1

The world leader in serving science

Dr. Detlef Jensen

Thermo Fisher Scientific

IC Sales Support Expert

November 12, 2014

Analysis of Disinfection Byproducts by Ion Chromatography

2

Disinfection Byproducts

• Chlorination Processes

• Generate chlorite, chlorate, halomethanes and haloacetic acids

• Ozonation Processes

• Generates bromate from bromide, and halomethanes and haloacetic

acids

• Highly regulated due to associated health issues

• Chlorite: nervous system, affect fetal development, anemia

• Haloacetic acids/bromate: carcinogenic

• Chlorate: produce gastritis, a late toxic nephritis, hemolysis,

methemoglobinemia, hemoglobinuria, and acute renal failure.

3

Analysis Using Ion Chromatography

Ion Chromatography is an analytical technique that

utilizes ion-exchange mechanisms to separate ionic

substances followed by detection utilizing conductivity,

amperometry, UV/Vis, fluorescence, mass

spectrometry or the new charge detectors.

Analyte classes include:

Anions

Cations

Organic Acids

Amines

Transition Metals

Carbohydrates & Amino Acids

4

Ion Chromatography: Anion-Exchange Mechanism

P2O33-

CO22-

+

Positively Charged

Polymeric Particle

Stationary

Phase

+

+

+

+

+

+

+

Mobile Phase

eluent, e.g., OH–

+

Cl–Analytes: anions

5

Typical Ion Chromatographic System - Anion Analysis

Eluent Bottle(CO3/HCO3

or NaOH)

Pump

Guard Column

Analytical Column

Suppressor

Conductivity Cell

Chromatography SoftwareIon

Exchange Separation

Post-Suppression Conductivity

Sample Injection

Regen Out

Data Handling and Instrument Control

Regen In

6

Advantages of Suppressed Conductivity

Time

F-

Cl - SO42-

F - Cl -SO4

2-

Time

-µS

µS

Without Suppression

With Suppression

Eluent (KOH)

Sample F-, Cl-, SO42-

Ion-Exchange

Separation Column

Anion Electrolytically

Regenerating

Suppressor

in H2O

KF, KCI, K2SO4

in KOH

Injection Valve

Counter ions

HF, HCI, H2SO4

7

Data Management

ConductivityDetector

High-Pressure Non-Metallic Pump

Eluent Generator

(OH– or H+)

Waste

Sample Inject(Autosampler)

Recycle Mode

Detection

Water/Eluent

CR-TC

CellEffluent

Electrolytic Eluent

Suppressor

Separation Column

Reagent-Free Ion Chromatography System (RFIC)

RFIC™, Innovation

and Ease-of-Use

Behind the Curtain

8

ISO & U.S. EPA Regulations

9

Determination of dissolved bromate – method by liquid chromatography of ions

ISO 15061 – Water Quality

10

Working Range for the Analytical Method - ISO 15061

• Concentrations between 0,5 µg/L and 1 000 µg/L

• Direct Injection or Preconcentration

• Chromatographic resolution: R ≥ 1.3

• Direct injection: LOD of 2.5 µg/L

• Matching reporting limit of 10 µg/L of EU-regulations

11

EPA Method 300.1 (B)

• Revision 2.1 Parts A and B published in 1993

• Outlines the method for determination of inorganic anions by ion

chromatography

• Specifies use of suppressed conductivity for determination of:

• Bromide (Part A) • Ortho-Phosphate-P (Part A)

• Chloride (Part A) • Sulfate (Part A)

• Fluoride (Part A) • Bromate (Part B)

• Nitrate (Part A) • Chlorate (Part B)

• Nitrite (Part A) • Chlorite (Part B)

• Applies to:

• Drinking water

• Ground and surface water

• Wastewater (domestic and industrial)

• Raw water (unfinished drinking water

12

U.S. EPA Regulations - Oxyhalides

• 1998 – U.S. EPA, Stage I disinfectants/disinfection

byproducts (D/DBP) rule • MCL 10 µg/L bromate

• MCL Goal (MCLG) of zero

• 2006 – U.S. EPA, Stage II D/DBP rule • Leaves MCL at 10 µg/L bromate

• Maximum Contamination Level (MCL)

• Chlorite: 1 mg/L (ppm)

• Chlorate: 500 µg/L (ppb)

• Bromate: Most toxic, lowest allowed level, 5 to 10 µg/L (ppb)

13

0 5 10 15 20 25 30-0.1

0.3

µS

679 10

23

4 5 8 111

–50

500

µS

0 5 10 15 20 25 30Minutes

11

4

81 67 95

Determination of Trace Concentrations of Bromate Using Prepared Eluents (Isocratic)

Columns: Thermo Scientific™ Dionex™ IonPac™

AG23/AS23, 4 mm

Eluents: 4.5 mM Sodium carbonate/

0.8 mM Sodium bicarbonate

Temperature: 30 C

Flow Rate: 1.0 mL/min

Inj. Volume: 200 µL

Detection: Suppressed conductivity,

Thermo Scientific™ Dionex™ ASRS™

Anion Self-Regenerating Suppressor,

4 mm, AutoSuppression™,

external water mode

Peaks: 1. Fluoride 1.0 mg/L (ppm)

2. Chlorite 0.01

3. Bromate 0.005

4. Chloride 50

5. Nitrite 0.1

6. Chlorate 0.01

7. Bromide 0.01

8. Nitrate 10

9. Carbonate 50

10. Phosphate 0.1

11. Sulfate 50

LOD

(µg/L)

1.63

14

Bromate in Simulated Drinking Water

System: Thermo Scientific™ Dionex™

ICS-5000+ HPIC™ system

Column: Dionex IonPac AS19-4µm + guard

(4 250 mm)

Eluent : 10 mM KOH from 0 to 10 min,

10–45 mM KOH from 10 to 25 min

Eluent Source: Thermo Scientific Dionex

EGC 500 KOH Cartridge

Flow Rate: 1.0 mL/min

Inj. Volume: 200 µL

Temperature: 30 ˚C

Detection: Suppressed Conductivity,

Thermo Scientific™ Dionex™

AERS™ 500 Electrolytically

Regenerated Suppressor, 4 mm

AutoSuppression, recycle mode

Sample: Simulated drinking water

Peaks: 1. Fluoride 1.0 mg/L

2. Chlorite 0.005

3. Bromate 0.005

4. Chloride 50.0

5. Nitrite 0.005

6. Chlorate 0.005

7. Bromide 0.005

8. Nitrate 10.0

9. Carbonate 25.0

10. Sulfate 50.0

11. Phosphate 0.20

-0.2

0

0.5

µS

1

23

4

5 67

8 9 10 11

0 8 16 24 32-50

500

µS

Minutes

123

4

5 678

9

10

11

LOD

(µg/L)

0.32

15

Comparing Hydroxide vs. Carbonate Eluents

AnalyteRange

(µg/L)Linearity (r2)

Retention Time

Precision

(%RSDb,c)

Peak Area

Precision

(%RSD)

MDL

Standard

(µg/L)

MDL

Calculated

(µg/L)

Dionex IonPac AS19 Column – Hydroxide Eluent

Chlorite 2-50 0.9999 0.04 1.20 1.0 0.18

Bromate 1-25 0.9995 0.03 1.40 2.0 0.31

Chlorate 2-50 0.9999 0.01 0.54 1.0 0.28

Dionex IonPac AS23 Column – Carbonate/Bicarbonate Eluent

Chlorite 10-50 0.9999 0.07 2.20 5.0 1.02

Bromate 5-25 0.9998 0.07 2.63 5.0 1.63

Chlorate 10-50 0.9998 0.11 2.48 9.0 2.05

aSee Application Note 184 for conditions; bRSD = relative standard deviation, n = 7cQuality control standard contained 10 ppb each of chlorite, chlorate, and bromide and 5 ppb bromate

RFIC Technology with Hydroxide Eluents is > 5x More Sensitive

16

ISO 11206 – Scope of the Method

• Method for the determination of dissolved bromate e.g. drinking

water, mineral water, raw water, surface water, partially treated

water or swimming pool water.

• Appropriate pretreatment of the sample allows a determination of

bromate ≥ 0,5 μg/L.

• This method may be combined with ISO 10304-1, ISO 10304-4

and ISO 15061.

17

System Configuration 1 for ISO 11206

Eluant

Autosampler

ICSDionex IonPac

AG9-HC Column

Dionex IonPac

AS9-HC

Column

Dionex AMMS

Suppressor

Conductivity

Detector

Mixing

Tee

Absorbance

Detector

Knitted RX Coil

PCH-2 Heater @ 80 oC

Effluent to waste

Dionex AMMS

Suppressor

PC10 PCR

Reservoir /

Pump

18

Analysis of Ozonated Bottled Water - ISO 11206

Column: Dionex IonPac AG9-HC/AS9-HC

Eluant: 9 mmol/L Na2CO3

Temperature: 30 C

Flow rate: 1.30 mL/min

Inj. volume: 225 µL

Detection: A) Suppressed conductivity

B) Absorbance, 352 nm

PCR: Acidified KI

PCR flow rate: 0.4 mL/min

Post-column heater: 80 C

Peaks: (A) 1. Chlorite n.d. µg/L2. Bromate 1.523. DCA*4. Bromide 1.125. Chlorate 1.08

(B) 2. Bromate 1.84 µg/L

* DCA = Dichloroacetate Quality control surrogate

Note: Iodate can be detected in the same run in UV-0.001

0.004

26.1

27.1

0 5 10 2015

0 5 10 2015Minutes

(A)

(B)

AU

2

3

4

5

2

µS

19

System Configuration - ISO 11206 (II)

Eluant

Autosampler

ICS Guard Separator

PC10 /

ICS Pump

Mixing

Tee

Absorbance

Detector

Knitted RX Coil

Ambient Temp.

Effluent to waste

20

Bromate Determination with Acidic Eluent - ISO 11206

Column: Thermo Scientific™ Dionex™

CarboPac™ PA1

(4 250 mm)

Eluent: 200 mmol/L MSA

Flow: 1 mL/min

Injection vol.: 500 µL

Detection: UV 352 nm (after PCR)

Temperature: 30 °C

PCR:

Solution A: 0.27 mol/L KI, 0.05 mmol/L

(NH4)6Mo7O24 · 4H2O

Flow 0.3 mL/min

Reaction coil: 375 µL

Bromate 1.2 µg/L

Reaction at Ambient Temp., and no Interferences from Chlorite

Cycle Time: 18 minutes

0.0 2.0 4.0 6.0 8.0 10.0

-5.0

-2.5

0.0

2.5

5.0

Bromate

mAU

LOD

(µg/L)

LOQ

(µg/L)

0.07 0.22

Minutes

21

Bromate U.S. EPA Method Summary

IC TechniqueEPA

MethodDionex IonPac Columns Eluent

Thermo Scientific

Application

MDL (ppb)

Suppressed Cond. 300.0 (B)Dionex IonPac AS9-HC or

Dionex IonPac AS23

Dionex IonPac AS-19

Carbonate

Hydroxide

AN167,

AN184

CD

5.0, 1.63

0.32

Suppressed Cond. 300.1Dionex IonPac AS9-HC or

Dionex IonPac AS23

Dionex IonPac AS-19

Carbonate

Hydroxide

AN167,

AN184

5.0, 1.63

0.32

2D-IC Suppressed

Cond. 302.0

4 mm Dionex IonPac AS19 to 2 mm Dionex IonPac AS24

4 mm Dionex IonPac DionexIonPac AS19 to 0.4 mm

Dionex IonPac AS20

Hydroxide

Hydroxide

AN187

---

0.036,

~ 0.004

Suppressed Cond. +

Postcolumn ODA317.0

Dionex IonPac AS9-HC

Dionex IonPac AS-19

Carbonate

HydroxideAN168

UV/vis

0.14

Suppressed Cond. +

Postcolumn acidified

KI

326.1Dionex IonPac AS9-HC

Dionex IonPac AS-19

Carbonate

Hydroxide

AN171

---0.17

IC-ICP/MS 321.8 Dionex CarboPac PA100Nitric Acid + Ammonium

nitrate

MS

0.010

22

DETERMINATION OF DISSOLVED ANIONS BY LIQUID CHROMATOGRAPHY (LC) - PART 4:

Determination of chlorate, chloride and chlorite in slightly polluted wastewater

ISO 10304-4 – Water Quality

23

ISO 10403-4 – Working Ranges and Detection

AnionWorking Range

(mg/L)Detection

Chlorate 0.03 to 10 Suppressed CD

Chloride 0.1 to 50 Suppressed CD

Chlorite

0.05 to 1 Suppressed CD

0.1 to 1 UV, (207 to 220 nm)

0.01 to 1 Amp, DC, 0.4 to 1.0 V

24

Column: Dionex IonPac AS23 with guard Sample: Dilution with DI Water ( 1:25)

Flow: 0.25 mL/min

Eluent: 4.5 mmol/L Na2CO3 + 1.5 mmol/L NaHCO3

Detection: Suppressed Conductivity, Dionex ASRS suppressor

Injection Vol.: 25 µL

0,0 5,0 10,0 15,0 20,0 25,0

0

100

200

300

400

500

600

0

1

2

3

4

5

µS µS

Anion mg/L

Chloride 8693

Chlorate 3,90

Nitrate 6,41

Phosphate /

Sulphate 17,2

Anion Determinations of a Disinfected Swimming Pool Water Sample

25

Alternative Approaches

26

Analysis Using IC×IC (2D-IC)

Large Loop

Suppressor

Cell 1Pump EG4 mm

Column

Injection ValveCRD

Switching Valve

EG

0.4 mm

Column

Dionex IonPac UTAC-ULP1

Concentrator

PumpCell 2

Suppressor

Thermo Scientific

Dionex CRD

Carbonate Removal Device

1st Dimension

Large-loop injection

Partially resolve matrix Intermediate Step

Remove time segment

Trap and concentrate ions of interest

2nd Dimension

Resolve on smaller column

Sensitivity enhancement

Different selectivity optional

27

—— Sample A (54 ng/L)

—— 100 ng/L bromate in deionized water

—— 30 ng/L bromate in deionized water

—— Deionized water

17 20 -0.3

0.5

µS

Minutes

1

Bro

mate

A. 1st Dimension

Column: Dionex IonPac AG19/AS19,

4 mm

Flow rate: 1 mL/min

Eluent: 10-60 mmol/L KOH (EG)

Suppressor: Thermo Scientific™ Dionex™

SRS™ 300 Self-Regenerating

Suppressor (4 mm)

Inj. volume: 1000 µL

Temperature: 30 °C

B. 2nd Dimension

Column: Dionex IonPac AS20 (0.4 mm)

Flow rate: 10 µL/min

Eluent: 35 mmol/L KOH (EG)

Suppressor: Thermo Scientific™ Dionex™

ACES™ 300 Anion Capillary

Electrolytic Suppressor

Temperature: 30 C

Concentrator: Capillary concentrator,

2500 µL of the suppressed

effluent from the 1st dimension

(7.5–10 min)

Trace Analysis of Bromate in Bottled Water by IC×IC

28

Haloacetic Acids in Drinking Water

● Simplified 2D IC solution

● Low MDLs

● Low operating costs

● Capillary, microbore and standard bore formats

A. First-Dimension ConditionsColumn: Dionex IonPac AG24/AS24, 4 mm Flow Rate: 1.0 mL/min Eluent: KOH Gradient - RFICTemperature:15 CDetection: Suppressed conductivity,

Dionex ASRS 300 suppressor, 4 mm, AutoSuppression, recycle mode

Inj. Volume: 1000 µL

Peak:

1. MCAA 6. TCAA

2. MBAA 7. BDCAA

3. DCAA 8. CDBAA

4. BCAA 9. TBAA

5. DBAA

First Dimension

µS1

3

45

2

97

6

8

3

010 20 30 40 600 50

65

Second Dimension1

34

52

9

7

6

8

15 30 45 600

3

0

µSB. Second-Dimension ConditionsColumn: Dionex IonPac AG26/AS26, 2 mmFlow Rate: 0.25 mL/minEluent: KOH Gradient - RFICTemperature: 15 CDetection: Suppressed conductivity,

Dionex ASRS 300 suppressor, 2 mm, AutoSuppression, recycle mode

Conc. Column: Dionex IonPac UTAC-LP2

29

Mass Spectrometry Methods

30

Determination of dalapon, trichloroacetic acid and selected haloacetic acids

– Method using gas chromatography (GC-ECD and/or GC-MS detection) after liquid-liquid extraction and derivatization

ISO 23631 Water quality, GC-MS

31

Haloacetic Acids with GC/MS after Derivatization

Instrumentation: Thermo Scientific ™

TRACE ™ Ultra Gas Chromatography

system

Carrier gas: Helium

Split flow: 10 mL/minute

Column flow: 1.2 mL/minute, Constant flow

Oven temperature: 40 C (1 minute),

5 C/minute, 135 C (1 minute)

Injector type: Split/Splitless

Injector mode: Split

Split ratio: 10:1

Injector temp.: 220 C

Injection conditions

Instrumentation: Thermo Scientific™ TriPlus™

Autosampler

Injection Volume: 2 µL

Pre and Post Injection

Dwell Time: 5 s

MS Conditions

Instrumentation: Thermo Scientific™ ISQ™

Quadrupole GC-MS system

Transfer Line Temp: 260 C

Source Temp.: 230 C

Ionization: EI

Electron energy: 70 eV

Solvent delay: 3.2 min

Scan range: 35-300 amu full scan

dwell time 0.25 s6 7 8 9 10 11 12 13 14 15 16 17 18

Time (min)

10

20

30

40

50

60

70

80

90

1

2

3

4

5

6

78

910

Peak Compound Abbrev

1 Monochloroacetic acid MCAA

2 Monobromoacetic acid MBAA

3 Dichloroacetc acid DCAA

4 Dalapon -

5 Trichloroacetic acid TCAA

6 Bromochloroacetic Acid BCAA

7 Dibromoacetic acid DBAA

8 Bromodichloroacetic acid BDCAA

9 Chlorodibromoacetic acid CDBAA

SIM at 250 ng/mL at m/z 59

100

32

MSQ Plus Low Mass Single Quadrupole MS

• Thermo Scientific Dionex suppressors allow the use of

standard IC elements and methods

• Eluent generators allows fast, clean switching between

anions and cations in IC-MS

• The Thermo Scientific™ MSQ™ Mass Spectrometer is

designed for the needs of IC

• Single quadrupole, low mass detector

• Universal detection and characterization of analytes in the 17–2000 m/z

range

• Enhanced low-mass response for analytes below 60 m/z

• High-sensitivity, self-cleaning ESI/APCI ionization source

• Outstanding sensitivity and noise reduction

33

IC-MS Flow Diagram

High-Pressure Non-Metallic Pump

Eluent Generator

(OH– or H+)

Waste

Sample Inject(Autosampler)

CR-TC

Electrolytic Eluent

Suppressor

Separation Column

Thermo Scientific™

MSQ Plus™

MS

Conductivity Detector

0.31 µS

Data Management

34

Oxyhalides and Bromide by IC-MS

counts

SIM_01 (Chlorite): m/z 67SIM_02 (Bromide): m/z 79SIM_03 (Bromate): m/z 129SIM_04 (Iodate): m/z 175

0.0 5.0 10.0 15.0 20.0 25.0 30.0 35.0-5 e3

2 e4

4.5 e4

Minutes

4

Iod

ate

3

Bro

ma

te

21

Ch

lori

te

0.0 5.0 10.0 15.0 20.0 25.0 30.0 35.0-2 e3

1 e4

2.2 e4

counts

4

Iod

ate

Bro

ma

te2

Bro

mid

e

1,3C

hlo

rite

Column: Dionex IonPac AS19,guard

(2 mm)

Eluent: KOH gradient

Flow: 0.25 mL/min

Detection: Suppressed conductivity

Background

Signal:

< 3 S/cm

Suppressor: Dionex ASRS suppressor, 2 mm

(external water mode)

Temperature: 30 °C

Inj. Volume: 250 µL

Ionisation: ESI in negative mode

Needle

Voltage:

3 kV

Cone Voltage: 50 V (chlorite) and 75 V

(iodate, bromate and bromide)

Probe Temp.: 400 °C

S.I.M: 67, 79, 129, 175 m/z

35

IC-MS (SIM) of Haloacetic Acids

Time (min)

Rela

tive A

bundance

0 2 4 6 8 10 12 14 16 18 20 22 240

50

1000

50

1000

50

100

0

50

1000

50

100

0

50

100

0

50

100

0

50

100 1

2

3

4

5

6

87

98

7

m/z 93

MCAA

m/z 139

MBAA

m/z 127

DCAA

m/z 173

BCAA

m/z 217

DBAA

m/z 161 TCAA (6) +

DCBAA (minus CO2)

m/z 207 DCBAA (7) +

DBCAA (minus CO2)

m/z 251 DBCAA (8) +

TBAA (minus CO2)

Column: Dionex IonPac AG16/AS16,

2 mm

EG Eluent: KOH gradient from 5 to 70

mM

Flow Rate: 0.25 mL/min

Suppressor: Dionex ASRS suppressor,

external water mode

Concentrator: Dionex IonPac TAC-LP1

Sample Volume: 5 mL

Detector: MSQ system, -ESI mode

Probe: -2.5 kV, 350 C

Cone voltage: 25 to 50 V

Peaks: HAAs at 1–3 µg/L (ppb)

36

IC-MS of Haloacetic Acids in 0.16% NaCl

Column: Dionex IonPac AS16, 2 250 mm

Sample Prep: Thermo Scientific™

Dionex™ OnGuard™ II Ag/H

Suppressor: Dionex ASRS suppressor, 2 mm,

external water mode

EG50 Eluent: 10 mM KOH

Flow Rate: 0.25 mL/min

Injection loop: 5 µL

Detector: MSQ system, -ESI, full scan

Probe: -3 kV, 350 oC

Source: 25 V

Analytes: 1. Chloroacetic acid 8 mg/L

2. Bromoacetic acid 7 mg/L

Minutes

m/z 137-139.2

m/z 92.5 - 93.5

TIC

40

100

0 2 4 6 8 10 12 14

Re

l. A

bu

nd

an

ce

0

100

0

100

1

2

2

1

37

Summary

• Toxic regulated disinfection byproducts are generated through

water treatment chlorination and ozonation processes

• Ion chromatography methods are ideal to determine DBPs

• Regulated ISO Methods 15061, 11206, and 11403

• Regulated U.S. EPA Methods 300.0 (B), 300.1, 302.0, 317.0, 326.1,

321.8

• Thermo Scientific, the leader and innovator in ion chromatography,

has worked hand-in-hand with ISO and U.S. EPA to develop and

provide these IC methods now and in the future

• Ion-exchange separations with

• Suppressed conductivity

• Post-column derivitization and UV absorbance

• MS, ICP-MS

38

Thank you!

WS71422-EN 1114S