HPLC COLUMNS - Continuing the Legacy of HPLC Column Performance

Continuing the Legacy of

 

[ HPLC COLUMNS ]

 

HPLC Column Performance

Creating Exceptional Chromatography 

Waters® reputation is based on chromatography, but we do not create 

chromatography — you do. Innovative thinking within your laboratory 

creates the chromatographic methods and assays that sustain your 

business. The metric of your success is monitored by the methods and 

results that you produce, and the HPLC column that you choose today  

needs to support your success for the future. Waters full line of  

state-of-the-art, reversed-phase and HILIC HPLC columns are chosen 

by scientists who understand that performance and innovation are linked 

and their success depends on them

.

HPLC Columns

2

CORTECS

XBridge

XSelect

Atlantis

SunFire

Symmetry

XTerra

Waters Spherisorb

Nova-Pak

Resolve

Delta-Pak

µBondaPak/BondaPak

µPorasil/Porasil

VanGuard

Waters Analytical  
Standards and Reagents 

3

O

Si

O

O

O

Si

O

O

O

Si

O

O

O

Si

O

O

* Expected or approximate values.

All CORTECS Columns are available in UPLC particle sizes.

O

Si

CH

Polar Group

3

CH 3

O

Si

O

O

Solid-core particle packing materials combine a fully-porous surface layer  
that has been bonded to a solid-core substrate. This combination creates a 
highly efficient particle substrate that maintains chromatographic resolution 
while offering the advantage of lower column back pressures.

CORTECS® 2.7 µm Solid-Core Particle Columns maximize resolution and peak  
capacity for all LC separations and are optimized to extend HPLC or UHPLC 
instrument performance. The innovative solid-core technology and bonding 
chemistry used in CORTECS Columns helps you by:

■

■

Reducing Operational Backpressure: Lower backpressure without  
sacrificing efficiency

■

■

Increasing Sensitivity: Improved signal-to-noise performance for  
LC-MS applications

■

■

Simplifying Method Transfers: Compatible with a wide range  
of chromatographic systems

The selection of CORTECS 2.7 µm Columns in both reversed-phase and HILIC 
phases gives you the flexibility to rapidly separate a wide range of compound 
classes. The improved efficiency of CORTECS 2.7 µm Solid-Core Columns 
produces sharper, narrower peaks compared to columns using fully-porous 
substrates and allows you to run faster flow rates to improve sample throughput.

CORTECS

C18+

C18

T3

Shield RP18

C8

Phenyl

HILIC

Ligand Type

Trifunctional C18

Trifunctional C18

Trifunctional C18

Monofunctional 

Embedded Polar

Trifunctional C8

Trifunctional C6 

Phenyl

None

Ligand Density*

2.4 µmol/m2

2.7 µmol/m2

1.6 µmol/m2

3.2 µmol/m2

3.4 µmol/m2

3.2 µmol/m2

N/A

Carbon Load*

5.7%

6.6%

4.7%

6.4%

4.5%

5.9%

Unbonded

End-capped

Proprietary

Proprietary

Proprietary

Proprietary

Proprietary

Proprietary

No

pH Range

2–8

2–8

2–8

2–8

2–8

2–8

1–5

Low pH Temp. Limit

45 °C

45 °C

45 °C

45 °C

45 °C

45 °C

45 °C

High pH Temp. Limit

45 °C

45 °C

45 °C

45 °C

45 °C

45 °C

45 °C

Pore Diameter

90Å

90Å

90Å

90Å

90Å

90Å

90Å

Surface Charge Modification

+

None

None

None

None

None

None

USP Classification

 L1

 L1

 L1   

 L1   

 L7

 L11 

 L3

4

Reduced Backpressure

 
CORTECS Columns reduce operational backpressure allowing you to run methods using conventional LC instrumentation without 
compromising efficiency or resolution. Additionally, longer columns can be used to improve resolution for co-eluting peaks in complex 
sample mixtures.

Backpressure Advantages of CORTECS 2.7 µm Columns

Flow 

Rate 

(mL/min)

2.1 x 50 mm

0.17

0.35

0.52

0.69

0.87

1.04

3.0 x 50 mm

0.35

0.71

1.06

1.41

1.77

2.12

4.6 x 50 mm

0.83

1.66

2.49

3.32

4.15

4.99

CORTECS 2.7 µm Columns offer a 25% reduction in operating backpressure— 
without sacrificing efficiency. Data conditions—Columns: 2.1 x 50 mm;  
Mobile phase: water/acetonitrile (25/75, v/v); Column temperature: 30 ˚C.

Comparative separations may not be representative in all applications.

Efficiency Advantages of CORTECS 2.7 µm Columns

Flow 

Rate  

(mL/min)

2.1 x 50 mm

0.17

0.35

0.52

0.69

0.87

1.04

3.0 x 50 mm

0.35

0.71

1.06

1.41

1.77

2.12

4.6 x 50 mm

0.83

1.66

2.49

3.32

4.15

4.99

CORTECS 2.7 µm Columns exhibit excellent efficiency compared to similarly-sized, 
fully-porous and solid-core particle columns. Data conditions—Columns: 2.1 x 50 mm; 
Mobile phase: water/acetonitrile (25/75, v/v); Column temperature: 30 ˚C; Compounds: 
acenaphthene (200 µg/mL), octanophenone (100 µg/mL).

Comparative separations may not be representative in all applications.

Increased Sensitivity

 
Charged surface technology improves peak shape and compound loading when using low-ionic strength mobile phases such as formic acid. 
The permanent low-level surface charge used during the C18+ bonding process enhances signal-to-noise performance by eliminating  
ion-pairing reagents and additives that would otherwise negatively impact LC-MS applications.
 

 
Superior Peak Shape for Low Level Impurity Analysis

HPLC-UV/MS analysis of the low-ionic basic antidepressant imipramine reveals a low level impurity. Using a CORTECS C18+, 2.7 µm Column designed for use with low ionic 

strength acidic mobile phases results in narrower peaks and improved signal-to-noise.

Comparative separations may not be representative in all applications.

LC Conditions
Columns:  

3.0 x 50 mm

Mobile phase A: 

0.1% formic acid in water

Mobile phase B: 

0.1% formic acid in acetonitrile

Gradient:  

25 to 35% B in 4.6 min

Flow rate: 

0.8 mL/min

Column temp.: 

30 °C

Detection: 

254 nm and ESI+ MS

Injection volume:  10 μL 

Compounds 

 

Imipramine (0.5 mg/mL)  
Impurity at 0.1% spike (0.5 μg/mL)

%

0

100

1: SIR of 1 Channel ES+

TIC (Impurity at 0.1% spike)

3.68e7

1: SIR of 1 Channel ES+

TIC (Impurity at 0.1% spike)

3.68e7

0.4

0.6

0.8

1.0

1.2

1.4

1.6 min

 (1) PDA Ch1 [email protected]

Impurity at 0.1% spike
PW13.4%: 0.089 min 

S/N Ratio: 180 

S/N Ratio: 113 

0.4

0.6

0.8

1.0

1.2

1.4

1.6 min

0.4 0.6

0.8

1.0

1.2

1.4

1.6 min

0.4

0.6

0.8

1.0

1.2

1.4

1.6 min

AU

0.0

5.0e-3

1.0e-2

1.5e-2

2.0e-2

2.5e-2

3.0e-2

3.5e-2

4.0e-2

 (1) PDA Ch1 [email protected]

Imipramine 

Imipramine 

Impurity at 0.1% spike
PW13.4%: 0.132 min 

Competitor Solid-Core C18, 2.6 µm 
Pressuremax: 3250 psi

CORTECS C

18+, 2.7 µm 

Pressuremax: 2750 psi

60% Increase

in S/N Ratio

35% Decrease 

in Peak Width

5

www.waters.com/cortecs

The CORTECS Family

 
A dedicated selection of 7 phases can be used to separate a wide array of compound classes. CORTECS C18 provides a balanced retention 
profile for acidic, basic, and neutral compounds. CORTECS C18+ gives the best peak shape and increased sensitivity of basic analytes when 
using low ionic strength mobile phases such as formic acid. CORTECS T3 is an excellent phase to use when separating compounds of 
various polarity.  The lower C18 ligand density provides balance retention for both polar and nonpolar compounds and the 120Å pore 
diameter allows for the use of 100% aqueous mobile phase. CORTECS C8, being less hydrophobic than a typical C18 bonded phase, is 
an excellent choice for the separation of strongly hydrophobic compounds. CORTECS Phenyl offers alternative selectivity to C8 and C18 
due to analyte interactions with the benzyl ring; selectivity differences for this phase are particularly noticed for aromatic compounds 
especially when using methanol as the organic modifier. The CORTECS Shield RP18 also provides alternative selectivity over typical C8 
and C18 bonded phases due to the embedded polar group, and is a great choice for method development, especially for phenolic and basic 
compounds. The orthogonal unbonded CORTECS HILIC phase provides superior peak shape and retention of polar analytes. With particle 
sizes that are compatible with HPLC, UPLC,® and UHPLC platforms, any method that you develop can be simply and seamlessly transferred 
without limitation to particle size, column configuration, or instrument manufacturer. 

Methods developed on 5 µm fully-porous columns can be scaled and transferred to shorter 2.7 µm columns.  
For further efficiency gains and productivity improvements, sub-2-µm UPLC columns can be used, enabling  
greater flexibility in method consistency when transitioning between laboratories within an organization  
or to contract partners.

1

2

3

4

5

AU

0.00

0.01

0.02

AU

0.00

0.01

0.02

AU

0.00

0.01

0.02

12.0

14.0

16.0

18.0

20.0

22.0

24.0

26.0

28.0

30.0 min

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

8.0 min

AU

0.00

0.01

0.02

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

8.0 min

1.8

2.0

2.2

2.4

2.6

2.8

3.0

3.2

3.4

3.6 min

Column: Fully-porous C

18, 5 µm, 4.6 x 150 mm

Flow rate: 1.00 mL/min

Injection volume: 8 µL

LC system: Alliance® HPLC 

Column: CORTECS C

18, 2.7 µm, 4.6 x 75 mm

Flow rate: 1.85 mL/min

Injection volume: 4 µL

LC system: Alliance HPLC 

 

Column: CORTECS C

18, 2.7 µm, 4.6 x 75 mm

Flow rate: 1.85 mL/min

Injection volume: 4 µL

LC system: ACQUITY® Arc

Column: CORTECS UPLC C

18, 1.6 µm, 2.1 x 50 mm

Flow rate: 0.65 mL/min

Injection volume: 0.6 µL

LC system: ACQUITY UPLC® H-Class

Original

 method

Transfer to 2.7 µm

4x faster method

2x less solvent

Compatible with 

UHPLC systems

4x less solvent

Transfer to 1.6 µm

9x faster method

15x less solvent

USP Method Transfer of Abacavir with Time and Solvent

Comparative separations may not be representative in all applications.

LC Conditions
Mobile phase A: 

0.1% trifluoroacetic acid in water

Mobile phase B: 

85% methanol in water

Column A: 

Fully-Porous C18, 5 μm, 4.6 x 150 mm

Column B: 

CORTECS C18, 2.7 μm, 4.6 x 75 mm

Column C: 

CORTECS C18, 2.7 µm, 3.0 x 75 mm

Column D:  

CORTECS C18, 1.6 µm, 2.1 x 50 mm

Geometrically-scaled gradients (i.e., same column volumes 
per gradient step):
Column A: 

5 to 30% B in 23.6 min and  

 

 

30 to 90% B in 14.8 min

Column B: 

5 to 30% B in 6.4 min and  

 

 

30 to 90% B in 4.0 min

Column C: 

5 to 30% B in 6.4 min and  

 

 

30 to 90% B in 4.0 min

Column D: 

5 to 30% B in 2.5 min and  

 

 

30 to 90% B in 1.6 min

Compounds 
1. Dicyclopropyl Abacavir 
2. Abacavir 
3. 1R,4R trans-Abacavir 
4. o-(4-Chloro-2,5-diaminopyimidnyl)-abacavir 
5. o-t-Butyl-abacavir

6

7

* Expected or approximate value.

Based on BEH Technology
Ethylene Bridged Hybrid (BEH) Technology synthesis creates particles 
that ensure extreme column performance and long column lifetimes 
under harsh operating conditions. The particle is prepared from two 
high purity monomers: tetraethoxysilane (TEOS)  and bis(triethoxysilyl)
ethane (BTEE), which results in a highly stable, pH resistant, and 
mechanically strong particle.

Particle Synthesis

Tetraethoxysilane

(T EOS)

Polyethoxysilane

(BPEOS)

Bis(triethoxysilyl)ethane

(BTE E)

+

4

Si

Et O

Et O

OEt

Et O

Si

Et O

Et O

Et O

Si

OEt

OEt

OEt

Si

Et O

O

CH 2 CH2

CH2

CH2

Si

O Si

Et O

OE t

Si

O

O

OEt

Si

O

Si

O

O

Et

O

O

O

Et

Et

Et

n

* US Patents 6,686,035; 7,223,473; 
7,250,214 Refer to “Ethylene-Bridged [BEH 
Technology] Hybrids and Their Use in Liquid 
Chromatography” whitepaper (720001159EN) 
for further detail.

XBridge® BEH HPLC Columns are designed for one purpose,  
to maximize your productivity. Whether your goal is to create  
a quality control method or develop a leading edge LC-MS assay, 
XBridge BEH Columns are designed to help you by: 

■

■

Improving pH Stability: Increased column lifetime

■

■

Improving Column Reliability: Assay ruggedness

■

■

Maximizing Particle Efficiency: Unmatched peak  
shape and peak capacity 

With a selection of 10 general purpose and application specific 
sorbents in the widest range of particle sizes available, no other  
HPLC column family gives you the tools you need for the most 
demanding chromatographic challenges. Whether you require 
robust HPLC methods, seamless UPLC transferability, or 
preparative scaling for product isolation, you can count on  
the versatility of an XBridge Column.

O

Si

O

O

O

Si

O

O

O

Si

O

O

O

Si

CH

Polar Grou

p

3

CH3

O

Si

O

O

O

Si

O

O

O

Si

O

O

O

Si

CH

Polar Grou

p

3

CH3

O

Si

O

O

O

Si

O

O

O

Si

O

O

O

Si

CH

Polar Grou

p

3

CH3

O

Si

O

O

O

Si

O

O

O

Si

O

O

O

Si

CH

Polar Grou

p

3

CH3

XBridge

C18

C8

Shield RP18

Phenyl

HILIC

Amide

Peptide BEH C18, 130Å

Peptide BEH C18, 300Å

Protein BEH C4, 300Å

Oligo BEH C18

SEC

Ligand Type

Trifunctional C18

Trifunctional C8

Monofunctional 

Embedded Polar 

Trifunctional 

Phenyl-Hexyl

Unbonded  

BEH Particle

Amide

Trifunctional C18

Trifunctional C18

Monofunctional C4

Trifunctional C18

SEC

Ligand Density*

3.1 µmol/m2

3.2 µmol/m2

3.3 µmol/m2

3.0 µmol/m2

N/A

7.5 µmol/m2

3.1 µmol/m2

3.1 µmol/m2

2.4 µmol/m2

3.1 µmol/m2

N/A

Carbon Load*

18%

13%

17%

15%

Unbonded

12%

18%

12%

8%

18%

12%

End-capped

Proprietary

Proprietary

TMS

Proprietary

No

No

Proprietary

Proprietary

No

Proprietary

No

USP Classification

L1

L7

L1

L11

L3

N/A

L1

L1

L26

L1

L33

pH Range

1–12

1–12

2–11

1–12

1–9

2–1

1–12

1–12

1–10

1–12

1–8

Low pH Temp. Limit

80 °C

60 °C

50 °C

80 °C

45 °C

90 °C

80 °C

80 °C

80 °C

80 °C

45 °C

High pH Temp. Limit

60 °C

60 °C

45 °C

60 °C

45 °C

90 °C

60 °C

60 °C

50 °C

60 °C

45 °C

Pore Diameter*

130Å

130Å

130Å

130Å

130Å

130Å

130Å

300Å

300Å

130Å

125, 200, 450Å

Surface Area*

185 m2/g

185 m2/g

185 m2/g

185 m2/g

185 m2/g

185 m2/g

185 m2/g

90 m2/g

90 m2/g

185 m2/g

220 m2/g

Particle Size

2.5, 3.5, 5, 10 µm

2.5, 3.5, 5, 10 µm

2.5, 3.5, 5, 10 µm

2.5, 3.5, 5 µm

2.5, 3.5, 5 µm

2.5, 3.5, 5 µm

3.5, 5, 10 µm

3.5, 5, 10 µm

3.5 µm

2.5 µm

3.5 µm

8

Column Reliability

 
Much of the cost when developing a chromatographic method is associated with the rigorous testing and validation of the final 
method. We understand that revalidation of your method is not an option, so we thoroughly test each batch of sorbent and final 
column product to ensure that you get the most reproducible columns available. With an XBridge BEH Column, you have the 
confidence that the method you develop today will be repeatable for the lifetime of your assay. 
 
 

XBridge Family USP Tailing Factors

1

2

3

C

8

Phenyl

Shield RP18

C

18

Zorbax®

SB-C18

Desipramine

Amitriptyline

pH 3

pH 7

Nortriptyline

Amitriptyline

Tailing Factor (

T

f)

The combination of excellent particle and ligand stability as well as high  
chromatographic efficiencies makes XBridge BEH Columns an ideal choice for  
low and intermediate pH methods.

Comparative separations may not be representative in all applications.

O 

Si 

O 

O

O 

NH

2

Linker

O

Si

O

O

O

Si

O

O

O

Si

O

O

O

Si

CH

Polar Grou

p

3

CH3

O

Si

O

O

O

Si

O

O

O

Si

O

O

O

Si

CH

Polar Grou

p

3

CH3

O

Si

CH

3

CH3

O

Si

CH

3

C 4

C 4

H 9

H 9

CH3

O

Si

O

O

O

Si

O

O

O

Si

O

O

O

Si

CH

Polar Grou

p

3

CH3

Particle Efficiency

 
BEH Technology™ offers many advantages over conventional 
silica-based particles, including the ability to control the silanol 
activity with great precision. By controlling the silanol activity, 
you control and reduce unwanted silanol interactions that 
increase peak tailing.

XBridge

C18

C8

Shield RP18

Phenyl

HILIC

Amide

Peptide BEH C18, 130Å

Peptide BEH C18, 300Å

Protein BEH C4, 300Å

Oligo BEH C18

SEC

Ligand Type

Trifunctional C18

Trifunctional C8

Monofunctional 

Embedded Polar 

Trifunctional 

Phenyl-Hexyl

Unbonded  

BEH Particle

Amide

Trifunctional C18

Trifunctional C18

Monofunctional C4

Trifunctional C18

SEC

Ligand Density*

3.1 µmol/m2

3.2 µmol/m2

3.3 µmol/m2

3.0 µmol/m2

N/A

7.5 µmol/m2

3.1 µmol/m2

3.1 µmol/m2

2.4 µmol/m2

3.1 µmol/m2

N/A

Carbon Load*

18%

13%

17%

15%

Unbonded

12%

18%

12%

8%

18%

12%

End-capped

Proprietary

Proprietary

TMS

Proprietary

No

No

Proprietary

Proprietary

No

Proprietary

No

USP Classification

L1

L7

L1

L11

L3

N/A

L1

L1

L26

L1

L33

pH Range

1–12

1–12

2–11

1–12

1–9

2–1

1–12

1–12

1–10

1–12

1–8

Low pH Temp. Limit

80 °C

60 °C

50 °C

80 °C

45 °C

90 °C

80 °C

80 °C

80 °C

80 °C

45 °C

High pH Temp. Limit

60 °C

60 °C

45 °C

60 °C

45 °C

90 °C

60 °C

60 °C

50 °C

60 °C

45 °C

Pore Diameter*

130Å

130Å

130Å

130Å

130Å

130Å

130Å

300Å

300Å

130Å

125, 200, 450Å

Surface Area*

185 m2/g

185 m2/g

185 m2/g

185 m2/g

185 m2/g

185 m2/g

185 m2/g

90 m2/g

90 m2/g

185 m2/g

220 m2/g

Particle Size

2.5, 3.5, 5, 10 µm

2.5, 3.5, 5, 10 µm

2.5, 3.5, 5, 10 µm

2.5, 3.5, 5 µm

2.5, 3.5, 5 µm

2.5, 3.5, 5 µm

3.5, 5, 10 µm

3.5, 5, 10 µm

3.5 µm

2.5 µm

3.5 µm

9

www.waters.com/xbridge

LC Conditions 
LC system: 

ACQUITY UPLC with TUV Detector  

Columns:  

XBridge BEH C18, 5 μm, 2.1 x 150 mm

 

 

XBridge BEH C18, 3.5 μm, 2.1 x 100 mm

 

 

XBridge BEH C18, XP, 2.5 μm, 2.1 x 75 mm

   ACQUITY UPLC BEH C18, 1.7 μm,  
 

 

2.1 x 50 mm  

Mobile phase A: 

0.1% formic acid in water

Mobile phase B: 

0.1% formic acid in acetonitrile

Isocratic:   

95% A:5% B

Sample conc.:   

25 µg/mL

Column temp.:   38 °C
Detection: 

 

280 nm

1.7 µm – 50 mm

Injection: 1.7 µL 

Flow rate: 0.6 mL/min

5 µm – 150 mm 

Injection: 5.0 µL 

Flow rate: 0.2 mL/min

2.5 µm – 75 mm

Injection: 2.5 µL 

Flow rate: 0.5 mL/min

3.5 µm – 100 mm

Injection: 3.3 µL 

Flow rate: 0.3 mL/min

Methods Transfer Using     2.5 µm Columns

 
All XBridge and XSelect® HPLC Columns (discussed on the next page) are offered in eXtended Performance [XP] 2.5 µm UHPLC 
column formats to help you transfer methods from HPLC to UPLC instrumentation. The XP 2.5 µm columns improve the performance  
of your current HPLC and UHPLC instrumentation and provide you with a pathway to gain maximum separation efficiency using  
sub-2-µm ACQUITY® UPLC Technology.

Columns of different lengths and particle sizes were used to successfully reduce run times  
and maintain resolution.

Scalable Separations

Accelerated High pH Stability Test of Competitive Columns

0

50

100

150

200 hours

Hours in 50 mM TEA, pH 10, 50 °C

Analyte:     Acenaphthene

30

50

70

90

110

% Initial 

N

5s

XBridge BEH C

18

XTerra® MS C

18

Gemini™ C

18

Luna® C18(2)

YMC™ Pro C

18

Zorbax® Extend C

18

Chromatograms, run at regular intervals during the high-pH lifetime study, verify that 
86% of the original XBridge Column efficiency remains after 300 hours at pH 10 and 
elevated temperature, with little change in peak shape or retention time. 

pH Stability

 
XBridge BEH Columns have been specifically designed to 
contain the most chemically-stable chromatographic sorbent 
available, allowing you to explore the full benefits of a wide  
pH (1–12) mobile-phase range. Chemical stability, especially  
for the extremes of pH, is built into the particle during 
the synthesis process and it cannot be duplicated using a 
conventional silica-based bonding process. No other column 
can match the chemical stability of an XBridge Column. 

0.0

2.0

4.0

6.0

8.0

10.0 min  

0.0

0.2

0.4

0.6

0.8

1.0

1.1 min

 

 

0.0

  0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 min  

0.0

0.2

0.4

0.6

0.8

1.0

1.2

1.4

1.6

1.8

2.0 min  

10

[ FOLIO INSIDE HEADER ]

11

XSelect HPLC Columns are designed for the method development 
scientist who demands the most diverse selection of sorbents to 
easily separate the most difficult analyte co-elutions. XSelect 
Columns are tools that are:

■

■

Designed for Selectivity: Improve your ability to separate 
closely eluting peaks

■

■

Intended for Isolation and Purification: Highest analyte 
mass loading available

■

■

Ideal for Rapid Method Development: Reduce the time  
and cost spent developing methods

The XSelect HPLC Column family features two base particles with 
a unique blend of optimized ligands to provide highly selective 
chromatographic phases while maintaining the reproducibility 
expected from modern high performance LC columns. With more 
than 500 column configurations that combine 8 selectivity-
optimized bonded phases and 3 scalable particles sizes, XSelect 
Columns are your first choice for method development.   

O

Si

F

F

F

F

F

O

O

O

Si

O

O

O

Si

O

O

O

Si

O

O

O

Si

O

O

O

Si

O

O

O

Si

F

F

F

F

F

O

O

O

Si

CN

* Expected or approximate value.

Charged Surface Hybrid (CSH™) particles incorporate a low level  
surface charge that improves sample loading and peak symmetry 
when using low ionic strength mobile phases. The CSH particle is 
the next evolution of hybrid particle technology that maintains the 
mechanical and chemical stability inherent in BEH particle technology.

Unbonded BEH 

Particle

Apply Controlled 

Surface Charge

Bond and  

Endcap

The Charged-Surface Particle

Many silica-based particles do not have the mechanical stability 
to withstand the high operational pressures used with modern LC 
instrumentation. High Strength Silica (HSS) is the first and only 100% 
silica-based particle substrate that has been designed and tested for 
mechanical stability up to 18,000 psi (1240 bar).

OEt

Si

O

O

EtO

Si

OEt

OEt

OEt

Si

OEt

EtO

EtO

EtO

Si

EtO

OEt

EtO

Polyethoxysilane

(PEOS)

Tetraethoxysilane

(TEOS)

XSelect

CSH C18

CSH  

Phenyl-Hexyl

CSH  

Fluoro-Phenyl

HSS T3

HSS C18

HSS C18 SB

HSS PFP

HSS CN

Ligand Type

Trifunctional  

C18

Trifunctional  

C6 Phenyl

Trifunctional  

Propylfluoro-

phenyl

Trifunctional 

C18

Trifunctional 

C18

Trifunctional 

C18

Trifunctional  

Pentafluoro-

phenyl

Monofunctional  

Cyano-propyl

Ligand Density*

2.3 µmol/m2

2.3 µmol/m2

2.3 µmol/m2

1.6 µmol/m2

3.2 µmol/m2

1.6 µmol/m2

3.2 µmol/m2

2.0 µmol/m2

Carbon Load*

15%

14%

10%

11%

15%

8%

7%

5%

End-capped

Proprietary

Proprietary

No

Proprietary

Proprietary

No

No

No

USP Classification

L1

L11

L43

L1

L1

L1

L43

L10

pH Range

1–11

1–11

1–8

2–8

1–8

2–8

2–8

2–8

Low pH Temp. Limit

80 ˚C

80 ˚C

60 ˚C

45 ˚C

45 ˚C

45 ˚C

45 ˚C

45 ˚C

High pH Temp. Limit

45 ˚C

45 ˚C

45 ˚C

45 ˚C

45 ˚C

45 ˚C

45 ˚C

45 ˚C

Pore Diameter*

130Å

130Å

130Å

100Å

100Å

100Å

100Å

100Å

Surface Area*

185 m2/g

185 m2/g

185 m2/g

230 m2/g

230 m2/g

230 m2/g

230 m2/g

230 m2/g

Particle Size

2.5, 3.5, 5, 10 µm

2.5, 3.5, 5 µm

2.5, 3.5, 5 µm

2.5, 3.5, 5 µm

2.5, 3.5, 5 µm

2.5, 3.5, 5 µm

2.5, 3.5, 5 µm

2.5, 3.5, 5 µm

12

Comparative separations may not be representative in all applications.

XSelect

CSH C18

CSH  

Phenyl-Hexyl

CSH  

Fluoro-Phenyl

HSS T3

HSS C18

HSS C18 SB

HSS PFP

HSS CN

Ligand Type

Trifunctional  

C18

Trifunctional  

C6 Phenyl

Trifunctional  

Propylfluoro-

phenyl

Trifunctional 

C18

Trifunctional 

C18

Trifunctional 

C18

Trifunctional  

Pentafluoro-

phenyl

Monofunctional  

Cyano-propyl

Ligand Density*

2.3 µmol/m2

2.3 µmol/m2

2.3 µmol/m2

1.6 µmol/m2

3.2 µmol/m2

1.6 µmol/m2

3.2 µmol/m2

2.0 µmol/m2

Carbon Load*

15%

14%

10%

11%

15%

8%

7%

5%

End-capped

Proprietary

Proprietary

No

Proprietary

Proprietary

No

No

No

USP Classification

L1

L11

L43

L1

L1

L1

L43

L10

pH Range

1–11

1–11

1–8

2–8

1–8

2–8

2–8

2–8

Low pH Temp. Limit

80 ˚C

80 ˚C

60 ˚C

45 ˚C

45 ˚C

45 ˚C

45 ˚C

45 ˚C

High pH Temp. Limit

45 ˚C

45 ˚C

45 ˚C

45 ˚C

45 ˚C

45 ˚C

45 ˚C

45 ˚C

Pore Diameter*

130Å

130Å

130Å

100Å

100Å

100Å

100Å

100Å

Surface Area*

185 m2/g

185 m2/g

185 m2/g

230 m2/g

230 m2/g

230 m2/g

230 m2/g

230 m2/g

Particle Size

2.5, 3.5, 5, 10 µm

2.5, 3.5, 5 µm

2.5, 3.5, 5 µm

2.5, 3.5, 5 µm

2.5, 3.5, 5 µm

2.5, 3.5, 5 µm

2.5, 3.5, 5 µm

2.5, 3.5, 5 µm

0.0

0.5

1.0

1.5

2.0

2.5

3.0

3.5 min

1

1

2

2

2

3

3

4

4

5,6

7

5,6

7

7

3

4

3

4

5

6

7

1

2

3

4

5

6

7

1

2

3

4

5

6

7

5,6

1

2

3

4

3

4

5,6

7

7

HSS C

18 SB

HSS C

18

HSS T3

HSS CN

HSS PFP

CSH Fluoro-Phenyl

CSH Phenyl-Hexyl

CSH C

18

1

2

1

5 6

1

2

LC Conditions 
LC system: 

ACQUITY UPLC with

 

 

 

ACQUITY UPLC PDA Detector

Columns:  

2.1 x 50 mm 

Mobile phase A: 

10 mM ammonium formate, pH 3.0

Mobile phase B: 

Methanol 

Flow  rate:  

0.4 mL/min  

Injection volume:  1 μL
Sample diluent:  

Water

Column temp.:   

30 °C

Gradient:

Time
(min)

%A

%B

0.00

70

30

3.00

15

85

3.50

15

85

3.51

70

30

4.50

70

30

Detection: 

 

260 nm

Enhanced Selectivity

 
Selectivity and retentivity are the most powerful tools a method developer has to 
influence chromatographic behavior. The XSelect family offers a diverse range of 
reversed-phase C18 columns (e.g., CSH C18, HSS C18, HSS C18 SB) for general purpose 
separations; as well as columns that offer improved polar retention (T3) and greater 
selectivity options (phenyl-hexyl, fluoro-phenyl, and cyano) for method development.

XSelect Columns Provide Diverse Analyte Selectivity

AU

0.00

0.02

0.04

0.06

0.08

Imipramine 

Imipramine

 

2.0% 

1.0% 

0.5% 

0.1% 

2.0% 

1.0% 

0.5% 

0.1% 

Amitriptyline 

Imipramine concentration held 

constant at 0.5 mg/mL 

Amitriptyline 

0.0

0.2

0.4

0.6

0.8

1.0

1.2

1.4

1.6

1.8

2.0 min

0.0

0.2

0.4

0.6

0.8

1.0

1.2

1.4

1.6

1.8

2.0 min

ACQUITY UPLC CSH C

18

AU

0.00

0.02

0.04

0.06

0.08

Kinetex® C

18

LC Conditions
LC system: 

ACQUITY UPLC H-Class with 

 

 

ACQUITY UPLC PDA Detector

Columns: 

2.1 x 50 mm

Mobile phase A: 

Water

Mobile phase B: 

Acetonitrile 

Mobile phase C: 

2% formic acid in water

Gradient:

Time
(min)

Flow

(mL/min) %A

%B

%C 

Initial

0.6

70

25

5

2.0

0.6

60

35

5

5.0

0.6

70

25

5

Injection volume: 

5 μL

Sample diluent: 

Water

Sample conc.: 

Imipramine: 0.5 mg/mL; amitriptyline: 

 

as indicated (% of imipramine) 

Column temp.: 

40 °C

Detection: 

254 nm

Isolation and Purification

 
High mass loading applications like compound purification, impurity profiling,  
and dissolution testing demand superior column performance. For these types of 
applications, column loading is limited by its inability to maintain symmetrical peak 
shape. This manifests itself as severe broadening of the main compound peak, which often 
overwhelms the trace impurities that you are trying to remove in the purification. XSelect 
CSH Columns consistently provide narrow peaks under high loading conditions that allow  
the chromatographer the ability to separate trace-level impurities or degradants giving you  
more loading capacity with less time and solvent.

Maintaining Peak Shape with High Mass Loading

Observed selectivity differences for a mixture of basic analytes. Compounds: [1] aminopyrazine, [2] pindolol, [3] 
quinine, [4] labetalol, [5] verapamil, [6] diltiazem, [7] amitriptyline.

 
T he improved mass loading of XSelect Columns permits the separation, identification, and quantification 
of closely eluting impurities or degradants

.

13

Method Development and Transfer

 
When developing methods, skilled chromatographers realize that any method 
developed using uniquely selective columns must be easily transferable across 
laboratories, independent of the LC system platform used. XSelect Columns are 
engineered for method development and are fully compatible with all modern 
detection modes.

LC Conditions
LC system: 

ACQUITY UPLC with   

 

 

ACQUITY UPLC PDA Detector  

Columns:  

2.1 x 50 mm

Flow rate: 

0.5 mL/min 

Mobile phase A: 

15.4 mM ammonium formate, pH 3.0

Mobile phase B: 

Acetonitrile

Gradient:  

5 to 90% B linear in 5 minutes

Injection volume:  5 μL
Column temp.: 

30 °C

Detection: 

254 nm

Compounds 
1. Thiourea
2. Resorcinol
3. Metoprolol
4. 3-Nitrophenol
5. 2-Chlorobenzoic acid
6. Amitriptyline
7. Diethylphthalate
8. Fenoprofen
9. Dipropylphthalate
10. Pyrenesulfonic acid

Reproducible and Scalable Separations

3.5 

0.0 

0.5 

1.0 

1.5 

2.0 

2.5 

3.0 

4.0 min 

1 

2 

3 

4 

5 

6 

7 

8 

9 

10 

XSelect CSH Fluoro-Phenyl, 5 µm 

XSelect CSH Fluoro-Phenyl, 3.5 µm

ACQUITY UPLC CSH Fluoro-Phenyl, 1.7 µm 

 
Reproducibility and scalability for gradient separations on 2.1 x 50 mm columns containing nine  
different batches of CSH fluoro-phenyl representing three (1.7,- 3.5,- and 5-µm) particle sizes.

www.waters.com/xselect

14

15

* Expected or approximate value.

O

Si

O

O

Atlantis

T3

dC18

HILIC Silica

Ligand Density*

1.6 µmol/g

1.6 µmol/g

N/A

Carbon Load*

14%

12%

N/A

End-capped

Proprietary

Proprietary

No

USP Classification

L1

L1

L3

pH Range

2–8

3–7

1-5

Low pH Temp. Limit

45 ˚C

45 ˚C

45 ˚C

High pH Temp. Limit

45 ˚C

45 ˚C

45 ˚C

Pore Diameter*

100Å

100Å

100Å

Surface Area*

330 m2/g

330 m2/g

330 m2/g

Particle Size

3, 5, 10 µm

3, 5, 10 µm

3, 5, 10 µm

 
Atlantis® HPLC Columns provide exceptional performance, 
versatility, and retention for polar compounds, while also 
affording balanced retention for broad analyte mixtures.

 

   
Compatibility with 100% Aqueous 
Mobile Phases

 
To maximize polar compound retention in reversed-phase 
methods, it is possible to use Atlantis Reversed-phase HPLC 
Columns with highly aqueous mobile phases and buffers without 
the risk of pore dewetting and hydrophobic collapse of the 
stationary phase.

 

Long Column Lifetimes Using  
Low-pH Mobile Phases

 
Atlantis Columns resist ligand hydrolysis when using strongly 
acidic mobile phases, thus maintaining method efficiency, 
compound retention, and critical analyte selectivity.

0

 

10

20

30

40

50

60

70

80

90

20 Hour Exposure to 0.5% TFA at 60 °C

Atlantis T3 SunFire C

18

Phenomenex

®

 Synergi™ Hydro-RP

 Shiseido Capcell PAK

®

 AQ

Phenomenex

®  Synergi™ Polar-RP

Agilent Zorbax Eclipse

®  Plus

Atlantis dC

18

Phenomenex

®

 Aqua

YMC Hydrosphere™ C18

GL Sciences Inertsil

®

 ODS-SP

YMC-Pack

™  ODS AQ

™

During this accelerated test, the columns were exposed to low pH and high 
temperature conditions to determine the affect of ligand loss due to hydrolysis. 
The Atlantis T3 bonding resists ligand hydrolysis to maintain analyte retention 
using extremely harsh mobile-phase conditions.

% Retention Loss for Methylparaben

Polar Compound Retention

Comparative separations may not be representative in all applications.

16

Polar Compound Retention  
without Ion-Pairing Reagents

 
Eliminating ion-pairing reagents improves detection limits, 
method reproducibility, and robustness, while reducing instrument 

maintenance due to harsh mobile-phase environments. 

LC Conditions 
LC system: 

Alliance 2695 with 2487 Dual- 

 

 

Wavelength Absorbance Detector

Column:   

4.6 x 150 mm

Mobile phase: 

10 mM ammonium formate, pH 3.0

Flow rate: 

1.3 mL/min for 3 µm

Injection volume:  2.0 µL
Column temp.: 

30 °C

Detection: 

254 nm

Compounds
1. Thiourea
2. 5-Fluorocystine
3. Adenine
4. Guanosine-5’-monophosphate
5. Thymine

Polar Compound Retention

Separating highly polar analytes on the Atlantis T3 Column compared to 
competitive brands. Scientists rely on the uncompromised peak shape and  
retention that only Atlantis Columns provide. 

1

2

3 4

5

1

2

4

3,5

1

2

3,4

5

1 2

4

3

5

1

2

3

4

5

0

1

2

3

4

5

6

7

8 min

Atlantis T3

Agilent Zorbax® SB-AQ

Thermo Hypersil Gold AQ™

Phenomenex ® Synergi™ Hydro-RP

Shiseido Capcell PAK® C

18 AQ

Comparative separations may not be representative in all applications.

www.waters.com/atlantis

17

* Expected or approximate value.

O

Si

O

CH 3

O

Si

O

CH 3

Isocratic Separation
LC system: 

Alliance 2695 with 2487 Dual-  

 

Wavelength Absorbance Detector 

Mobile phase A:  35% 20 mM dipotassium phosphate/  
 

20 mM monopotassium phosphate pH 7.0 

Mobile phase B:  65% methanol
Wavelength: 

254 nm

Flow rate: 

1.0 mL/min

Injection vol.: 

14 µL

Column temp.: 

23 ˚C

Compounds
1. Uracil 
2. Propranolol 
3. Butylparaben 
4. Naphthalene

0

1

2

3

4

5

6

5

10

15

20

25

30

35

40 min

TUSP – 1.26

TUSP – 1.35

TUSP – 1.79

SunFire C18 

4.6 x 150 mm, 5 µm

Phenomenex® Luna® C18 (2) 

4.6 x 150 mm, 5 µm

ACT® Ace® C18 

4.6 x 150 mm, 5 µm

Comparative separations may not be representative in all applications.

SunFire

C8

C18

Silica

Ligand Density*

3.5 µmol/g

3.5 µmol/g

N/A

Carbon Load*

12%

16%

N/A

End-capped

Proprietary

Proprietary

No

USP Classification

L7

L1

L3

pH Range

2–8

2–8

2–8

Low pH Temp. Limit

40 ˚C

50 ˚C

55 ˚C

High pH Temp. Limit

40 ˚C

40 ˚C

45 ˚C

Pore Diameter*

100Å

100Å

100Å

Surface Area*

340 m2/g

340 m2/g

340 m2/g

Particle Size

2.5, 3.5, 5, 10 µm

2.5, 3.5, 10 µm

2.5, 3.5, 10 µm

 
 
SunFire™ Columns set the standard for state-of-the-art bonded 
C18- and C8- silica HPLC columns. Benefiting from years of 
research and product development, SunFire Columns represent 
the best in particle and bonding expertise and deliver industry- 
leading levels of chromatographic performance. 

Excellent Low-pH Stability

 
Under low-pH mobile-phase conditions, SunFire Columns exhibit 
superior column lifetimes that exceed many silica-based HPLC 
column brands.

 

   
High Efficiency

 
A synergistic combination of particle synthesis, packing 
technology, and hardware engineering is required for high 
efficiency. SunFire Intelligent Speed™ (IS™) and Optimum Bed 
Density (OBD™) Columns were developed specifically from  
this knowledge.

 

   
Superior Peak Shapes

 
SunFire Columns provide symmetrical peaks for improved 
resolution of acidic, neutral and basic compounds 
at low and moderate pH ranges (2–8).

 

Peak Shape Comparison of SunFire Columns

18

www.waters.com/sunfire

0

5

10

15

20

25

30

35

40 min

Batch 112 (2005)

Batch 118 (2007)

Batch 130 (2007)

Batch 134 (2008)

Batch-to-Batch Reproducibility of SunFire Columns

This excellent reproducibility is a result of our commitment to maintaining the 
tightest specifications in the HPLC column industry. SunFire Columns start with 
high purity raw materials, and are produced using controlled manufacturing 
processes and column packing procedures that provide today’s scientists with  
the best, most reproducible HPLC columns available. 

Batch-to-Batch Reproducibility

 
Waters is dedicated to maintain the tightest specifications in the 
HPLC industry. Controlled manufacturing processes and column 
packing procedures ensure that you receive the best, most 
reproducible HPLC column available. 

19

O

Si

CH 3

CH 3

O

Si

CH 3

CH 3

Si

CH 3

Polar Group

CH 3

O

Si

CH 3

Polar Group

CH 3

O

O

Si

CH 3

CH 3

O

Si

O

O

Symmetry

Symmetry and 
SymmetryPrep 

C18

Symmetry and 
SymmetryPrep 

 C8

SymmetryShield

 

 RP18

SymmetryShield 

SymmetryPrep

 

 RP8

Symmetry300 

 C18

Symmetry300 

 C4

Particle Size

3.5, 5, 7 µm

3.5, 5, 7 µm

3.5, 5, 7 µm

3.5, 5, 7 µm

3.5, 5 µm

3.5, 5 µm

Particle Shape

Spherical

Spherical

Spherical

Spherical 

Spherical

Spherical

Pore Size

100Å

100Å

100Å

100Å

300Å

300Å

Carbon Load

19%

12%

17%

15%

8.5%

2.8%

End-capped

Proprietary

Proprietary

Proprietary

Proprietary

Proprietary

Proprietary

Symmetry® Columns are manufactured 
using high purity silica and tightly  
controlled manufacturing processes to 
ensure that you receive a column that 
exceeds the standards for HPLC column  
performance. No other silica-based LC 
column brand can match the column-to- 
column and batch-to-batch reproducibility  
of the Symmetry family. Symmetry 
Columns are available in column,  
cartridge, and guard formats:

■

■

Symmetry and SymmetryPrep™ Columns: Deliver maximum reproducibility

■

■

SymmetryShield™ RP18 and RP8 Columns: Provide superior peak shape

■

■

Symmetry300™ C18 and C4 Columns: Offer high recoveries of peptides and proteins 

0.0

1

2

3

4

1.50

3.0

4.5

6.00

7.5

9.0

10.5

12.0

Batch 136

Batch 142

Batch 149

Batch 151

Batch 156

13.5

15.0 min

Unmatched year-to-year reproducibility.

 

Batch-to-Batch Reproducibility of Symmetry Columns

LC Conditions
Column:   

Symmetry C18, 5 µm, 

 

 

4.6 x 150 mm  

Mobile phase A: 

Water

Mobile phase B: 

Acetonitrile

Mobile phase C: 

pH 3.75; 100 mM 

 

 

ammonium formate in water 

Flow rate: 

1.4 mL/min

Isocratic:  

30% A; 60% B; 10% C 

 

    

RSD’s for retention times
1. Terbinafine HCI  0.7% 
2. Ibuprofen 

0.8%

3. Lovastatin 

0.6%

4. Simvastatin 

0.7%  

Symmetry Columns for Reproducibility

 
You can rely on a Symmetry HPLC Column for rugged and  
reproducible performance. Narrow column specification ranges 
minimize variation giving you the confidence that the methods  
you use today will continue to be used in the future.

 
 

 

Injection vol.: 

5.0 µL

Column temp.: 

30 °C

Detection: 233nm 

 

  

20

Symmetry

Symmetry and 
SymmetryPrep 

C18

Symmetry and 
SymmetryPrep 

 C8

SymmetryShield

 

 RP18

SymmetryShield 

SymmetryPrep

 

 RP8

Symmetry300 

 C18

Symmetry300 

 C4

Particle Size

3.5, 5, 7 µm

3.5, 5, 7 µm

3.5, 5, 7 µm

3.5, 5, 7 µm

3.5, 5 µm

3.5, 5 µm

Particle Shape

Spherical

Spherical

Spherical

Spherical 

Spherical

Spherical

Pore Size

100Å

100Å

100Å

100Å

300Å

300Å

Carbon Load

19%

12%

17%

15%

8.5%

2.8%

End-capped

Proprietary

Proprietary

Proprietary

Proprietary

Proprietary

Proprietary

Symmetry Columns for Superior 
Peak Shape

 
SymmetryShield Columns feature Waters’ patented Embedded Polar 
Group Technology that shields the silica’s residual silanols from 
highly basic analytes that improves overall peak shape. Additionally, 
by placing the embedded polar group close to the silica surface, 
the activity of the surface silanols is further reduced. This imparts 
selectivity and retention that is different compared to the Symmetry 
C18 ligand.

Unmatched year-to-year reproducibility.

 

Batch-to-Batch Reproducibility of Symmetry Columns

RSD’s for retention times
1. Terbinafine HCI  0.7% 
2. Ibuprofen 

0.8%

3. Lovastatin 

0.6%

4. Simvastatin 

0.7%  

Embedded Polar Group Technology improves chromatographic peak shape  
and selectivity. 

SymmetryShield Columns Deliver Unique Selectivity

USP Tailing Factor = 1.2

SymmetryShield RP18

USP Tailing Factor = 1.9

Symmetry C18

0

5

10

15

20

25

1

2

3

4

5

7

6

0

5

10

15

20

25

1

2

3

4

5

6

7

AU

AU

30 min

30 min

LC Conditions 
Columns: 

SymmetryShield RP18, 5 µm, 3.9 x 150 mm  

 

Symmetry C18, 5 µm, 3.9 x 150 mm

Mobile phase:  65% methanol; 35% 20 mM 

 

 

monopotassium phosphate/ 

 

 

dipotassium phosphate at pH 7 

Flow rate: 

1.0 mL/min

Detection: 

254 nm

Column temp.:  23 °C 

Compounds

 

1.

 Uracil

2. Propranolol
3. Butylparaben
4. Dipropyl phthalate
5. Naphthalene
6. Amitriptyline
7. Acenaphthene

 
 

 

Injection vol.: 

5.0 µL

Column temp.: 

30 °C

Detection: 233nm 

 

  

www.waters.com/symmetry

21

XTerra® MS, Shield RP, and Phenyl Columns combine the best  
properties of silica and polymeric bonded phases with patented 
Hybrid Particle Technology that replaces one out of every three 
silanols with a methyl group during particle synthesis. This can  
only be achieved during the initial particle synthesis and the  
inclusion of this methyl group is an integral part of the base  
particle backbone. The result is a mechanically strong particle  
that can be used for high pH separations that will improve loading 
and peak shapes for basic compounds.

The Efficiency of Silica with 
Stability of Polymers

 
The vast majority of reversed-phase HPLC separations take place 
on silica-based stationary phases. Silica has long enjoyed such 
attributes as high efficiency and mechanical strength. However, 

XTerra

MS C18

Shield 

RP18

Shield 

RP8

Phenyl

Particle Size

2.5, 3.5, 5, 

10 µm

3.5, 5, 

10 µm

 3.5, 5,  

10 µm

3.5, 5 µm

Particle Shape

Spherical

Spherical

Spherical

Spherical

Pore Size

125Å

125Å

125Å

125Å

Carbon Load

15.5%

15.0%

13.5%

12.0%

End-capped

Proprietary 

Proprietary 

Proprietary 

Proprietary 

Traditional Silica Manufacturing Process 
Bonded and End-capped Silica Particle

XTerra Manufacturing Process  
Much more than a surface modification 
Bonded and End-capped XTerra Particle

silica suffers from poor peak shape for bases and a limited pH 
range. One way that chromatographers have attempted to overcome 
these limitations is by turning to polymer-based stationary phases. 
Polymers, however, have not enjoyed the acceptance of silica due  
to poor efficiency, low mechanical strength, and unpredictable peak 
elution order when transferring methods from polymeric to silica-
based columns.

Hybrid Particle Technology overcomes these limitations and  
combines the best attributes of both these materials while  
overcoming each material’s weaknesses. The result is a rugged 
material that has high mechanical strength, high efficiency,  
excellent peak shape for bases, and easy scale-up from analytical 
to preparative chromatography.

Traditional Silica versus XTerra Manufacturing Process

Si

CH 3

Polar Group

CH 3

O

Si

CH 3

Polar Group

CH 3

O

O

Si

C H 3

C H

2

C H

C H

3

O

 

■

 Highest, most  

 

   Homogenous Coverage

 

■

 1/3 Less Silanols

 

■

 Superior Peak Shape

 

■

 pH Range 1-12

 

■

 Lowest Surface Coverage

 

■

 Poor Peak Shape

 

■

 pH Range 2–8

Unbonded

XTerra

Particle

Methypolyethoxysilane

(MPEOS)

Tetraethoxysilane

(TEOS)

Methyltriethoxysilane

(MTEOS)

Unbonded  

Silica Particle

Polythoxysilane

(PEOS)

Tetraethoxysilane

(TEOS)

* Expected or approximate value.

OH

Si

O

O

O

O

Si

O

OH

O

Si

O

OH

O

Si

O

O

O

Si

O

Si

O

OH

Si

OH

O

Si

O

O

Si

Si

Si OH

O

O

Si O

O

Si

Si

Si

Si

Si

Si

Si

HO

Si

Si

O

O

Si

O

O

Si O

HO

O

Si

Si

Si

Si

O

HO

O

Si

Si

O

HO

O

Si

O

HO

HO

Si

Si

O

Si

O

OH

O

Si

Si

OH

O

O

O

Si

OH

O

O

Si

Si

Si

OH

O

O

Si

O

O

Si OH

O

Si

Si

Si

Si

OH

Si

O

O

O

Si

O

O

Si

O

OH

O

Si O

O

O

Si

O

Si

O

OH

Si

O

Si

O

O

O

Si

Si

Si

O

O

Si O

O

Si

Si

Si

Si

Si

Si

Si

HO

Si

O

Si

O

O

Si

O

O

Si O

HO O

Si

Si

Si

Si

O

O

Si

Si

O

HO

O

Si

O

HO

Si

Si

O

Si

O

OH

O

Si

Si

OH

O

O

Si

O

O

Si

Si

Si OH

O

O

Si

O

O

Si OH

O

Si

Si

Si

Si

H3C

H3C

CH3

H3C

H3C

H3C

CH3

CH3

H3C

CH3

CH3

CH3

H3C

H3C

CH3

CH3

22

Silica Separations at Polymer pH

0

0.1

0.2

0.3

0.4

0.5

1

3

2

4

6

5

10

20

AU

6

5

4

3

2

1

AU

0

0.4

0.8

1.2

10

20

min

min

LC Conditions 
LC system: 

Alliance 2690 with 996 PDA Detector 

Mobile phase A:  20 mM ammonium hydroxide, pH 10.7 
Mobile phase B:  Acetonitrile
Flow rate: 

3 mL/min 

Gradient: 

Time   Profile   

 

(min)   %A %B

 

0.0 

 70 30

 

25.0   40 60

Injection vol.: 

5 µL 

Column temp.: 

Ambient 

Detection: 

220 nm 

Compounds

 

 

1.

 Codeine

2. Yohimbine 

 

3. Thebaine
4. Cocaine
5. Resperine
6. Methadone

XTerra Shield RP18: 4.6 x 150 mm 
pH 10.7

Polymer Column: 4.1 x 150 mm  
pH 10.7

Traditional Silica versus XTerra Manufacturing Process

0

0.1

0.2

0.3

0.4

0.5

1

3

2

4

6

5

10

20

AU

6

5

4

3

2

1

AU

0

0.4

0.8

1.2

10

20

min

min

www.waters.com/xterra

23

WATERS SPHERISORB COLUMNS

 
Waters Spherisorb® Columns are one of the most widely referenced HPLC columns in the scientific literature. There are over  
2,000 analytical abstracts published using Waters Spherisorb Columns, providing a tremendous range of validated methods and 
applications to assist in your method development process. 

Waters Spherisorb Columns are produced in a wide range of particle sizes (3-, 5-, and 10- µm) and bonded phases to meet your  
chromatographic needs. In addition, Waters Spherisorb Columns’ high quality bonded phases give many different and unique  
separation selectivities. Waters Spherisorb Analytical Columns are supplied with industry-standard Parker-style column  
end fittings. 

Ligand Type

Phenyl

CN (Nitrile)

OD/CN

W (Silica)

SCX

SAX

Particle Size

3, 5, 10 μm

3, 5, 10 μm

5 μm

3, 5, 10 μm

5, 10 μm

5, 10 μm

Surface Area

220 m2/g

220 m2/g

220 m2/g

220 m2/g

220 m2/g

220 m2/g

Particle Shape

Spherical

Spherical

Spherical

Spherical

Spherical

Spherical

Pore Size

80Å

80Å

80Å

80Å

80Å

80Å

Carbon Load

2.5%

3.1%

5%

N/A

4%

4%

Ligand Coverage

2.72 µmol/m2

3.29 µmol/m2

1.15 µmol/m2

N/A

N/A

N/A

End-capped

No

No

Proprietary 

No

No

No

 Spherisorb

Ligand Type

ODS2 (C18)

ODS1 (C18)

ODSB (C18)

C8

C6

C1

NH2 (Amino)

Particle Size

3, 5, 10 μm

3, 5, 10 μm

5 μm

3, 5, 10 μm

3, 5, 10 μm

3, 5, 10 μm

3, 5, 10 μm

Surface Area

220 m2/g

220 m2/g

220 m2/g

220 m2/g

220 m2/g

220 m2/g

220 m2/g

Particle Shape

Spherical

Spherical

Spherical

Spherical

Spherical

Spherical

Spherical

Pore Size

80Å

80Å

80Å

80Å

80Å

80Å

80Å

Carbon Load

11.5%

6.2%

11.5%

7.75%

4.7%

2.15%

1.9%

Ligand Coverage

2.98 µmol/m2

1.49 µmol/m2

2.98 µmol/m2

3.12 µmol/m2

3.36 µmol/m2

2.97 µmol/m2

2.64 µmol/m2

End-capped

Proprietary 

No

Proprietary 

Proprietary 

Proprietary 

No

No

www.waters.com/spherisorb

24

Ligand Type

Phenyl

CN (Nitrile)

OD/CN

W (Silica)

SCX

SAX

Particle Size

3, 5, 10 μm

3, 5, 10 μm

5 μm

3, 5, 10 μm

5, 10 μm

5, 10 μm

Surface Area

220 m2/g

220 m2/g

220 m2/g

220 m2/g

220 m2/g

220 m2/g

Particle Shape

Spherical

Spherical

Spherical

Spherical

Spherical

Spherical

Pore Size

80Å

80Å

80Å

80Å

80Å

80Å

Carbon Load

2.5%

3.1%

5%

N/A

4%

4%

Ligand Coverage

2.72 µmol/m2

3.29 µmol/m2

1.15 µmol/m2

N/A

N/A

N/A

End-capped

No

No

Proprietary 

No

No

No

 
NOVA-PAK COLUMNS

 
The bonded phases of Nova-Pak® Columns are available in 4 µm and 6 µm particle sizes that offer high resolution as well 
as faster and more efficient chromatography. The smaller particle size in conjunction with shorter column lengths can be used to 
reduce solvent consumption while maintaining resolution for complex mixtures. Analytical columns with 4 µm particle size packing 
are available in 75, 150, and 300 mm length steel columns. Semi-preparative Prep Nova-Pak HR Columns are packed with 6 µm 
particle-size packings and provide an unparalleled range of separation possibilities. The high-efficiency packing of Prep Nova-Pak HR 
Columns provides faster separations using less solvent with the added advantage of more concentrated fractions, all of which reduce 
preparative chromatography cost. All Nova-Pak Columns are packed to stringent QC procedures in our cGMP manufacturing facility to 
ensure batch-to-batch reproducibility.

 Spherisorb

Ligand Type

ODS2 (C18)

ODS1 (C18)

ODSB (C18)

C8

C6

C1

NH2 (Amino)

Particle Size

3, 5, 10 μm

3, 5, 10 μm

5 μm

3, 5, 10 μm

3, 5, 10 μm

3, 5, 10 μm

3, 5, 10 μm

Surface Area

220 m2/g

220 m2/g

220 m2/g

220 m2/g

220 m2/g

220 m2/g

220 m2/g

Particle Shape

Spherical

Spherical

Spherical

Spherical

Spherical

Spherical

Spherical

Pore Size

80Å

80Å

80Å

80Å

80Å

80Å

80Å

Carbon Load

11.5%

6.2%

11.5%

7.75%

4.7%

2.15%

1.9%

Ligand Coverage

2.98 µmol/m2

1.49 µmol/m2

2.98 µmol/m2

3.12 µmol/m2

3.36 µmol/m2

2.97 µmol/m2

2.64 µmol/m2

End-capped

Proprietary 

No

Proprietary 

Proprietary 

Proprietary 

No

No

Nova-Pak

Chemistry

C18

C8

Phenyl

CN

Silica

Prep HR C18

 Prep HR Silica

Particle Size

4 µm

4 µm

4 µm

4 µm

4 µm

6 µm

6 µm

Particle Shape

Spherical

Spherical

Spherical

Spherical

Spherical

Spherical

Spherical

Pore Size

60Å

60Å

60Å

60Å

60Å

60Å

60Å

Carbon Load

7%

4%

5%

2%

N/A

7%

N/A

End-capped

Proprietary 

Proprietary 

Proprietary 

Proprietary 

No

Proprietary 

No

www.waters.com/nova-pak

25

RESOLVE COLUMNS

 
The non-endcapped Resolve™ packing is significantly different from Waters other packing materials. The change in chromatographic 
behavior is most commonly noticed with polar compounds which are typically more retained. Basic compounds can be  
chromatographed using mobile phase modifiers, such as ion-pairing reagents, which reduce excessive tailing.

DELTA-PAK COLUMNS

 
Delta-Pak™ Columns are ideal for separation and isolation of peptides, proteins, and natural products and are available in  
two different pore sizes that are optimized for large molecule separations. Delta-Pak Columns are known for consistent and  
predicable scaling between column formats, allowing purification scientists the ability to isolate target compounds from  
the milligram to gram quantities. The highly stable Delta-Pak bonded silica is available in 5 μm and 15 μm  
particle sizes.

Resolve

Ligand Type

Silica

C18

C8

CN

Particle Size

5, 10 µm

5, 10 µm

10 µm

10 µm

Particle Shape

Spherical

Spherical

Spherical

Spherical

Pore Size

90Å

90Å

90Å

90Å

Carbon Load

10 %

10%

5%

3%

End-capped

No

No

No

No

Delta-Pak

Ligand Type

C18

C18

C4

C4

Particle Size

5, 15 μm

5, 15 μm

5, 15 μm

5, 15 μm

Particle Shape

Spherical

Spherical

Spherical

Spherical

Pore Size

100Å

300Å

100Å

300Å

Carbon Load

17%

7%

7%

3%

End-capped

Proprietary 

Proprietary 

Proprietary 

Proprietary 

www.waters.com/delta-pak

www.waters.com/resolve

26

Irregular Particle Technology

 
The first HPLC packing materials were comprised of non-spherical and irregularly shaped particles. Typically, these columns have 
reduced mechanical stability and lower efficiency compared to a column packed with spherical particles. However, even with these 
limitations, there are many methods that require the use of these sorbents. As a primary manufacture of sorbents and bonded 
materials, Waters has demonstrated consistent and reliable column performance for over 40 years and we will continue to support 
these brands for the future.

µBONDAPAK/BONDAPAK COLUMNS

 
If your method calls for a μBondapak® Column, there is only one column that contains μBondapak C18 packing material. Many  
companies claim “μBondapak-like” selectivity, but none have passed Waters stringent QC batch tests. μBondapak or BondaPak® 
packing materials have demonstrated reproducibility from year-to-year since 1973, allowing μBondapak Columns to be the one  
of the most widely referenced HPLC column brands.

 

 
µPORASIL/PORASIL COLUMNS

 
μPorasil™ and Porasil™ particles were one of the first commercially available fully porous packing materials used for LC  
separations. In contrast to the reversed-phase separation ability of μBondapak C18, the non-bonded, silica-based material  
in μPorasil Columns was produced to provide normal-phase separations for a wide array of sample types.

µBondapak/

Bondapak

Ligand Type

C18

Phenyl

CN

NH2

Particle Size

10 μm

10 μm

10 μm

10 μm

Particle Shape

Irregular

Irregular

Irregular

Irregular

Pore Size

125Å

125Å

125Å

125Å

Carbon Load

10%

8%

6%

3.5%

End-capped

Proprietary

Proprietary

Proprietary

No

www.waters.com/bondapak

µPorasil/Porasil

Ligand Type

Silica

Particle Size

10, 15-20 μm

Particle Shape

Irregular

Pore Size

125Å

Carbon Load

N/A

End-capped

No

www.waters.com/delta-pak

www.waters.com/resolve

www.waters.com/porasil

27

Column  
Performance Monitoring

Intended Use

Detector 

Performance Monitoring

Intended Use

Neutrals QC Reference 
Material

Provides chromatographic performance 
information under isocratic conditions  
using 3 neutral probes.

QDa QC Reference Material

Provides chromatographic and mass 
spectrometer information using an  
8 component mixture in an optimized  
format for the ACQUITY QDa® Detector.  
This solution contains 1 critical pair to 
measure chromatographic performance.

Reversed-Phase QC 
Reference Material

Provides reversed-phase chromatographic 
performance information under gradient 
conditions using 1 void marker, 3 neutral,  
1 acidic, and 2 basic probes.

Quad LCMS QC Reference 
Material

Provides chromatographic and mass 
spectrometer information using a 9 component 
mixture in a format optimized for quadrupole 
MS Systems. This solution contains 2 
critical pairs to measure chromatographic 
performance.

HILIC QC Reference Material

Provides chromatographic performance 
information inclusive of mobile-phase pH 
in HILIC mode using 1 void marker, 1 polar 
neutral, and 2 polar basic probes. 

LCMS QC Reference Material

Provides chromatographic and mass 
spectrometer information using a 9 component 
mixture in a format optimized for the highest 
resolution Tof/QTof MS Systems. This 
solution contains 2 critical pairs to measure 
chromatographic performance.

 

How Do You Know Your Chromatographic System is in Proper Working Order?

Quality Control (QC) Reference Materials contain mixtures of standards specifically chosen to provide an 
easy and reliable way to monitor the performance of any chromatographic system. By using a QC Reference 
Materials, you can be assured that your column and system are ready to analyze your samples. Regular use 
of QC Reference Materials also provides an opportunity to benchmark your chromatographic systems and 
trend performance over time, making it easier to proactively identify problems and resolve them faster.

Since chromatographic analyses are complex and depend on many different variables, such as mobile- 
phase composition, column type, and detection method, Waters has formulated specific QC Reference 
Material mixtures designed to test systems with these differences in mind.

To locate additional information for standards specific to calibration, qualification, and tuning of instruments 
and detectors, as well as a more comprehensive list of available standards and reagents, visit asr.waters.com

Extend Column Lifetime with VanGuard 
Column Protection Products
VanGuard™ Pre-columns and Cartridges are optimized to protect 
and prolong analytical column lifetimes without compromising 
chromatographic performance. They are available in a wide selection of 
particle sizes and stationary phases, making them ideally suited for the 
physical and chemical protection for all Waters analytical columns.

■

■

Removes particulates and chemical contamination

■

■

Maintains UPLC, UHPLC, and HPLC separation efficiency

■

■

Provides cost effective protection for all Waters analytical columns

28

Column  
Performance Monitoring

Intended Use

Detector 

Performance Monitoring

Intended Use

Neutrals QC Reference 
Material

Provides chromatographic performance 
information under isocratic conditions  
using 3 neutral probes.

QDa QC Reference Material

Provides chromatographic and mass 
spectrometer information using an  
8 component mixture in an optimized  
format for the ACQUITY QDa® Detector.  
This solution contains 1 critical pair to 
measure chromatographic performance.

Reversed-Phase QC 
Reference Material

Provides reversed-phase chromatographic 
performance information under gradient 
conditions using 1 void marker, 3 neutral,  
1 acidic, and 2 basic probes.

Quad LCMS QC Reference 
Material

Provides chromatographic and mass 
spectrometer information using a 9 component 
mixture in a format optimized for quadrupole 
MS Systems. This solution contains 2 
critical pairs to measure chromatographic 
performance.

HILIC QC Reference Material

Provides chromatographic performance 
information inclusive of mobile-phase pH 
in HILIC mode using 1 void marker, 1 polar 
neutral, and 2 polar basic probes. 

LCMS QC Reference Material

Provides chromatographic and mass 
spectrometer information using a 9 component 
mixture in a format optimized for the highest 
resolution Tof/QTof MS Systems. This 
solution contains 2 critical pairs to measure 
chromatographic performance.

 

Waters Part Selector and Selectivity Chart for iPad®

www.waters.com/apps

Electronic Tools

Waters Reversed-Phase Column Selectivity Chart

www.waters.com/selectivitychart

Waters Column Advisor

www.waters.com/columnadvisor

29

Waters, The Science of What’s Possible, CORTECS, UPLC, Alliance, ACQUITY UPLC, XBridge, XSelect, 
Atlantis, Symmetry, XTerra, Waters Spherisorb, Nova-Pak, BondaPak, μBondaPak, QDa, SunFire, and 
ACQUITY are registered trademarks of Waters Corporation. BEH Technology, CSH, Intelligent Speed,  
IS, OBD, SymmetryShield, Symmetry300, μPorasil, Porasil, Resolve, Delta-Pak, and SymmetryPrep  
are trademarks of Waters Corporation. All other trademarks are property of their respective owners.

©2014-16 Waters Corporation. Printed in the U.S.A. 
December 2016 720003995EN KP-SIG

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

www.waters.com/hplccolumns

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