Ultimate Efficiency Unleashed -...

©2017 Waters Corporation 1COMPANY CONFIDENTIAL

Ultimate Efficiency Unleashed:

Introducing CORTECS™ Solid-Core Particle Columns for

Maximizing Resolution and Throughput

Willem Joubert

Microsep

Waters Division


Agenda

Review of Solid-Core Particles

CORTECS® Solid-Core Particle Columns

Column Benefits for Improved Laboratory Performance

Practical Applications of CORTECS Columns

Summary



Solid-Core particles have been around since the 1970’s

– Waters Corasil I 30-80 µm particles

– Poor efficiency and poor loading capacity

Today's modern core-shell particles are prepared by the multi-layering of

silica sols around a solid silica core.

– Thicker shell, allows for better Loading capacity

– Better particle design, improvements in packing provide higher efficiency

FIB SEM ImagesSolid-Core

Core

Part

icle


The Solid-Core Particle

Only thin outer layer contains the pores

with the chromatographic surface

The center core is nonporous

The outer shell is typically “bumpy” not

pretty

The particle size distribution is narrower

CORTECSSolid-core

dcore = 1.1 µm

dp

= 1

.6 µ

m

Rho, r = 1.1/1.6 = 0.7

66% Porous Volume

ρ = 0 → fully porous particle

ρ = 1 → nonporous particle

ρ = core diameter / particle diameter

Compared to Fully Porous Particles

G. Guiochon, F. Gritti, J. Chromatogr. A 1218 (2011) 1915–1938 Omamogho et al., J. Chromatogr. A 1218 (2011) 1942-1953


Why are Modern Solid-Core Columns so

Popular?

Provide higher efficiency when compared to fully porous particles of

equivalent particle size.

Provide lower backpressure when compared to fully porous particles of

equivalent particle size.

Efficiency

Pressure


Achieving Optimal Efficiency

Impact of System Dispersion

The Effect of System Dispersion on Column Performance

– In 2004, key component of UPLC introduction was the relationship between

observed efficiency of small particle columns and the dispersion of the system

– As system dispersion decreases, observed efficiency of columns increases

– In order to realize full efficiency of CORTECS Columns, match the column

dimension and particle size with the instrument dispersion


Where Does System Dispersion Occur?

Band Spreading: 1) From the Injector2) Into, through and out

of the column3) Into the Detector

Extra ColumnWithin Column


Band Spreading, Peak Height, and

Resolution

LC systems (column and instrument) capable of producing narrower/sharper bands create narrower/sharper peaks

This results in better resolution, taller peaks and better sensitivity

Better separationMore concentrated “Bands”Higher Sensitivity

Both analytes (blue and red) are not separated [a partial co-elution –shown as a “purple” band]

System withMORE

Band Spreading

System with LESS

Band Spreading


System Dispersion Influence Across Different

Column i.d.s

AU

0.00

0.02

0.04

0.06

AU

0.00

0.02

0.04

0.06

AU

0.00

0.02

0.04

0.06

Minutes

0.00 0.10 0.20 0.30 0.40 0.50 0.60 0.70 0.80 0.90 1.00 1.10 1.20 1.30 1.40 1.50

4.6 x 50 mm

3.0 x 50 mm

2.1 x 50 mm

7200

4800

2900

Efficiency

3.6

2.5

1.4

Alliance HPLC: Bandspread 36 µL

Resolution

Using a larger column I.D. (higher column volume) mitigates the effect of system dispersion on a separation


Matching System Dispersion With Column I.D.

AU

0.00

0.02

0.04

0.06

AU

0.00

0.02

0.04

0.06

AU

0.00

0.02

0.04

0.06

Minutes

0.00 0.10 0.20 0.30 0.40 0.50 0.60 0.70 0.80 0.90 1.00 1.10 1.20 1.30 1.40 1.50

4.6 x 50 mm

3.0 x 50 mm

2.1 x 50 mm

HPLC: AllianceDispersion: >30 µL

UHPLC: ACQUITY ArcDispersion: 12-30 µL

UPLC: ACQUITY UPLC H-ClassDispersion: <12 µL

7200

7600

7300

Efficiency

3.6

Resolution

3.8

3.7

Matching column I.D. (column volume) to system dispersion helps to maintain column efficiency and resolution


Matching System Dispersion With Column I.D.

Dispersion > 30 µL

Columns accepted: • 3.0 – 4.6 mm ID• 3 - 10 µm particles

Recommended: • 4.6 mm ID, 3.5 or 5 µm

Typical operating pressure: • < 6,000 PSI

Dispersion 12 - 30 µL

Columns accepted: • 2.1 - 4.6 mm ID• 1.7 - 5 µm particles

Recommended: • 3.0 mm ID, 2.x µm

Typical operating pressure: • 6,000 – 15,000 PSI

Dispersion < 12 µL

Columns accepted: • 1.0 - 4.6 mm ID• 1.6 - 5 µm particles

Recommended: • 2.1 mm ID, 1.x µm

Typical operating pressure: • 9,000 – 15,000 PSI

Increased flexibility and sample characterization


What makes Solid-Core Particles Different from

Fully Porous?

Lets look into the science of how to get lower backpressure and higher

efficiency

– Backpressure equation

– Improved efficiency

o - the van Deemter equation

F. Gritti, G. Guiochon, J. Chromatogr. A 1221 (2012) 2– 40F. Gritti, Chromatography Today May/June (2012) 4-11



Fully Porous?


efficiency






Lower Backpressure? How?

Pre

ssure

(psi)

External or Interstitial Porosity, ee

3

2

22

)1(180

e

e

pdr

LFP

e

e

Kozeny-Carman Equation

The effect particle size, dp, and

the external porosity, ee , on

column backpressure

Fully porous sub-2-µm particle

columns tend to have ee <0.38

CORTECS solid-core columns tend

to have ee 0.39

A. Daneyko, Anal. Chem. 2011, 83, 3903–3910

Flow Rate (F) = 0.5 mL/min

1.7 µm BEH(Fully Porous)

1.6 µm CORTECS(Solid Core)


Similar Backpressure

0

2,000

4,000

6,000

8,000

10,000

12,000

14,000

0.00 0.25 0.50 0.75 1.00 1.25

Pre

ssure

(psi)

Flow Rate (mL/min)

SpecificPermeability

CORTECS UPLC C18+ 1.6 µm: k0 = 2.22E-15 m²

ACQUITY UPLC BEH C18 1.7 µm: k0 = 2.28E-15 m²

Conditions: Tested on an ACQUITY UPLC I-Class using 2.1 x 50 mm columns, 70% Acetonitrile at 30 °C



Fully Porous?


efficiency






More Efficient, How???

The van Deemter Equation the basics

The van Deemter equation describes empirically additive sources of

dispersion that result as a function of mobile phase velocity and particle

size.

ucu

baH

0

2

4

6

8

10

0 5 10 15 20 25 30

Linear Velocity u

H-u






size.

The a-term was thought to be a constant dp

and takes into account flow heterogeneity

ucu

baH

0

2

4

6

8

10

0 5 10 15 20 25 30

Linear Velocity u

0

2

4

6

8

10

0 5 10 15 20 25 30

Linear Velocity u

H-u

a-term






size.



The b-term is the longitudinal diffusion term

which diminishes at high linear velocity

ucu

baH

0

2

4

6

8

10

0 5 10 15 20 25 30

Linear Velocity u

0

2

4

6

8

10

0 5 10 15 20 25 30

Linear Velocity u

0

2

4

6

8

10

0 5 10 15 20 25 30

Linear Velocity u

H-u

a-term

b-term






size.



The b-term is the longitudinal diffusion term

which diminishes at high linear velocity

The c-term is the mass transfer term which

increases at high linear velocity

ucu

baH

0

2

4

6

8

10

0 5 10 15 20 25 30

Linear Velocity u

0

2

4

6

8

10

0 5 10 15 20 25 30

Linear Velocity u

0

2

4

6

8

10

0 5 10 15 20 25 30

Linear Velocity u

0

2

4

6

8

10

0 5 10 15 20 25 30

Linear Velocity u

H-u

a-term

b-term

c-term


What makes Solid–Core Particles more Efficient

than Fully Porous???

The one of the major differences for small molecules is in the b-term:

The b-term is the longitudinal diffusion term and is the

easiest to explain and examine:

– At the () optimum linear velocity the b-term

contribution is significant.

– The impervious solid-core at the center of these

particles decrease the volume available for diffusion thereby decreasing the b-

term. Higher Rho-values lead to lower b-terms.

– At higher linear velocities, convective flow dominates and the b-term no longer

plays a significant role.

– Independent measurement of the b-term can be made by peak parking

experiments.

F. Gritti, G. Guiochon, J. Chromatogr. A 1221 (2012) 2– 40K. Miyabe, ANALYTICAL SCIENCES MARCH 2013, VOL. 29

0

2

4

6

8

10

0.0 0.5 1.0 1.5Linear Velocity cm/sec

H-u


Reduced van Deemter Equation

h = A+ B/u + Cu

Reduced h for Heptanophenone k' = 3.5

Reduced Linear Velocity (ue)

hmin = 1.9

Solid-Core 1.6 µm CORTECS C18+

A = 0.48, B = 4.20, C = 0.070R² = 0.9994

Fully Porous 1.7 µm BEH C18

A = 0.30, B = 6.93, C = 0.093R² = 0.9997

0

2

4

0 5 10 15 20 25 30

htotal for ACQUITY UPLC 1.7 µm BEH C18

hEddy (A) hLong (B)

hmin = 1.6

htotal for CORTECS UPLC 1.6 µm C18+

0

2

4

0 5 10 15 20 25 30

hmass trans (C) + heat

Comparative separations may not be representative in all applications.


Competitive

Solid-Core Comparison

CORTECS 1.6 µm C18 +

Competitor Solid-Core 1.7 µm C18

Solid-Core 1.7 µm Competitor C18

A = 0.85, B = 4.72, C = 0.066R² = 0.9996

3,000

5,000

7,000

9,000

11,000

13,000

15,000

17,000

19,000

21,000

0.0 0.5 1.0 1.5

Pla

tes (

4 s

igm

a)

Flow Rate (mL/min)

-

34%higher

Reduced h for Heptanophenone k' = 3.5 and 3.3

hEddy (A) hLong (B) hmass trans (C) + heat

Solid-Core 1.6 µm CORTECS™ C18+

A = 0.48, B = 4.20, C = 0.070R² = 0.99940

2

4

0 5 10 15 20 25 30

htotal for Competitor Solid-Core 1.7 µm C18

hmin = 1.8

hmin = 1.6


0

2

4

0 5 10 15 20 25 30




CORTECS Efficiency Advantage

(small molecules)

The largest improvement in efficiency comes from the reduction of the b-

term (longitudinal diffusion)

– The impervious solid core prevents the analytical band from spreading through

the particle

A-term (Eddy dispersion) is a smaller contribution

– Influenced by how well the column bed is packed/arranged

C-term (mass transfer) is already small for small particle columns that

are either fully porous or superficially porous

– The story of a shorter diffusion path is negligible

S. Khirevich et al., J. Chromatogr A 1212 (2010) 4713F. Gritti, G. Guiochon, J. Chromatogr A 1221 (2012) 2– 40

F. Gritti, Chromatogr. Today, May/June (2012), 4-11


Agenda




Complete Confidence

Quality

– Set the industry standard in column

reproducibility

– Experienced primary manufacturer of silica

and hybrid particles

o Own every step of the manufacturing

process

o Exhibit control over our processes

• From synthesis to hardware production

to column packing

High quality, reproducible columns


Batch-to-Batch Reproducibility


1.00

1.50

2.00

2.50

3.00

3.50

4.00

4.50

5.00

5.50

6.00

0.0 5.0 10.0 15.0 20.0 25.0 30.0

Reduced P

late

Heig

ht,

h (

USP)

Linear Velocity (cm/min)

2.1 x 50 mm

3.0 x 50 mm

4.6 x 50 mm

Consistent packing qualityacross all three diameters

provides for improved scalability and smooth method

transfer

Column Packing Consistency

CORTECS Columns

This is difficult to achieve!


Challenges of Efficiently Packing Across

Different Column Diameters

1.00

1.50

2.00

2.50

3.00

3.50

4.00

0.0 5.0 10.0 15.0 20.0 25.0 30.0

Reduce

d P

late

Heig

ht,

h (

USP)

Linear Velocity (cm/min)

Kinetex C18, 2.6µm - van Deemter Performance Across Diameters

2.1 x 50 mm

3.0 x 50 mm

4.6 x 50 mm

Competitive sub-3-µm Solid-Core Columns

Variable efficiencies acrossinternal diameters (14.3% difference in efficiency)


E. Oláh, S. Fekete, J. Fekete, K. Ganzler, J Chrom. A 1217, (2010), 3642

2.1 mm

3.0 mm

4.6 mm


CORTECS Column Family

UPLC Columns featuring 1.6 µm solid-core silica particles

HPLC/UHPLC Columns featuring 2.7 µm solid-core silica particles

Key Benefits

– High Efficiency

o Resolution

o Speed

– Scalability UPLC HPLC/UHPLC

7 chemistries:

Phenyl

T3

HILIC

Shield RP18

C18+

C18

C8


Having Scalability Between Different Solid-Core

Particle Sizes

Solid-Core

Core

Part

icle

Attribute CORTECS

r 0.7

Particle Size 1.6 µm, 2.7 µm

Pore Volume 0.26 cm³/g

Pore Size 90 Å*

Surface Area 100 m²/g

FIB SEM Images

Solid-Core

Core

Part

icle

1.6 µm 2.7 µmRho (ρ) = 0 → fully porous

particle

ρ = 1 → nonporous particle

r = core diameter / particle diameter

Maintaining the rho value,Allows for scalability between the particle sizes

*T3 chemistry is 120 Å


Agenda





Performance Benefits

Particle Equivalency

CORTECS Solid-Core Particle Columns

– Efficiencies that are equivalent to that of smaller fully porous particles

– Backpressures equivalent to that of larger fully porous particles

CORTECS Particle Size

Equivalent Porous Particles

Efficiency Backpressure

1.6 µm 1.3 µm 1.8 µm

2.7 µm 2.2 µm 3.1 µm


Fully Porous C18, 3.5 µm

Psi: 2900

N peak 4: 17,600

High Efficiency Separation at HPLC

BackpressuresA

U

0.00

0.02

0.04

0.06

0.08

0.10

AU

0.00

0.02

0.04

0.06

0.08

0.10

AU

0.00

0.02

0.04

0.06

0.08

0.10

Minutes

1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50

1

2 34

Rs 4.9

Rs 6.3

Rs 8.2

Fully Porous C18, 5 µm

Psi: 1900

N peak 4: 10,400

CORTECS C18, 2.7 µm

Psi: 3200

N peak 4: 23,600

1. Estradiol2. Ethinyl estradiol3. Estrone4. Levonorgestrel

Configuration: 4.6 x 150 mm

Note: 2.5 µm fully porous Psi ~6000


Transferability

Method Transfer

– Scaled synthetic process

o Rho value scaled

– UPLC HPLC

o Seamless method transfer

o Future proofing

1.6 µm UPLC 2.7 µm HPLC/UHPLC


Agenda





– The right column for your challenging assays


The CORTECS Family

C18

C18+

HILIC

C8

Phenyl

T3

Shield RP18

General Purpose, balanced acidic, basic, neutral analyte retention

Best choice for use with MS detection and low ionic strength acidic mobile phases

Analysis of very polar analytes using HILIC mode

Lower retention; general purpose

Different selectivity particularly for aromatic analytes

Balanced retention of non-polar and polar compounds using RPLC

Different selectivity Improved peak shapes for basic analytes

vs. CORTECS C18


Method Transfer:

HPLC UHPLC UPLC

1

2

34 5

AU

0.00

0.01

0.02AU

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

3.5 4.0 4.5 5.0 5.5 6.0 6.5 7.0 7.5 8.0

AU

0.00

0.01

0.02

3.5 4.0 4.5 5.0 5.5 6.0 6.0 7.0 7.5 8.0

Minutes

1.8 2.0 2.2 2.4 2.6 2.8 3.0 3.2 3.4 3.6

Fully Porous C18, 5 µm4.6 x 150 mm1.00 mL/minAlliance® HPLC

CORTECS 2.7 µm C184.6 x 75 mm1.85 mL/minAlliance HPLC

CORTECS 2.7 µm C184.6 x 75 mm1.85 mL/minH-Class


Initial Method

Compatible with HPLC, UHPLC, and

UPLC systems

Transfer to 1.6 µm9X Faster method15X Less solvent



1

2

34 5

AU

0.00

0.01

0.02AU

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

3.5 4.0 4.5 5.0 5.5 6.0 6.5 7.0 7.5 8.0

AU

0.00

0.01

0.02

3.5 4.0 4.5 5.0 5.5 6.0 6.0 7.0 7.5 8.0

Minutes

1.8 2.0 2.2 2.4 2.6 2.8 3.0 3.2 3.4 3.6





Initial Method

Compatible with HPLC, UHPLC, and

UPLC systems



Method Transfer:

HPLC UHPLC UPLC


1

2

34 5

AU

0.00

0.01

0.02AU

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

3.5 4.0 4.5 5.0 5.5 6.0 6.5 7.0 7.5 8.0

AU

0.00

0.01

0.02

3.5 4.0 4.5 5.0 5.5 6.0 6.0 7.0 7.5 8.0

Minutes

1.8 2.0 2.2 2.4 2.6 2.8 3.0 3.2 3.4 3.6



CORTECS 2.7 µm C184.6 x 75 mm1.85 mL/minACQUITY UPLC H-Class


Initial Method

Compatible with UHPLC systems4X Less Solvent



AU

0.000

0.010

0.020

3.5 4.0 4.5 5.0 5.5 6.0 6.0 7.0 7.5 8.0

CORTECS 2.7 µm C183.0 x 75 mm0.79mL/minACQUITY Arc UHPLC

Method Transfer:

HPLC UHPLC UPLC


Agenda





Summary


Summary

CORTECS Solid Core columns set the new gold standard for efficiency,

reproducibility, and quality.

Built on a proven solid-core particle technology, CORTECS Solid-core

Columns provide more information in less time

CORTECS Columns are fully scalable between particle sizes, and can be

used with any UPLC, UHPLC, and HPLC system in your laboratory.

o Ultimate Efficiency = CORTECS 1.6 µm Columns

o Ultimate Utility = CORTECS 2.7 µm Columns


Premier Particle Technologies

CORTECS™ Solid-Core Technology

Highestefficiency

Optimized Backpressure

Seamless scalability

BEH Technology

Unparalleled pH stability

Mobile phase and temperature versatility

Seamless scalability

HSS Technology

Enhanced retention

Particle and ligand selectivity

Seamless Scalability

CSH™ Technology

Exceptional loading capacity

Superior basic peak shape

Seamless Scalability

Wide pH Range Wide Selectivity Range High EfficiencyPeak Capacity


Thank you for your attention

QUESTIONS


CORTECS Performance, when System and Column

Dispersion are not Matched

UHPLCDispersion 30 – 12 µL

UPLCDispersion <12 µL

HPLCDispersion >30 µL

AU

0.00

0.20

0.40

0.60

0.80

1.00

Minutes

0.50 1.00 1.50 2.00

AU

0.00

0.20

0.40

0.60

0.80

1.00

Minutes

0.50 1.00 1.50 2.00

AU

0.00

0.20

0.40

0.60

0.80

1.00

Minutes

0.50 1.00 1.50 2.00 2.50

2.1 mm ID1.6 μm50mm length

AU

0.00

0.20

0.40

0.60

0.80

1.00

Minutes

0.50 1.00 1.50 2.00

AU

0.00

0.20

0.40

0.60

0.80

1.00

Minutes

0.50 1.00 1.50 2.00

AU

0.00

0.20

0.40

0.60

0.80

1.00

Minutes

0.50 1.00 1.50 2.00 2.50

AU

0.00

0.20

0.40

0.60

0.80

1.00

Minutes

0.50 1.00 1.50 2.00

AU

0.00

0.20

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0.60

0.80

1.00

Minutes

0.50 1.00 1.50 2.00

AU

0.00

0.20

0.40

0.60

0.80

1.00

Minutes

0.50 1.00 1.50 2.00 2.50




CORTECS Performance, when System and Column

Dispersion are not Matched

UHPLC UPLCHPLC

AU

0.00

0.20

0.40

0.60

0.80

1.00

Minutes

0.50 1.00 1.50 2.00

AU

0.00

0.20

0.40

0.60

0.80

1.00

Minutes

0.50 1.00 1.50 2.00

AU

0.00

0.20

0.40

0.60

0.80

1.00

Minutes

0.50 1.00 1.50 2.00 2.50


AU

0.00

0.20

0.40

0.60

0.80

1.00

Minutes

0.50 1.00 1.50 2.00

AU

0.00

0.20

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1.00

Minutes

0.50 1.00 1.50 2.00

AU

0.00

0.20

0.40

0.60

0.80

1.00

Minutes

0.50 1.00 1.50 2.00 2.50

AU

0.00

0.20

0.40

0.60

0.80

1.00

Minutes

0.50 1.00 1.50 2.00

AU

0.00

0.20

0.40

0.60

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1.00

Minutes

0.50 1.00 1.50 2.00

AU

0.00

0.20

0.40

0.60

0.80

1.00

Minutes

0.50 1.00 1.50 2.00 2.50



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