Xbridge Amide Columns

[ product solution ]

XBridge Amide Columns

O O O Si Linker

O NH 2

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

Hydrophilic Interaction Chromatography XBridge Amide Analysis of Ginsenoside Rb1

Since 2003, Waters has developed innovative stationary phases Orento extract for Hydrophilic Interaction Chromatography [HILIC] to overcome the 0.010 challenge of retaining and separating extremely polar compounds. 0.008 Based on Waters novel ethylene bridged hybrid [BEH] particle 0.006 AU technology, NEW XBridge™ Amide columns utilize a chemically stable, 0.004

0.002 Ginsenoside Rb1 trifunctionally-bonded amide phase, enabling a new dimension in 0.000 stability and versatility for HILIC separations. XBridge Amide columns offer the same selectivity as ACQUITY UPLC® BEH Amide columns, 20 µg/mL Ginsenoside Rb1 standard allowing scientists to transfer their separations between HPLC and 0.010 ® UPLC technology platforms. 0.008

0.006

Designed to retain polar analytes and metabolites that are too polar AU 0.004 to retain by reversed-phase [RP] chromatography, XBridge Amide 0.002 columns facilitate the use of a wide range of mobile phase pH [2 – 11] 0.000 Ginsenoside Rb1 to facilitate the exceptional retention of polar analytes spanning a 0.00. 5 1.01. 5 2.02. 5 3.03. 5 4.04. 5 5.05. 5 6.06. 5 min wide range in polarity, structural moiety and pKa. Column: XBridge Amide, 3.5 µm, 4.6 x 150 mm Part Number: 186004869

Mobile Phase : 80/20 ACN/H2O In addition to enhanced retention of polar compounds, XBridge Flow Rate: 1.4 mL/min Inj. Vol.: 11.5 µL Sample Diluent: MeOH Amide columns provide increased mass spectrometry response, direct Column Temp.: 60 °C

Needle Wash: ACN/H2O 95/5 compatibility with sample preparation eluates [PPT, LLE and SPE] as Detection: UV @ 203 nm Sampling Rate: 20 points/sec Time Constant: 0.1 well as orthogonal selectivity compared to reversed-phase materials, Instrument: Waters Alliance® with 2998 PDA Detector making HILIC an attractive alternative separation technique. [ product solution ]

C arbohydrate Analysis XBridge Amide Analysis of Saccharides

One of the most abundant and diverse classes of compounds analyzed 3 by HILIC are carbohydrates [monosaccharides, disaccharides, oligosaccharides and polysaccharides].

4

The XBridge Amide column is an exceptional tool for the analysis of 6

2

7 carbohydrates, providing the following benefits: 1

8 . The 3.5 µm particle size enables high resolution, high speed analysis of 5 9 carbohydrates in complex sample matrices while maintaining or improving

0 2 46810 12 14 16 18 20 22 24 26 28 30 32 min chromatographic resolution. Column: XBridge Amide, 3.5 µm, 4.6 x 150 mm Part Number: 186004869 Mobile Phase A: 30/70 MeCN/H O with 0.10% NH OH . Increased chemical stability enables the use of high pH and high 2 4 Mobile Phase B: 80/20 MeCN/H2O with 0.10% NH4OH Flow Rate: 0.4 mL/min temperature to collapse anomers without the loss of reducing sugars. Gradient: Profile Time (min) %A %B Initial 0 100 30.6 60 40 . BEH particle technology, in combination with a trifunctionally-bonded 30.7 0 100 Inj. Vol.: 10 µL

amide phase, provides exceptional column lifetime, thus improving Sample Diluent: 50/50 MeCN/H2O Column Temp.: 35 °C

Needle Wash: ACN/H2O 75/25 assay robustness. Instrument: Waters ACQUITY UPLC with ACQUITY UPLC TQD Ionization Mode: ES- Capillary: 2.8 kV Unlike amine-based columns used for carbohydrate analysis, the Cone Voltage: 25 V (fructose, glucose); 15 V (sucrose, maltose) and 40 V (maltooligosaccharides [n=1 to 5]) . Source Temp.: 120 °C Desolv. Temp.: 350 °C XBridge Amide column is not susceptible to Schiff base formation, thus Desolv. Gas Flow: 500 L/Hr SIR m/z: 179.2 (fructose, glucose); 341.3 (sucrose, maltose); 503.4, 665.5, 827.6, 989.7, improving quantitation accuracy. 115 (maltooligosaccharides [n=1 to 5]) Dwell Time: 0.2 s Compounds: 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

Ordering Information

XBridge Amide Columns XBridge Amide Columns Chemistry Particle Size Dimension Type Part Number Chemistry Particle Size Dimension Type Part Number Amide 3.5 µm 1.0 x 50 mm Column 186004871 Amide 3.5 µm 3.0 x 100 mm Column 186004864 Amide 3.5 µm 2.1 x 10 mm Guard 1860048571 Amide 3.5 µm 4.6 x 20 mm Guard 1860048652 Amide 3.5 µm 2.1 x 30 mm Column 186004858 Amide 3.5 µm 4.6 x 30 mm Column 186004866 Amide 3.5 µm 2.1 x 50 mm Column 186004859 Amide 3.5 µm 4.6 x 50 mm Column 186004867 Amide 3.5 µm 2.1 x 100 mm Column 186004860 Amide 3.5 µm 4.6 x 100 mm Column 186004868 Amide 3.5 µm 2.1 x 150 mm Column 186004861 Amide 3.5 µm 4.6 x 150 mm Column 186004869 Amide 3.5 µm 3.0 x 30 mm Column 186004862 Amide 3.5 µm 4.6 x 250 mm Column 186004870

Amide 3.5 µm 3.0 x 50 mm Column 186004863 1 Requires Universal Sentry Guard Holder - 2.1 x 10 mm WAT097958 2 Requires Universal Sentry Guard Holder - 4.6 x 20 mm WAT046910

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

Waters Corporation 34 Maple Street © 2009 Waters Corporation. Waters, The Science of What’s Possible, Milford, MA 01757 U.S.A. XBridge, ACQUITY UPLC, UPLC and Alliance are trademarks of Waters Corporation. All other trademarks are the property of their respective owners. T: 1 508 478 2000 F: 1 508 872 1990 October 2009 Printed in the U.S.A. 720003232EN KK-FP www.waters.com