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Normal Phase Hydrophilic Interaction Chromatography

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. 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.

By using an amide or amino bonded phase column, polar compounds can be retained by a normal phase or hydrophilic interaction chromatography (HILIC; see references) 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. - Anomer formation can be prevented by raising mobile phase temperature up to 80°C.	- NH2-100 bonded phase is more stable than conventional amino phases due to a special endcapping prior to introduction of aminoalkyl groups. - 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 - polyols (polyalchols) - drug metabolites - peptides - melanine & cyanuric acid - oligonucleotides - polar compounds in - fermentation broth cystosine	- saccharides - polyols (polyalchols) - methotrexate polyglutamate derivatives - pyridylaminated oligosaccharides - water-soluble vitamins - nucleic acid fragments

Literature:

• Instruction Manual Annex for 2µm

• Instruction Manual

Particle Size (mean):	2 µm
Pore Size (mean):	10 nm
Functional Group:	carbamoyl
pH Range:	2.0 - 7.5
Max. temperature :	50 ºC
Surface Area (m²/g):	450

P/N	Description	Particle size	Housing material	ID (mm)	Length (cm)
0023454	TSKgel Amide-80 2µm	2 µm	Stainless Steel	2	5
0023455	TSKgel Amide-80 2µm	2 µm	Stainless Steel	2	10
0023456	TSKgel Amide-80 2µm	2 µm	Stainless Steel	2	15
0023457	TSKgel Amide-80 2µm	2 µm	Stainless Steel	3	5
0023458	TSKgel Amide-80 2µm	2 µm	Stainless Steel	3	10
0023459	TSKgel Amide-80 2µm	2 µm	Stainless Steel	3	15
0023460	TSKgel guardclumn Amide-80 2μm	2 µm	Stainless Steel	2	1

Literature:

• Catalog

• HILIC Column Products Table

• Instruction Manual

Particle Size (mean):	3 µm
Pore Size (mean):	10 nm
Functional Group:	carbamoyl
pH Range:	2.0 - 7.5
Max. temperature :	50 ºC
Surface Area (m²/g):	450

P/N	Description	Particle size	Housing material	ID (mm)	Length (cm)
0021864	TSKgel Amide-80 3µm	3 µm	Stainless Steel	2	5
0021865	TSKgel Amide-80 3µm	3 µm	Stainless Steel	2	15
0022850	TSKgel Amide-80 3µm	3 µm	Stainless Steel	3	5
0022851	TSKgel Amide-80 3µm	3 µm	Stainless Steel	3	10
0022852	TSKgel Amide-80 3µm	3 µm	Stainless Steel	3	15
0021862	Guard Cartridge for 2 and 3mmID columns, 3pk	3 µm	Stainless Steel	2	1
0019308	Guard Cartridge Holder for P/N 0021862		Stainless Steel	2	1
0021866	TSKgel Amide-80 3µm	3 µm	Stainless Steel	4.6	5
0022849	TSKgel Amide-80 3µm	3 µm	Stainless Steel	4.6	10
0021867	TSKgel Amide-80 3µm	3 µm	Stainless Steel	4.6	15
0021863	Guard Cartridge for 4.6mmID columns, 3pk	3 µm	Stainless Steel	3.2	1.5
0019018	Guard Cartridge Holder for P/N 0021863		Stainless Steel	3.2	1.5

Literature:

• Separation Report No. 55

• Separation Report No. 79

• Catalog

• HILIC Column Products Table

• Instruction Manual

Particle Size (mean):	5 and 10 µm
Pore Size (mean):	10 nm
Functional Group:	carbamoyl
pH Range:	2.0 - 7.5
Max. temperature :	80 ºC
Surface Area (m²/g):	450

P/N	Description	Particle size	Housing material	ID (mm)	Length (cm)
0020009	TSKgel Amide-80	5 µm	Stainless Steel	1	5
0020010	TSKgel Amide-80	5 µm	Stainless Steel	1	10
0021486	TSKgel Amide-80	5 µm	Stainless Steel	1	15
0021487	TSKgel Amide-80	5 µm	Stainless Steel	1	25
0019694	TSKgel Amide-80	5 µm	Stainless Steel	2	5
0019695	TSKgel Amide-80	5 µm	Stainless Steel	2	10
0019696	TSKgel Amide-80	5 µm	Stainless Steel	2	15
0019697	TSKgel Amide-80	5 µm	Stainless Steel	2	25
0021941	Guard Cartridge for 2mm ID columns, 3 pk, Replaces P/N 0042140	5 µm	Stainless Steel	2	1
0019308	Guard Cartridge holder for P/N 0021941		Stainless Steel	2	1
0019532	TSKgel Amide-80	5 µm	Stainless Steel	4.6	5
0019533	TSKgel Amide-80	5 µm	Stainless Steel	4.6	10
0013071	TSKgel Amide-80	5 µm	Stainless Steel	4.6	25
0019021	Guard Column for 4.6mm ID columns	5 µm	Stainless Steel	4.6	1
0019010	Guard Column for 4.6mm ID columns, 3 pk	5 µm	Stainless Steel	3.2	1.5
0019018	Guard Cartridge holder for P/N 0019010		Stainless Steel	3.2	1.5
0014459	TSKgel Amide-80	10 µm	Stainless Steel	7.8	30
0014460	TSKgel Amide-80	10 µm	Stainless Steel	21.5	30
0014461	Guard column for P/N 0014460	10 µm	Stainless Steel	21.5	7.5

Literature:

• Catalog

• HILIC Column Products Table

• Instruction Manual

• Instruction Manual for NH₂-100 DC

Particle Size (mean):	3 µm
Pore Size (mean):	10 nm
Functional Group:	alkylamine
pH Range:	2.0 - 7.5
Max. temperature:	50 ºC
Surface Area (m²/g):	450

P/N	Description	Particle size	Housing material	ID (mm)	Length (cm)
0021967	TSKgel NH₂-100	3 µm	Stainless Steel	2	5
0021968	TSKgel NH₂-100	3 µm	Stainless Steel	2	15
0021971	Guard Cartridge for 2mmID columns, 3pk	3 µm	Stainless Steel	2	1
0019308	Guard Cartridge Holder for P/N 21971		Stainless Steel	2	1
0021999	TSKgel NH₂-100 DC	3 µm	Stainless Steel	4.6	5
0021969	TSKgel NH₂-100	3 µm	Stainless Steel	4.6	5
0021970	TSKgel NH₂-100	3 µm	Stainless Steel	4.6	15
0021972	Guard Cartridge for 4.6mmID columns, 3pk	3 µm	Stainless Steel	3.2	1.5
0019018	Guard Cartridge Holder for P/N 21972		Stainless Steel	3.2	1.5