Selective method for the analysis of perchlorate in drinking waters at nanogram per liter levels, using two-dimensional ion

chromatography with suppressed conductivity detection

CHEM 699

Izzat w. Kazi
1

outline
Introduction

Experimental
Instrumentation Result

& Discussion Conclusion Recommendation

Introduction

Perchlorate can interfere with the uptake of iodide by the thyroid gland Health effects associated with the presence of perchlorate in various food products and drinking water is a cause of concern since certain populations are particularly susceptible, such as pregnant women and infants

High doses of perchlorate can affect metabolism, growth and development of the body.
Early in 2005, the National Academy of Sciences recommended a maximum perchlorate dose of 0.7 mg kg1 body weight per day from all sources, including water, milk, vegetables and prepared food
Office of Environmental Health Hazard Assessment, March 11, 2004,http://www.oehha.ca.gov/public info/facts/faqperchlorate.html. National Academy of Science, January 10, 2005, http://www4.nationalacademies.org/news.nsf/isbn/0309095689?OpenDocument 3

Introduction

contd

Until recently, U.S. EPA method 314.0 was used for determining the level of perchlorate in drinking and groundwaters Excellent utility at 4 mg l1 or higher concentration levels, But Requires offline sample pretreatment using solid phase extraction (SPE) cartridges Difficult to determine perchlorate at low levels.
High levels of matrix ions chloride, sulfate or bicarbonate

Labor intensive and cumbersome sample pretreatment

4

Safe Drinking Water Act Analytical Methods and Laboratory Certification

Introduction
In

contd

order to eliminate these more labor intensive procedures, EPAs Office of Ground Water and Drinking Water (OGWDW) and National Exposure Research Laboratory, initiated method development projects to increase sensitivity and selectivity using direct injection techniques for the analysis of perchlorate in drinking water and in simulated high ionic strength matrices.
5

Introduction
Two-dimensional

contd

ion chromatography (2DIC) with suppressed conductivity detection met all OGWDW requirements for
selectivity sensitivity precision accuracy and method robustness
6

Introduction

contd

This is essentially an automated heart-cutting, column concentration and matrix elimination technique with second column confirmation In the first dimension, a large sample volume is injected onto a first separation column and the matrix ions are diverted to waste, while the perchlorate and a small amount of other anions that elute within the cut window are diverted and trapped onto a concentrator column In the second dimension, the contents of the concentrator column are eluted off and diverted onto a second analytical, smaller diameter (higher resolution) column for separation and quantitation of perchlorate.
7

Introduction

contd

Dionex Corporation and OGWDW worked in partnership to develop a 2D-IC method in order to simplify the Method Development of this new
highly sensitive and selective 2D-IC, suppressed conductivity method

Drinking water Lowest Concentration Minimum Reporting Level (LCMRL) of 55 ng/L, and has
comparable sensitivity and selectivity simpler and more economical than IC-MS or ICMSMS techniques.

Revisions to the Unregulated Contaminant Monitoring Regulation for Public Water Systems, Proposed Rule, EPA document #815R-05-006, November 2004 (http://www.epa.gov/safewater/methods/pdfs/method lcmrl.pdf). 8

Experimental
Reagents

All solutions and all dilutions were prepared using puried reagent water (RW) 18 MW
ltered

with a 0.2 mm lter and did not contain any measurable quantity of the target analyte or interfering compound with 1000 mg/L of chloride, bicarbonate and

laboratory synthetic sample matrix (LSSM)

fortied

sulfate This solution was used to simulate a drinking water matrix containing a total of 3000 mg/L of these anions

 Standard

and sample preparation

Calibration standards, Continuing calibration check standards and Perchlorate spiking solutions
prepared

using a 1000 mg/L perchlorate (ClO4) standard stock solution prepared from solid sodium perchlorate (Sigma, Milwaukee, WI).

10

Instrumentation
Dionex

Model ICS-3000 dual system

Dual pump (DP) module Eluent generator (EG) module Detector/chromatography (DC) module Autosampler (AS) Continuously-regenerated anion trap columns, Suppressors Carbonate removal devices Dual conductivity detectors, and Computer-based data acquisition and control system.

11

Instrumental conditions and equipment for first dimensions

Ion chromatograph Sample loop Load volume Eluent generator Eluent Eluent flow Dionex ICS-3000 dual system 2.0 or 4.0 mL 4.7 mL Dionex ICS-3000 eluent generator (EG) with dual channel EluGen Catridges Isocratic 35 mM potassium hydroxide ramped to 100 mM potassium hydroxide following the elution of perchlorate (when injection valve system #2 is switched to the inject position) 1.0 mL/min Autosampler module (AS) with sequential injection, sample preparation options and a large volume (8.2 mL) sample needle assembly Dionex IonPac AG 20 (4 mm 50 mm) Dionex IonPac AS 20 (4 mm 250 mm)

Autosampler

Columns Typical system back2500 psi pressure Conductivity Dionex ASRS Ultra II, 4-mm with 4-mm carbonate removal device (CRD) suppressor Chromatography DC-Module maintained at 30 C module Detector Conductivity detector (CD) with integrated cell held at 35 C Total analysis time 45 min

12

Instrumental conditions and equipment for second dimensions

Ion chromatograph Dionex ICS-3000 dual system Sample loop Load volume Eluent generator Eluent Eluent flow Autosampler Columns Dionex UTAC ULP1, 5 mm 23 mm, concentrator column Cut-window time set per procedures Dionex ICS-3000 eluent generator (EG) with dual channel EluGen Catridges Isocratic 65 mM KOH 0.25 mL/min Autosampler module (AS) Dionex IonPac AG 16 (2 mm 50 mm) Dionex IonPac AS 16 (2 mm 250 mm)

Typical system back2500 psi pressure Conductivity Dionex ASRS Ultra II, 2-mm with 2-mm carbonate removal device (CRD) suppressor Chromatography DC-Module maintained at 30 C module
Detector Total analysis time Conductivity detector (CD) with integrated cell held at 35 C 45 min
13

Fig. 1 Scheme of 2-D IC. Acronyms defined in above Table

14

Results and discussion

Two

complete IC systems are integrated to provide a single IC

Automated analyses that incorporate heartcutting, column concentration Matrix elimination techniques Single injection for the analysis can accommodate a large sample injection volume (up to 4.0-mL)
greatly

increases sensitivity
15

refocus the perchlorate peak that is partially resolved in the first

Setting the cut window

window

must be set to ensure that the perchlorate is completely eluted from the column and that the amount of other potentially interfering species, which may also elute during this time, are kept to a minimum Ideally, a surrogate would be included in the method to monitor and ensure that the cut window is functioning properly
16

Setting the start time for the cut window in the rst dimension
Since

4 mL)

2D-IC utilizes a large volume injection (up to

high ionic matrix concentration on the retention time is a critical factor address when setting the cut window injection volume is also a critical factor
retention

time for perchlorate determined in 1000 mg/L (LSSM)

to set the start time of the cut window and
RW

to set the stop time of the cut window

17

 The

start time for the cut window

2.0 mL of a 50 mg/L perchlorate standard prepared in the 1000 mg/L LSSM

injection

valve #2 on system #2 is switched to the load position

pressure spike baseline deflection occurs after about 30 s

start

time for the cut window

18

1 min before the rise in baseline for the perchlorate peak in the 1000 mg/L LSSM.

Fig 2: start time for the cut window using a 2.0-mL injection volume of a 50 mg/L ClO4 fortification in the 1000 mg/L LSSM.

19

 To

ensure the start time is set using the perchlorate peak, not this earlier baseline deflection
2.0-mL injection of the blank 1000 mg/L LSSM (without the perchlorate) also be injected.

Retention

time

19.5 min and, therefore, the cut window start time was set at 18.5 min.
20

Setting the stop time for the cut window

The

stop time for the cut window

2.0 mL of a 25 mg/L perchlorate standard prepared in the RW

stop

time for the cut window is also the time at which injection valve #2 on system #2 is switched to the inject position
elution of perchlorate must be completed at least 60 s before injection valve #2 is switched into the inject position perchlorate peak returned to baseline at 22.5 min and the stop time for the cut window was established at 23.5 min. 21

Fig. 3. Setting the stop time for the cut window using a 2.0-mL injection volume of a 25mg/L ClO4 fortification in RW.

22

Fig. 4. First- and second-dimension chromatogram using a 2.0-mL injection volume of a 25mg/L ClO4 fortification in RW.

23

Incorporating QC standards to monitor the cut window

Column

performance can deteriorate as chromatographic columns age

Should monitored on a daily basis, with the use of QC standards

Highest

calibration standard should be prepared in the 1000 mg/L LSSM as a continuing calibration check standard (CCC)
to ensure the perchlorate is being trapped within the set cut window.

. Demonstration of method performance using a 2.0-mL injection volume

Instrument
0.025,

calibration

standards prepared in RW
0.050, 0.10, 0.30, 0.50,1.0, 3.0, 5.0 and 10g/L correlation coefficient was 0.99956

LCMRL

determination
50 and 150% recovery with 99% confidence

LCMRL is the lowest true concentration

between 0.050,

seven individual, sterile filtered replicates

0.10, 0.20 and 0.30g/L perchlorate fortified RW samples
25

LCMRL of 0.055g/L

Precision and accuracy (P&A) data

Table 2 Precision and accuracy (P&A) data (n = 7) using 2.0mL injection volume Native concentration Fortified Precision (g/L) concentration (g/L) (%RSD) Matrix Reagent <0.055 0.50, 5.0 4.5, 1.8 water LSSMa <0.055 0.50, 5.0 6.9, 1.5 Ground water Surface water-1 Surface water-2 0.2 0.5 0.50, 5.0 0.50, 5.0 5.8, 1.8 4.2, 1.8

Accuracy (%recovery) 94.7, 96.1 94.6, 95.0 92.4, 97.0 95.3, 97.1

0.74

0.50, 5.0

4.3, 1.2

91.8, 96.2
26

Table 3 Precision and accuracy (P&A) data (n = 7) using 4.0-mL injection volume Native Fortified concentratio concentration Precision Accuracy n (g/L) (g/L) Matrix (%RSD) (%recovery) Reagent water <0.041 0.50, 5.0 4.2, 0.59 97.2, 98.6

LSSMa

<0.041

0.50, 5.0

4.6, 1.3

97.2, 97.0

Ground water

0.2

0.50, 5.0
0.50, 5.0 0.50, 5.0

4.2, 2.1
3.4, 1.1 1.2, 0.85

97.3, 97.2
110, 97.4 92.8, 97.5
27

Surface water-1 0.49 Surface water-2 0.74

a Laboratory synthetic sample matrix containing 1000 mg/L each of chloride, bicarbonate and sulfate.

Demonstration of method performance using a 4.0-mL injection volume

Larger

injection volume increases sensitivity. cut window using a 4.0- mL

dramatic effect on retention

Resetting

injection

start time for the cut window was established at 20.0 min stop time was set at 26.0 min
LCMRL

is 0.041g/L
28

Fig. 5. Setting the start time for the cut window in the first dimension using a 4.0-mL injection volume of a 25mg/L ClO4 fortification in the 1000 mg/L LSSM.

29

Fig. 6. Setting the stop time for the cut window in the first dimension using a 4.0-mL injection volume of a 10mg/L ClO4 fortification in RW

30

Precision and accuracy data

Table 3 Precision and accuracy (P&A) data (n = 7) using 4.0-mL injection volume Native Fortified concentration concentration Precision Accuracy (g/L) (g/L) Matrix (%RSD) (%recovery) Reagent water <0.041 0.50, 5.0 4.2, 0.59 97.2, 98.6

LSSMa

<0.041

0.50, 5.0

4.6, 1.3

97.2, 97.0

Ground water

0.2

0.50, 5.0
0.50, 5.0 0.50, 5.0

4.2, 2.1
3.4, 1.1 1.2, 0.85

97.3, 97.2
110, 97.4 92.8, 97.5
31

Surface water-1 0.49 Surface water-2 0.74

a Laboratory synthetic sample matrix containing 1000 mg/L each of chloride, bicarbonate and sulfate.

Comparison of 2.0 and 4.0 mL injection volumes

Peak

areas for the 4.0-mL injection volume

mid and high-level CCC standards (n 5)

2.0 to 2.1 times the peak area than the 2.0-mL injection volume for the
low,

LCMRL

was also slightly improved native concentrations in the three drinking water matrices
ranged from 0.199 to 0.742mg/L
statistically

equivalent (95% confidence interval) in both injection volumes

32

Conclusions
The

new two-dimensional IC method with suppressed conductivity detection

excellent sensitivity Selectivity precision accuracy and method robustness

Simpler

and more economical than IC-MS or IC- MSMS techniques

33

Recommendation
Apply

method for the determination of other ion like

Bromate (elutes just before chloride) Nitrate (elutes just after chloride) etc.

In sea water or high matrix water Simplification of the instrumentation

Replacing two-6port valve by single 10port valve Eliminating CD-1
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