Here's Waters 6508 method that was approved for drinking water. Aren't

you limited to methods that are listed as approved for drinking water?

Based on the info at the top of the method, I'm guessing this may now be

an ASTM method. It was evaluated under the ATP program, so EPA was

given the method prior to the ASTM process. I don't know this for sure

and it will be something I investigate as we begin putting together our

next Expedited Methods Approval FR action. (If it is the same method,

we'll probably list it in Appendix A.)

(See attached file: Waters Method D 6508, Rev

2_EPA-HQ-OW-2003-0070-0063.pdf)

As for the other method, it was approved prior to the 2007 methods rule.

I don't have a copy of it, because I wasn't involved in the earlier

methods rules. However, I have asked our ATP coordinator to see if it

is in the ATP file. When I hear back from him, I'll let you know.

Hope this helps,

Pat

"Mike

McCambridge"

<mccambridge@ipc?

To

b.state.il.us>?

Pat Fair/Cl/USEPA/US@EPA

CC

07/08/2008 03:29

PM

?

?

Subject

Re: Waters Methods

Whatever you can do for me when you get back. I have continued to look

into this today. I am convinced that Method 6500 added to SW-846 in

Update IV in the end of 2007 is the method you have called "Method

D6508" from Waters. See 73 Fed. Reg. 486 (Jan. 3, 2008);

http://www.epa.gov/SW-846/pdfs/6500.pdf. If this is true, I will likely

cite the SW-846 version of the method, since it is much easier to obtain

than the method from Waters. As described, Waters initially acted like

I spoke a foreign language when I asked for "Method D6508." As for

From:

To:

Date:

Subject:

Mike,

<Fair.Pat@epamail.epa.gov>

MCCAMBM@ipcb.state.il.us

7/10/2008 7:46:26 AM

Re: Waters Methods

RECEIVED

CLERK'S OFFICE

AUG 0

2008

STATE

OF ILLINOIS

Pollution Control Board

---

Method B-1011, it seems to distill down to me needing the title to the

document in which the method appears.

Talk to you when you return.

Michael J. McCambridge

Attorney

Illinois Pollution Control Board

312-814-6924

>>> <Fair.Pat@epannail.epa.gov> 7/8/2008 2:18 PM >>>

Mike,

I'm working off site today, so I don't have access to the references I

need to answer your questions. I should have copies of the methods that

were added to 40 CFR 141 as part of the 2007 methods update rule. If

these Waters methods are prior to that, I might not be able to help you.

Unfortunately, I don't know who might have

.

them other than Waters.

I'll see what I can find tomorrow and get back to you.

Pat

"Mike McCambridge" <mccambridge@ipcb.state.il.us> wrote:

To: Pat Fair/CUUSEPA/US@EPA

From: "Mike McCambridge" <mccambridge@ipcb.state.il.us>

Date: 07/08/2008 01:57PM

Subject: Waters Methods

I have tried to obtain copies of the two Waters methods referenced in 40

C.F.R. 141.23(k)(1) for fluoride and nitrite/nitrate using the contact

information included in the rule. At first, the Waters rep could not

locate anything based on the EPA descriptions included in the rule.

This morning I received two documents that purport to be the methods.

The documents raise questions that you might answer for me.

The copy of Method B-1011 sent me by Waters is nearly identical to one

that I found on the USEPA website. The only difference between the two

is that the method from the USEPA website is headed "Waters." The

document it appears to include pages 13 through 17 from some reference.

It is undated, which means that I cannot use it for an incorporation by

reference. Do you have a dated copy of Method B-1011 or a fuller copy

of the posted reference that would include the date? It appears that

the method is just one cited out of a fuller reference, and I should

cite to that fuller reference by its own title. I will also approach

Waters with this request.

Your rule cites "Waters Method D6508, Rev. 2," entitled "Test Method for

Determination of Dissolved Inorganic Anions in Aqueous Matrices Using

Capillary Ion Electrophoresis and Chromate Electrolyte." Waters sent me

a document marked "Method 6500," "revision 0," and dated February 2007,"

and entitled "Dissolved Inorganic Anions In Aqueous Matrices By

Capillary Ion Electrophoresis." That document appears to be Method 6500

from SW-846. Is "Waters Method D6508, Rev. 2" the same as Method 6500,

---

rev. 0 from SW-846? If so, why did USEPA cite this as "D6508"? If not,

can you forward me a copy of Method D6508 or give me enough information

to identify the method to Waters, that I might obtain a copy of the

right method?

Michael J. McCambridge

Attorney

Illinois Pollution Control Board

312-814-6924

---

Page 1 of 5

Mike McCambridge - Re: Waters Methods

From:

<Fair.Pat epamail.epa.gov>

To: "Mike

McCambridge" <mccambridge@ipcb.state.il.us>

Date:

7/10/2008 9:31 PM

Subject:

Re: Waters Methods

Mike,

I haven't done a one-to-one check of the ASTM method against the Waters method, so I can't say

for sure that they are the same. My comment was meant to let you know that I would do that

BEFORE we issue the next set of method approvals. If they are the same or only have insignificant

differences, then we will include the ASTM method as an approved method. Legally, it won't be an

approved drinking water method until we publish a notice in the Federal Register.

It's my opinion that if the Waters methods aren't easily available from Waters, then you can easily

justify not including them in your state regulations. Our ATP coordinator wasn't able to find a copy

of the nitrate/nitrite method in his files. However, he is still checking on it.

I have the GA Tech method. I can email it to you on Monday. If you need it before then, you can

go to the e-docket for the 2007 Methods Update Rule. I know the method is in the docket,

because I put it there and it is available for download through the docket site.

I will see if I can find out how we should be referencing the GA Tech method. I thought our

information was correct when we went final on the rule.

Hope this helps.

Pat

"Mike McCambridge" <mccambridge@ipcb.state.il.us> wrote:

To: Pat Fair/Cl/USEPA/US@EPA

From: "Mike McCambridge" <mccambridge@ipcb.state.il.us>

Date: 07/10/2008 06:07PM

Subject: Re: Waters Methods

Thank you. That nails it down. I will cite it as an ASTM method.

I have another method problem. I have been trying

:

to obtain a copy

of that Ra-226/Ra-228 method by gamma-ray spectometry developed by

Georgia Insitute of Technology. The "Environmental Resources

Center" has been disbanned or something, so that the number at 40

C.F.R. 141.74 is no longer valid. It may have become the

Environmental Radiation Center or something. I have placed several

calls and e-mails with Bernd Kahn and the Center in an attmpt to

locate the method, but no luck so far.

Can you help on this one too?

file://C:\Documents and Settings\McCambM\Local Settings\Temp\GW100001.HTM 8/4/2008

---

Page 2 of 5

Michael J. McCambridge

Attorney

Illinois Pollution Control Board

312-814-6924

>>> <Fair.Pat@epamail.epa.gov> 7/10/2008 7:43 AM >>>

Mike,

Here's Waters 6508 method that was approved for drinking water.

Aren't

you limited to methods that are listed as approved for drinking

water?

Based on the info at the top of the method, I'm guessing this may

now be

an ASTM method. It was evaluated under the ATP program, so EPA was

given the method prior to the ASTM process. I don't know this for

sure

and it will be something I investigate as we begin putting together

our

next Expedited Methods Approval FR action. (If it is the same

method,

we'll probably list it in Appendix A.)

(See attached file: Waters Method D 6508, Rev

2 EPA-HQ-0W-2003-0070-0063.pdf)

As for the other method, it was approved prior to the 2007 methods

rule

I don't have a copy of it, because I wasn't involved in the earlier

methods rules. However, I have asked our ATP coordinator to see if

it

is in the ATP file. When I hear back from him, I'll let you know.

Hope this helps,

Pat

To

CC

"Mike

McCambridge"

<mccambridge@ipc

b.state.il.us>

Pat Fair/Cl/USEPA/US@EPA

file://C:\Documents and Settings\MeCambM\Local Settings\Temp\GW100001.HTM

8/4/2008

---

Page 3 of 5

07/08/2008 03:29

PM

Subject

Re: Waters Methods

Whatever you can do for me when you get back. I have continued to

look

into this today. I am convinced that Method 6500 added to SW-846 in

Update IV in the end of 2007 is the method you have called "Method

D6508" from Waters. See 73 Fed. Reg. 486 (Jan. 3, 2008);

http://www.epa.gov/SW-846/pdfs/6500.pdf. If this is true, I will

likely

cite the SW-846 version of the method, since it is much easier to

obtain

than the method from Waters. As described, Waters initially acted

like

I spoke a foreign language when I asked for "Method D6508." As for

Method B-1011, it seems to distill down to me needing the title to

the

document in which the method appears.

Talk to you when you return.

Michael J. McCambridge

Attorney

Illinois Pollution Control Board

312-814-6924

>>> <Fair.Pat@epamail.epa.gov> 7/8/2008 2:18 PM >>>

Mike,

I'm working off site today, so I don't have access to the references

I

file://C: \Documents and Settings\McCambM\Local Settings\Temp\GW}00001.HTM 8/4/2008

---

Page 4 of 5

need to answer your questions. I should have copies of the methods

that

were added to 40 CFR 141 as part of the 2007 methods update rule.

If

these Waters methods are prior to that, I might not be able to help

you.

Unfortunately, I don't know who might have them other than Waters.

I'll see what I can find tomorrow and get back to you.

Pat

"Mike McCambridge" <mccambridge@ipcb.state.il.us> wrote:

To: Pat Fair/Cl/USEPA/US@EPA

From: "Mike McCambridge" <mccambridge@ipcb.state.il.us>

Date: 07/08/2008 01:57PM

Subject: Waters Methods

I have tried to obtain copies of the two Waters methods referenced

in 40

C.F.R. 141.23(k)(1) for fluoride and nitrite/nitrate using the

contact

information included in the rule. At first, the Waters rep could

not

locate anything based on the EPA descriptions included in the rule.

This morning I received two documents that purport to be the

methods.

The documents raise questions that you might answer for me.

The copy of Method B-1011 sent me by Waters is nearly identical to

one

that I found on the USEPA website. The only difference between the

two

is that the method from the USEPA website is headed "Waters." The

document it appears to include pages 13 through 17 from some

reference.

It is undated, which means that I cannot use it for an incorporation

by

reference. Do you have a dated copy of Method B-1011 or a fuller

copy

of the posted reference that would include the date? It appears

that

the method is just one cited out of a fuller reference, and I should

cite to that fuller reference by its own title. I will also

approach

Waters with this request.

Your rule cites "Waters Method D6508, Rev. 2," entitled "Test Method

file://C:\Documents and Settings\MeCambM\Local Settings\Temp\GW}00001.HTM 8/4/2008

---

Page 5 of 5

for

Determination of Dissolved Inorganic Anions in Aqueous Matrices

Using

Capillary Ion Electrophoresis and Chromate Electrolyte." Waters

sent me

a document marked "Method 6500," "revision 0," and dated February

2007,"

and entitled "Dissolved Inorganic Anions In Aqueous Matrices By

Capillary Ion Electrophoresis." That document appears to be Method

6500

from SW-846. Is "Waters Method D6508, Rev. 2" the same as Method

6500,

rev. 0 from SW-846? If so, why did USEPA cite this as "D6508"? If

not,

can you forward me a copy of Method D6508 or give me enough

information

to identify the method to Waters, that I might obtain a copy of the

right method?

Michael J. McCambridge

Attorney

Illinois Pollution Control Board

312-814-6924

file://C:\Documents and Settings\McCambM\Local Setfings\Temp\GW100001.HTM

?

8/4/2008

---

00---09,1b--001°--1966,3

This ASTM D6508, Rev2 method document has been reviewed

by EPA

Office of Drinking

Water and Wastewater for EPA Tier 3 approval. Added updated QC criteria based upon

statistical analysis

by Dyncorp.

ATP Case #:

Draft #:

Date:

Method Author:

Telephone:

FAX:

N00-0002 and D00-0002

Second draft with EPA Modifications: ASTM D6508, Rev

2

December 2000

Jim Krol

508/482-2131

508/482-3625

Test Method for

Determination of Dissolved Inorganic Anions

in Aqueous Matrices Using

Capillary Ion Electrophoresis

1 Scope

and Chromate Electrolyte

1.1 This test method covers the determination of the inorganic anions fluoride,

bromide, chloride, nitrite, nitrate, ortho-phosphate, and sulfate in drinking water,

wastewater, and other aqueous matrices using capillary ion electrophoresis

(CIE) with indirect UV detection. See Fig. 1 through 6.

1.2 The test method uses a chromate-based electrolyte and indirect UV detection

at 254 nm. It is applicable for the determination of inorganic anions in the

range of 0.2 to 50 mg/L except for fluoride whose range is 0.2 to 25 mg/L.

1.3 It is the responsibility of the user to ensure the validity of this test method for

other anion concentrations and untested aqueous matrices.

Note 1: The highest accepted anion concentration submitted for P&B extend the anion

concentration range for the following anions; Chloride to 93 mg/L, Sulfate to 90 mg/L,

Nitrate to 72 mg/L, and ortho-phosphate to 58 mg/L.

1.4 This method does not purport to address all of the safety problems, if any,

associated with its use. It is the responsibility of the user of this standard to

establish appropriate safety and health practices and determine the

applicability of regulatory limitations prior to use. For specific hazard

statements, see sec. 9.

---

2

2 Referenced Documents

2.1 ASTM Standards

D 1066 Practice for Sampling Steam'

D

1129 Terminology Relating to Water'

D 1193 Specification for Reagent Water'

D

2777 Practice for Determination of Precision and Bias of Applicable

Methods of Committee D-19 on Water'

D 3370 Practices for Sampling Water'

D

3856 Guide for Good Laboratory Practices in Laboratories Engaged in

Sampling and Analysis of Water'

D

5810 Standard Practice of Spiking Samples'

D

5847 Standard Practice for Writing Quality Control Specifications for

Standard Test Methods for Water Analysis'

D

5905 Standard Specification for Substitute Wastewater'

F 488 Test Method for Total Bacterial Count in Water2

2.2 EPA 40 CFR Ch.1 (7-1-92 Edition), Pt 136, App. B, page 565 — 567: Definition

and Procedure for the Determination of the Method Detection Limit-Revision 1.11.

2.3 Draft Protocol for EPA Approval of New Methods for Organic and Inorganic

Analytes in Wastewater and Drinking Water, dated Mar 1999, EPA-821-B-98-003.

3 Terminology

3.1 Definitions - For definitions of terms used in this test method, refer to Terminology

D1129.

3.2 Description of Terms Specific to This Test Standard:

3.2.1 Capillary Ion Electrophoresis -- an electrophoretic technique in which an UV

absorbing electrolyte is placed in a 50 pm to 75 pm fused silica capillary.

Voltage is applied across the capillary causing electrolyte and anions to

migrate towards the anode and through the capillary's UV detector window.

Anions are separated based upon the their differential rates of migration in the

electrical field. Anion detection and quantitation are based upon the principles

of indirect UV detection.

3.2.2 Electrolyte -- combination of a UV absorbing salt and an electroosmotic flow

modifier placed inside the capillary, used as a carrier for the analytes, and for

detection and quantitation. The UV absorbing portion of the salt must be anionic

and have an electrophoretic mobility similar to the analyte anions of interest.

3.2.3 Electroosmotic Flow (EOF) -- the direction and velocity of electrolyte solution

flow within the capillary under an applied electrical potential (voltage); the

velocity and direction of flow is determined by electrolyte chemistry, capillary

wall chemistry, and applied voltage.

3.2.4 Electroosmotic Flow Modifier (OFM) -- a cationic quaternary amine in the electrolyte

that dynamically coats the negatively charged silica wall giving it a net positive

charge. This reverses the direction of the electrolyte's natural electroosmotic flow

and directs it towards the anode and detector. This modifier augments anion

migration and enhances speed of analysis. Its concentration secondarily effects

anion selectivity and resolution. See Fig. 7.

1)

Annual Book of ASTM Standards, Vol. 11.01

2)

Annual Book of ASTM Standards, Vol. 11.02

---

3

3.2.5 Electrophoretic Mobility -- the specific velocity of a charged analyte in the

electrolyte under specific electroosmotic flow conditions. The mobility of an

analyte is directly related to the analyte's equivalent ionic conductance and

applied voltage, and is the primary mechanism of separation.

3.2.6 Electropherogram -- a graphical presentation of UV detector response versus

time of analysis; the x axis is migration time which is used to qualitatively identify

the anion, and the y axis is UV response which can be converted to time

corrected peak area for quantitation.

3.2.7 Hydrostatic Sampling -- a sample introduction technique in which the capillary

with electrolyte is immersed in the sample, and both are elevated to a specific

height, typically 10 cm, above the receiving electrolyte reservoir for a preset

amount of time, typically less than 60 s. Nanolitres of sample are siphoned into

the capillary by differential head pressure and gravity.

3.2.8 Indirect UV Detection -- a form of UV detection in which the analyte displaces an

equivalent net charge amount of the highly UV absorbing component of the

electrolyte causing a net decrease in background absorbance. The magnitude

of the decreased absorbance is directly proportional to analyte concentration.

Detector output polarity is reversed in order to obtain a positive mV response.

3.2.9.Midpoint of Peak Width -- CIE peaks are typically asymmetrical with the peak

apex shifting with increasing concentration, and peak apex may not be

indicative of true analyte migration time. Midpoint of peak width is the midpoint

between the analyte peak's start and stop integration, or the peak center of

gravity.

3.2.10 Migration Time -- the time required for a specific analyte to migrate through the

capillary to the detector. The migration time in capillary ion electrophoresis is

analogous to retention time in chromatography.

3.2.11 Time Corrected Peak Area -- normalized peak area; peak area divided by

migration time. CE principles state that peak area is dependent upon migration

time, i.e. for the same concentration of analyte, as migration time increases

(decreases) peak area increases (decreases). Time corrected peak area

accounts for these changes.

4 Summary of Test Method

4.1 Capillary ion electrophoresis, see Fig. 7 through Fig. 10, is a free zone

electrophoretic technique optimized for the determination of anions with molecular

weight less than 200. The anions migrate and are separated according to their

mobility in the electrolyte when an electrical field is applied through the open

tubular fused silica capillary. The electrolyte's electroosmotic low modifier

dynamically coats the inner wall of the capillary changing the surface to a net

positive charge. This reversal of wall charge reverses the natural EOF. The

modified EOF in combination with a negative power supply augments the mobility

of the analyte anions towards the anode and detector achieving rapid analysis

times. Cations migrate in the opposite direction towards the cathode and are

removed from the sample during analysis. Water and other neutral species move

toward the detector at the same rate as the EOF. The neutral species migrate

slower than the analyte anions and do not interfere with anion analysis. See Fig.

7 and 8.

---

4

4.2 Sample is introduced into the capillary using hydrostatic sampling. The inlet of the

capillary containing electrolyte is immersed in the sample and the height of the

sample raised 10 cm for 30 s where low nanolitre volumes are siphoned into the

capillary. After sample loading, the capillary is immediately immersed back into

the electrolyte. The voltage is applied initiating the separation process.

4.3 Anion detection is based upon the principles of indirect UV detection. The UV

absorbing electrolyte anion is displaced charge-for-charge by the separated

analyte anion. The analyte anion zone has a net decrease in background

absorbance. This decrease in UV absorbance is quantitatively proportional to

analyte anion concentration. See Fig. 9. Detector output polarity is reversed to

provide positive mV response to the data system, and to make the negative

absorbance peaks appear positive.

4.4 The analysis is complete once the last anion of interest is detected. The capillary

is automatically vacuum purged by the system of any remaining sample, and

replenished with fresh electrolyte. The system is now ready for the next analysis.

5 Significance and Use

5.1 Capillary ion electrophoresis provides a simultaneous separation and

determination of several inorganic anions using nanolitres of sample in a single

injection. All anions present in the sample matrix will be visualized yielding an

anionic profile of the sample.

5.2 Analysis time is less than 5 min with sufficient sensitivity for drinking water, and

wastewater applications. Time between samplings is less than 7 minutes allowing

for high sample throughput.

5.3 Minimal sample preparation is necessary for drinking water and wastewater

matrices. Typically only a dilution with water is needed.

5.4 This test method is intended as an alternative to other multi-analyte methods and

various wet chemistries for the determination of inorganic anions in water and

wastewater. Compared to other multi-analyte methods the major benefits of CIE

are speed of analysis, simplicity, and reduced reagent consumption and operating

costs.

6 Interferences

6.1 Analyte identification, quantitation, and possible comigration occur when one

anion is in significant excess to other anions in the sample matrix. For two

adjacent peaks, reliable quantitation can be achieved when the concentration

differential is less than 100:1. As the resolution between two anion peaks

increase so does the tolerated concentration differential. In samples containing

1000

mg/L CI,

1 mg/L SO

4

can be resolved and quantitated, however, the high CI

will interfere with Br and NO

2

quantitation.

6.2 Dissolved carbonate, detected as HCO3

1

, is an anion present in all aqueous

samples, especially alkaline samples. Carbonate concentrations greater than 500

mg/L will interfere with

PO4

quantitation.

6.3 Monovalent organic acids, except for formate, and neutral organics commonly

found

in wastewater migrate later in the electropherogram, after carbonate, and

do not interfere. Formate, a common organic acid found in environmental

samples, migrates shortly after fluoride but before phosphate. Formate

'concentrations greater than 5 mg/L will interfere with fluoride identification and

quantitation. Inclusion of 2 mg/L formate into the Mixed Anion Working Solution

aids in fluoride and formate identification and quantitation.

---

5

6.4 Divalent organic acids usually found in wastewater migrate after phosphate. At

high concentrations, greater than 10 mg/L, they may interfere with phosphate

identification and quantitation.

6.5 Chlorate also migrates after phosphate and at concentrations greater than 10

mg/L will interfere with phosphate identification and quantitation. Inclusion of 5

mg/L chlorate into the Mixed Anion Working Solution aids in phosphate and

chlorate identification and quantitation.

6.6 As analyte concentration increases, analyte peak shape becomes asymmetrical.

If adjacent analyte peaks are not baseline resolved, the data system will drop a

perpendicular between them to the baseline. This causes a decrease in peak

area for both analyte peaks and a low bias for analyte amounts. For optimal

quantitation, insure that adjacent peaks are fully resolved, if they are not, dilute

the sample 1:1 with water.

6.7 Samples containing high levels of TOC, total organic carbon, may effect the

observed analyte migration times. The TOC binds to the capillary surface

decreasing the EOF and increasing analyte migration times. Refer to Figure 7.

However, the change in EOF does not effect analyte selectivity. Analytes are

identified using normalized analyte migration times with respect to a reference

peak, chloride, always the first peak in the electropherogram. The surface can be

regenerated with a 5 minute wash with 500 mM Na0H.

7

.Apparatus

7.1 Capillary Ion Electrophoresis System -- the system consists of the following

components, as shown in Fig. 10, or equivalent:3

7.1.1

High Voltage Power Supply --

capable of generating voltage (potential)

between 0 and minus 30 kV relative to ground with the capability working in a

constant current mode.

7.1.2

Covered Sample Carousel --

to prevent environmental contamination of the

samples and electrolytes during a multi-sample batch analysis.

7.1.3

Sample Introduction Mechanism –

capable of hydrostatic sampling technique,

using gravity, positive pressure, or equivalent.

7.1.4

Capillary Purge Mechanism --

to purge the capillary after every analysis with

fresh electrolyte to eliminate any interference from the previous sample matrix,

and to clean the capillary with other reagents, such as sodium hydroxide.

7.1.5 UV Detector --

having the capability of monitoring 254 nm, or equivalent, with a

time constant of 0.3 s.

7.1.6

Fused Silica Capillary --

a 75 pm (inner diameter) x 375 pm (outer diameter) x

60 cm (length) having a polymer coating for flexibility, and a non-coated

section to act as the cell window for UV detection.''

7.1.7

Constant Temperature Compartment --

to keep the samples, capillary, and

electrolytes at constant temperature.

3) Available from Waters, 34 Maple St., Milford, Ma., 01757, 800/252-4752.

---

6

7.2 Data System -- computer system that can acquire data at 20 points per second

minimum, express migration time in minutes to 3 decimal places, use midpoint of

the analyte peak width, or center of gravity, to determine the analyte migration

time, use normalized migration times with respect to a reference peak for

qualitative identification, use time corrected peak area response for analyte

quantitation, and express results in concentration units.3

Note 2: It is recommended that integrators or standard chromatographic data processing not be used with

this test method.

7.3 Anion Exchange Cartridges in the Hydroxide form.4

7.4 Plastic Syringe -- 20 mL, Disposable.

7.5 Vacuum Filtration Apparatus -- capable for filtering 100 mL of reagent through a

0.45 gm aqueous filter.

8 Reagents and Materials

8.1 Purity of Reagents: -- Unless otherwise indicated, it is intended that all reagents

shall conform to the reagent grade specification of the Analytical Reagents of the

American Chemical Society, where such specifications are available. Other

grades may be used, provided it is first ascertained that the reagent is of sufficient

high purity to permit its use without lessening the performance or accuracy of the

determination. Reagent chemicals shall be used for all tests.

Note 3: Calibration and detection limits of this method are biased by the purity of the reagents.

8.2 Purity of Water:-- Unless otherwise indicated, references to water shall be

understood to mean Type I reagent water conforming or exceeding specification

D1193. Freshly drawn water should be used fcr preparation of all stock and

working standards, electrolytes, and solutions. Performance and detection limits

of this method are limited by the purity of reagent water, especially TOC.

8.3 Reagent Blank: Reagent Water or any other solution used to preserve or dilute

the sample.

8.4 Individual Anion Solution, Stock:

Note 4: It is suggested that certified individual

1000

mg/L anion standards be purchased for use

with this test method.

Note 5: All weights given are for anhydrous or dried salts. Must account for reagent purity to

calculate true value concentration. Certify against NIST traceable standards.

8.4.1 Bromide Solution, Standard (1.0 mL = 1.00

MCI

Bromide):

Dry approximately 0.5 g of sodium bromide (NaBr) for 6 h at 150°C and cool in

a desiccator. Dissolve 0.128 g of the dry salt in a 100 mL volumetric flask with

water, and fill to mark with water.

4)

Available from Alltech Associates, P/N 30254, 2051 Waukegan Rd, Deerfield IL, 60015, 847/948-

8600.

5)

Reagent Chemicals, American Chemical Society Specifications,

Am. Chem. Soc., Washington, DC

For suggestions on the testing of reagents not listed by the American Chemical Society, see

Analar

Standards for Laboratory Chemicals,

BDH Ltd., Poole, Dorset. U.K., and the United States

Pharmacopeia and National Formulary, U.S.

Pharmacopoeia Convention, Inc. (USPC), Rockville,

Md.

6)

Although the reagent water may exceed

D1193 specification, the reagent water needs to be

periodically tested for bacterial contamination. Bacteria and their waste products may adversely

affect system performance. As a guide, ASTM type IA water specifies a total bacteria count of 10

colonies/L. Refer to Test Method F 488 for analysis procedure.

---

7

8.4.2 Chloride Solution, Standard (1.0 mL = 1.00 ma Chloride):

Dry approximately 0.5 g of sodium chloride (NaCI) for 1 h at 100°C and cool in

a desiccator. Dissolve 0.165 g of the dry salt in a 100 mL a volumetric flask

with water, and fill to mark with water.

8.4.3 Fluoride Solution, Standard (1.0 mL = 1.00 mq Fluoride):

Dry approximately 0.5 g of sodium fluoride (NaF) for 1 h at 100°C and cool in,a

desiccator. Dissolve 0.221 g of the dry salt in a 100 mL volumetric flask with

water, and fill to mark with water.

8.4.4 Formate Solution, Standard (1.0 mL = 1.00 mg Formate):

Dissolve 0.151 g of sodium formate in a 100 mL volumetric flask with water, and fill

to mark with water.

8.4.5 Nitrate Solution, Standard (1.0 mL = 1.00

ma

Nitrate):

Dry approximately 0.5 g of sodium nitrate (NaNO3)

for 48 h at 105°C and cool

in a desiccator. Dissolve 0.137 g of the dry salt in a 100 mL volumetric flask

with water, and fill to mark with water.

8.4.6 Nitrite Solution, Standard (1.0 mL = 1.00 mg Nitrite):

Dry approximately 0.5 g of sodium nitrite (NaNO2)

for 24 h in a desiccator

containing concentrated sulfuric acid. Dissolve 0.150 g of the dry salt in a 100

mL volumetric flask with water, and fill to mark with water. Store in a sterilized

glass bottle. Refrigerate and prepare monthly.

Note 6: Nitrite is easily oxidized, especially in the presence of moisture. Use only fresh

reagent.

Note 7: Prepare sterile bottles for storing nitrite solutions by heating for 1 h at 170

0C

in an air

oven.

8.4.7 Ortho-Phosphate Solution, Standard (1.0 mL = 1.00 ma o-Phosphate):

Dissolve 0.150 g of anhydrous dibasic sodium phosphate (Na

2HPO4)

in a 100

mL volumetric flask with water, and fill to mark with water.

8.4.8 Sulfate Solution, Standard (1.0 mL = 1.00 mg Sulfate):

Dry approximately 0.5 g of anhydrous sodium sulfate (Na

2SO4)

for 1 h at

110°C and cool in a desiccator. Dissolve 0.148 g of the dry salt in a 100 mL

volumetric flask with water, and fill to mark with water

8.5 Mixed Anion Solution, Working: Prepare a 0.2 mg/L and at least 3 different

working standards concentrations for the analyte anions of interest bracketing the

desired range of analysis, typically between 0.2 and 50 mg/L, and add 2 mg/L

formate to all standards. Add an appropriate aliquot of Individual Anion Stock

Solution (8.4) to a pre-rinsed 100 mL volumetric flask, and dilute to 100 mL with

water.

Note 8: Use 100 p.1_ of Individual Anion Stock Solution (8.4) per 100 mL for 1 mg/L anion.

Note 9: Anions of no interest may be omitted.

Note 10: The mid-range Mixed Anion Solution, Working may be used for the determination of

migration times and resolution described in 12.1.

8.6 Calibration Verification Solution (CVS): A solution formulated by the laboratory of

mixed analytes of known concentration prepared in water. The CVS solution must

be prepared from a different source to the calibration standards.

8.7 Performance Evaluation Solution (PES): A solution formulated by an independent

source of mixed analytes of known concentration prepared in water. Ideally, the

PES solution should be purchased from an independent source.

---

8

8.8 Quality Control Solution (QCS): A solution of known analyte concentrations added

to a synthetic sample matrix such as Substitute Wastewater that sufficiently

challenges the Test Method.

8.9 Buffer Solution (100 mM CHES / 1 mM Calcium Gluconate): Dissolve 20.73 g of

CHES (2-[N-Cyclohexylamino]-Ethane Sulfonic Acid) and 0.43 g of Calcium

Gluconate in a 1 L volumetric flask with water, and dilute to 1 L with water. This

concentrate may be stored in a capped glass or plastic container for up to 1year.

8.10 Chromate Concentrate Solution (100 mM Sodium Chromate): Dissolve 23.41 g of

sodium chromate tetrahydrate (Na

2Cr04•4 H

2

0) in a 1 L volumetric flask with

water, and dilute to 1 L with water. This concentrate may be stored in a capped

glass or plastic container for up to 1 year.

8.11 OFM Concentrate Solution (100 mM Tetradecyltrimethyl Ammonium Bromide):

Dissolve 33.65 g of Tetradecyltrimethyl Ammonium Bromide (TTABr) in a 1 L

volumetric flask with water, and dilute to 1 L with water. Store this solution in a

capped glass or plastic container for up to 1 year.

Note 11: TTABr needs to be converted to the hydroxide form

7(TTAOH)

for use with this test method.

TTAOH is commercially available as 100 mM TTAOH which is an equivalent substitute.

8.12 Sodium Hydroxide Solution (500 mM Sodium Hydroxide)-- Dissolve 20 g

- of sodium

hydroxide (NaOH) in a 1 L

plastic volumetric flask with water, and dilute to 1 L with

water.

8.13 Electrolyte Solution, Working (4.7 mM Chromate / 4 mM TTAOH / 10 mM CHES /

0.1 mM Calcium Gluconate) : Wash the anion exchange cartridge in the hydroxide

form (7.3) using the 20 mL plastic syringe (7.4) with 10 mL of 500 mM NaOH

(8.12) followed by

10

mL of water. Discard the washings. Slowly pass 4 mL of the

100 mM TTABr Solution (8.11) through the cartridge into a 100 mL volumetric

flask. Rinse the cartridge with 20 mL of water, adding the washing to the

volumetric flask.

Note 12: The above procedure is used to convert the TTABr to TTAOH, which is used in the

electrolyte. If using commercially available 100 mM TTAOH, the above conversion step is not

necessary; substitute 0.5 mL of 100 mM TTAOH and continue below

Into the 100 mL volumetric flask add 4.7 mL of Chromate Concentrate Solution

(8.10) and 10 mL of Buffer solution (8.9). Mix and dilute to 100 mL with water.

The natural pH of the electrolyte should be 9 ± 0.1. Filter and degas using the

vacuum filtration apparatus. Store the any remaining electrolyte in a capped

glass or plastic container at ambient temperature. The electrolyte is stable for 1

year.

7)

Available from Waters Corp. as lonSelect 100mM OFM Hydroxide Concentrate, 100 mM

TTAOH, P/N 49387.

8)

Availiable from Waters Corp. as lonSelect High Mobility Anion Electrolyte, P/N 49385.

---

9

9 Precautions

9.1 Chemicals used in this test method are typical of many useful laboratory

chemicals, reagents and cleaning solutions, which can be hazardous if not

handled properly. Refer to Guide D 3856.

9.2 It is the responsibility of the user to prepare, handle, and dispose of chemical

solutions in accordance with all applicable federal, state, and local regulations.

9.3

Warning --

This capillary electrophoresis method uses high voltage as a means

for separating the analyte anions, and can be hazardous if not used properly.

Use only those instruments that have all proper safety features.

10 Sampling

10.1 Collect samples in accordance with Practice D 3370.

10.2 Rinse samples containers with sample and discard to eliminate any

contamination from the container. Fill to overflowing and cap to exclude air.

10.3 Analyze samples as soon as possible after collection. For nitrite, nitrate, and

phosphate refrigerate the sample at 4°C after collection. Warm to room

temperature before dilution and analysis.

10.4 At the lab, filter samples containing suspended solids through a pre-rinsed 0.45

pm aqueous compatible membrane filter before analysis.

10.5 If sample dilution is required to remain within the scope of this Test Method,

dilute with water only.

11 Preparation of Apparatus

11.1 Set up the CE and data system according to the manufacturer's instructions.

11 2 Program the CE system to maintain a constant temperature of 25° ± 0.5°C; or

5°C above ambient laboratory temperature. Fill the electrolyte reservoirs with

fresh chromate electrolyte working solution (8.13), and allow 10 min for thermal

equilibration.

11.3 Condition a new capillary (7.1.6) with 500 mM NaOH Solution (8.12) for 5 min

followed by water for 5 min. Purge the capillary with electrolyte (8.13) for 3 min.

11.4 Apply 15 kV of voltage and test for current. The current should be 14 ± 1 IA. If

no current is observed, then there is a bubble and/or blockage in the capillary.

Degas the chromate electrolyte working solution and retry. If still no current,

replace the capillary.

11.5 Set the UV detector to 254 nm detection, or equivalent. Zero the detector to

0.000 absorbance. UV offset is less then 0.1 AU.

11.6 Program the CE system for constant current of 14 gA.

11.7 Program the CE system for a hydrostatic sampling of 30 s. Approximately 37nL

of sample is siphoned into the capillary. Different sampling times may be used

provided that the samples and standards are analyzed identically.

11.8 Program the CE system for a 1 min purge with the chromate electrolyte working

• solution between each analysis. Using a 15 psi vacuum purge mechanism, one

60 cm capillary volume can be displaced in 30 s.

---

10

11.9 Program the data system for an acquisition rate of at least 20 points per s.

Program the data system to identify analyte peaks based upon normalized

migration time using CI as the reference peak, and to quantitate analyte peak

response using time corrected peak area.

Note 13: Under the analysis conditions CI is always the first peak in the electropherogram, and

can be used as a migration time reference peak.

12 Calibration

12.1 Determination of Migration Times-- Calibrate Daily. The migration time of an

anion is dependent upon the electrolyte composition, pH, capillary surface and

length, applied voltage, the ionic strength of the sample, and temperature. For

every fresh electrolyte determine the analyte migration time, in min to the third

decimal place, of the mid-range mixed anion standard working solution (8.5),

described in Sec 11. Use the mid-point of analyte peak width as the determinant

of analyte migration time.

Note 14: Analyte peak apex may be used as the migration time determinant, but potential analyte

misidentification may result with asymmetrical peak shape at high analyte concentrations.

12.2 Analyze the blank (8.3), a 0.2 mg/L, and at least 3 working mg/L solutions (8.5),

using the set-up described in sec 11. For each anion concentration (X-axis) plot

time corrected peak area response (Y-axis). Determine the best linear

calibration line through the data points, or use the linear regression calibration

routine (1/X Weighting and Linear Through Zero) available in the data system.

Note 15: Do not use peak height for calibration. Peak area is directly related to migration time, i.e.

for the same analyte concentration, increasing migration time gives increasing peak area.

Note 16: EPA recommends calibration at the minimum concentration of 0.2 mg/L plus 3 additional

points.

The r2

(coefficient of determination) values should be greater than 0.995; typical

r2

values obtained from the interlaboratory collaborative are given in Table A2.

12.3 Calibrate daily and with each change in electrolyte, and validate by analyzing the

CVS solution (8.6) according to procedure in Sec16.4.

12.4 After validation of linear multiple point calibration, a single point calibration

solution can be used between 0.2 and 50 mg/L for recalibration provided the

quality control requirements in Sec 16.4 are met.

13 Procedure

13.1 Dilute the sample, if necessary with water, to remain within the scope (Sec 1.2,

1.3) and calibration of this test method. Refer to A1.5.1.

13.2 Analyze all blanks (8.3), standards (8.5), and samples as described in Sec 11

using the quality control criteria described in Sec 16.5 to 16.9. Refer to Fig. 1

through 5 for representative anion standard, detection limit standard, substitute

wastewater, drinking water, and wastewater electropherograms.

13.3 Analyze all blanks, calibration standards, samples, and quality control solutions

in singlicate. Perform at least one matrix spike analysis in duplicate as part of

the QC protocol, Sec 16.7. Optional: Duplicate analyses are preferred due to

short analysis times.

Note 17: Collaborative data was acquired, submitted and evaluated as the average of duplicate

samplings.

---

11

13.4 After 20 sample analyses, or batch, analyze the QCS solution (8.8). If

necessary, recalibrate using a single mixed anion standard working solution

(8.5), and replace analyte migration time.

Note 18: A change in analyte migration time of the mixed anion standard working solution by more

than +5% suggests that components in the previously analyzed sample matrices have

contaminated the capillary surface. Refer to

sec

6.7. Continue but wash the capillary with

NaOH solution (8.12) before the next change in electrolyte.

14 Calculation

14.1 Relate the time corrected peak area response for each analyte with the

calibration curve generated in section 12.2 to determine mg/L concentration of

analyte anion. If the sample was diluted prior to analysis, then multiply mg/L

anion by the dilution factor to obtain the original sample concentration, as

follows:

Original Sample mg/L Analyte = (A x SF) where;

A = analyte concentration determined from the calibration curve, in mg/L,

SF = scale or dilution factor.

15 Report Format

15.1 The sample analysis report should contain the sample name, analyte anion

name, migration time reported to 3 decimal places, migration time ratio, peak

area, time corrected peak area, sample dilution, and original solution analyte

concentration. Optional: Report analysis method parameters, date of sample

data acquisition, and date of result processing for documentation and validation

purposes.

16 Quality Control

16.1 Before this test method is applied to the analysis of unknown samples, the

analyst should establish quality control according to procedures recommended in

Practice D5847, and Guide D5810.

16.2 The laboratory using this test must perform an initial demonstration of laboratory

capability according to procedures outlined in Standard Practice D5847, and

Appendix C.

Note 19: Certified Performance Evaluation Solutions (PES) and QC Solutions (QCS and CVS) are

commercially available, and recommended.

16.3 Initial Demonstration of Performance: Analyze seven replicates of a Performance

Evaluation Solution (PES, 8.7). Calculate analyte concentration mean and

standard deviation of the seven replicates and compare to the precision and

Initial %Recovery for the analyte in reagent water given in Table 8.

16.3.1 Repeat the 7 replicate analysis protocol before using a freshly prepared QVS

solution (8.6) and QCS solution (8.8) for the first time. Calculate the standard

deviation and compare with previous results using the student t-test. If no

significant difference is noted then use the combined standard deviation to

determine the QC limits, for the QVS and QCS solutions.

16.4 Calibration Verification: After calibration, verify the calibration linearity and

acceptable instrument performance using a Calibration Verification Solution (8.6)

treated as an unknown. If the determined CVS concentrations (8.6) are not

within ± 3 standard deviations of the known true values as described in 16.3.1,

the calibration solutions may be out of control. Reanalyze, and if analyte

concentration still falls outside the acceptable limits, fresh calibration solutions

(8.5) are required. Successful CVS analyte concentration must be confirmed

after recalibration before continuing with the Test Method.

---

12

16.5 Analyze a reagent blank (8.3) with each batch to check for contamination

introduced by the laboratory or use of the Test Method.

16.6 Quality Control Solution: Analyze one QCS (8.8) after 20 samples, or batch. The

analyte concentrations for the QCS should fall within the lower limit (LL) and

upper limits (UL) given in Table 8.

16.7 Matrix Spike Recovery: One Matrix Spike (MS) must be analyzed in duplicate

with each batch of samples to test method recovery and relative %difference

between them. Spike a portion of one sample from each batch with a known

concentration of analyte, prepared in accordance with Guide D3856. The

cY0

recovery of the spike should fall within the MS/MSD lower and upper limits, and

the Relative %Difference given in Table 8 for the appropriate sample matrix. If it

does not, an interference may be present and the data for the set of similar

samples matrices must be qualified with a warning that the data are suspect, or

an alternate test method should be used. Refer to Guide D5810.

16.7.1 If the known analyte concentration is between 15 and 50 mg/L, then spike the

sample solution to increase analyte concentration by 50%.

16.7.2 If the known analyte concentration is between 2 mg/L and 15 mg/L, then

spike the sample solution to increase analyte concentration by 100%, but not

less than 2 mg/L.

16.7.3 If the known analyte concentration is less than 2 mg/L, then spike the sample

solution with 1 mg/L, 5 times the ML.

16.7.4 Calculate the percent recovery of the spike using the following formula:

% Recovery = 100 [A (Vs + V) - B Vs] / C V where

A = Analyte Concentration (mg/L) in Spiked Sample

B = Analyte Concentration (mg/L) in Unspiked Sample

C = Concentration (mg/L) of Analyte in Spiking Solution

Vs = Volume (mL) of Sample Used

V = Volume (mL) Added with Spike.

Evaluate performance according to Practice D5847.

16.8 Method Precision: One unknown sample should be analyzed in triplicate with

each batch to test method precision. Calculate the standard deviation and use

the F-Test to compare with the single operator precision given in Tables 1

through 7 for the equivalent analyte concentration and matrix type. Evaluate

performance according to Practice D5847.

16.9 The laboratory may perform additional quality control as desired or appropriate.

17 Precision and Bias

17.1 The precision and bias data presented in this test method meet the requirements

of Practice 2777-98, and are given in Tables 1 through 7. The full Research

Report, RR# D19-1165, can be obtained from ASTM Headquarters.

17.2 This test method interlaboratory collaborative was performed by 11 laboratories

using one operator each. Four Youden Pair spike concentrations for the 7

analytes anions yielding 8 analyte concentration levels. Test data was submitted

for 11 Reagent Waters, 11 Substitute Wastewaters, 15 Drinking Waters, and 13

Wastewater sample matrices.

---

13

17.3 All data given in this method was quantitated using non-weighted linear

calibration through zero, except where noted.

17.4 The precision, bias, and matrix recovery of this test method per anion analyte in

the 4 tested sample matrices are based upon the analyte true value, calculated

using weight, volume, and purity. True value spiking solution concentrations are

given in Table A4.

17.5 The bias and matrix recovery statements for less than 2 mg/L of chloride, sulfate,

and nitrate in naturally occurring sample matrices may be misleading due to

spiking of small analyte concentration into a high naturally occurring analyte

concentration observed with the matrix blank. The commonly occurring analyte

concentrations observed in the sample matrix blanks for the naturally occurring

tested matrices are given in Table A5.

17.6 The high nitrate bias and %recovery noted for the 0.5 mg/L NO

3

spike solution

are attributed to the spiking solution containing 50 mg/L nitrite and 0.5 mg/L

nitrate. Refer to Appendix Table A4, Solution 3. Some of the nitrite converted to

nitrate prior to analysis. Similar NOx conversion effect is observed with the 2

mg/L nitrate and 2 mg/L nitrite spike, Solution 7.

17.7 All collaborative participants used the premade Chromate electrolyte, (lOnSelect

High Mobility Anion Electrolyte, available from Waters Corp.) Ten laboratories

used a Waters CIA Analyzer with Millennium Data Processing Software, and one

laboratory used a Agilent CE System with Diode Array Detector that provided

equivalent results, although different sampling and detection conditions were

necessary for equivalent performance.

Note 20: Refer to reference B1.16 and Agilent (the former HP Company) website for

recommended operating conditions.

18 Key Words

Anion

Capillary Electrophoresis

Drinking Water

Ion Analysis

Reagent Water

Substitute Wastewater

Wastewater

---

14

Appendix A

Mandatory Information

A1.1 All data presented in the following Tables conform and exceed the requirements of

D2777-98. Data from eleven reagent waters, eleven substitute wastewater, fifteen

Drinking Water, and thirteen wastewater sample matrices, were tested using a set

of 4 Youden Pair concentrations for 7 analyte anions. All submitted individual data

points are the average of duplicate samplings.

A1.2 Calibration Linearity

A1.2.1 All laboratories used a provided set of 4 certified, mixed anion calibration

solutions in concentrations between 0.5 mg/L and 50 mg/L, formulated in

random concentrations given in Table A1. They were prepared from certified,

individual 1000 mg/L Stock Standards obtained from APG, Inc, Belpre, Ohio.

No dilution was necessary.

Table Al: Collaborative Calibration Standard, mg/L Concentrations

Analyte Anion

Standard 1

Standard 2

Standard 3

Standard 4

Chloride

50

25

0.5

10

Bromide

0.5

25

10

50

Nitrite

25

0.5

50

10

Sulfate

10

25

0.5

50

Nitrate

25

0.5

50

10

Fluoride

5

0.5

10

25

Phosphate

50

25

0.5

10

A1.2.2 A Linear Through Zero; no weighting regression was used to calculate the

calibration curve. The range coefficient of determination (r

2 ) values obtained

from the collaborative is shown in Table A2

Table A2: Expected Range of (r

2) Coefficient of Determination

Anion

?

/

Average, n=29

Lowest

Highest

Chloride

0.99987

0.99959

0.99997

Bromide

0.99953

0.99878

0.99996

Nitrite

0.99983

0.99961

0.99999

Sulfate

0.99976

0.99901

0.99999

Nitrate

0.99957

0.99840

0.99999

Fluoride

0.99972

0.99797

0.99999

Phosphate

0.99982

0.99942 0.99999

A1.2.3 EPA requires that 1/X weighting be used for calibration. The P & B data

were derived using unweighted calibration. Table A2a shows there is no

significant difference in r2

linearity between these 2 calibration routines.

Table A2a Coefficient of Determination r

2

from a Single Calibration

Analyte

Anion

No Weighted 1/x Weighted

Calibration?

Calibration

Chloride?

0.99994

Bromide

?

0.99942

Nitrite?

0.99975

Sulfate?

0.99971

Nitrate?

0.99975

Fluoride

?

0.99986

Phosphate

?

0.99999

0.99996

0.99923

0.99981

0.99974

0.99974

0.99967

0.99999

---

15

A1.3 Quality Control Solution Preparation

A1.3.1 The Quality Control Solution (QCS) was also used as the Calibration

Verification Solution (CVS).

A1.3.2 Quality Control Solution (QCS) was manufactured, analyzed using ion

chromatography, and certified by APG as 100X concentrate, to replicate typical

Drinking Water concentrations. Required 1:100 dilution with water before

analysis. The QCS analyte concentrations, required control limits, and

interlaboratory determined control limits based upon n# analyses are given in

Table A3.

Table A3: Quality Control Acceptance Limits

Analyte

Anion

True Value

mg/L

Certified

Value

mg/L

Required

99%

Confidence

Interval

Determined

QCS

Mean ± Std Dev,

n = 82

Chloride

Bromide

48.68

0.00

48.61 ± 0.12

0.00

43.99 - 52.96

0.00

47.64

±1.53

0.00

Nitrite

2.87

2.90 ± 0.07

2.39 - 3.26

2.88 ± 0.19

Sulfate

35.69

35.63

±

0.25

29.54 - 40.53

35.02

± 1.21

Nitrate

15.76

15.78 ± 0.15

12.80

-18.39

15.33 ±

4.35

Fluoride

1.69

1.68 ± 0.01

1.49

-1.87

1.67 ± 0.09

Phosphate

5.47

5.55

± 0.12

4.78

- 6.20

5.58

± 0.28

A1.3.3 A single day's QCS was reprocessed using a 1/X weighting linear

calibration and remained within the QC Acceptance Limits.

Table A3a QC Standard Results: Reprocessed Using 1/x Calibration

Analyte

Anion

No Weighted

Calibration

1/x Weighted?

QC Acceptance

Calibration?

99%Conf Interval

Chloride

48.64 ± 1.06

48.77 ± 1.07

43.99

- 52.96

Nitrite

2.93

± .03

2.82

± .03

2.39

- 3.26

Sulfate

34.49 ± .79

34.64 ± .79

29.54

- 40.53

Nitrate

15.28

± .15

15.23

± .18

12.80

-18.39

Fluoride

1.74 ± .02

1.63

± .02

1.49

-1.87

Phosphate

5.75 ± .15

5.77

± .15

4.78

- 6.20

A1.4 Youden Pair Spiking Solution Preparation

A1.4.1 Eight mixed anion, 100X concentrate, spiking solutions were prepared in

accordance with Sec 8.3 (Reagents and Materials) of the test method using

anhydrous sodium salts. The mg/L concentrations of the eight standards

followed the approved Youden Pair design - 0.5 & 0.7, 2 & 3, 15 & 20, 40 & 50

mg/I for all anions except fluoride, which is 0.5 & 0.7, 2 & 3, 7& 10, 20 &

25mg/L. The analyte true value concentrations were randomized among the

eight spiking solutions as described in Table A4.

A1.4.2 A ninth solution containing approximately 10 mg/L of each analyte was

diluted 1:50 with water, and was used for method detection limit calculations.

---

16

Table A4: True Value Youden Pair Spiking mg/L Concentrations

Anion / TV

1

2

3

4

5

6

7

89

Chloride

0.71

2.00

2.98

14.92

39.81

19.91

49.76

0.50

10.20

Bromide

2.00

3.01

14.93

39.81

19.91

49.77

0.70

0.51

10.49

Nitrite

2.98

39.61

19.81

14.86

49.52

0.50

2.00

0.70

9.94

Sulfate

39.60

49.51

0.49

0.70

1.98

2.98

14.86

19.81

10.23

Nitrate

14.92

19.19

39.87

49.78

0.50

0.70

2.00

2.98

10.35

Fluoride

2.00

0.71

0.50

3.00

9.99

6.99

19.98

24.99

10.40

Phosphate

49.51

39.60

19.90

0.50

2.98

1.99

0.69

14.86

10.48

These solutions, kept at ambient temperature, were analyzed before and during

the collaborative to monitor for accuracy and stability. The mg/L True Value in

was used to determine bias, matrix recovery, and the single operator and

interlaboratory precision in the P & B tables per the requirement of D 2777.

Solution 3 and 7 exhibited some conversion of nitrite to nitrate before analysis.

This conversion is evident in the bias and % Recovery for 0.5 mg/L and 2 mg/I

nitrite and nitrate.

A1.5 Sample Matrix Preparation

A1.5.1 All participating laboratories provided and tested reagent water, substitute

wastewater, naturally occurring drinking water, and naturally occurring

wastewater. Before matrix spiking with the Youden Pair solutions, the sample

matrix was evaluated, then appropriately diluted to give the highest anion

concentration below 50 mg/L. The diluted sample matrix was used to dilute

each Youden Pair spiking solution 1:100.

A1.5.2 Reagent Water was used as-is. Substitute wastewater was diluted 1:20

with water. Naturally occurring drinking water was used as-is or diluted 1:5 with

water. Naturally occurring wastewater was diluted between 1:3 and 1:20, except

one which required a 1:1000 dilution due to high chloride.

A1.5.3 Due to the anion content of the naturally occurring drinking water and "real"

wastewater matrices, some of the reported spike matrix results exceeded the

scope of this test method. Linearity and matrix recovery data obtained from the

collaborative indicated that these data are acceptable, and extended the useful

range of this test method.

A1.5.4 Due to the anion content of the naturally occurring sample matrices given

in Table A5, the low concentration bias and recovery may be misleading

because of spiking a low anion concentration increment into a large naturally

occurring concentration of the same anion.

Table A5: Blank Analvte Concentrations for Naturally Occurring Sample Matrices

Data in mg/L

Chloride

Sulfate

Nitrate

Drinking Water

0.7 to 41.9

0.5 to 33.6

0.2 to 6.5

Substitute Wastewater

20.5 to 25.5 3.2 to 4.0

Not Detected

"Real"

0.9 to 43.4

0.5 to 50.4

0.3 to 23.0

Wastewater

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17

A1.6 Test Method Detection Limits:

A1.6.1 Spiking Solution #9, containing 10 mg/L of each analyte, was diluted 1:50 with

water and was used for detection limit calculations. Ten laboratories performed

seven replicate samplings, and the mean and standard deviation from each

laboratory was calculated. The mean time corrected peak area response for the

7 replicates was given the true value of the solution #9, and from a simple

proportion, the standard deviation was calculated as mg/L.

Std Dev, mg/L = (True Value Conc Sol'n #9, mo/L)(Response Std Dev)

Ave Response of Sol'n #9

A1.6.2 Method detection limits (MDL) were derived using "pooled" EPA protocol and

the student t-test at 6 degrees of freedom, as follows;

The method detection limit (MDL) .(3.14)(Std Dev, mg/L).

A1.6.3 The upper and lower confidence limits were calculated as;

95% Confidence Interval:

?

LCL (Lower Confidence Limit) = 0.64 x MDL

UCL (Upper Confidence Limit) = 2.20 x MDL

A1.6.4 Method Detection Limits are given in Table A6.

Table A6: Method Detection Limits

Anion

mg/L Solution?

Method Detection I 95% Confidence interval

Concentration?

MDL, mg/L?

mg/L

Chloride

0.204

0.075

0.048 to 0.165

Bromide

0.210

0.120

0.077 to 0.264

Nitrite

0.199

0.103

0.066 to 0.227

Sulfate

0.205

0.065

0.042 to 0.143

Nitrate

0.207

0.076

0.049 to 0.167

Fluoride

0.208

0.032

0.020 to 0.070

Phosphate

0.210

0.097

0.062 to 0.213

---

18

Table 1

Precision, Bias, and Matrix Recovery for Chloride

Matrix

?

# of

?

True?

Mean

?

Bias vs?

Recovery

?

Interlab

?

Interlab?

Single?

Analyst

Values?

Value?

Result

?

True?

vs True?

Std Dev

?

%RSD

?

Operator

?

%RSD

?

Value?

Value

?

S(t)

?

Std Dev, S(o)

Reagent

?

9?

0.50?

0.55?

0.05?

110.0?

0.11?

19.8

Water?

10

?

0.71?

0.69?

-0.02?

97.2?

0.08

?

11.5?

0.05?

7.5

?

10

?

2.00?

1.97

?

-0.03?

98.5?

0.14?

6.8

?

9

?

2.98?

2.97?

-0.01?

99.7?

0.11?

3.8

?

0.05?

2.1

?

10?

14.92?

14.76

?

-0.16?

98.9?

0.61?

4.2

?

10?

19.91?

19.81?

-0.10

?

99.5?

0.81?

4.1

?0.48?

2.8

?

10?

39.81

?

38.58?

-1.23?

96.9?

1.43?

3.7

?

10?

49.76

?

48.70?

-1.06?

97.9?

1.94?

4.0?

1.36?

3.1

Substitute?

9?

0.50

?

0.46?

-0.04?

92.0?

0.51?

111.1

Wastewater

?

9?

0.71?

0.43?

-0.28?

60.6?

0.69?

160.7

?

0.42?

93.8

9

?

2.00?

1.52?

-0.48?

76.0?

0.68?

45.0

?

9?

2.98?

2.58

?

-0.40

?

86.6?

0.63?

24.5?

0.50?

24.3

?

9?

14.92?

14.29?

-0.63?

95.8

?

1.02?

7.1

?

9

?

19.91

?

18.93?

-0.98?

95.1?

1.24?

6.6

?

0.60?

3.6

?

9?39.81

?

37.34?

-2.47?

93.8?

5.44?

14.6

?

9?

49.76?

47.54?

-2.22?

95.5?

3.13?

6.6

?

4.43?

10.4

Drinking

?

12?0.50

?

0.63?

0.13?

126.0

?

0.67?

106.1

Water?

12

?

0.71?

0.75?

0.04?

105.6?

0.34?

45.5?

0.40?

57.2

?

12?

2.00?

2.15?

0.15?

107.5

?

0.51?

23.6

?

12?

2.98?

2.95?

-0.03?

99.0?

0.39?

13.1?

0.47

?

18.5

?

12?

14.92?

14.54

?

-0.38?

97.5?

0.71?

4.9

?

12?

19.91?

19.09?

-0.82?

95.9?

1.11

?

5.8

?

0.37?

2.2

?

12?

39.81

?

38.38?

-1.43?

96.4?

1.56?

4.1

49.76

?

47.97?

-1.79?

96.4?

2.19?

4.6?

1.26?

3.9

°Real"

?

9

?0.50?

0.42?

-0.08?

84.0

?

0.34?

81.0

Wastewater

?

10?

0.71?

0.47?

-0.24?

66.2?

0.34?

72.6?

0.26?

59.3

?

10

?2.00?

1.56?

-0.44

?

78.0?

0.51?

32.7

?

9?

2.98?

2.78

?

-0.20?

93.3

?

0.19?

6.8?

0.37?

17.3

?

10

?

14.92

?

14.29?

-0.63?

95.8

?

0.63?

4.4

?

10?

19.91?

18.83?

-1.08?

94.6?

0.78?

4.1

?0.46?

2.8

?

9?39.81?

37.01

?

-2.80?

93.0?

2.78?

7.5

?

10?

49.76

?

48.24?

-1.52?

96.9

?

3.15?

6.5?

2.54?

6.0

---

19

Table 2

Precision, Bias, and Matrix Recovery for Bromide

Matrix

?

# of?

True?

Mean?

Bias vs

?

Recovery

?

Interlab

?

Interlab?

Single?

Analyst

Values?

Value

?

Result?

True

?

vs True

?

Std Dev

?

%RSD

?

Operator

?

%RSD

Value?

Value?

S(t)

?

Std Dev, S(o)

Reagent?

10

?

0.51?

0.60?

0.09?

117.6?

0.19

?

31.0

Water?

10?

0.70?

0.83?

0.13?

118.6

?

0.23?

28.2?

0.10?

14.6

?

10?

2.00?

2.06?

0.06?

103.0

?

0.14?

6.6

?

10

?3.01?

2.88?

-0.13?

95.7?

0.23?

7.9?

0.15?

6.3

?

10

?

14.93

?

15.00

?

0.07?

100.5

?

0.58?

3.9

?

10

?

19.91

?

19.32

?

-0.59?

97.0?

0.97?

5.0

?

0.75?

4.4

?

10

?

39.81?

39.66

?

-0.15?

99.6

?

1.24?

3.1

?

10?

49.77

?

50.04

?

0.27?

100.5?

2.94?

5.9?

1.61?3.6

Substitute

?

9?

0.51?

0.67?

0.16?

131.4?

0.19?

28.8

Wastewater

?

9?

0.70?

0.96

?

0.26

?

137.1?

0.21?

21.8?

0.08?

9.3

?

9

?

2.00?

2.14?

0.14?

107.0

?

0.22

?

10.2

?

9

?

3.01?

2.72?

-0.29?

90.4?

0.35?

12.8?

0.17

?

7.0

?

9?

14.93

?

14.70?

-0.23?

98.5

?

0.58?

3.9

?

9

?

19.91

?

18.91

?

-1.00

?

95.0?

2.62?

13.8

?

1.63

?

9.7

?

9?

39.81

?

38.76?

-1.05

?

97.4

?

1.11?

2.9

?

9

?

49.77

?

48.81?

-0.96?

98.1?

1.52?

3.1

?

0.48

?

1.1

Drinking?

13

?

0.51

?

0.58?

0.07?

113.7?

0.25?

43.4

Water

?

13?

0.70?

0.83?

0.13

?

118.6

?

0.22?

26.5?

0.14?

19.9

?

13

?

2.00?

1.98?

-0.02?

99.0?

0.25

?

12.5

?

13

?3.01?

2.56?

-0.45?

85.0?

0.25?

9.7?

0.15?

6.8

?

13

?

14.93

?

14.63?

-0.30?

98.0

?

0.50

?

3.4

?

13?

19.91

?

19.22

?

-0.69

?

96.5?

1.10?

5.7?

0.77?

4.6

?13

?

39.81?

38.97?

-0.84?

97.9?

1.99

?

5.1

?

13?

49.77

?

48.74?

-1.03?

97.9?

1.49?

3.1?1.13?

2.6

"Real"

?

11

?

0.51

?

0.59?

0.08?

115.7?

0.11?

19.3

Wastewater

?

12

?0.70?

0.78?

0.08?

111.4

?

0.19?

24.4?

0.10?

14.0

?

11

?2.00?

2.08?

0.08?

104.0

?

0.13?

6.3

?

12

?3.01?

2.70?

-0.31?

89.7?

0.41?

15.1?

0.27?

11.5

?

12

?

14.93

?

15.16?

0.23?

101.5?

0.90

?

6.0

?

11

?

19.91

?

19.46?

-0.45

?

97.7?

1.63?

8.4?

1.09?

6.3

?

12

?

39.81

?

40.24?

0.43?

101.1?

2.27?

5.7

?

12

?

49.77?

49.97

?

0.20

?

100.4?

2.52?

5.0?

0.91?

2.0

---

20

Table 3

Precision, Bias, and Matrix Recovery for Nitrite

Matrix

?

# of?

True?

Mean

?

Bias vs

?

Recovery?

Interiab?

Interlab?

Single

?

Analyst

Values

?

Value?

Result

?

True?

vs True

?

Std Dev?

%RSD?

Operator?

%RSD

Value?

Value?

S(t)?

Std Dev,

S(o)

Reagent?

9?

0.50?

0.62?

0.12?

124.0

?

0.16?

26.1

Water

?

9?

0.70?

0.72?

0.02

?

102.9

?

0.08?

10.5?

0.05?

7.1

?

10

?

2.00?

1.31?

-0.69?

65.5?

0.25?

19.2

?

10?

2.98?

3.11?

0.13?

104.4?

0.17?

5.4

?

0.13?

6.0

?

10

?

14.86

?

14.70

?

-0.16?

98.9?

0.47?

3.2

?

10

?

19.81?

19.88?

0.07?

100.4

?

0.70?

3.5?

0.27?

1.5

?

10?

39.61

?

39.90?

0.29?

100.7

?

0.88?

2.2

?

10?

49.52?

48.24

?

-1.28?

97.4?

1.34?

2.8

?

1.25?

2.8

Substitute?

9?

0.50?

0.37?

-0.13?

74.0?

0.22?

59.7

Wastewater

?

9?

0.70?

0.59?

-0.11?

84.3?

0.28?

48.1?

0.21?

43.2

?

10?

2.00?

1.25?

-0.75?

62.5?

0.38?

30.8

?

9?2.98?

2.62?

-0.36?

87.9?

0.82?

31.4?

0.43

?

22.1

?

9?

14.86?

14.40

?

-0.46?

96.9?

0.58?

4.0

?

10?

19.81

?

19.50?

-0.31?

98.4?

1.66?

8.5

?

0.81?4.8

?

10?

39.61

?

39.97?

0.36?

100.9

?

2.02

?

5.0

?

9?

49.52?

49.09?

-0.43

?

99.1?

3.03?

6.2

?

2.11?4.7

Drinking

?

11?

0.50

?

0.52

?

0.02?

104.0

?

0.08?

14.4

Water?

12?

0.70?

0.74?

0.04?

105.7?

0.17?

23.3

?

0.09?

13.5

?

12

?

2.00?

1.30

?

-0.70

?

65.0?

0.21?

15.9

?

12?

2.98?

2.97?

-0.01?

99.7?

0.14?

4.6

?

0.16?

7.4

?

11?

14.86?

14.60

?

-0.26?

98.3?

0.40?

2.8

?

11?

19.81

?

19.82?

0.01?

100.1

?

0.59?

3.0

?

0.26?

1.5

?

11?

39.61

?

39.35

?

-0.26?

99.3?

0.99?

2.5

?

12?

49.52

?

49.14?

-0.38?

99.2?

1.93

?

3.9?

0.64?

1.5

"Real"

?

9?0.50?

0.55?

0.05?

110.0?

0.13?

24.5

Wastewater

?

10

?0.70?

0.73

?

0.03

?

104.3?

0.24?

32.9?

0.07?

10.8

?

9?

2.00?

1.27?

-0.73?

63.5?

0.18?

14.2

?

10?

2.98?

2.99?

0.01

?

100.3?

0.19?

6.2

?

0.15?

7.0

?

10

?

14.86?

14.55?

-0.31?

97.9?

0.46?

3.1

?

10?

19.81

?

19.68

?

-0.13?

99.3?

0.71?3.6

?

0.38?

2.2

?

9?

39.61

?

39.21

?

-0.40?

99.0?

1.03?

2.6

?

9?

49.52?

47.27

?

-2.25?

95.5?

3.50?

7.4

?

2.40?

5.6

---

21

Table 4

Precision, Bias, and Matrix Hecovery for Sulfate

Matrix?

# of

?

True?

Mean

?

Bias vs

?

Recovery?

Interlab

?

Interlab?

Single?

Analyst

Values

?

Value?

Result?

True

?

vs True

?

Std Dev

?

%RSD

?

Operator?

%RSD

Value?

Value

?

S(t)?

Std Dev,

S(o)

Reagent

?

9

?

0.49?

0.49?

0.00

?

100.0

?

0.18?

37.5

Water?

10?

0.70?

0.71?

0.01?

101.4

?

0.20?

29.2?

0.05?

8.3

?

10?

1.98?

2.04?

0.06?

103.0

?

0.19?

9.7

?

10?

2.98?

3.09?

0.11?

103.7

?

0.24?

7.9?

0.06?

2.5

?

10

?

14.86

?

14.67?

-0.19?

98.7?

0.57?

4.0

?

10

?

19.81?

19.67?

-0.14?

99.3

?

0.73?

3.8?

0.44

?

2.6

?

10?

39.60?

39.66?

0.06?

100.2

?

0.92?

2.4

?

10

?

49.51?

49.27

?

-0.24

?

99.5

?

1.26?

2.6?

0.49?

1.1

Substitute

?

9?

0.49?

0.38

?

-0.11?

77.6?

0.25?

66.9

Wastewater?

9?

0.70?

0.51?

-0.19?

72.9?

0.08?

16.4

?

0.18?

39.3

?

9?

1.98?

1.83?

-0.15?

92.4?

0.29?

16.2

?

9?

2.98?

2.86?

-0.12?

96.0?

0.31?

11.2?

0.20?

8.6

9?14.86?

14.19

?

-0.67

?

95.5?

1.06?

7.7

?

9

?

19.81

?

19.23?

-0.58?

97.1?

0.97?

5.2

?

0.46?

2.8

?

9?39.60

?

38.45

?

-1.15?

97.1?

1.33?

3.6

?

9

?49.51

?

47.75

?

-1.76?

96.4?

1.43

?

3.1

?0.75?

1.8

Drinking

?

12

?

0.49?

0.41?

-0.08?

83.7?

0.21?

52.8

Water?

12

?

0.70

?

0.41?

-0.29?

58.6?

0.20?

50.3?

0.14

?

34.3

?

13

?1.98?

1.77?

-0.21?

89.4

?

0.53?

30.3

?

13?

2.98

?

2.68?

-0.30?

89.9?

0.42?

16.2?

0.27?

12.1

?

13?

14.86

?

14.25

?

-0.61?

95.9?

1.11?

8.0

?

12?

19.81

?

19.31?

-0.50?

97.5?

1.39?

7.4

?

1.48?

8.9

?

12?

39.60?

38.58

?

-1.02?

97.4

?

1.96?

5.2

?

13?

49.51?

48.43?

-1.08?

97.8?

2.04?

4.3?

1.44

?

3.3

'Real"

?

10?

0.49?

0.37?

-0.12?

75.5?

0.39?

106.4

Wastewater

?

11?

0.70?

0.16

?

-0.54?

22.9?

1.19?

765.2

?

0.47?

179.6

?

11?

1.98?

1.57?

-0.41?

79.3?

0.87?

55.4

?

11

?

2.98

?

2.53?

-0.45?

84.9?

0.64?

25.4?

0.24?

11.9

?

11

?

14.86

?

14.69?

-0.17?

98.9?

1.26

?

8.6

?

10

?

19.81

?

19.38

?

-0.43?

97.8?

0.90?

4.6?

0.57?

3.4

?

11

?

39.60?

38.74?

-0.86?

97.8?

1.71

?

4.4

?

10?

49.51

?

48.36

?

-1.15?

97.7?

1.51?3.1?0.47?

1.1

---

22

Table 5

Precision, Bias, and Matrix Recovery for Nitrate

Matrix?

# of?

True?

Mean

?

Bias vs

?

Recovery

?

interlab

?

Interlab

?

Single?

Analyst

Values?

Value

?

Result?

True?

vs True?

Std Dev?

%RSD?

Operator

?

%RSD

?

Value

?

Value

?

S(t)

?

Std Dev, S(o)

Reagent?

10?

0.50?

1.02?

0.52?

204.00?

0.08?

7.4

Water

?

10

?

0.69?

0.71?

0.02?

102.90?

0.08?

11.6?

0.06?

6.4

?

11

?1.99?

2.83

?

0.84?

142.21

?

0.23?

8.1

?

11?

2.97

?

2.89?

-0.08?

97.31

?

0.18?

6.4?

0.14?

5.0

?

11?

14.91?

14.77

?

-0.14

?

99.06

?

0.44?

3.0

?

11

?

19.18?

19.77?

0.59?

103.08?

0.64

?

3.2

?

0.24?

1.4

?

10?

39.86

?

39.09

?

-0.77?

98.07

?

1.43?

3.7

?

10?

49.77

?

48.93

?

-0.84?

98.31

?

1.72?

3.5?

0.62?

1.4

Substitute?

11?

0.50?

1.18?

0.68?

236.00?

0.41?

34.9

Wastewater?

10?

0.69?

0.55?

-0.14

?

79.71

?

0.30?

55.3?

0.42?

4.9

?

10?

1.99

?

2.70?

0.71?

135.68?

0.42?

15.4

?

10

?

2.97?

2.33

?

-0.64

?

78.45

?

1.10?

47.3?

0.39?

15.4

?

9

?

14.91

?

14.29

?

-0.62?

95.84?

0.78?

5.4

?

10

?

19.18

?

18.69

?

-0.49?

97.45?

1.46?

7.8?

025?

1.5

?

11?

39.86

?

37.70?

-2.16?

94.58?

1.93

?

5.1

?

11?

49.77?

47.78

?

-1.99?

96.00?

2.18?

4.6

?

1.62?

3.8

Drinking?

11?

0.50

?

1.06 .?

0.56?

212.00?

0.19?

18.1

Water

?

11?

0.69?

0.65?

-0.04?

94.20?

0.06?

8.7

?

0.12?

14.4

?

12?

1.99?

3.05?

1.06?

153.27?

0.39?

12.8

?

11?

2.97?

3.01?

0.04?

101.35?

0.22?

7.2

?

0.33

?

10.8

?

12

?

14.91

?

14.69

?

-0.22?

98.52?

0.62?

4.2

?

12?

19.18

?

20.05

?

0.87

?

104.54?

0.88?

4.4

?

0.46?

2.7

?

12?

39.86

?

39.31

?

-0.55?

98.62?

1.67?

4.3

?

12?

49.77?

48.93

?

-0.84?

98.31?

1.43?

2.9

?

0.78?

1.8

"Rear

?

11?

0.50

?

0.94?

0.44?

188.00?

0.80?

84.7

Wastewater

?

10?

0.69?

0.69?

0.00?

100.00?

0.09?

13.3?

0.39?

47.6

?

10

?

1.99?

3.00?

1.01?

150.75?

0.38?

12.7

?

10

?

2.97?

3.01?

0.04?

101.35?

0.20?

6.6?

0.23?

7.8

?

11?

14.91

?

14.52

?

-0.39?

97.38?

0.66

?

4.6

?

11?

19.18

?

19.26

?

0.08

?

100.42?

0.77?

4.0?

0.77?

4.6

?

11?

39.86

?

39.13?

-0.73?

98.17?

1.78

?

4.6

?

11?

49.77?

49.17

?

-0.60

?

98.79?

2.26?

4.6

?

0.93?

2.1

---

23

Table 6

Precision, Bias, andNigi1371<

ecovery for Fluoride

Matrix?

# of?

True

?

Mean

?

Bias vs

?

Recovery?

Interlab

?

Intertab?

Single

?

Analyst

Values?

Value

?

Result

?

True

?

vs True

?

Std Dev?

%RSD?

Operator?

%RSD

?

Value?

Value

?

S(t)?

Std Dev, S(o)

Reagent

?

10?

0.50?

0.51?

0.01?

102.00

?

11.00?

11.4

Water

?

10?

0.71?

0.73

?

0.02?

102.82

?

7.90?

8.1?

0.02?

2.9

?

10

?2.00?

2.05?

0.05

?

102.50

?

3.60?

3.7

?

10?

3.00?

2.96?

-0.04?

98.67

?

4.40?

4.6

?0.09?

3.4

?

10?

6.99?

7.02?

0.03?

100.43?

5.40?

5.6

?

10?

9.99?

9.79?

-0.20?

98.00

?

4.60?

4.8?0.13?

1.6

?

10

?

19.98

?

19.60

?

-0.38?

98.10?

3.80?

3.9

?

10?

24.99

?

24.51

?

-0.48?

98.08?

4.80?

4.9

?0.74?

3.4

Substitute?

10

?0.50?

0.50?

0.00?

100.00?

0.09?

18.0

Wastewater

?

10?

0.71?

0.71?

0.00?

100.00?

0.09?

12.0?

0.01

?

2.3

?

10?

2.00?

1.98?

-0.02?

99.00

?

0.12?

6.0

?

10?3.00?

2.94?

-0.06?

98.00?

0.10?

3.4?

0.06?

2.6

?

10?

6.99?

6.92?

-0.07?

99.00

?

0.28?

4.1

?

9?

9.99

?

9.94?

-0.05?

99.50?

0.46?

4.7?

0.28?

3.3

?

10?

19.98

?

19.67?

-0.31?

98.45

?

0.94?

4.8

?

10?

24.99

?

24.78?

-0.21?

99.16?

1.09?

4.4?

0.63?

2.8

Drinking?

13?

0.50

?

0.48?

-0.02?

96.00

?

0.06?

12.9

Water

?

13

?0.71?

0.68?

-0.03?

95.77?

0.06?

9.5?

0.02?

3.4

?

13?

2.00?

1.96?

-0.04?

98.00

?

0.08?

3.9

?

13?

3.00?

2.90?

-0.10?

96.67

?

0.10?

3.4

?

0.08?

3.5

?

13

?6.99?

6.91?

-0.08?

98.86?

0.25?

3.6

?

13?

9.99?

9.91?

-0.08?

99.20

?

0.37?

3.7?

0.18?

2.2

?

13

?

19.98?

19.94

?

-0.04?

99.80?

0.68?

3.4

?

12?

24.99

?

24.27?

-0.72?

97.12?

1.63?

6.7

?1.30?

5.9

"Rear

?

11

?0.50?

0.47?

-0.03?

94.00?

0.08?

16.9

Wastewater

?

11

?

0.71

?

0.68?

-0.03?

95.77

?

0.08?

11.7?

0.04?

7.6

?

11

?2.00?

1.96?

-0.04

?

98.00?

0.12?

6.3

?

11

?

3.00

?

2.93?

-0.07?

97.67?

0.18?

6.2

?

0.09?

3.5

?

11

?

6.99?

6.85

?

-0.14

?

98.00

?

0.26?

3.8

?

10?

9.99?

9.56?

-0.43?

95.70?

0.73?

7.7?

0.44?

5.3

?

11?

19.98?

20.06

?

0.08?

100.40

?

1.23?

6.1

?

11

?

24.99?

25.12?

0.13?

100.52?

1.34?

5.3?

0.32?

1.4

---

24

Table 7

Precision, Bias, and Matrix Recovery for o-Phosphate

Matrix

?

# of?

True

?

Mean?

Bias vs

?

Recovery?

Interlab?

Interlab

?

Single

?

Analyst

Values?

Value?

Result

?

True

?

vs True?

Std Dev?

%RSD?

Operator

?

%RSD

?

Value?

Value?

S(t)?

Std Dev, S(o)

Reagent

?

10

?

0.50?

0.41?

-0.09?

82.00

?

0.12

?

29.6

Water?

9?

0.69

?

0.51?

-0.18?

73.91

?

0.13?

26.6?

0.03?

7.2

10?

1.99?

1.88?

-0.11?

94.47?

0.16?

8.3

10?

2.98?

2.76?

-0.22?

92.62

?

0.14

?

4.9?

0.08?

3.2

10?

14.86?

14.93

?

0.07?

100.47?

0.64?

4.3

9

?

19.80?

19.76

?

-0.04?

99.80?

1.00?

5.1

?

0.85?

4.9

10?

39.60?

39.79

?

0.19?

100.48?

1.38?

3.5

10?

49.51

?

50.10?

0.59?

101.19

?

1.76?

3.5?

0.72?

1.6

Substitute

?

11

?

0.50?

0.49?

-0.01?

98.00?

0.15?

30.0

Wastewater

?

10

?

0.69

?

0.59?

-0.10?

85.51?

0.17?

28.8?

0.13?

24.4

11?

1.99?

1.92?

-0.07?

96.48?

0.28?

14.6

10?

2.98?

2.89?

-0.09?

96.98

?

0.22?

7.6

?

0.18?

7.5

11?

14.86

?

15.31

?

0.45?

103.03

?

1.74?

11.4

11

?

19.80?

19.78

?

-0.02?

99.90

?

1.16?

5.9?

0.84?

4.8

11

?

39.60?

39.58?

-0.02?

99.95

?

2.72?

6.9

11?

49.51

?

49.19?

-0.32?

99.35

?

3.98?

8.1

?

2.18?

4.9

Drinking

?

12?

0.50?

0.46?

-0.04?

92.00

?

0.14?

30.0

Water?

13?

0.69

?

0.55

?

-0.14?

79.71

?

0.20?

36.3?

0.07?

13.4

13

?

1.99?

1.89?

-0.10?

94.97?

0.22?

11.9

13?

2.98

?

2.87?

-0.11?

96.31?

0.24?

8.5

?

0.07?

2.8

12

?

14.86?

15.09

?

0.23?

101.55?

0.91?

6.1

13

?

19.80

?

20.28

?

0.48?

102.42?

0.96?

4.7?

1.06?

6.0

13?

39.60

?

40.37?

0.77?

101.94?

2.15?

5.3

13

?

49.51

?

50.75?

1.24?

102.50?

3.14?62?

1.03?

2.3

'Real°

?

11

?0.50?

0.43?

-0.07?

86.00?

0.17?

39.1

Wastewater?

11

?0.69?

0.53

?

-0.16?

76.81?

0.24?

46.5?

0.12?

25.8

11?

1.99?

1.72?

-0.27?

86.43

?

0.27?

15.8

11?

2.98?

2.52?

-0.46?

84.56

?

0.48?

19.2?

0.30?

14.0

11

?

14.86

?

14.93?

0.07?

100.47

?

0.91?

6.1

11?

19.80?

19.90?

0.10?

100.51?

1.35?

6.8

?

0.91?

5.2

11?

39.60

?

38.98?

-0.62

?

98.43?

1.45?

3.7

10?

49.51

?

48.26?

-1.25?

97.48?

1.80?

3.7?

0.82?

1.9

---

25

Table 8

QC Acceptance Criteria

Analyte

Matrix

Precision

% RSD

Average

%Recovery

Initial

LL - UL

Ongoing

LL - UL

MS/MSD

LL - UL

MS/MSD

RPD

Chloride

RW

6.30

98.5

90.8 - 106.2

88.7 - 108.3

89.4 - 107.5

12.0

DW

10.00

97.0

84.0 - 110.0

81.1 - 113.0

81.9 - 112.5

18.6

WW

7.00

92.8

83.0 -102.6

81.4 - 104.2

81.8 - 103.8

13.2

Bromide

RW

10.10

99.7

92.2 - 107.2

86.7 - 112.7

88.5 - 111.0

19.2

DW

12.70

95.8

85.9 -105.6

79.8 - 111.8

81.8 - 109.8

23.3

WW

14.40

99.2

87.2 - 111.2

80.1 -118.3

82.4 - 116.0

26.9

Nitrite

RW

6.40

100.6

95.1 - 106.0

91.9 - 109.2

92.8 - 108.3

12.1

DW

4.30

99.6

92.4 - 106.7

91.5 - 107.7

91.8 - 107.4

8.1

WW

4.90

98.9

91.3 - 106.5

90.2 - 107.6

90.5 - 107.3

9.2

Sulfate

RW

9.40

100.4

90.9 -109.9

86.9 - 113.9

88.2 - 112.6

17.9

DW

16.1

95.6

82.6 - 108.6

74.9 - 116.2

77.5 - 113.7

29.7

WW

19.60

95.3

78.9 - 111.7

70.1 - 120.5

72.6 - 118.0

36.9

Nitrate

RW

8.40

99.5

93.1 - 105.9

88.6 - 110.4

90.0 - 108.9

15.9

DW

9.40

100.2

93.0 -107.4

88.0 -112.4

89.4 - 111.0

17.4

WW

6.70

99.1

90.7 - 107.6

88.6 -109.7

89.2 - 109.1

12.4

Fluoride

RW

7.90

99.5

92.2 - 106.7

88.7 - 110.3

89.8 - 109.1

14.9

DW

4.86

98.3

91.9 - 104.8

90.5 - 106.2

90.9 - 105.7

9.0

WW

7.90

98.5

90.0 - 107.1

87.0 - 110.1

88.0 - 109.1

14.7

Phosphate

RW

10.60

98.2

91.9 - 104.5

85.4 - 111.0

87.4 - 109.0

20.1

DW

9.40

100.2

89.3 - 111.1

85.8 - 114.6

87.0 - 113.4

17.4

WW

16.90

94.6

81.5 - 107.7

73.5 - 115.8

76.1 - 113.1

31.5

All data determined as spike recovery from ASTM method validation and EPA Tier 3 Criteria

Reagent water (RW) data between 0.5 and 50 mg/L, except Fluoride 0.5 and 25 mg/L

consisting of 4 Youden Pairs

Drinking (DW) and Wastewater (WW) data between 2 and 50 mg/L except Fluoride 2 and 25 mg/L

consisting of 3 Youden Pairs

RSD = %Relative Standard Deviation; (std dev / mean)(100)

LL = Lower Limit of %Recovery

UL = Upper Limit of %Recovery

RPD = Relative % Difference between MSD

---

26

Appendix B

(Non-mandatory Information)

B.1 Suggested Background References

B1.1 EPA Method 6500, "Dissolved Inorganic Anions in Aqueous Matrices by

Capillary Ion Electrophoresis", SW846, Rev 0, January 1998.

B1.2 Method 4140, "Inorganic Anions by Capillary Ion Electrophoresis", Standard

Methods for the Examination of Water and Wastewater, 20 Edition, 1998, p 4-12

to 4-20.

B1.3 Krol, Benvenuti, and Romano, "Ion Analysis Methods for IC and CIA and

Practical Aspects of Capillary Ion Analysis Theory", Waters Corp, Lit Code WT-139,

1998.

B1.4 Jandik, P., Bonn, G., "Capillary Electrophoresis of Small Molecules and Ions",

VCH Publishers, 1993

B1.5 Romano, J., Krol, J, "Capillary Ion Electrophoresis, An Environmental Method

for the Determination of Anions in Water", J. of Chromatography, Vol. 640, 1993, p.

403.

B1.6 Romano, J., "Capillary Ion Analysis: A Method for Determining Ions in Water

and Solid Waste Leachates", Amer. Lab., May 1993, p. 48.

B1.7 Jones, W., "Method Development Approaches for Ion Electrophoresis", J. of

Chromatography, Vol. 640, 1993, p. 387.

B1.8 Jones, W., Jandik, P., "Various Approaches to Analysis of Difficult Sample

Matrices for Anions using Capillary Electrophoresis", J. of Chromatography, Vol.

608, 1992, p. 385.

B1.9 Bondoux, G., Jandik, P., Jones, W., "New Approaches to the Analysis of Low

Level of Anions in Water", J. of Chromatography, Vol. 602, 1992, p. 79.

B1.10 Jandik, P., Jones, W., Weston, A., Brown, P.,"Electrophoretic Capillary Ion

Analysis: Origins, Principles, and Applications", LC•GC, Vol. 9, Number 9, 1991, p.

634.

B1.11 Romano, J., Jackson, P., "Optimization of Inorganic Capillary Electrophoresis

for the Analysis of Anionic Solutes in Real Samples", J.

of

Chromatography, Vol.

546, 1991, p. 411.

B1.12 Jandik, P., Jones, W., "Optimization of Detection Sensitivity in the Capillary

Electrophoresis of Inorganic Anions", J

of

Chromatography, Vol. 546, 1991, p. 431.

B1.13 Jandik, P., Jones, W., "Controlled Changes of Selectivity in the Separation

of

Ions by Capillary Electrophoresis", J. of Chromatography, Vol. 546, 1991, p 445.

B1.14 Foret, R., et.al., "Indirect Photometric Detection in Capillary Zone

Electrophoresis", J. of Chromatography, Vol. 470, 1989, p. 299.

B1.15 Hjerte'n, S. et. al., "Carrier-free Zone Electrophoresis, Displacement

Electrophoresis and Isoelectric Focusing in an Electrophoresis Apparatus", J. of

Chromatography, Vol. 403, 1987, p. 47.

B1.16 Serwe, M., "New ASTM Standard: Recommended Operating Conditions for the

Agilent CE", Agilent Technologies Application Brief, Publication Number 5968-

8660E.

---

27

Appendix C

Capillary Ion Electrophoresis

Initial Demonstration of Performance

Single Operator

General Inorganic Anion

&

Organic Acid Analysis with Indirect UV Detection

Basis for EPA Method 6500, ASTM D6508, and Standard Methods 4140

The performance data given in this appendix was provided in the collaborative instruction

booklet to evaluate initial demonstration of performance required by the collaborative design.

E

ce)

2

4

5

6

7

11

PPM Standards

1

2

3

4

5

6

7

8

9

10

11

Chloride

Bromide

Nitrite

Sulfate

Nitrate

Oxalate

Fluoride

Formate

Phosphate

Bicarbonate

Acetate

= 2

= 4

= 4

= 4

= 4

= 5

= 1

= 5

= 4

= 5

10

■.)

I?

'

?

I

? I

3.000

?

3.500

Minutes

?

4.000

?

4.500

Analysis Conditions:

Electrolyte:

Capillary:

Temperature:

Power Supply:

Voltage:

Current:

Sampling:

Detection:

Time Constant:

Sampling Rate:

Analyte MT:

Quantitation:

lonSelect High Mobility Anion Electrolyte, P/N 49385

75 gm (id) x 375 gm (od) x 60 cm (length)

25°C (5°C Above Ambient)

Negative

15 kV

14 ± 1 pA (Use Constant Current for Analysis)

Hydrostatic for 30 Seconds

Indirect UV at 254 nm, Hg Lamp, 185 or 254 nm Window

0.3 Seconds, or less

20 Data Points per Second

Mid-Point of Analyte Peak Width at Baseline

Time Corrected Peak Area (Peak Area / MT)

---

Millennium Data Processing Method:

28

CIE Processing

Integration

Calibration

Method using Mid-Point of Peak Width for Migration Time

Peak Width = 2.25 - 3.00 Threshold = 100 ± 25

Min Area = 100

?

Min Height = 50

Inhibit lntg. = 0 to 3 min

Averaging?

= None?MT Window = 2%

Update MT?

= Average Standards

Peak Match

?

= First for Chloride

(CI is always first in the pherogram, use as a ref peak)

CI MT Window = 10%

Other Analytes = Closest

Quantitate By = Time Corrected Peak Area

Fit Type

?

= Linear Through Zero

Report

Analyte Name

Analyte Migration Time

Analyte Migration Time Ratio (respect to CI Ref Peak)

Peak Area

Time Corrected Peak Area

Amounts

Use fresh electrolyte daily; recalibrate with every change in electrolyte.

Clear previous calibration (in Quick Set Page) before recalibration.

Do Not use analyte peak height for quantitation due to asymmetrical peak shapes.

Method Validation:

The single operator performance given below using the ASTM validation design is intended as

a basis to evaluate Initial Demonstration of Performance.

Individual Youden Pair Standard, in ppm

12345678

CI

0.7

2.0

3.0

15.0

40.0

20.0

50.0

0.5

Br

2.0

3.0

15.0

40.0

20.0

50.0

0.7

0.5

NO2

3.0

40.0

20.0

15.0

50.0

0.5

2.0

0.7

SO4

40.0

50.0

0.5

0.7

2.0

3.0

15.0

20.0

NOs

15.0

20.0

40.0

50.0

0.5

0.7

2.0

3.0

F

2.0

0.7

0.5

3.0

10.0

7.0

20.0

25.0

PO4

50.0

40.0

20.0

0.5

3.0

2.0

0.7

15.0

O

5,

C

---

CI

SO4

CI?

R2= 0.9996

SO. R 2 = 0.9998

Br?

R2= 0.9995

Br

3 Data Points per Concentration

Using Validation Standards

Method Linearity:

10

—

7

.5

r.n

5

.c0

2.5

—

—

NO a

3 Data Points per Concentration

Using Validation Standards

0

0

20

ppm Anion

30

40

50

NO 3

R2

= 0.9992

29

0

?

10

?

20?

30

?

40

?

50

ppm Anion

10 —

F?

R2= 0.9985

?

F

PO4

PO4?

R2= 0.9996

7.5 —

2.5

3 Data Points per Concentration

Using Validation Standards

0

?

0

?

10

?

2 Op

pm Anion

0

?

40

?

50

10 —

?

7.5 —

?

NO 2

?

R2= 0.9996

---

SO4

CI

PO4

30

Method Detection Limits:

100 ppb Anion Standard

E

I

?

I?

I

3.200

?

3.400

?

3.600

?

3.800?

4.000

M in ute s

Seven replicates of the above 100 ppb anion standard were used to calculate time corrected

peak area precision. Using EPA and Standard Methods protocols, the detection limits, as

ppb, for these analytes are:

Chloride = 46

Nitrate = 84

Bromide = 90

Fluoride = 20

Nitrite = 72

Phosphate = 41

Sulfate = 32

This method has been validated between 0.1 to 50 ppm. Quantitation below 0.1 ppm is not

advised.

---

31

Migration Time Reproducibility:

Use mid-point of analyte peak width at the baseline as the analyte migration time determinant.

Data given as average absolute migration time for each validation standard analyzed in

triplicate.

Analyte

CI

Br

NO2

SO4

NO3

F

PO4

1

3.132

3.226

3.275

3.405

3.502

3161

3.906

2

3.147

3.239

3.298

3.431

3.517

3.779

3.931

as

03

-0

3.138

3.231

3.283

3.411

3.497

3.771

3.925

cco

?

4

3.158

3.244

3.307

3.434

3.510

3.781

3.963

u)co?

5

3.184

3.271

3.331

3.435

3.551

3.787

3.981

fs

6

3.171

3.260

3.312

3.418

3.537

3.776

3.964

Vs'

>

?

7.

3.191

3.272

3.315

3.437

3.544

3.773

3.978

8

3.152

3.248

3.294

3.418

3.526

3.739

3.954

Std Dev

0.021

0.015

0.018

0.012

0.20

0.015

0.027

%RSD

0.67%

0.46%

0.55%

0.36%

0.56%

0.40%

0.68%

Average Standard Deviation = 0.018 min = 1.1 sec

Average %RSD of Analyte Migration Time = 0.53%

Quantitation Precision:

Time Corrected Peak Area Precision, given as %RSD, based upon 3 samplings per

concentration.

Analyte

CI

Br

NO2

SO4

NO3

F

PO4

0.1

12.36

18.89

16.19

13.25

23.13

9.82

14.00

0.5

10.51

20.00

3.90

2.25

2.18

2.03

7.71

c

....m.?

0.7

1.23

13.36

2.01

2.95

0.37

2.72

4.41

Li

a'

0.32

3.76

4.14

1.79

2.17

0.73

1.91

0

c

3

Q

0.63

1.80

1.72

1.70

0.58

0.98

2.70

0

L 15

0.43

0.27

0.48

0.07

0.36

0.15

1.37

0...

20

0.45

0.66

0.17

0.13

0.88

0.16

0.81

40

0.36

0.56

0.36

0.46

0.58

0.47

50

0.45

0.51

0.48

0.16

0.46

0.46

---

32

Quantitation Accuracy:

Used a Certified Performance Evaluation Standard diluted 1:100 with DI water. Amounts

based upon multi-point calibration curve prepared from certified standards.

Analyte

CI

NO2

SO4

NO3

F

PO4

Performance

True

Evaluation

Standard

Value

in ppm

43.00

1.77

37.20

15.37

2.69

6.29

Official

Measured

43.30

1.77

37.00

15.42

2.75

6.38

Anion

Mean

Methods

Measured

3.09

0.07

2.24

1.15

0.26

0.21

Wet Chem & IC

Std

Dev

CIA Using

Chromate

Ave CIA

n=18

43.34

1.64

37.11

14.41

2.64

6.34

Electrolyte

CIA/Mean

1.003

0.927

1.003

0.935

0.959

0.993

CIA/TrueValue

1.008

0.927

0.996

0.938

0.981

1.008

A CIA/True Value, or Mean = 1.000 indicates perfect agreement between CIA and official

anion methods.

Method Recovery:

A Certified Performance Evaluation Standard (PES) was diluted 1:100 with Typical Drinking

Water (DW). Amounts based upon multi-point calibration curve prepared from certified

standards.

Analyte

CI

NO2

SO4

NO3

F

PO4

Drinking Water

n=3, as ppm

24.72 ±

0.18

Not

Detected

7.99 ±

0.07

0.36 +

0.05

Not

Detected

Not

Detected

Amount %RSD

0.73%

0.91%

13.3%

Performance

Evaluation Std

43.00

1.77

37.20

15.37

2.69

6.29

DW + PES

n=3; as ppm

66.57±

0.34

1.74±

0.03

45.19±

0.17

15.42±

0.12

2.62±

0.07

5.55±

0.31

Amount %RSD

0.51%

1.85%

0.38%

0.79%

2.69%

5.52%

% Recovery

97.9%

98.3%

100.2%

98.1%

97.4%

88.2%

---

1

2

Li

L.

Anions in mg/L. No Dlution

1 Chloride = 93.3

2 Nitrite = 0.46

3 Sulfate =

60.3

4 Nitrate =

498

5 Carbonate = Natural

•'?

I?

•?"?

•?

•

3 ODD?

3'500

Minutes

4.000

'

•?

I?

'

4.500

'

•?

•?

•

?

•?"?

I

?

•

Minutes

4.000?

4.500

• I?

•

3.500

33

Fig. 1 Electropherogram of Mixed Anion Working Solution

and Added Common Organic Acids

Anion Standard in me&

1 Chloride = 2 7

Fluoride

/

4

?

I

2 Bromide = 4 8

Formate = 5

3 Mlle =4 9

Phosphate = 4

8?

4 Sulfate = 4 10

Carbonate

5 Nitrate = 4 11

Acetate = 5

9?

11

Fig. 2 Electropherogram of 0.2 mg/L Anions

Used to Determine MDL

6

1 Chloride

5

Nitrate

2 Brorride

6

Fluoride

3 Nitrite

7 Phosphate

4 Sulfate

4

3

5

3

E

4

C0.

0

7

2

\ON

,/ \A-vkilL

\41,-..,,Av■Yetv

I • " •?

•

3 000?

3.500?

4.000?

4.500

Minutes

1?1?1?1?

1?1?

1?1

?

I?I?

1?1?

f

?

i

3

030

3.200

3.400 3.600

3.800

Minutes

0)

Fig. 3 Electropherogram of Substitute Wastewater

fiknions n moil–

120 Dilution

1 Chloride =24.2

2 Sulfate = 3.77

3 Phosphate = 0.89

4 Carbonate = Nahral

2

3

Fig. 4 Electropherogram of Drinking Water

Anions In moil-, No Didion

1 Chloride = 20.2

2 Sulfate = 7.5

3 Nitrate =

1.6

4 RtnriCe

=

ace

5 Carbonate = Natural

3

3.000

11

?1?1

3.500

?

4/.003

Knutes

3.000

• I

?

•?

•?

•

3.500?

Minlite3

• I?•

4.003

'?

I?

•

4.500

Fig. 5 Electropherogram of Municipal Wastewater

?

Fig. 6 Electropherogram of Industrial Wastewater

Treatment Plant Discharge

3

E

0

Anions

in

moil_ Dilution

1 Chloride = 2.0

2 Nitrite =

1.6

3 Sulfate =34.7

4 Nitrate =

1&5

5 Formate < 0.05

6 Phosphate = 12.3

7 Carbonate = Nahral

4

L.

3.000

---

?

INorpmc

?

avnlent .

ANC,

:'5

Or

.

Adds,

?

' -

Ci; Br„

:

r

?

Oxyrn.r.?tals

?

NO:,,

S

C4

?

'

'

F

,

PO4,

?

No., ,,l_?

CIO C..10.

?

SO ; S

20 3?

':FanTlate,

Migration Time

High Mobility?

Low Mobility MT > 7 mini,

Anions

?

Anions

.4

MT= 0

O.?

1'

0.

?

?

9'

0.

?

51.

0

Y-1 9' 9'

N+? N? N

.

?

O.

?

O_

?

OH

N?

N

Anode

Matyte Ion (A) displaces electrolyte ion (e)

charge for charge or-transfer ratio causing

a net decrease n battleground absotbance.

The charge in absorbance is directly

related to Am:1sta concentration.

75

pm x 60 en7"..'..\\

Slim Capillary

CapPiary with Polyirnide

Coating Removal,

CNI Window

UV Detector

at 254 nm

Vane= Purge

rE

lectrolyranism

Constant

Temperature

Compartment,

25.30° C

•

•

!ta

& Samples

ndard: •

•

High Voltage Power Supply 0 to -30 kV

34

Fig. 7 Pictorial Diagram of Anion Mobility and

?

Fig. 8 Selectivity Diagram of Anion Mobility Using

ElectroOsomotic Flow Modifier

?

Capillary Ion Electrophoresis

HO

Mobility

Anion Q --••■■•••••••÷

Low Mobility Anion?

Cicotnto

•iE

?

-

Nurtnnis

(

?

&

W4or

* )

1"›

Donn:lion

Side

N1.14'.1""t4

Cathodo

0

miscues

Sid.

Fig. 9 Pictorial Diagram of Indirect UV Detection

e eeeeeAA Aeeeeeeeee

e

eeeeAAA Aeeeeeeee

e eeeeeAAA Aeeeeeeee

e

eeeeAAA Aeeeeeeee

Fig. 10 General Hardware Schematic of a

Capillary Ion Electrophoresis System