Chromatography Theory

and

Applications

Chromatography Theory

u

Overview

u

Separation Modes

F

Anion Exchange Chromatography

F

Cation Exchange Chromatography

F

Ion Exclusion Chromatography

F

Ligand Exchange Chromatography

F

Reversed Phase Chromatography

u

Chemical Suppression

u

Detection Modes

Overview

Separator Column

Suppressor

Injector

Detector

Chromatography (LC) is the separation of
species by various separation modes
followed by suppressed conductivity, UV
or some other type of detection

Separation Modes

Separator Column

Suppressor

Injector

Detector

Chromatography (LC) is the separation of
species by various separation modes
followed by suppressed conductivity, UV
or some other type of detection

Substrates: Polymers Used

Most common Polymer used is a co-polymer of
polystyrene and divinylbenzene (PS/DVB)

CH

2

CH

CH

2

CH

CH

2

CH

CH

CH

2

CH

CH

2

CH

2

CH

CH

2

CH

2

CH

2

Features of PS/DVB

u

Rugged Cross-linked polymer

F pH range 0-14

F solvent compatible with many solvents

F Temperature stable

F Able to control shrink/swell with X-linking

u

Control particle size and porosity

u

Well defined polymerization and functionalization

chemistries

Separation Modes

u

Ion Exchange

F Anion Exchange

F Cation Exchange

u

Ion Exclusion

u

Ligand Exchange

u

Reversed Phase (Ion Pairing)

Anion Exchange

u

Transgenomic Bead Characteristics

F Cross-linked PS/DVB core
F Coating
F Quaternary Ammonium Exchange

Groups

u

Common Buffers

F Sodium Hydroxide
F Sodium Carbonate/Bicarbonate
F Sodium Chloride

Mechanism for Anion Exchange

Resin-NR

3

+

OH

-

+ A

-

Resin-NR

3

+

A

-

+ OH

-

K

eq

u

Common Applications

F Inorganic Anions
F Organic Acids
F Oxyhalides
F Amino Acids
F Proteins/Peptides
F DNA/Oligos
F Transition Metals

Anion Exchange IC Applications

1. Fluoride

2. Chloride

3. Nitrite

4. Bromide

5. Nitrate

6. Phosphate

7. Sulfate

Conductivity

0

2

4

6

8

10

Minutes

1

2

3

4

5

6

7

Oxyhalides by Anion Exchange

1

2

3

4

5

6

7

8

9

10

5

1 5

20

25

1 0

0

30

Time

Conductivit

y

COMPONENT

1. Fluoride
2. Chlorite
3. Bromate
4. Chloride
5. Nitrite
6. Chlorate
7. Nitrate
8. Bromide
9. Phosphate

10. Sulfate

Cation Exchange

u

Transgenomic Bead Characteristics

F Cross-linked PS/DVB core
F Sulfonic, Phosphonic, Carboxylic

Acid Functional Groups

u

Common Eluants

F Organic Acids (Citrate, Oxalate)
F Methane Sulfonic Acid
F Buffers
F NaCl

Mechanism for Cation Exchange

Resin-SO

3

-

H

+

+ A

+

Resin-SO

3

-

A

+

+ H

+

K

eq

u

Common Applications

F Grp I & II Cations

F Ammonium Species

F Transition Metals

F Amino Acids

F Proteins/Peptides

Cation Exchange Applications

asp

thr ser

glu

gly

ala

phe

his

arg

lys

tyr

leu

met

val

iso-leu

0

10

20

30

40

50

60

70

80

Hydrolysate Amino Acid Analysis using a Beckman

Sodium AAA Column

Cation IC Applications

1. Copper

2. Nickel

3. Zinc

4. Iron

5. Manganese

6. Cadmium

7. Lead

0

5

10

20

25

30

35

15

Minutes

1

2

3

4

5

6

7

Ion Exclusion Chromatography

u

Bead Characteristics

F Cross-linked PS/DVB core
F Macroporous
F Highly Sulfonated

u

Common Eluants

F Strong Acids (HCL, Sulfuric)
F Organic Acids (OSA, Perchloric,

Perfluorobutyric Acid)

Mechanism for Ion Exclusion

RCOOH (retained)

RCOO

-

(excluded)

pKa

u

Common Applications

F Organic Acids

F Borate/Carbonate

F Alcohols

Ion Exclusion Applications

1. Citric

2. Tartaric

3. Glucose

4. Malic

5. Fructose

6. Lactic

7. Glycerol

8. Acetic

9. Methanol

10. Ethanol

1

2

3

5

4

0 5

10

15

20

25

30

35

Minutes

6

7

8

9

10

Krebs Cycle Acids

Reversed Phase (Ion Pairing)
Chromatography

u

Transgenomic Bead Characteristics

F Cross-linked PS/DVB core
F Macroporous or Nonporous
F May be alkylated (-CO, -C18)

u

Common Mobile Phases

F Acetonitrile/Water
F Methanol/Water

u

Common Ion Pairing Reagents

F TEAA
F TMAOH
F TBAOH

Mechanism for Ion Pairing

TBA

+

A

-

(retained)

TBA

+

+ A

-

(moves)

u

Common Applications

F Surfactants

F DNA

F ethylamines

F Aliphatic Acids

F Transition Metals

Ion Pairing Reversed Phase

Reversed Phase Applications

1. Physostigmine

2. Acetophenone

3. Ethyl Phenone

4. Butyl Phenone

5. Valerphenone

Minutes

0

2

4

6

8

10

12

14

1

2

3

4

5

20-mer on an OLIGOSep Column

20mer

19mer

18mer

Time

0

5

10

15

20

UV Response

-5.0

0.0

5.0

10.0

15.0

Separation of 20-mer from n-1, n-2 failures

Ligand Exchange Chromatography

u

Bead Characteristics

F Cross-linked PS/DVB core
F Macroporous
F Highly Sulfonated
F Metal Loaded (Pb, Ca, etc.)

u

Common Eluants

F Water
F Sodium Hydroxide

Mechanism for Ligand Exchange

Resin-SO

3

-

M

+ +

H

2

O + A

-

K

eq

u

Common Applications

F Alcohols

F Carbohydrates

F Weak Acids

Resin-SO

3

-

M

+ +

A

-

+ H

2

O

Ligand Exchange Application

0

4

8

12

16

suc

gal

sor

man

fru

raf

sta

glu

min

Monosaccharides on a CARBOSep CHO-620

Chemical Suppression

Separator Column

Suppressor

Injector

Detector

Used Only In IC: Ion Chromatography
(IC) is the separation of ionic species by
ion exchange chromatography coupled
with suppressed conductivity detection

Chemical Suppression

u

Purpose

Reduce background conductivity from eluant and

increase signal from analytes to make high

sensitivity ion analysis possible

u

Types of Suppressors Available

F Dionex: self regenerating suppressor,

membrane suppressors, fiber suppressors,

packed bed suppressors

F Altech: packed bed suppressor

How Chemical Suppression Works

Waste

H

+

H

+

H

+

H

+

H

+

H

+

H

+

Eluant

Na

+

Na

+

Na

+

Na

+

Na

+

Na

+

Regenerant

Regenerant

Waste

Na

+

Na

+

OH

-

OH

-

OH

-

H

+

H

+

OH

-

H

2

O

H

2

O

H

2

O

H

2

O

Detector Cell

Chemical Suppression Reactions

u

Anion IC

H

+

Na

+

Na

+

+ OH

-

H

2

O

H

+

Na

+

Na

+

+ HCO

3

-

H

2

O + CO

2

H

2

CO

3

H

+

Na

+

Na

+

+ A

-

H

+

+ A

-

Chemical Suppression Reactions

u

Cation IC

OH

-

MSA

-

H

+

+ MSA

-

H

2

O

OH

-

Cl

-

A

+

+ Cl

-

A

+

+ OH

-

LC Detection Modes

Separator Column

Suppressor

Injector

Detector

Chromatography (LC) is the separation of
species by various separation modes
followed by suppressed conductivity, UV
or some other type of detection

LC Detection Modes

u

UV/Vis - Most common for LS

F DIrect UV (diode array)

F Indirect UV - Refractive Index

F Direct UV w/ Post Column Reaction

u

Conductivity - Most Common for IC

u

Amperometry - For Carbohydrates

u

Other (Mass Sepc, ELS, etc.)

Agenda

u

Introduction

u

Chromatography Theory

u

Cetac IC Products

u

Transgenomic Life Science Products

u

Competitive Analysis

u

Conclusion

Transgenomic

Ion Chromatography

Products

ICSep Ion Exchange Columns

u

ICSep Columns for Anion Analysis

l ICSep AN1

l ICSep AN1-SC

l ICSep AN2

u

ICSep Columns for Cation Analysis

l

ICSep CN2

l ICSep AN300
l ICSep AN300b
l ICSep ANSC

Features and Benefits

of

Cetac IC Columns

Features of the ICSep AN1 Column

u

Separates 7 Standard Ions in 15 minutes

u

Fluoride separated from water dip with

Carb/Bicarb Eluant

u

Runs in both suppressed and nonsuppressed

formats

u

High Capacity for dirty samples

u

Direct replacement for the Dionex AS11

u

Works with hydroxide, PHBA or Carbonate

eluants

Conditions
Eluant: 1.8mM Carb/1.7mM
Bicarb
Flow 1.0mL/min
Detect: Suppressed Cond

Features of the ICSep AN300 Column

u

Separates 7 Standard Ions in 8 minutes

u

Designed and Verified equivalent for EPA

method 300.0(a)

u

Fluoride completely resolved from water dip

u

Works with hydroxide or Carbonate eluants

u

Direct Replacement for AS-4A column running

anions in water

Conditions
Eluant: 1.8mM Carb/1.7mM
Bicarb
Flow 2.0mL/min
Detect: Suppressed Cond

Features of the ICSep AN300b Column

u

Designed to separate 7 Standard Ions as well as
oxyhalides and organic acids

u

Designed and Verified equivalent for EPA method
300.0(b)

u

Solvent compatible, can clean with even organic
solvents (i.e. methanol)

u

Works with hydroxide or Carbonate eluants

u

Very High Capacity - Direct Replacement for AS9-
HC column running disinfection agents in water

Features of the ICSep ANSC Column

u

Same selectivity as the Dionex AS-4ASC

u

Compatible with Organic Solvents

u

7 Standard Anions in 11 minutes

u

Compatible with both hydroxide and

carbonate-based eluants

Conditions
Eluant: 1.8mM Carb/1.7mM
Bicarb
Flow 2.0mL/min
Detect: Suppressed Cond

Features of the ICSep AN1-SC

u

Same features as the AN1

u

Direct Replacement for systems using the Dionex

AS11/AS12

u

Now Compatible with up to 100% Organic

Solvents***

Conditions
Eluant: 1.8mM Carb/1.7mM
Bicarb
Flow 1.0mL/min
Detect: Suppressed Cond

Features of the New ICSep AN2

Very high Capacity

Separates organic acids and ions in same run

Compatible with Organic Solvents

Direct replacement for systems running Dionex

AS14 columns (1mM HCO

3

/3.5mM CO

3

)

Compatible with both hydroxide and carbonate-

based eluants

COMPONENTS:

1. FLUORIDE
2. ACETATE
3. CHLORIDE
4. NITRITE
5. BROMIDE
6. NITRATE
7. PHOSPHATE
8. SULFATE

ICSep AN2

Features of the ICSep CN2 Column

u

Separates Group I and Group II Cations as

well as Transition series metals

u

Separates Ammonium Species

u

Compatible with all common Cation
exchange eluants

u

Works in both suppressed and
nonsuppressed modes

OEM Vendors for IC Columns

u

Latchatt - Special AN300 to Latchatt specs

u

Metrohm -

F Metrosupp 1 : AN300b

F Metrosupp 2 : AN1

F Metrosupp 3 : AN1-SC

Symptoms of Column Contamination

u

Inorganic Contamination - peaks tail, retention time

decreases, pressure increases

u

Metals - Phosphate disappears

u

Organic Contamination - increase pressure, nitrate

tails, etc.

u

Notes: Always recommend guard column, if

pressure high change frit.

Care of Transgenomic IC Columns

u

Long Term Storage - either water or 10mM NaOH

u

Inorganic Contamination

F 40mM EDTA @ 0.5mL/min for 1 hour (for metals)
F 0.1M NaOH @ 1.0mL/min for 3 hours

u

Organic Contamination

F Solvent Compatible Columns flush 90% ACN or MeOH

@ 1.0mL/min for 2 hours (ANSC, AN1-SC, AN300b, AN2)

F Non-SC columns flush with 10% MeOH @ 0.5mL/min

overnight (AN1, AN300)

F Follow with Inorganic Cleanup

Agenda

u

Introduction

u

Chromatography Theory

u

Cetac IC Products

u

Transgenomic Life Science Products

u

Competitive Analysis

u

Conclusion

Transgenomic

Chromatography Products

for Life Science

Applications

Amino Acid

Columns

Amino Acid Analysis Columns

u

AMINOSep Columns for Amino Acid Analysis

l AA 911

l AA 903

u

Transgenomic Columns for the 63/7300

l

Transgenomic Lithium

l Transgenomic Sodium AA column for System Gold

l AA 511

l Transgenomic Sodium

Features of Transgenomic AAA Columns

u

Both Sodium and Lithium Cation Exchange Forms

u

Separates both hydrolysates and physiological fluid

amino acids

u

Very tight particle size range for high efficiency

u

Standard for Beckman and Pharmacia AAA

Systems (we supply 95% of the worlds use for IEX

amino acid analysis columns/resin)

u

Rugged, long-lasting design

Transgenomic Lithium Column for 63/7300

Transgenomic Sodium Column for 63/7300

Sodium Column for System Gold

OEM Vendors for AAA Columns

u

Beckman - System Gold, Na+ and Li+

u

Waters - Na+ Cation Exchange

u

Merck - Na+ Cation Exchange

u

Phamacia - Resin

u

Pickering - Resin

Symptoms of Column Contamination

u

Peaks Co-Elute

u

Retention Times Increase

u

Irreproducible Ammonia Peak

u

Peaks begin to tail

u

Notes: this is a really complex analysis and

columns may need to be regenerated frequently

Care of Transgenomic AAA Columns

u

Long Term Storage - either C Buffer or 10mM NaOH

u

Column Contamination

F First flush with 20% IPA or MeOH @ 0.5mL/min for 30

minutes

F Next, If Lithium form column use 0.1M LiOH if Sodium

form column use 0.1M NaOH

F Flush @ 0.5mL/min for 1 - 3 hours
F Flush with DI Water for 15 minutes
F Reequilibrate on A Buffer

Follow with Inorganic Cleanup

F NOTE: Column regeneration buffer available

Carbohydrate

Analysis

Columns

Carbohydrate Analysis Columns

u

CARBOSep Columns for Carbohydrate Analysis

l CHO-411

l CHO-611 (Na)

l CHO-620 (Ca)

l CHO-682 (Pb)

l CHO-820

l CHO-882

l COREGEL 87C
l COREGEL 87H

l COREGEL 87K
l COREGEL 87N
l COREGEL 87P

Features of Transgenomic Carbohydrate Columns

u

Separates sugars and alcohols

u

Compatible with water and hydroxide mobile phases

u

Longest history with ligand-exchange gels

u

Separates both mono and oligosaccharides up to DP7

u

Compatible with USP methods

u

Very rugged, long lasting design

u

Highest efficiency ligand-exchange columns available

Component

CHO-620

CHO-682

CHO-820

COREGEL

87C

COREGEL

87P

COREGEL

87K

ICSep

COREGEL

87H

Stachyose

5.94

11.84

8.46

7.85

11.35

6.32

6.94

Raffinose

6.56

12.01

9.01

8.31

14.41

6.96

7.65

Maltotriose

6.68

12.63

9.16

8.35

15.24

7.36

7.18

Sucrose

7.48

13.51

9.94

9.18

15.77

8.08

ND

Cellobiose

7.36

13.53

9.79

9.01

15.65

NA

7.76

Trehalose

7.32

NA

NA

9.14

16.05

8.22

8.00

Maltose

7.59

14.43

10.06

9.24

16.68

8.56

7.78

Melibiose

7.67

15.25

NA

9.43

17.70

NA

7.88

Lactose

7.84

15.09

10.49

9.51

17.44

8.72

8.13

Lactulose

8.53

18.93

11.49

10.24

20.77

NA

NA

Maltitol

9.15

NA

NA

12.29

30.45

8.16

NA

Glucose

9.36

16.09

12.09

11.22

19.21

11.20

10.11

Sorbose

10.22

19.45

NA

12.90

22.45

13.16

9.90

Xylose

10.31

17.96

NA

12.37

20.71

12.24

10.33

Rhamnose

10.41

19.53

NA

12.93

22.63

13.37

11.20

Mannose

10.51

20.39

14.06

12.83

25.57

12.48

9.98

Fructose

11.14

22.59

15.61

13.68

25.90

12.16

10.39

Fucose

11.33

27.93

15.41

13.89

24.23

11.98

12.05

Arabinose

11.63

21.73

15.76

14.00

24.02

13.44

11.23

Mannitol

12.76

34.51

20.54

17.89

40.07

10.08

NA

Arabitol

13.23

33.98

20.94

18.43

39.80

10.80

NA

Sorbitol

14.91

50.76

25.61

21.41

55.56

10.64

8.32

Xylitol

15.06

44.76

25.03

22.03

51.14

11.36

8.35

Sorbose

16.46

45.59

26.38

21.89

54.33

14.16

9.21

CARBOSep CHO-620 (Calcium Form)

CARBOSep CHO-682 (Lead Form)

CARBOSep CHO-820 (Calcium Form)

CARBOSep CHO-611 (Sodium Form)

Features of COREGEL Columns

u

9 micron particle size

u

8% crosslinked

u

Very tight particle size range for highest efficiency

u

Available in Calcium, Sodium, Lead, Hydrogen,

Silver and Potassium

u

Direct substitution for BioRad AMINEX columns

and any other AMINEX equivalent

CARBOSep COREGEL 87C (Ca Form)

CARBOSep COREGEL 87K (K Form)

CARBOSep COREGEL 87N (Na Form)

CARBOSep COREGEL 87P (Pb Form)

OEM Vendors for Carbohydrate Columns

u

Waters - SugarPak

u

Merck - Polysphere

u

MetaChem - Metacarb

u

Machery Nagel

u

Varian Chrompack

Symptoms of Column Contamination

u

Peaks Broaden

u

Retention Time Shifts

u

Pressure Increases

u

Irrepudicible Retention Times

u

Note: Always recommend a guard column with

carbohydrate columns

Care of Transgenomic Carbo Columns

u

Long Term Storage - DI Water

u

Column Contamination

F These columns are very hard to clean and usually

require replacement.

F For Organic Contamination: NEVER EXCEED 10%

ACN or MeOH because these resins swell very easily,
flush at 0.3mL/m for several hours, then flush with DI
Water. Watch column pressure.

Care of Transgenomic Carbo Columns

u

Column Contamination - Inorganics and
Biocontamination

F Select cleaning solvent very carefully - it should match

the ionic form of the column.

F Use 0.1M NaOH for CHO-611/OH and COREGEL 87N
F Use 0.1M CaOH for CHO-820 and COREGEL 87C
F Use 0.1M KOH for COREGEL 87K

F Flush column at 0.3mL/min overnight then flush with

water for several hours

F For CHO-682 or COREGEL 87P use 0.1M NaOH

overnight followed by 0.1M PbCl for 3 hours @
0.3mL/min

Ion Exclusion

Columns

Ion Exclusion Columns

u

ICSep Columns for Organic Acid, Carbonate

and Alcohol Analysis

l ICSep ICE-ION-300
l ICSep ICE-ION-310
l ICSep ICE-ARH-601
l ICSep ICE-ORH-801
l ICSep ICE-COREGEL 64H
l ICSep ICE-COREGEL 87H

Features of the ICSep ICE-ION 300

u

Separates Organic Acids, Sugars and

Alcohols with VERY high resolution

u

Small Particle Size provide high efficiency

u

Designed for use with normal matrices

u

Recommend for Wine Analysis

u

Designed for High Resolution Applications

Features of the ICSep ICE-ORH 801

u

Separates Organic Acids, Sugars and

Alcohols with good resolution

u

Intermediate resolution between ION-300

and Coregel 87H

u

Blended to provide optimum resolution with

good ruggedness

u

Can also run inorganic anions such as

sulfite, azide, fluoride and arsenic species

u

Designed as general purpose column

Features of the ICSep ICE-COREGEL 87H

u

The most rugged column (8% cross linked)

u

Larger bed volume, bead size, lower pressure

allows the column to be run at fast flow rates

u

Separates Sugars, Alcohols and Organic Acids

all on one column in same run

u

Designed for use with tough matrices

u

Direct replacement for BioRad Aminex Columns

Features of the ICSep ICE-ION 310

u

Designed for fast run organic acid analysis

u

High sensitivity, rapid analysis of borate and

bicarbonate

u

Rugged column

u

Designed for use with tough matrices

u

Choose this column when speed is the

primary concern

Features of the ICSep ICE-ARH 601

u

Designed for analysis of Aromatic Organic

Acids

u

Eluants employ only Sulfuric Acid and Water,

no organic modifiers are required

u

Only ICE column available designed

especially for Aromatic Organic Acids

Features of the ICSep ICE-COREGEL 64H

u

Larger bed volume, bead size, lower pressure

allows the column to be run at fast flow rates

u

Separates Sugars, Alcohols and Organic Acids

all on one column in same run

u

Direct replacement for BioRad Aminex Column

OEM Vendors for Ion Exclusion Columns

u

Metachem - MetaCarb

u

Alltech

u

Merck

u

Machery Nagel

Symptoms of Column Contamination

u

Inorganic Contamination - peaks tail, retention time

decreases, pressure increases

u

Metals - Irreproducible results

u

Organic Contamination - increased pressure

u

Notes: Always recommend guard column this will

help protect the separator.

Care of Transgenomic Ion Exclusion
Columns

u

Long Term Storage - 10mM Sulfuric Acid

u

Inorganic Contamination

F 40mM EDTA @ 0.4mL/min for 1 hour (for metals)

F 0.1M Sulfuric Acid @ 0.4mL/min for 6 hours

F flush with DI-water then reequilibrate

u

Organic Contamination

F NEVER EXCEED 10% ACN or MeOH and watch

pressure carefully, flush at 0.4mL/min for 3 hours then
flush thoroughly with water.

Reversed Phase

Columns for

Ion Pairing

Chromatography

RPSep Reversed Phase/Ion Pairing Columns

u

RPSep Columns for Reversed Phase and

Ion Pairing Chromatography

l RPSep PRX-1

l RPSep ACT-1 C18

l RPSep PolyRP CO

l OLIGOSep - 1

Column Options

Two Options: non-functionalized or
alkylated PS/DVB

Non-Functionalized

5

µ

m PS/DVB

C18 Alkylated

5

µ

m PS/DVB

OLIGOSep



, PRX,

PolyRP C0

ACT-1

Features of the RPSep PolyRP-CO Column

u

CO functionalization provides hydrophillic

surface for separation of less polar compounds

u

Proprietary, patented, alkylation technology

u

Small particle size with tight particle size range

for high efficiency

Features of the RPSep PRX-1 Column

u

Nonfunctionalized PS/DVB ideal for separation

of both polar and nonpolar species

u

Fine particle size range for high efficiency

u

Exceptionally rugged design

u

Direct replacement for Dionex NS1 or Hamilton

PRP-1 columns

Features of the RPSep ACT-1 C18 Column

u

Alkylated with C18 groups, provides hydrophobic

surface ideal for the separation of polar species

u

pH Stable over range of 0-14

u

Proprietary, patented, technology

u

Very high efficiency

Care of Transgenomic Reversed Phase
Columns

u

Long Term Storage - 40% ACN and Water

u

Inorganic Contamination

F 40mM EDTA @ 1.0mL/min for 1/2 hour (for metals)

F 0.1M Sodium Hydroxide @ 1.0mL/min for 1 hours

F flush with DI-water then reequilibrate

u

Organic Contamination

F Flush with 95% ACN or MeOH @ 1.0mL/minute for 3

hours

Agenda

u

Introduction

u

Chromatography Theory

u

Cetac IC Product Launch

u

Transgenomic Life Science Products

u

Competitive Analysis

u

Conclusion

Competitive

Positioning and

Analysis

IC Products

Goals for Transgenomic

IC Products

Product Positioning

Cetac Technologies Offers a

Complete Line of Chromatography

Columns that Provide Superior

Solutions for the Analysis of Ionic

Species at a lower cost than

Competitive Alternatives

Summary:

Make Better Columns at a

Lower Cost

Key Competitor in

the IC Market--

Dionex Corporation

Dionex - Background

u

Founded 1975

u

Recognized leader in Ion Chromatography

u

> 20,000 IC Systems Installed

u

~$165M in Annual Sales

u

~$37M Annual Sales in IC Columns

Dionex - Advantages

u

Corporate IC Focus

u

Large Installed Base

u

Many Applications Developed

u

Approved Methods (E.P.A.)

u

Complete IC Product Offering

u

Proprietary Suppressor Technology

u

Protected Market

Dionex - Disadvantages

u

Gouge Customers

u

Single Expertise - Not good at other fields

u

Not very flexible

u

Poor Service

Transgenomic Strategy

Offer the IC customer a compatible
replacement for his Dionex column at a
lower cost

Cetac Column

ICSep AN2

ICSep AN1

ICSep AN1-SC

ICSep AN300

ICSep AN300B

ICSep ANSC

ICSep CN2

Dionex Compatible

IonPac AS14

IonPac AS11/AS12

IonPac AS11/AS12

IonPac AS4A

IonPac AS9-HC

IonPac AS4A-SC

IonPac CS3

Transgenomic - Advantages

u

Compatible Selectivities

u

Superior Fluoride Retention

u

Covalently Bound Surface Chemistries - Long Life

u

Lower Cost

u

Employ Guard Disc to provide added protection

Covalent and Pellicular

Anion Exchangers

Covalent Anion Exchangers

♦

May Have Pores (“holes”) or be NonPorous

♦

All ion exchange groups attached covalently
to the surface of the bead

Porous

NonPorous

Characteristics of Covalent IC Resins

u

Larger surface area for porous

u

Higher Capacity

u

Covalently bound (grafted) functional groups

u

Can control “exclusion”of molecules by pore size

u

Lower column pressures

u

Can be used for both anion and cation columns

Pellicular Anion Exchangers

u

Latex microspheres ionically attached to
sulfonated core

u

Ion exchange groups on the surface of the
latex microbead

PS/DVB

Latex Microsphere

NR

3

+

NR

3

+

NR

3

+

NR

3

+

NR

3

+

NR

3

+

NR

3

+

NR

3

+

NR

3

+

NR

3

+

NR

3

+

Characteristics of Pellicular IC Resins

u

All interactions are on the surface of the

microbead

u

High efficiency from small microspheres

u

Use larger beads for less backpressure

u

Less secondary interactions

u

Core bead sulfonated, latex ionically attached

Benefits of Covalent and

Pellicular Anion

Exchangers

Covalent Anion Exchangers

Advantages of Covalent Ion Exchangers

u

Higher capacity - can handle very dirty samples

u

More rugged and will last longer - covalently

bound chemistries won’

t come off

u

Easily cleaned

u

Fluoride can be retained

u

Can take advantage of surface interactions

u

Less hardware requirements - lower running

pressures

Disadvantages of Covalent Ion Exchanger

u

Materials can “hide”in pores

u

More surface interactions - nitrate/iodide tails

u

Must use smaller particles for high efficiency

Anions on a Cetac AN1-SC

1

Time

0.0

4.0

8.0

12.0

5.0

10.0

15.0

20.0

25.0

30.0

35.0

40.0

45.0

Response (mV)

ICSep AN1SC Column

4

3

2

5

6

7

Fluoride
Retained

Nitrate Tails

Pellicular

Anion Exchangers

Advantages of Pellicular Ion Exchangers

u

High Efficiency - Sharper Peaks

u

Less surface interaction - Nitrate tailing minimal

u

Use larger core bead for less backpressure

Disadvantages of Pellicular Ion Exchangers

u

Fluoride not easily retained

u

Susceptible to metals contamination -

phosphate results inconsistent

u

Lower capacities - trouble with dirty samples

u

Ionically bound chemistries - shorter column

lifetime

u

Can contaminate irreversibly

Anions on a Dionex AS4A-SC

0

2.00

4.00

6.00

8.00

10.00

Minutes

0

2.0

4.0

6.0

8.0

10.0

12.0

14.0

µ

S

7 Anions on a Dionex AS4A

1

2

3

4

5

6

7

Less Nitrate Tailing

Fluoride not retained

Comparison of Efficiency Loss from Metal Contamination

1000

1500

2000

2500

3000

3500

4000

4500

5000

5500

6000

0

20

40

60

80

100

Run #

Efficiency

AN1-SC

AS6

Reason

u

Latex microspheres ionically attached to
sulfonated core

u

Metals bind Ion Exchange Sites Irreversibly

PS/DVB

NR

3

+

NR

3

+

NR

3

+

NR

3

+

NR

3

+

NR

3

+

NR

3

+

NR

3

+

NR

3

+

NR

3

+

NR

3

+

SO

3

-

SO

3

-

SO

3

-

SO

3

-

Cu

+2

Cu

+2

Cu

+2

Selling Hints

u

Target Dionex customers with “Clone”approach

u

ALWAYS recommend a guard cartridge

u

Cetac Columns designed to Last Longer

u

Promote compatible separation at lower cost

u

Focus on Cetac specialty - Inorganic Analysis

u

Highlight New Chemistries provide better separation

Comparison Cetac AN2 & the Dionex AS14

0

2.00

4.00

6.00

8.00

10.00

Minutes

0

2.00

4.00

6.00

8.00

µ

S

1

2

3

4

5

6

7

7

8

5

6

1

2

4

Time

0.0

5.0

10.0

15.0

20.0

Response

60.0

80.0

100.0

120.0

140.0

160.0

180.0

200.0

3

1. Fluoride
2. Acetate
3. Chloride
4. Nitrite
5. Bromide
6. Nitrate
7. Phosphate
8. Sulfate

1. Fluoride
2. Chloride
3. Nitrite
4. Bromide
5. Nitrate
6. Phosphate
7. Sulfate

Comparison Cetac AN1 & the Dionex AS12

1

Time

0.0

4.0

8.0

12.0

5.0

10.0

15.0

20.0

25.0

30.0

35.0

40.0

45.0

Response (mV)

4

3

2

5

6

7

2.00

4.00

6.00

8.00

10.00

12.00

14.00

Minutes

-0.50

0

0.50

1.00

1.50

2.00

2.50

µ

S

1

2

3

4

5

6

7

1. Fluoride
2. Chloride
3. Nitrite
4. Bromide
5. Nitrate
6. Phosphate
7. Sulfate

1. Fluoride
2. Chloride
3. Nitrite
4. Bromide
5. Nitrate
6. Phosphate
7. Sulfate

Comparison Cetac ANSC & Dionex AS4A-SC

0

2.00

4.00

6.00

8.00

10.00

Minutes

0

2.0

4.0

6.0

8.0

10.0

12.0

14.0

µ

S

1

2

3

4

5

6

7

Time

2

3

4

5

6

7

1

0.0

2.0

4.0

6.0

8.0

10.0

12.0

14.0

16.0

Conductivity

1. Fluoride
2. Chloride
3. Nitrite
4. Bromide
5. Nitrate
6. Phosphate
7. Sulfate

1. Fluoride
2. Chloride
3. Nitrite
4. Bromide
5. Nitrate
6. Phosphate
7. Sulfate

Comparison Cetac AN300 & Dionex AS4A

0

2.00

4.00

6.00

8.00

10.00

Minutes

0

2.0

4.0

6.0

8.0

10.0

12.0

14.0

µ

S

1

2

3

4

5

6

7

Minutes

Conductivity

0

2

4

6

8

10

1

2

3

4

5

6

7

1. Fluoride
2. Chloride
3. Nitrite
4. Bromide
5. Nitrate
6. Phosphate
7. Sulfate

1. Fluoride
2. Chloride
3. Nitrite
4. Bromide
5. Nitrate
6. Phosphate
7. Sulfate

Comparison Cetac AN300B & Dionex A9HC

0

5.00

10.00

15.00

20.00

25.00

Minutes

0

0.50

1.00

1.50

2.00

2.50

3.00

3.50

4.00

µ

S

1

2

3

4

5

6

7

8

9

10

1

2

3

4

5

6

7

8

9

10

5

15

20

25

10

0

30

Time

Conductivity

1. Fluoride
2. Chlorite
3. Bromate
4. Chloride
5. Nitrite

6. Chlorate
7. Nitrate
8. Bromide
9. Phosphate
10. Sulfate

1. Fluoride
2. Chlorite
3. Bromate
4. Chloride
5. Nitrite

6. Phosphate
7. Bromide
8. Chlorate
9. Nitrate
10. Sulfate

Competitive

Positioning and

Analysis

Life Science Products

Life Science Position

Transgenomic supplies polymeric

chromatography products which

provide superior solutions for target

biological applications

Summary:

Focus on the development of

superior columns for niche

applications for the life science

market place

Conclusion

IC Product Positioning

Cetac Technologies Offers a

Complete Line of Chromatography

Columns that Provide Superior

Solutions for the Analysis of Ionic

Species at a lower cost than

Competitive Alternatives

Life Science Product

Position

Transgenomic supplies polymeric

chromatography products which

provide superior solutions for target

biological applications