Chromatography Theory and Applications - Instituto de Qu...

Chromatography Theory and

Applications

Chromatography Theory

u Overviewu Separation ModesF Anion Exchange ChromatographyF Cation Exchange ChromatographyF Ion Exclusion ChromatographyF Ligand Exchange ChromatographyF Reversed Phase Chromatography

u Chemical Suppressionu Detection Modes

Overview

Separator Column Suppressor

InjectorDetector

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

Separation Modes


InjectorDetector


Substrates: Polymers Used

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

CH2 CH CH2 CH CH2 CH

CH CH2 CH CH2 CH2 CHCH2 CH2 CH2

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-NR3+ OH- + A- Resin-NR3

+ A- + OH-Keq

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

Con

duct

ivit

y

0 2 4 6 8 10

Minutes

1

2

3

45

6

7

Oxyhalides by Anion Exchange

1

23

4

5

67

89

10

5 1 5 20 251 00 30

Time

Con

duct

ivit

y

COMPONENT

1. Fluoride 2. Chlorite 3. Bromate4. Chloride5. Nitrite6. Chlorate 7. Nitrate 8. Bromide9. Phosphate

10. Sulfate

Cation Exchange

u Transgenomic Bead Characteristics


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-SO3- H+ + A+ Resin-SO3

- A+ + H+ Keq


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

hisarg

lys

tyr

leumet

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 3515

Minutes

1

23 4 5

6

7

Ion Exclusion Chromatography

u Bead Characteristics

FCross-linked PS/DVB core

FMacroporous

FHighly Sulfonated

u Common Eluants

FStrong Acids (HCL, Sulfuric)

FOrganic Acids (OSA, Perchloric, Perfluorobutyric Acid)

Mechanism for Ion Exclusion

RCOOH (retained) RCOO- (excluded)pKa


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

12

35

4

0 5 10 15 20 25 30 35

Minutes

6

7

8

910

Krebs Cycle Acids

Reversed Phase (Ion Pairing) Chromatography

u Transgenomic Bead CharacteristicsFCross-linked PS/DVB coreFMacroporous or NonporousFMay be alkylated (-CO, -C18)

u Common Mobile PhasesFAcetonitrile/WaterFMethanol/Water

uCommon Ion Pairing ReagentsF TEAAF TMAOHF TBAOH

Mechanism for Ion Pairing

TBA+A- (retained) TBA+ + A- (moves)


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

Minutes0 2 4 6 8 10 12 14

1

2

3

4

5

20-mer on an OLIGOSep Column

20mer

19mer

18mer

Time0 5 10 15 20

UV

Res

pons

e

-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 Macroporous

F Highly Sulfonated

F Metal Loaded (Pb, Ca, etc.)

u Common Eluants

F Water

F Sodium Hydroxide

Mechanism for Ligand Exchange

Resin-SO3- M + + H2O + A-

Keq


F Alcohols

F Carbohydrates

FWeak Acids

Resin-SO3- M + + A- + H2O

Ligand Exchange Application

0 4 8 12 16

suc

gal sormanfruraf

sta glu

min

Monosaccharides on a CARBOSep CHO-620

Chemical Suppression


InjectorDetector

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-

H2O

H2O

H2OH2O Detector Cell

Chemical Suppression Reactions

u Anion ICH+

Na+Na+ + OH- H2O

H+

Na+Na+ + HCO3

- H2O + CO2H2CO3

H+

Na+Na+ + A- H+ + A-

Chemical Suppression Reactions

u Cation ICOH-

MSA-H+ + MSA- H2O

OH-

Cl-A+ + Cl- A+ + OH-

LC Detection Modes


InjectorDetector


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

TransgenomicIon 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 Eluantu Runs in both suppressed and nonsuppressed

formatsu High Capacity for dirty samplesu Direct replacement for the Dionex AS11u Works with hydroxide, PHBA or Carbonate

eluants

ConditionsEluant: 1.8mM Carb/1.7mM BicarbFlow 1.0mL/minDetect: 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

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 eluantsu 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

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***

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 HCO3/3.5mM CO3)

Compatible with both hydroxide and carbonate-

based eluants

COMPONENTS:

1. FLUORIDE2. ACETATE3. CHLORIDE4. NITRITE5. BROMIDE6. NITRATE7. PHOSPHATE8. 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 ContaminationF 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 Cetac IC Products



u Conclusion

Transgenomic Chromatography Products

for Life Science Applications

Amino AcidColumns

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


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

CarbohydrateAnalysisColumns

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


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 BiocontaminationF 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 87NF Use 0.1M CaOH for CHO-820 and COREGEL 87CF 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 Analysisl 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


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 PhaseColumns forIon 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

5µm PS/DVB

C18 Alkylated

5µm 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 Cetac IC Product Launch



u Conclusion

CompetitivePositioning and

Analysis IC Products

Goals for TransgenomicIC 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 inthe 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 PellicularAnion 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

NR3+

NR3+

NR3+

NR3+

NR3+

NR3+NR3

+

NR3+

NR3+

NR3+

NR3+

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 12.05.0

10.0

15.0

20.0

25.0

30.0

35.0

40.0

45.0

Res

pons

e (m

V)

ICSep AN1SC Column

4

32

5

6

7

Fluoride Retained

Nitrate Tails

PellicularAnion 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

µS7 Anions on a Dionex AS4A

1

2

3

45

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 #

Eff

icie

ncy

AN1-SCAS6

Reason

u Latex microspheres ionically attached to sulfonated core

u Metals bind Ion Exchange Sites Irreversibly

PS/DVB NR3+

NR3+

NR3+

NR3+

NR3+

NR3+NR3

+

NR3+

NR3+

NR3+

NR3+

SO3-

SO3-

SO3-

SO3-

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

µS

1

2

3 4 5

6

7

7

8

5 6

1

2

4

Time0.0 5.0 10.0 15.0 20.0

Res

pons

e

60.0

80.0

100.0

120.0

140.0

160.0

180.0

200.03 1. Fluoride

2. Acetate3. Chloride4. Nitrite5. Bromide6. Nitrate7. Phosphate8. Sulfate

1. Fluoride2. Chloride3. Nitrite4. Bromide5. Nitrate6. Phosphate7. Sulfate

Comparison Cetac AN1 & the Dionex AS12

1

Time0.0 4.0 8.0 12.0

5.0

10.0

15.0

20.0

25.0

30.0

35.0

40.0

45.0

Res

pons

e (m

V)

4

32

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

µS

1

2

34 5

6

7



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

µS

1

2

34

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

Con

duct

ivity



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

µS

1

2

34

5

6

7

Minutes

Con

duct

ivity

0 2 4 6 8 10

1

2

3

4 5

6

7



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

µS

1

23

45

67

8

9

10

1

23

4

5

6

7

89

10

5 15 20 25100 30

Time

Con

duct

ivity

1. Fluoride2. Chlorite3. Bromate4. Chloride5. Nitrite

6. Chlorate7. Nitrate8. Bromide9. Phosphate10. Sulfate

1. Fluoride2. Chlorite3. Bromate4. Chloride5. Nitrite

6. Phosphate7. Bromide8. Chlorate9. Nitrate10. Sulfate

CompetitivePositioning 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

Chromatography Theory and Applications - Instituto de Qu...

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