X B ri dg e Am i d e Co lum ns

HydropHiliC int erACt ion CHromAtogrApHy

Since 2003, Waters has developed innovative stationary phases

for Hydrophilic Interaction Chromatography [HILIC] to overcome the

challenge of retaining and separating extremely polar compounds.

Based on Waters novel ethylene bridged hybrid [BEH] particle

technology, NEW XBridge™ Amide columns utilize a chemically stable,

trifunctionally-bonded amide phase, enabling a new dimension in

stability and versatility for HILIC separations. XBridge Amide columns

offer the same selectivity as ACQUITY UPLC® BEH Amide columns,

allowing scientists to transfer their separations between HPLC and

UPLC® technology platforms.

Designed to retain polar analytes and metabolites that are too polar

to retain by reversed-phase [RP] chromatography, XBridge Amide

columns facilitate the use of a wide range of mobile phase pH [2 – 11]

to facilitate the exceptional retention of polar analytes spanning a

wide range in polarity, structural moiety and pKa.

In addition to enhanced retention of polar compounds, XBridge

Amide columns provide increased mass spectrometry response, direct

compatibility with sample preparation eluates [ PPT, LLE and SPE] as

well as orthogonal selectivity compared to reversed-phase materials,

making HILIC an attractive alternative separation technique.

[ product solution ]

O Si O O

O NH2

Linker

Particle Type: Ethylene Bridged Hybrid [BEH]

Ligand Type: Trifunctional Amide

Particle Size: 3.5 µm

Ligand Density: 7.5 µmol/m2

Carbon Load: 12%

Endcap Style: None

pH Range: 2-11

Pressure Tolerance: 400 bar

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Gin

seno

side

Rb1

Gin

seno

side

Rb1

Orento extract

20 µg/mL Ginsenoside Rb1 standard

AUAU

Column: XBridge Amide, 3.5 µm, 4.6 x 150 mmPart Number: 186004869Mobile Phase : 80/20 ACN/H2OFlow Rate: 1.4 mL/min Inj. Vol.: 11.5 µL Sample Diluent: MeOHColumn Temp.: 60 °C Needle Wash: ACN/H2O 95/5Detection: UV @ 203 nm Sampling Rate: 20 points/secTime Constant: 0.1Instrument: Waters Alliance® with 2998 PDA Detector

X Bridge Amide Analysis of ginsenoside rb1

XBridge Amide Columns

Chemistry Particle Size Dimension Type Part Number

Amide 3.5 µm 1.0 x 50 mm Column 186004871

Amide 3.5 µm 2.1 x 10 mm Guard 1860048571

Amide 3.5 µm 2.1 x 30 mm Column 186004858

Amide 3.5 µm 2.1 x 50 mm Column 186004859

Amide 3.5 µm 2.1 x 100 mm Column 186004860

Amide 3.5 µm 2.1 x 150 mm Column 186004861

Amide 3.5 µm 3.0 x 30 mm Column 186004862

Amide 3.5 µm 3.0 x 50 mm Column 186004863

ordering informAtion

Austria and European Export (Central South Eastern Europe, CIS and Middle East) 43 1 877 18 07, Australia 61 2 9933 1777, Belgium 32 2 726 1000, Brazil 55 11 4134 3788, Canada 1 800 252 4752 x2205, China 86 21 6879 5888, CIS/Russia +497 727 4490/290 9737, Czech Republic 420 2 617 1 1384, Denmark 45 46 59 8080, Finland 358 9 5659 6288, France 33 1 30 48 72 00, Germany 49 6196 400600, Hong Kong 852 29 64 1800, Hungary 36 1 350 5086,

India and India Subcontinent 91 80 2837 1900, Ireland 353 1 448 1500, Italy 39 02 265 0983, Japan 81 3 3471 7094, Korea 82 2 6300 4800, Mexico 52 55 5524 7636, The Netherlands 31 76 508 7200, Norway 47 6 384 60 50, Poland 48 22 833 4400, Puerto Rico 1 787 747 8445, Singapore 65 6593 7100, Spain 34 93 600 9300, Sweden 46 8 555 11 500, Switzerland 41 56 676 70 00, Taiwan 886 2 2543 1898, United Kingdom 44 208 238 6100,

All other countries: Waters Corporation U.S.A. 1 508 478 2000/1 800 252 4752

© 2009 Waters Corporation. Waters, The Science of What’s Possible, XBridge, ACQUITY UPLC, UPLC and Alliance are trademarks of Waters Corporation. All other trademarks are the property of their respective owners.

October 2009 Printed in the U.S.A. 720003232EN KK-FP

Waters Corporation 34 Maple Street Milford, MA 01757 U.S.A. T: 1 508 478 2000 F: 1 508 872 1990 www.waters.com

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Column: XBridge Amide, 3.5 µm, 4.6 x 150 mmPart Number: 186004869Mobile Phase A: 30/70 MeCN/H2O with 0.10% NH4OH Mobile Phase B: 80/20 MeCN/H2O with 0.10% NH4OH Flow Rate: 0.4 mL/min Gradient: Profile Time (min) %A %B Initial 0 100 30.6 60 40 30.7 0 100 Inj. Vol.: 10 µL Sample Diluent: 50/50 MeCN/H2OColumn Temp.: 35 °C Needle Wash: ACN/H2O 75/25Instrument: Waters ACQUITY UPLC with ACQUITY UPLC TQDIonization Mode: ES-Capillary: 2.8 kVCone Voltage: 25 V (fructose, glucose); 15 V (sucrose, maltose) and 40 V (maltooligosaccharides [n=1 to 5])Source Temp.: 120 °CDesolv. Temp.: 350 °CDesolv. Gas Flow: 500 L/HrSIR m/z: 179.2 (fructose, glucose); 341.3 (sucrose, maltose); 503.4, 665.5, 827.6, 989.7, 115 (maltooligosaccharides [n=1 to 5])Dwell Time: 0.2 sCompounds: 1. Fructose 10 µg/mL 4. Maltose 10 µg/mL 7. Maltopentaose 10 µg/mL 2. Glucose 10 µg/mL 5. Maltotriose 10 µg/mL 8. Maltohexahose 10 µg/mL 3. Sucrose 10 µg/mL 6. Maltotetraose 10 µg/mL 9. Maltoheptaose 10 µg/mL

X Bridge Amide Analysis of saccharidesCArBoHydrAt e AnAlysis

One of the most abundant and diverse classes of compounds analyzed

by HILIC are carbohydrates [monosaccharides, disaccharides,

oligosaccharides and polysaccharides].

The XBridge Amide column is an exceptional tool for the analysis of

carbohydrates, providing the following benefits:

The 3.5 µm particle size enables high resolution, high speed analysis of

carbohydrates in complex sample matrices while maintaining or improving

chromatographic resolution.

Increased chemical stability enables the use of high pH and high

temperature to collapse anomers without the loss of reducing sugars.

BEH particle technology, in combination with a trifunctionally-bonded

amide phase, provides exceptional column lifetime, thus improving

assay robustness.

Unlike amine-based columns used for carbohydrate analysis, the

XBridge Amide column is not susceptible to Schiff base formation, thus

improving quantitation accuracy.

XBridge Amide Columns

Chemistry Particle Size Dimension Type Part Number

Amide 3.5 µm 3.0 x 100 mm Column 186004864

Amide 3.5 µm 4.6 x 20 mm Guard 1860048652

Amide 3.5 µm 4.6 x 30 mm Column 186004866

Amide 3.5 µm 4.6 x 50 mm Column 186004867

Amide 3.5 µm 4.6 x 100 mm Column 186004868

Amide 3.5 µm 4.6 x 150 mm Column 186004869

Amide 3.5 µm 4.6 x 250 mm Column 186004870

1 Requires Universal Sentry Guard Holder - 2.1 x 10 mm WAT0979582 Requires Universal Sentry Guard Holder - 4.6 x 20 mm WAT046910