgel Amide-80, 3µm

Ordering information:
To order or get a price, please contact Søren Jensen, sorenj@md-scientific.dk, Mobile 2421 2559.

P/N	Description	Particle size	Housing material	ID (mm)	Length (cm)
21982	TSKgel Amide-80 HR	5µm	Stainless Steel	4.6	25
21864	TSKgel Amide-80	3µm	Stainless Steel	2	5
21865	TSKgel Amide-80	3µm	Stainless Steel	2	15
21862	Guard Cartridge for 2mmID columns, 3pk	3µm	Stainless Steel	2	1
19308	Guard Cartridge Holder for P/N 21862		Stainless Steel	2	1
21866	TSKgel Amide-80	3µm	Stainless Steel	4.6	5
21867	TSKgel Amide-80	3µm	Stainless Steel	4.6	15
21863	Guard Cartridge for 4.6mmID columns, 3pk	3µm	Stainless Steel	3.2	1.5
19018	Guard Cartridge Holder for P/N 21863	3µm	Stainless Steel	3.2	1.5
20009	TSKgel Amide-80	5µm	Stainless Steel	1	5
20010	TSKgel Amide-80	5µm	Stainless Steel	1	10
21486	TSKgel Amide-80	5µm	Stainless Steel	1	15
21487	TSKgel Amide-80	5µm	Stainless Steel	1	25
19694	TSKgel Amide-80	5µm	Stainless Steel	2	5
19695	TSKgel Amide-80	5µm	Stainless Steel	2	10
19696	TSKgel Amide-80	5µm	Stainless Steel	2	15
19697	TSKgel Amide-80	5µm	Stainless Steel	2	25
21941	Guard Cartridge for 2mm ID columns, 3 pk, Replaces P/N 42140	5µm	Stainless Steel	2	1
19308	Guard Cartridge holder for P/N 21941		Stainless Steel	2	1
19532	TSKgel Amide-80	5µm	Stainless Steel	4.6	5
19533	TSKgel Amide-80	5µm	Stainless Steel	4.6	10
13071	TSKgel Amide-80	5µm	Stainless Steel	4.6	25
19021	Guard Column for 4.6mm ID columns	5µm	Stainless Steel	4.6	1
19010	Guard Column for 4.6mm ID columns, 3 pk	5µm	Stainless Steel	3.2	1.5
14459	TSKgel Amide-80	10µm	Stainless Steel	7.8	30
14460	TSKgel Amide-80	10µm	Stainless Steel	21.5	30
14461	Guard column for P/N 14460	10µm	Stainless Steel	21.5	7.5

TSKgel NH2-100

TSKgel NH2-100 amino columns expand the range of TSK-GEL columns for hydrophilic interaction chromatography (HILIC). Offering a different selectivity from the well known TSK-GEL Amide-80 columns, these novel amino-bonded phase columns stand out by providing much improved chemical stability, a prerequisite for achieving reproducible and reliable results.

TSK-GEL NH2-100 columns are packed with 3µm silica particles containing 100Å pores. A novel bonding strategy was adopted to improve chemical stability of the bonded phase. First the silica is reacted with a trimethylsilane endcapping reagent at a low stoichiometric ratio, before reacting residual and accessible silanol groups with trifunctional alkylaminosilane reagent. The resulting bonded phase provides a better safeguard against hydrolysis of the underlying silica.

TSK-GEL NH2-100 columns are unique in that these columns contain not only amino groups but also tertiary amino groups. Anionic compounds are retained on the column by ionic interaction. This allows for the use of gradients in salt concentration, in addition to gradient elutions with acetonitrile. Since the TSK-GEL NH2-100 columns have cationic sites, the columns can be used as mixed mode columns under some conditions.

TSK-GEL NH2-100 columns are well suited for the analysis of all types of hydrophilic compounds, including carbohydrates and peptides. Due to a high ligand density and large surface area, these columns show stronger retention of polar compounds than TSK-GEL Amide-80 columns.