METHOD 552.3 DETERMINATION OF HALOACETIC ACIDS AND DALAPON IN 

DRINKING WATER BY LIQUID-LIQUID MICROEXTRACTION, 

DERIVATIZATION, AND GAS CHROMATOGRAPHY WITH 

ELECTRON CAPTURE DETECTION 

EPA 815-B-03-002 

Revision 1.0 

July 2003 

M. M. Domino and B.V. Pepich (Shaw Environmental and Infrastructure, Inc.) 

D.J. Munch and P.S. Fair (US EPA, Office of Ground Water and Drinking Water) 

Y. Xie (Penn State University) 

D.J. Munch, J.W. Munch (US EPA, Office of Ground Water and Drinking Water) and A. M. PawleckiVonderheide (ICI) Method 552.2, Revision 1.0 (1995) 

J.W. Hodgeson (USEPA), D. Becker (Technology Applications, Inc.) Method 552.1, (1992) 

J.W. Hodgeson (USEPA), J. Collins and R. E. Barth (Technology Applications, Inc.) Method 552.0, 

(1990) 

TECHNICAL SUPPORT CENTER

OFFICE OF GROUND WATER AND DRINKING WATER

U. S. ENVIRONMENTAL PROTECTION AGENCY

CINCINNATI, OHIO 45268

552.3-1
METHOD 552.3 

DETERMINATION OF HALOACETIC ACIDS AND DALAPON IN DRINKING WATER BY 

LIQUID-LIQUID MICROEXTRACTION, DERIVATIZATION, AND GAS 

CHROMATOGRAPHY WITH ELECTRON CAPTURE DETECTION 

1. SCOPE AND APPLICATION 

1.1 This is a gas chromatography (GC) method for the determination of haloacetic acids and 

dalapon in drinking waters. Accuracy, precision, and Detection Limit (DL) data are 

presented for the following compounds in reagent water and finished ground and surface 

waters. 

Analyte 

Bromochloroacetic acid (BCAA)

Bromodichloroacetic acid (BDCAA)

Chlorodibromoacetic acid (CDBAA)

Dalapon

Dibromoacetic acid (DBAA)

Dichloroacetic acid (DCAA)

Monobromoacetic acid (MBAA)

Monochloroacetic acid (MCAA)

Tribromoacetic acid (TBAA)

Trichloroacetic acid (TCAA)

Chemical Abstracts Service

(CAS)

Registry Number

5589-96-8 

71133-14-7 

5278-95-5 

75-99-0 

631-64-1 

79-43-6 

79-08-3 

79-11-8 

75-96-7 

76-03-9 

1.2 Detection Limits are compound, instrument, and matrix dependent. The DL is defined as 

the statistically calculated minimum amount that can be measured with 99% confidence 

that the reported value is greater than zero.(1) Detection Limits for the above listed 

analytes are provided in Section 17, Table 5. The DL differs from, and is lower than, the 

Minimum Reporting Level (MRL) (Sect. 3.17). The concentration range for target 

552.3-2
analytes in this method was evaluated between 0.5 μg/L and 30 μg/L for a 40-mL sample. 

Precision and accuracy data are presented in Section 17, Tables 6-11. 

1.3 This method is restricted to use by or under the supervision of analysts skilled in liquidliquid extractions, derivatization procedures and the use of GC and interpretation of gas 

chromatograms. 

2. SUMMARY OF METHOD 

2.1 A 40-mL volume of sample is adjusted to a pH of 0.5 or less and extracted with 4 mL of either 

methyl tert-butyl ether (MTBE) or tert-amyl methyl ether (TAME) containing an internal 

standard. The haloacetic acids that have been partitioned into the organic phase are then 

converted to their methyl esters by the addition of acidic methanol followed by heating for 2 

hours. The solvent phase containing the methylated haloacetic acids is separated from the acidic 

methanol by adding 7 mL of a concentrated aqueous solution of sodium sulfate. The aqueous 

phase is discarded. The extract is then neutralized with a saturated solution of sodium 

bicarbonate and the solvent layer is removed for analysis. The target analytes are identified and 

quantified by capillary column gas chromatography using an electron capture detector 

(GC/ECD). Analytes are quantified using procedural standard calibration. 

3. DEFINITIONS 

3.1 EXTRACTION BATCH – A set of up to 20 Field Samples (not including calibration 

standards and QC samples) extracted together by the same person(s) during a work day 

using the same lots of solvents, surrogate solution, and fortifying solutions. Required QC 

samples include a Laboratory Reagent Blank, Laboratory Fortified Matrix, either a Field 

Duplicate or Laboratory Fortified Matrix Duplicate and an appropriate number of 

Continuing Calibration Checks. 

3.2 ANALYSIS BATCH – A set of samples that is analyzed on the same instrument during a 

24-hour period that begins and ends with the analysis of the appropriate Continuing 

Calibration Check standards (CCC). Additional CCCs may be required depending on the 

length of the analysis batch and/or the number of Field Samples. 

552.3-3
3.3 INTERNAL STANDARD (IS) – A pure analyte added to an extract or standard solution 

in a known amount and used to measure the relative responses of other method analytes 

and surrogates. The internal standard must be an analyte that is not a sample component. 

3.4 SURROGATE ANALYTE (SUR) – A pure analyte, which chemically resembles target 

analytes and is extremely unlikely to be found in any sample. This analyte is added to a 

sample aliquot in a known amount before extraction or other processing, and is measured 

with the same procedures used to measure other sample components. The purpose of the 

SUR is to monitor method performance with each sample. 

3.5 LABORATORY REAGENT BLANK (LRB) – An aliquot of reagent water or other 

blank matrix that is treated exactly as a sample including exposure to all glassware, 

equipment, solvents, reagents, sample preservatives, internal standards, and surrogates 

that are used in the extraction batch. The LRB is used to determine if method analytes or 

other interferences are present in the laboratory environment, the reagents, or the 

apparatus. 

3.6 LABORATORY FORTIFIED BLANK (LFB) – An aliquot of reagent water or other 

blank matrix to which known quantities of the method analytes and all the preservation 

compounds are added. The LFB is analyzed exactly like a sample, and its purpose is to 

determine whether the methodology is in control, and whether the laboratory is capable of 

making accurate and precise measurements. 

3.7 LABORATORY FORTIFIED SAMPLE MATRIX (LFSM) – An aliquot of a Field 

Sample to which known quantities of the method analytes and all the preservation 

compounds are added. The LFSM is processed and analyzed exactly like a sample, and 

its purpose is to determine whether the sample matrix contributes bias to the analytical 

results. The background concentrations of the analytes in the sample matrix must be 

determined in a separate aliquot and the measured values in the LFSM corrected for 

background concentrations. 

3.8 LABORATORY FORTIFIED SAMPLE MATRIX DUPLICATE (LFSMD) – A second 

aliquot of the Field Sample used to prepare the LFSM which is fortified, extracted and 

analyzed identically to the LFSM. The LFSMD is used instead of the Field Duplicate to 

access method precision and accuracy when the occurrence of a target analyte is 

infrequent. 

552.3-4
3.9 LABORATORY DUPLICATES (LD1 and LD2) – Two aliquots of the same sample split 

in the laboratory and analyzed separately with identical procedures. Analyses of LD1 and 

LD2 indicate precision associated with laboratory procedures, but not with sample 

collection, preservation, and storage procedures. 

3.10 FIELD DUPLICATES (FD1 and FD2) – Two separate samples collected at the same 

time and place under identical circumstances, and treated exactly the same throughout 

field and laboratory procedures. Analyses of FD1 and FD2 give a measure of the 

precision associated with sample collection, preservation, and storage, as well as with 

laboratory procedures. 

3.11 STOCK STANDARD SOLUTION (SSS) – A concentrated solution containing one or 

more method analytes prepared in the laboratory using assayed reference materials or 

purchased from a reputable commercial source. 

3.12 PRIMARY DILUTION STANDARD (PDS) SOLUTION – A solution containing 

method analytes prepared in the laboratory from stock standard solutions and diluted as 

needed to prepare calibration solutions and other needed analyte solutions. 

3.13 CALIBRATION STANDARD (CAL) – A solution prepared from the primary dilution 

standard solution and/or stock standard solutions and the internal standards and surrogate 

analytes. The CAL solutions are used to calibrate the instrument response with respect to 

analyte concentration. 

3.14 CONTINUING CALIBRATION CHECK (CCC) – A calibration standard containing the 

method analytes, internal standard(s) and surrogate(s), which is analyzed periodically to 

verify the accuracy of the existing calibration for those analytes. 

3.15 QUALITY CONTROL SAMPLE (QCS) –A sample prepared using a PDS of method 

analytes that is obtained from a source external to the laboratory and different from the 

source of calibration standards. The second source PDS and the surrogate PDS are used 

to fortify the QCS at a known concentration. The QCS is used to check calibration 

standard integrity. 

3.16 DETECTION LIMIT – The minimum concentration of an analyte that can be identified, 

measured and reported with 99% confidence that the analyte concentration is greater than 

zero (Sect. 9.2.4). This is a statistical determination of precision. Accurate quantitation 

is not expected at this level.(1) 

552.3-5
3.17 MINIMUM REPORTING LEVEL (MRL) – The minimum concentration that can be 

reported as a quantified value for a target analyte in a sample following analysis. This 

defined concentration can be no lower than the concentration of the lowest continuing 

calibration standard for that analyte, and can only be used if acceptable quality control 

criteria for this standard are met. 

3.18 PROCEDURAL STANDARD CALIBRATION – A calibration method in which 

aqueous calibration standards are prepared and processed (e.g., extracted, and/or 

derivatized) in exactly the same manner as the samples. All steps in the process from 

addition of sampling preservatives through instrumental analyses are included in the 

calibration. Using procedural standard calibration compensates for any inefficiencies in 

the processing procedure. 

3.19 MATERIAL SAFETY DATA SHEET (MSDS) – Written information provided by 

vendors concerning a chemical’s toxicity, health hazards, physical properties, fire, and 

reactivity data including storage, spill, and handling precautions. 

4. INTERFERENCES 

4.1 All glassware must be meticulously cleaned. Wash glassware with detergent and tap 

water, rinse with tap water followed by reagent water. A final rinse with solvents may be 

needed. In place of a solvent rinse, non-volumetric glassware can be muffled at 400 °C 

for 2 hours. Volumetric glassware should not be heated in an oven above 120 °C. Store 

glassware inverted or capped with aluminum foil. 

4.2 Method interferences may be caused by contaminants in solvents, reagents (including 

reagent water), sample bottles and caps, and other sample processing hardware that lead 

to discrete artifacts and/or elevated baselines in the chromatograms. All items used in the 

method must be routinely demonstrated to be free from interferences (less than 1

/3 the 

MRL for each target) under method conditions by processing and analyzing laboratory 

reagent blanks as described in Section 9. Subtracting blank values from sample 

results is not permitted. 

4.2.1 Sodium sulfate can be a source of method interferences. After screening several 

brands, it was found that a grade suitable for pesticide residue analysis had the 

lowest amount of method interferants. If the suitability of the available sodium 

552.3-6
sulfate is in question, extract and analyze a laboratory reagent blank (Sect. 3.5) to 

test for interferences prior to sample processing. 

4.2.2 The ester of bromochloroacetic acid (BCAA) coelutes with a low-level interferant 

on both the primary and confirmation column. This interferant is present in the 

LRB and believed to be dimethyl sulfide that originates in the sodium sulfate. 

Because of difference in peak shape, the BCAA ester tends to “ride on” the 

interferant allowing accurate quantitation. Concentrations of BCAA at or below 

2:g/L may require manual integration of the BCAA. 

4.2.3 TAME, which has higher methylation efficiencies for the trihaloacetic acids than 

MTBE, is currently only available at 97% purity. Different lots of TAME should 

be carefully evaluated for potential interferants. Several lots of TAME contained 

an interferant that coeluted with the ester of monochloroacetic acid (MCAA) on 

the primary column. 

4.3 The esters of the surrogate used in Method 552.2 (2,3-dibromopropanoic acid) and 

dichlorobromoacetic acid are not completely resolved under the chromatographic 

conditions for the confirmation column listed in Tables 3 and 4. The current surrogate, 

2-bromobutanoic acid, is fully resolved from all method compounds on both columns 

under the conditions listed in Tables 1-4 (Figs. 1-4). 

4.4 Matrix interferences may be caused by contaminants that are extracted from the sample. 

The extent of matrix interferences will vary considerably from source to source, 

depending upon the water sampled. 

4.5 Interferences by phthalate esters can pose a major problem in analysis when using an 

electron capture detector (ECD). These compounds generally appear in the chromatogram 

as large peaks. Common flexible plastics contain varying amounts of phthalates that are 

easily extracted or leached during laboratory operations. Cross contamination of clean 

glassware routinely occurs when plastics are handled during extraction steps, especially 

when solvent-wetted surfaces are handled. Interferences from phthalates can best be 

minimized by avoiding the use of plastics in the laboratory. Exhaustive purification of 

reagents and glassware may be required to eliminate background phthalate 

contamination.(2,3) 

552.3-7
5. SAFETY 

5.1 The toxicity or carcinogenicity of each reagent used in this method has not been precisely 

defined. Each chemical compound should be treated as a potential health hazard, and 

exposure to these chemicals should be minimized. The laboratory is responsible for 

maintaining a current awareness file of OSHA regulations regarding the safe handling of 

the chemicals specified in this method. A reference file of MSDSs should also be made 

available to all personnel involved in the chemical analysis. Additional references to 

laboratory safety are available.(4-6) 

5.2 Pure standard materials and stock standards of these compounds should be handled with 

suitable protection to skin and eyes, and care should be taken not to breathe the vapors or 

ingest the materials. 

5.3 The toxicities of the extraction solvents, MTBE or TAME, have not been well defined. 

Susceptible individuals may experience adverse effects upon skin contact or inhalation of 

vapors. Therefore protective clothing and gloves should be used and MTBE or TAME 

should be used only in a chemical fume hood or glove box. The same precaution applies 

to pure standard materials. 

6. EQUIPMENT AND SUPPLIES 

6.1 SAMPLE CONTAINERS – Amber glass bottles, at least 50-mL, fitted with PTFE 

(polytetrafluoroethylene) lined screw caps. 

6.2 EXTRACTION VIALS – 60-mL clear glass vials with PTFE-lined screw caps. 

6.3 AUTOSAMPLER VIALS – 2.0-mL amber vials with screw or crimp cap and a PTFEfaced seal. 

6.4 STANDARD SOLUTION STORAGE CONTAINERS – 10- to 20-mL amber glass vials 

with PTFE-lined screw caps. 

6.5 GRADUATED CONICAL CENTRIFUGE TUBES WITH PTFE-LINED SCREW 

CAPS – 15-mL with graduated 1-mL markings (Fisher Cat. #: 05-538-30A or 

equivalent). 

552.3-8
6.6 PASTEUR PIPETTES – Glass, disposable. 

6.7 PIPETTES – Class A, 2.0-, 3.0-, 4.0-, and 7.0-mL glass, or adjustable volume dispensers. 

6.8 VOLUMETRIC FLASKS – Class A, suggested sizes include 5-mL, 10-mL, 100-mL. 

6.9 MICRO SYRINGES – Various sizes. 

6.10 HEATER (BLOCK,or SAND BATH or WATER BATH) – A block heater (Thermolyne 

Model DB16525, Fisher Cat. #: 11-716-50 or equivalent) capable of maintaining 

regulated, elevated temperature equipped with a sand bath or heating block (Fisher Cat. #: 

11-716-27 or equivalent) capable of holding screw cap conical centrifuge tubes in Section 

6.5 was used for method development. A thermostated water bath equipped with a 

surface layer of small plastic spheres (Fisher Cat. #. 14-220-31 or equivalent) to retard 

evaporation and subsequent heating of tube walls may also be used. 

6.11 pH PAPER - With a pH range of 0 - 1.5 (Fisher Cat. #. 14-853-55 or equivalent). 

6.12 BALANCE – Analytical, capable of weighing to the nearest 0.0001 g. 

6.13 VORTEXER – Used to mix sample extracts (VWR Vortex-Genie, Cat. #:14216-184 or 

equivalent). 

6.14 GAS CHROMATOGRAPH – Capillary GC, with a low volume (150 μL) micro ECD 

(Agilent Model 6890 or equivalent). The injector system should not allow analytes to 

contact hot stainless steel or other metal surfaces that promote decomposition. The 

performance data in Section 17 was collected using hot, splitless injection using a 2-mm 

i.d. quartz liner. Other injection techniques such as temperature programmed injection, 

cold on-column injection, and large volume injection may be used if the QC criteria of 

Sections 9 and 10 are met. Alternate detectors which have equivalent or greater 

selectivity for the target compounds may be used. 

6.15 AUTOSAMPLER – Agilent Model 7683 or equivalent. 

6.16 PRIMARY GC COLUMN – DB-1701, 30-meter length, 0.25-mm i.d., 0.25-μm film, 

fused silica capillary with chemically bonded (14% 

cyanopropylphenyl-methylpolysiloxane), or equivalent bonded, fused silica column. 

552.3-9
6.17 CONFIRMATION GC COLUMN – DB-5.625, 30-meter length, 0.25-mm i.d., 0.25-μm 

film, fused silica capillary with chemically bonded (“equivalent to” 5% phenylmethylpolysiloxane), or equivalent bonded, fused silica column. 

7. REAGENTS AND STANDARDS 

7.1 REAGENTS AND SOLVENTS – Reagent grade or better chemicals should be used in 

all analyses. Unless otherwise indicated, it is intended that all reagents shall conform to 

the specifications of the Committee on Analytical Reagents of the American Chemical 

Society, where such specifications are available. Other grades may be used, provided it is 

first determined that the reagent is of sufficiently high purity to permit its use without 

lessening the quality of the determination. 

7.1.1 REAGENT WATER – Purified water which does not contain any measurable 

quantities of any target analytes or interfering compounds greater than 1/3 the 

MRL for each compound of interest. 

7.1.2 METHYL tert-BUTYL ETHER (MTBE) – High purity, demonstrated to be free 

from analytes and interferences (HPLC grade or better). 

7.1.3 Tert-AMYL METHYL ETHER (TAME) – High purity, demonstrated to be free 

from analytes and interferences. 

7.1.3.1 Currently, TAME is available in > 97% purity. Several manufacturers’ 

products were evaluated and found to contain a compound that elutes with 

MCAA on the primary column. Lots evaluated from Fluka (Riedel-de 

Haen), stored over molecular sieves, were free from this interferant. 

7.1.4 SODIUM SULFATE, Na2SO4 – Pesticide grade, granular, anhydrous. 

Interferences have been observed when lower quality grades have been used. If 

interferences are observed, it may be necessary to heat the sodium sulfate in a 

shallow tray at 400 °C for up to 4 hr to remove phthalates and other interfering 

organic substances. Store in a capped glass bottle rather than a plastic container. 

7.1.5 SODIUM BICARBONATE, NaHCO3 – ACS reagent grade. 

7.1.6 AMMONIUM CHLORIDE, NH4Cl – ACS reagent grade. 

552.3-10
7.1.7 SULFURIC ACID – Concentrated, ACS reagent grade. Substitution of 

hydrochloric acid (HCl) is not allowed. Solutions of HCl can contain trace 

levels of bromide, which can promote the formation of brominated HAAs. 

7.1.8 HELIUM (or HYDROGEN)– 99.999% pure or better, GC carrier gas. 

7.1.9 ARGON 95%/METHANE 5% (or NITROGEN) – 99.999% pure or better, ECD 

make-up gas. 

7.1.10 SODIUM SULFATE SOLUTION – Prepare an aqueous solution of sodium 

sulfate in reagent water at a concentration of 150 g/L. Substitution of sodium 

chloride is not allowed. Sodium chloride can contain trace levels of bromide, 

which can promote the formation of brominated HAAs. 

7.1.11 SODIUM BICARBONATE SOLUTION, SATURATED – Add sodium 

bicarbonate to a volume of water, mixing periodically until the solution has a 

small amount of undissolved sodium bicarbonate that does not disappear upon 

further mixing. 

7.1.12 10% SULFURIC ACID IN METHANOL SOLUTION – Add 10 mL of sulfuric 

acid dropwise (due to heat evolution) to 50-60 mL of methanol contained in a 

100-mL volumetric flask that has been placed in a cooling bath. Mix, let cool, 

and dilute to the 100-mL mark with methanol. 

7.2 STANDARD SOLUTIONS – Standard Solutions may be prepared from certified, 

commercially available solutions or from neat compounds. When preparing from neat 

material, compound purity needs to be 96% or greater. When a compound purity is 

assayed to be 96% or greater, the weight can be used without correction to calculate the 

concentration of the stock standard. Solution concentrations listed in this section were 

used to develop this method and are included as an example. Standards for sample 

fortification generally should be prepared in the smallest volume that can be accurately 

measured to minimize the addition of organic solvent to aqueous samples. Laboratories 

should use standard QC procedures to determine when Standard Solutions described in 

this section need to be replaced. 

7.2.1 STANDARD PREPARATION AND STORAGE TECHNIQUES – All Standard 

Solutions are prepared from either neat or solid materials following the general 

procedure outlined below. This procedure assumes that the standard stock 

552.3-11
solutions are prepared in 10-mL volumetric flasks. The procedure should be 

scaled accordingly if larger volumetric flasks are used. Most standards in this 

method are made up in MTBE, whether the extracting solvent is MTBE or 

TAME. This is because MTBE is more water-soluble than TAME. 

7.2.1.1 For analytes which are solids in their pure form, prepare stock standard 

solutions by accurately weighing the suggested amount (refer to the 

sections below for suggested weights) of pure material into a clean, tared 

10-mL volumetric flask. Dilute the flask to volume with MTBE and mix 

thoroughly. 

7.2.1.2 For analytes which are liquid in their pure form at room temperature, place 

about 9.8 mL of MTBE into a 10-mL volumetric flask. Allow the flask to 

stand, unstoppered, for about 10 minutes to allow solvent film to evaporate 

from the inner walls of the volumetric, and weigh to the nearest 0.1 mg. 

Using a 10-μL (or gastight) syringe add the desired volume of the neat 

standard material to the flask by keeping the syringe needle just above the 

surface of the MTBE. Be sure that the standard material falls dropwise 

directly into the MTBE without contacting the inner wall of the 

volumetric. Record the weight and calculate the concentration of the stock 

standard from the net gain in weight. Dilute to volume, stopper, then mix 

by inverting the flask several times. 

7.2.1.3 Transfer the stock solutions to amber glass vials or amber bottles with 

PTFE-lined caps and store at # 0 °C. 

7.2.2 INTERNAL STANDARD (IS) SOLUTIONS – 1,2,3-trichloropropane (99+%) is 

used as an internal standard for the method. This compound has been shown to be 

an effective internal standard for the method analytes, but other compounds may 

be used if the QC requirements in Section 9 are met. 

7.2.2.1 INTERNAL STANDARD STOCK SOLUTION (2.0 mg/mL) – Prepare 

an internal standard stock solution by accurately transferring 

approximately 0.0200 g of neat 1,2,3-trichloropropane (weighed to the 

nearest 0.1mg) into a 10-mL volumetric flask containing methanol as 

described in Section 7.2.1. The resulting concentration of the stock 

internal standard solution will be approximately 2.0 mg/mL. 

552.3-12
7.2.2.2 INTERNAL STANDARD PRIMARY DILUTION STANDARD 

(100μg/mL) – Prepare an internal standard fortification solution at 

100μg/mL (or other suitable concentration) by the addition of 500 μL (or 

other appropriate volume) of the stock standard to a 10-mL volumetric 

flask containing MTBE. Dilute to volume, mix thoroughly and transfer to 

an amber glass vial with a PTFE-lined screw cap and store at # 0°C. 

7.2.2.3 MTBE (or TAME) EXTRACTION SOLVENT WITH INTERNAL 

STANDARD (1.00 μg/mL) – The internal standard 1,2,3-trichloropropane 

is added to the extraction solvent prior to analyte extraction to compensate 

for any volumetric differences encountered during sample processing. 

Have enough working solvent available to extract all calibration standards, 

QC samples and field samples in each extraction batch. The volume of 

fortified solvent should be determined by the sample workload. Never use 

two different lots of working solvent for one extraction batch. The 

following example illustrates preparation of 100 mL of fortified solvent. 

Add 1 mL of the primary dilution standard (100 μg/mL) to a 100-mL 

volumetric flask containing MTBE (or TAME), dilute to volume and mix 

thoroughly. Transfer the standard to an amber bottle for storage. This 

results in a final internal standard concentration of 1.00 μg/mL. This 

solution is used to extract the samples (Sect. 11.1). 

7.2.3 SURROGATE (SUR) ANALYTE STANDARD SOLUTION – 2-bromobutanoic 

acid (99+%) is used as a surrogate compound in this method to evaluate the 

extraction and derivatization procedures. This compound has been shown to be 

an effective surrogate for the method analytes and is well resolved from all target 

analytes and common interferants on both chromatographic columns. The Method 

552.2 surrogate, 2,3-dibromopropanoic acid, can be used with this method, but 

requires a longer chromatographic run to separate it from DCBAA (Sect. 4.3). 

Other SUR analytes may be used as long as they are halogenated carboxylic acids 

and the QC requirements of Section 9.7 are met. All surrogates must be added as 

free acids. Alternate candidates should be confirmed to have adequate 

esterification efficiencies (e.g. $ 80%) by comparison to a commercially available 

premethylated standard. Lower esterification efficiencies may lead to poor SUR 

precision. 

Note: 2-bromo-2-methylpropanoic acid was ruled out as a potential SUR because 

it was found to degrade in the Field and QC Samples if they were not processed 

552.3-13
immediately. Nearly 50% of the fortified concentration was lost when samples 

were fortified and then allowed to set for 3 hours prior to acidification. Alternate 

surrogate compounds MUST be carefully evaluated. 

7.2.3.1 SURROGATE STOCK SOLUTION (10 mg/mL) – Prepare surrogate 

stock standard solutions of 2-bromobutanoic acid [80-58-0] by accurately 

transferring approximately 0.100 g of the neat material (weighed to the 

nearest 0.1 mg.) into a clean, tared 10-mL volumetric flask as described in 

Section 7.2.1. The resulting concentration of the Surrogate Stock Solution 

will be approximately 10 mg/mL. 

7.2.3.2 SURROGATE PRIMARY DILUTION STANDARD (20 μg/mL) – 

Prepare a primary dilution standard (PDS) at a concentration of 20 μg/mL 

(or other suitable concentration) by adding 50 μL (or suitable volume) of 

the SUR stock standard to a 25-mL volumetric flask containing MTBE. 

Dilute the flask to volume, and mix thoroughly. Transfer the SUR PDS to 

an amber glass vial with a PTFE-lined screw cap and store at # 0 °C. 

Addition of 20 μL of the primary dilution standard to the 40-mL aqueous 

sample results in a surrogate concentration of 10 ng/mL. 

7.2.4 ANALYTE STANDARD SOLUTIONS – Obtain the analytes listed in the table 

in Section 1.1 as neat or solid free acid standards or as commercially prepared 

ampulized solutions from a reputable standard manufacturer. The use of 

premethylated standards is not allowed for the preparation of analyte standards. 

Prepare the Analyte Stock and Primary Dilution Standards as described below. 

7.2.4.1 ANALYTE STOCK SOLUTION – Prepare separate stock standard 

solutions for each analyte of interest at a concentration of 1-5 mg/mL in 

MTBE. Method analytes are available as neat materials or ampulized 

solutions (> 99% purity) from a number of commercial suppliers. 

7.2.4.1.1 For analytes which are solids in their pure form, prepare stock 

standard solutions by accurately weighing approximately 0.01 

to 0.05 grams of pure material in a 10-mL volumetric flask 

using the technique described in Section 7.2.1.1. 

552.3-14
7.2.4.1.2 For analytes which are liquid in their pure form at room 

temperature, add 10.0 μL of standard material to the flask using 

the technique described in Section 7.2.1.2. 

7.2.4.2 ANALYTE PRIMARY DILUTION STANDARD (PDS)– Prepare the 

Analyte PDS solution by combining and diluting the Analyte Stock 

Solutions prepared above (Sect. 7.2.4.1) with MTBE. This solution will 

be used to prepare the Analyte Secondary Dilution Standard. As a 

guideline to the analyst, the concentration used in the primary dilution 

standard solution during method development was 100 μg/mL for all ten 

analytes. 

7.2.4.3ANALYTE SECONDARY DILUTION STANDARD (SDS)– The Analyte 

SDS is used to fortify reagent water for calibration standards. Prepare at 

least one Analyte SDS by diluting the Analyte PDS with methanol. Two 

Analyte SDSs were used during method development. The first was 

prepared at a concentration of 5.00 μg/mL (1/20 dilution of the Analyte 

PDS) and the second at a concentration of 16.7 μg/L (1/6 dilution of the 

Analyte PDS). The SDS should be made daily. It should not be stored, 

since methanol will derivatize the acids that it contains. The lowest 

calibration standard concentration must be at or below the MRL of each 

analyte. The concentrations of the other standards should span the range of 

concentration expected in the Field Samples or the working range of the 

instrument. 

7.2.5 CALIBRATION STANDARDS – At least 5 calibration standards are required to 

prepare the initial calibration curve (Sect. 10.2). Fortify an appropriate number of 

reagent water solutions with varying amounts of the Analyte PDS over the 

concentration range of interest. During method development, the Calibration 

Standards typically ranged from 1.0 μg/L to 20 μg/L. The lowest standard must 

be at or below the MRL, which will depend upon instrument sensitivity. Because 

this method employs a procedural calibration technique , these standards must be 

treated like samples and require the addition of all preservation and other 

reagents. They are extracted by the procedure described in Section 11. 

552.3-15
8. SAMPLE COLLECTION, PRESERVATION, AND STORAGE 

8.1 SAMPLE BOTTLE PREPARATION 

8.1.1 Grab samples must be collected in accordance with conventional sampling 

practices(7) using amber glass containers with PTFE-lined screw caps and 

capacities of at least 50 mL. 

8.1.2 Prior to shipment to the field, add crystalline or granular ammonium chloride 

(NH4Cl) to the sample containers to produce a concentration of 100 mg/L in the 

Field Sample. For a typical 50-mL sample, this requires 5 mg of ammonium 

chloride. 

Note: Enough ammonium chloride must be added to the sample to convert the 

free chlorine residual in the sample matrix to combined chlorine. Chloramines, 

formed by the reaction of hypochlorite with the ammonium ion, do not react 

further to produce additional haloacetic acids at significant concentrations and 

protect against microbiological degradation.(8) This concentration of ammonium 

chloride was determined to convert 8 mg/L of free chlorine residual to combined 

chlorine. 

8.2. SAMPLE COLLECTION 

8.2.1 Fill sample bottles but take care not to flush out the ammonium chloride. Because 

the target analytes of this method are not volatile, it is not necessary to ensure that 

the sample bottles are completely headspace free. 

8.2.2 When sampling from a water tap, remove the aerator. Open the tap and allow the 

system to flush until the water temperature has stabilized (usually about 3-5 

minutes). Collect samples from the flowing system. 

8.2.3 When sampling from an open body of water, fill a 1-quart wide-mouth bottle or 

1 liter beaker with sample from a representative area, and carefully fill sample 

vials from the container. 

8.2.4 After collecting the sample, seal the bottle and agitate by hand for 15 seconds. 

552.3-16
8.3 SAMPLE SHIPMENT AND STORAGE – All samples must be chilled during shipment 

and must not exceed 10 °C during the first 48 hours after collection. Samples must be 

confirmed to be at or below 10 °C when they are received at the laboratory. Samples 

stored in the lab must be held at or below 6 °C and protected from light until extraction. 

Samples should not be frozen. 

8.4 SAMPLE AND EXTRACT HOLDING TIMES – Chlorinated Field Samples that are 

preserved according to the method should not exhibit biological degradation of analytes 

during the allotted 14-day storage time. Samples must be extracted within 14 days. 

Unchlorinated Field Samples should be extracted as soon as possible to prevent 

biological degradation of analytes. Extracts must be stored at # -10 °C or less and 

protected from light in glass vials with PTFE-lined caps. MTBE extracts must be 

analyzed within 21 days of extraction. TAME extracts must be analyzed within 28 days 

of extraction. 

8.4.1 The brominated trihaloacetic acids can exhibit degradation during storage. 

Tribromoacetic acid (TBAA), the least stable HAA ester, is degraded to 

bromoform. This is thought to occur as a result of peroxide contamination in the 

solvent. Low concentrations of peroxides that were barely detectable with the 

commercially available peroxide test strips were found to generate significant 

levels of bromoform in standard solutions. Bromoform can be chromatographed 

under method conditions (see Tables 1-4), and can be monitored as an indication 

of high peroxide levels in the solvent. If large bromoform peaks are observed in 

HAA-fortified reagent water samples (or calibration standards), the ether solvent 

should be purified or replaced. Storing these solvents under nitrogen minimizes 

peroxide formation. 

8.4.2 Extracts that were prepared using the TAME procedure showed much lower 

tendency to form bromoform during the method development studies. This is 

thought to be associated with lower levels of peroxides. This may be due to 

TAME being stored over molecular sieves (Sect. 7.1.3). 

9. QUALITY CONTROL 

9.1 Each laboratory using this method is required to operate a formal Quality Control (QC) 

program. The requirements of this program consist of an Initial Demonstration of 

Capability (IDC) , and subsequent analysis in each analysis batch of a Laboratory 

552.3-17
Reagent Blank (LRB), Continuing Calibration Check Standards (CCCs), a Laboratory 

Fortified Sample Matrix (LFSM), and either a Laboratory Fortified Sample Matrix 

Duplicate (LFSMD) or a Field Duplicate Sample. This section details the specific 

requirements for each QC parameter. The QC criteria discussed in the following sections 

are summarized in Section 17, Tables 15 and 16. These criteria are considered the 

minimum acceptable QC criteria, and laboratories are encouraged to institute additional 

QC practices to meet their specific needs. 

9.1.1 Process all quality control samples through all steps of Section 11, including 

methylation. Sample preservatives as described in Section 8.1 must be added 

prior to extracting and analyzing the quality control samples. 

9.2 INITIAL DEMONSTRATION OF CAPABILITY (IDC) – Requirements for the Initial 

Demonstration of Capability are described in the following sections and summarized in 

Section 17, Table 15. 

9.2.1 INITIAL DEMONSTRATION OF LOW SYSTEM BACKGROUND – Before 

any samples are analyzed, it must be demonstrated that a laboratory reagent blank 

(LRB) is reasonably free of contamination. 

9.2.2 INITIAL DEMONSTRATION OF ACCURACY – Prior to the analysis of the 

IDC samples, verify calibration accuracy with the preparation and analysis of a 

mid-level QCS as defined in Section 9.10. If the analyte recovery is not + 30% of 

the expected value, the accuracy of the method is unacceptable. The source of the 

problem must be identified and corrected. After the accuracy of the calibration 

has been verified, prepare and analyze 4-7 replicate LFBs (or CCCs in this 

method) fortified at 10 μg/L, or near the mid-range of the initial calibration curve, 

according to the procedure described in Section 11. Sample preservatives as 

described in Section 8.1.2 must also be added to these samples. The average 

recovery of the replicate values must be within ± 20% of the expected value. 

9.2.3 INITIAL DEMONSTRATION OF PRECISION – Using the same set of replicate 

data generated for Section 9.2.2, calculate the standard deviation and relative 

standard deviation of the replicate recoveries. The relative standard deviation 

(RSD) of the results of the replicate analyses must be less than 20%. 

9.2.4 DETECTION LIMIT (DL) DETERMINATION – Prepare, extract and analyze at 

least seven replicate LFBs at a concentration estimated to be near the Detection 

552.3-18
Limit over a period of at least three days (both extraction and analysis should be 

conducted over at least three days) using the procedure described in Section 11. 

The fortification level may be estimated by selecting a concentration with a signal 

of 2 to 5 times the noise level. The appropriate concentration will be dependent 

upon the sensitivity of the GC/ECD system being used. Sample preservatives as 

described in Section 8.1.2 must be added to these samples. Calculate the 

Detection Limit using the equation 

DL = St(n - 1, 1 -" = 0.99) 

where 

t(n - 1, 1 - " = 0.99) = Students t value for the 99% confidence level with n-1 

degrees of freedom ( 3.143 for 7 replicates) 

n = number of replicates, and 

S = standard deviation of the replicate analyses. 

NOTE: Do not subtract blank values when performing DL calculations. The DL 

is a statistical determination of precision only.(1) If the DL replicates are fortified 

at a low enough concentration, it is likely that they will not meet precision and 

accuracy criteria, and may result in a calculated DL that is higher than the fortified 

concentration. Therefore no precision and accuracy criteria are specified for the 

DL. 

9.2.5 Minimum Reporting Level (MRL) – The MRL is the threshold concentration of 

an analyte that a laboratory can expect to accurately quantitate in an unknown 

sample. The MRL should be established at an analyte concentration that is at 

least 3 times the DL or at a concentration which would yield a signal-to-noise 

(S/N) ratio of greater than or equal to five. Depending upon the study’s data 

quality objectives it may be set at a higher concentration. The concentration of 

the lowest calibration standard must be at or below the MRL. 

9.2.6 METHOD MODIFICATIONS – The analyst is permitted to modify GC columns, 

GC conditions (see Tables 1-4), and internal standards (see Sect. 7.2.2) or 

surrogate standards (see Sect. 7.2.3). Other detectors (see Sect. 6.14) may be used 

if they have equivalent or better selectivity and have sufficient sensitivity. Each 

time such method modifications are made, the analyst must repeat the procedures 

of the IDC (Sect. 9.2). 

552.3-19
9.3 LABORATORY REAGENT BLANK (LRB) – An LRB is required with each extraction 

batch (Sect. 3.1) of samples to determine the background system contamination. If the 

LRB produces a peak within the retention time window of any analyte that would prevent 

the determination of that analyte, determine the source of contamination and eliminate the 

interference before processing samples. Background contamination must be reduced to 

an acceptable level before proceeding. Background from method analytes or 

contaminants that interfere with the measurement of method analytes must be below 1/3 

of the MRL. If the target analytes are detected in the LRB at concentrations equal to or 

greater than this level, then all data for the problem analyte(s) must be considered invalid 

for all samples in the extraction batch. 

9.4 CONTINUING CALIBRATION CHECK (CCC) – CCC Standards are prepared in the 

same extraction batch as the samples of interest. They must contain all compounds of 

interest, and they are extracted in the same manner as the Field Samples and calibration 

solutions used to prepare the initial calibration curve. Calibration checks, prepared with 

the samples being analyzed, are required at the beginning of each analysis batch, after 

every ten samples, and at the end of the analysis batch. See Section 10.3 for 

concentration requirements, frequency requirements, and acceptance criteria. 

9.5 LABORATORY FORTIFIED BLANK (LFB) – Since this method utilizes procedural 

calibration standards, which are fortified reagent waters, there is no difference between 

the LFB and the continuing calibration check standard. Consequently, the analysis of an 

LFB is not required; however, the acronym LFB is used for clarity in the IDC. 

9.6 INTERNAL STANDARDS (IS) – The analyst must monitor the peak area of each 

internal standard in all injections during each analysis day. The IS response (as indicated 

by peak area) for any chromatographic run must not deviate by more than ± 50% from the 

average area measured during the initial calibration for that IS. A poor injection could 

cause the IS area to exceed these criteria. Inject a second aliquot of the suspect extract to 

determine whether the failure is due to poor injection or instrument response drift. 

9.6.1 If the reinjected aliquot produces an acceptable internal standard response, report 

results for that aliquot. 

9.6.2 If the internal standard area for the reinjected extract deviates greater than 50% 

from the initial calibration average, the analyst should check the continuing 

calibration check standards that ran before and after the sample. If the continuing 

calibration check fails the criteria of Section 10.3, recalibration is in order per 

552.3-20
Section 10. If the calibration standard is acceptable, extraction of the sample 

should be repeated provided the sample is still within holding time. Otherwise, 

report results obtained from the reinjected extract, but annotate as suspect. 

9.7 SURROGATE RECOVERY – The surrogate standard is fortified into the aqueous 

portion of all samples, LRBs, CCCs, LFSMs, and LFSMDs prior to extraction. It is also 

added to the calibration standards. The surrogate is a means of assessing method 

performance from extraction to final chromatographic measurement. Calculate the 

recovery (R) for the surrogate using the equation 

where 

A = calculated surrogate concentration for the QC or Field Sample, and 

B = fortified concentration of the surrogate. 

9.7.1 When surrogate recovery from a sample, blank, or CCC is less than 70% or 

greater than 130%, check (1) calculations to locate possible errors, (2) standard 

solutions for degradation, (3) contamination, and (4) instrument performance. 

Correct the problem and reanalyze the extract. 

9.7.2 If the extract reanalysis meets the surrogate recovery criterion, report only data for 

the reanalyzed extract. 

9.7.3 If the extract reanalysis fails the 70-130% recovery criterion, the analyst should 

check the calibration by injecting the last calibration standard that passed. If the 

calibration standard fails the criteria of Section 9.7.1, recalibration is in order per 

Section 10.2. If the calibration standard is acceptable, extraction of the sample 

should be repeated provided the sample is still within the holding time. If the reextracted sample also fails the recovery criterion, report all data for that sample as 

suspect/surrogate recovery to inform the data user that the results are suspect due 

to surrogate recovery. 

9.8 LABORATORY FORTIFIED SAMPLE MATRIX (LFSM) – Analysis of an LFSM 

(Sect. 3.7) is required in each extraction batch and is used to determine that the sample 

matrix does not adversely affect method accuracy. If the occurrence of target analytes in 

the samples is infrequent, or if historical trends are unavailable, a second LFSM or 

552.3-21
LFSMD must be prepared, extracted, and analyzed from a duplicate field sample used to 

prepare the LFSM to assess method precision. Extraction batches that contain LFSMDs 

do not require the analysis of a Field Duplicate (Sect. 9.9). If a variety of different 

sample matrices are analyzed regularly, for example, drinking water from groundwater 

and surface water sources, method performance should be established for each. Over 

time, LFSM data should be documented for all routine sample sources for the laboratory. 

9.8.1 Within each extraction batch, a minimum of one Field Sample is fortified as an 

LFSM for every 20 samples extracted. The LFSM is prepared by spiking a 

sample with an appropriate amount of Analyte PDS (Sect. 7.2.4.2). Select a 

spiking concentration that is greater than or equal to the matrix background 

concentration, if known. Use historical data and rotate through the designated 

concentrations when selecting a fortifying concentration. 

9.8.2 Calculate the recovery (R) for each analyte using the equation 

where 

A = measured concentration in the fortified sample, 

B = measured concentration in the unfortified sample, and 

C = fortification concentration. 

9.8.3 Analyte recoveries may exhibit matrix bias. For samples fortified at or above 

their native concentration, recoveries should range between 70 and 130%, except 

for low-level fortification near or at the MRL (within a factor of 2-times the MRL 

concentration) where 50 to 150% recoveries are acceptable. If the accuracy of any 

analyte falls outside the designated range, and the laboratory performance for that 

analyte is shown to be in control in the CCCs, the recovery is judged to be matrix 

biased. The result for that analyte in the unfortified sample is labeled 

suspect/matrix to inform the data user that the results are suspect due to matrix 

effects. 

9.8.3.1 Because HAAs are disinfection by-products, many Field Samples will 

contain a number of HAAs at varying concentrations. Field Samples that 

have native HAA concentrations above the DL but below the MRL and 

are fortified at concentrations at or near the MRL should be corrected for 

552.3-22
the native levels in order to obtain meaningful R values. This is the only 

permitted use of analyte results below the MRL. 

9.9 FIELD DUPLICATE OR LABORATORY FORTIFIED SAMPLE MATRIX 

DUPLICATE (FD or LFSMD) – Within each extraction batch, a minimum of one Field 

Duplicate (FD) or Laboratory Fortified Sample Matrix Duplicate (LFSMD) must be 

analyzed. Duplicates check the precision associated with sample collection, preservation, 

storage, and laboratory procedures. HAAs are typically found in waters disinfected with 

chlorine; however, if target analytes are not routinely observed in Field Samples, an 

LFSMD should be periodically analyzed rather than an FD. 

9.9.1 Calculate the relative percent difference (RPD) for duplicate measurements (FD1 

and FD2) using the equation 

9.9.1.1 RPDs for Field Duplicates should fall in the range of ± 30%. Greater 

variability may be observed when Field Duplicates have analyte 

concentrations that are within a factor of 2 of the MRL. At these 

concentrations Field Duplicates should have RPDs that fall in the range of 

± 50%. If the RPD of any analyte falls outside the designated range, and 

the laboratory performance for that analyte is shown to be in control in the 

CCC, the recovery is judged to be matrix biased. The result for that 

analyte in the unfortified sample is labeled suspect/matrix to inform the 

data user that the results are suspect due to matrix effects. 

9.9.2 If an LFSMD is analyzed instead of a Field Duplicate, calculate the relative 

percent difference (RPD) for duplicate LFSMs (LFSM and LFSMD) using the 

equation 

9.9.2.1 RPDs for duplicate LFSMs should fall in the range of ± 30% for samples 

fortified at or above their native concentration. Greater variability may be 

observed when LFSMs are fortified at analyte concentrations that are 

within a factor of 2 of the MRL. LFSMs fortified at these concentrations 

552.3-23
should have RPDs that fall in the range of ± 50% for samples fortified at 

or above their native concentration. If the RPD of any analyte falls outside 

the designated range, and the laboratory performance for that analyte is 

shown to be in control in the CCC, the recovery is judged to be matrix 

biased. The result for that analyte in the unfortified sample is labeled 

suspect/matrix to inform the data user that the results are suspect due to 

matrix effects. 

9.10 QUALITY CONTROL SAMPLES (QCS) – As part of the IDC (Sect. 9.2), each time a 

new Analyte PDS (Sect. 7.2.4.2) is prepared, and at least quarterly, analyze a QCS 

sample from a source different from the source of the calibration standards. If a second 

vendor is not available then a different lot of the standard should be used. The QCS 

should be prepared and analyzed just like a CCC. Acceptance criteria for the QCS is 

identical to the CCCs; the calculated amount for each analyte must be + 30% of the 

expected value. If measured analyte concentrations are not of acceptable accuracy, check 

the entire analytical procedure to locate and correct the problem. 

10. CALIBRATION AND STANDARDIZATION 

10.1 An acceptable initial calibration must be established during the IDC and prior to 

analyzing Field or QC Samples. After initial calibration is successful, a Continuing 

Calibration Check (CCC) is required at the beginning and the end of each analysis batch, 

and after every tenth sample (Sect. 10.3). Because this is a procedural standard method, 

the analyst will need to make a decision to include either an appropriate number of CCCs 

or an entire initial calibration curve with each extraction batch. Initial calibration must be 

repeated each time a major instrument modification is made or maintenance is performed. 

Failure to meet CCC criteria may also require recalibration. 

10.2 INITIAL CALIBRATION – This method uses the procedural calibration technique to 

compensate for incomplete methylation of some of the target compounds. This is most 

pronounced for the brominated trihaloacetic acids. Many of the QC criteria throughout 

this method are expressed in terms of percent recovery. It should be noted that these 

recoveries are relative to the initial procedural curve rather than absolute recoveries. 

10.2.1 Establish GC operating parameters equivalent to the suggested specifications in 

Section 17, Table 1 (or Table 2). The GC system must be calibrated using the 

552.3-24
internal standard (IS) technique. Other columns or conditions may be used if 

equivalent or better performance can be demonstrated. 

10.2.2 Prepare a set of at least 5 calibration standards as described in Section 7.2.5. The 

lowest concentration calibration standard must be at or below the MRL, which 

will depend on system sensitivity. It is recommended that at least four of the 

calibration standards be at concentrations greater than the MRL. 

10.2.3 CALIBRATION – Use the GC data system software to generate a linear 

regression or quadratic calibration curve using the internal standard. The analyst 

may choose whether or not to force zero to obtain a curve that best fits the data. 

Examples of common GC system calibration curve options are: 1) Ax /Ais versus 

Qx /Qis; and 2) RRF versus Ax /Ais. Establish a relative response factor by using 

the equation 

where 

Ax = integrated peak area of the analyte, 

Ais = integrated peak area of the internal standard, 

Qx = quantity of analyte injected in ng or concentration units, 

Qis = quantity of internal standard injected in ng or concentration 

units, and 

RRF = relative response factor. 

10.2.4 Acceptance criteria for the calibration of each analyte is determined by calculating 

the concentration of each analyte and surrogate in each of the analyses used to 

generate the calibration curve. Each calibration point, except the lowest point, for 

each analyte should calculate to be 70-130% of its expected value. The lowest 

point should calculate to be 50-150% of its expected value. Laboratories that 

have difficulty achieving these criteria will have trouble meeting the QC 

requirements summarized in Section 10.3.2. These laboratories should reanalyze 

the calibration standards, restrict the range of calibration, or select an alternate 

method of calibration. 

552.3-25
10.3 CONTINUING CALIBRATION CHECK (CCC) – An appropriate number of CCCs 

must be prepared with each extraction batch. The CCC verifies the initial calibration at 

the beginning and the end of each analysis batch, and after every 10th sample during 

analyses. In this context, a “sample” is considered to be a Field Sample. LFSMs, 

LFSMDs, FSDs, and CCCs are not counted as samples. The beginning CCC for each 

analysis batch must be at or below the MRL in order to verify instrument sensitivity prior 

to any analyses. If standards have been prepared such that all analytes are not in the same 

Calibration Standard, it may be necessary to analyze two Calibration Standard solutions 

to meet this requirement. Subsequent CCCs should alternate between a medium and a 

high concentration standard. 

10.3.1 Inject an aliquot of the appropriate concentration calibration check standard 

solution prepared with the extraction batch and analyze using the same conditions 

used during the initial calibration. 

10.3.2 Calculate the concentration of each analyte and surrogate in the check standard. 

The calculated amount for each analyte for medium and high level CCCs must be 

± 30% of the expected value. The calculated amount for the lowest calibration 

level for each analyte must be within ± 50% of the expected value. If these 

conditions do not exist, then all data for the problem analyte must be considered 

invalid, and remedial action should be taken which may require recalibration. 

Any Field Sample extracts that have been analyzed since the last acceptable 

calibration verification should be reanalyzed after adequate calibration has been 

restored. For Extraction Batches with more than 10 Field Samples the analyst 

may either extract and analyze a third CCC or reanalyze the mid-level or highlevel CCC as the final CCC. 

11. PROCEDURE 

11.1 SAMPLE EXTRACTION 

11.1.1 Remove the samples from storage (Sect. 8.3) and allow them to equilibrate to 

room temperature. 

11.1.2 Place 40 mL of the water sample into a precleaned 60-mL glass vial with a PTFElined screw cap using a clean, graduated cylinder for each sample. 

552.3-26
11.1.3 Add 20 μL of surrogate standard (20 μg/mL of 2-bromobutanoic acid in MTBE 

per Section 7.2.3.2) to the aqueous sample. 

NOTE: When fortifying an aqueous sample with components that are contained in 

MTBE solution, be sure that the needle of the syringe is well below the surface of 

the water. After injection, cap the sample and invert once. This will insure that 

the standard solution is mixed well. 

11.1.4 Adjust the pH to less than or equal to 0.5 by adding up to 2 mL of concentrated 

sulfuric acid. Cap, mix and then check the pH with narrow range pH paper (Sect. 

6.11). Substitution of other acids is not allowed. 

11.1.5 Add approximately 18 g of muffled sodium sulfate (Sect. 7.1.4) and immediately 

shake until almost all is dissolved. Substitution of other salts for sodium 

sulfate is not allowed. Sodium sulfate is added to increase the ionic strength of 

the aqueous phase and thus further drive the haloacetic acids into the organic 

phase. The addition of salt also decreases the solubility of MTBE or TAME in 

the aqueous phase and allows greater volumetric recovery of the extraction 

solvent. The addition of this salt should be done immediately after the addition of 

the sulfuric acid so that the heat generated from the addition of the acid (Sect. 

11.1.4) will help dissolve the salt. 

11.1.6 Add exactly 4.0 mL MTBE with IS or TAME with IS (Sect. 7.2.2.3) and shake 

vigorously for three minutes by hand. This may be accomplished for a entire 

Extraction Batch using a test tube rack. 

11.1.7 Allow the phases to separate for approximately 5 minutes. 

11.2 SAMPLE METHYLATION WITH ACIDIC METHANOL 

11.2.1 Using a pasteur pipet, transfer 3 mL of the upper MTBE or TAME layer to a 

15-mL graduated conical centrifuge tube. 

11.2.2 Add 3 mL of 10% sulfuric acid in methanol (Sect. 7.1.12) to each centrifuge tube. 

Cap the tube. 

11.2.3 Methylation of the method analytes is accomplished during this step. Careful 

control of both reaction time and reaction temperature are critical to method 

552.3-27
precision and accuracy.(9) Place the tubes in a heating block (or sand bath) at 

50 ± 2 °C (for MTBE) or 60 ± 2 °C (for TAME) and heat for 2 hours (± 10 min). 

The tubes should fit snugly into the heating block to ensure proper heat transfer. 

Verify the reaction temperature by placing a thermometer into a tube containing 

water rather than inserting it into the block to ensure an accurate reading. Placing 

the thermometer directly in the heating block well or sand bath will give a higher 

temperature reading than the actual sample temperature. The MTBE reaction 

temperature is set at its highest practical limit (50 °C), since MTBE boils at 55 °C. 

Similarly, the TAME reaction temperature has been set at 60 °C, since methanol 

boils at 65 °C. This helps prevent solvent loss from the reaction tubes. 

11.2.3.1 Even at 2 hours, methylation for some of the more sterically hindered 

compounds like TBAA, CDBAA and BDCAA is not complete. 

Shortening reaction time will decrease methylation efficiencies for 

compounds and result in lower precision. 

11.2.3.2 Methylation efficiencies are increased with increasing temperature (and 

time). The TAME procedure has higher methylation efficiencies than 

MTBE procedure for the sterically hindered HAAs like the brominated 

trihaloacetic acids (Sect. 11.2.3.1) and is more precise for these targets. 

Methylation temperature, however, should not be increased above 

the recommended reaction temperatures (Sect. 11.2.3). 

11.2.3.3 Lower reaction temperatures or times are not allowed. Care should 

be taken to ensure that the calibration standards are heated identically to 

Field Samples in the extraction batch. 

11.2.3.4 Methylation may be accomplished by heating the reaction tubes with a 

water bath, provided the water bath is not covered in a way as to cause 

the entire tube to be heated. If tube walls are heated, the tubes can lose 

some of their contents leading to higher variability in analytical results. 

A water bath that is covered with a layer of small, floating plastic 

spheres may be used (Sect. 6.10). 

11.2.4 Remove the centrifuge tubes from the heating source and allow them to cool 

before removing their caps. 

552.3-28
11.2.5 Add 7 mL of a 150 g/L sodium sulfate solution (Sect. 7.1.10) to each centrifuge 

tube. Vortex each tube to ensure full equilibration between the phases. Allow the 

two phases to settle fully, but do not allow the tubes to sit more than a few 

minutes. Adding the sodium sulfate solution may cause some loss of the formed 

HAA-esters through acid-catalyzed hydrolysis over prolonged periods. 

11.2.6 Remove and discard the lower (acidic aqueous methanol) phase from each tube 

with a long pasteur pipet. Leave no more than 0.3 mL of aqueous phase to ensure 

complete neutralization in the following step. 

11.2.7 Add 1 mL of saturated sodium bicarbonate solution (Sect. 7.1.11). Vortex each 

centrifuge tube for several seconds at least four times to complete the 

neutralization reaction. Loosen the tube caps after the first agitation to release the 

evolved CO2. 

11.2.8 Transfer 1 mL of the upper ether layer to an auto-sampler vial. A duplicate vial 

should be filled using the excess extract. 

11.2.9 Analyze the samples as soon as possible. Store the extracts at #-10 °C (Sect. 8.4). 

11.3 GAS CHROMATOGRAPHY 

11.3.1 The instrument used in the development of this method was equipped with a low 

volume (150 μL) micro electron capture detector (ECD). Other configurations are 

allowed as described in Section 6.14. 

11.3.2 COLUMN SELECTION AND INSTALLATION – Strict attention should be paid 

to established column installation guidelines with regard to the proper cutting and 

placement of the capillary columns within the instrument. The brominated 

trihaloacetic acids are particularly sensitive to the condition of the injection port 

and GC column. If the response for these or other method analytes diminish, 

trimming approximately 0.5 m from the head of the column and replacing the GC 

inlet liner often restores the response for these analytes. Quartz inlet liners and 

inlet liners with standard deactivation were found to require less frequent 

maintenance than those with SiltekTM deactivation for this method. If conditions 

in the laboratory necessitate frequent column trimming, a guard column is 

recommended. 

552.3-29
11.3.3 Method development was conducted with the GC set in constant pressure mode. 

If constant velocity mode is used by the analyst, linear velocities should be 

reduced from those listed in Tables 1-4 to obtain similar resolution. 

11.4 ANALYSIS OF EXTRACTS 

11.4.1 Establish operating conditions as described in Section 17, Table 1 or Table 2 

(Table 3 or 4 if performing confirmation). Confirm that retention times, 

compound separation, and resolution are similar to those summarized in Tables 1-

4 and Figures 1- 4. 

11.4.2 Establish an appropriate relative retention time window for each target and 

surrogate to identify them in the QC and Field Samples. This should be based on 

measurements of actual relative retention time variation for each compound in 

standard solutions analyzed on the GC over the course of time. Plus or minus 

three times the standard deviation of the relative retention time observed for each 

compound while establishing the initial calibration can be used to calculate a 

suggested window size; however, the experience of the analyst should weigh 

heavily on the determination of the appropriate retention window. 

11.4.3 Calibrate the instrument as described in Section 10.2 or confirm the calibration is 

still valid by analyzing CCCs as described in Section 10.3. Begin analyzing Field 

and QC Samples at their appropriate frequency by injecting aliquots under the 

same conditions used to establish the initial calibration. 

11.4.4 The analyst must not extrapolate beyond the established calibration range. If an 

analyte result exceeds the range of the initial calibration curve, the extract may be 

diluted with MTBE or TAME containing the internal standard (Sect. 7.2.1.3), and 

the diluted extract injected. Acceptable surrogate performance (Sect. 9.7) should 

be determined from the undiluted sample extract. Incorporate the dilution factor 

into final concentration calculations. The dilution will also affect analyte MRLs. 

12. DATA ANALYSIS AND CALCULATION 

12.1 Identify the method analytes in the sample chromatogram by comparing the retention 

time of the suspect peak to retention time of an analyte peak in a calibration standard. 

552.3-30
Surrogate retention times should be confirmed to be within acceptance limits (Sect. 

11.4.2) even if no target compounds are detected. 

12.2 Calculate the analyte concentrations using the initial calibration curve generated as 

described in Section 10.2. Quantitate only those values that fall between the MRL and 

the highest calibration standard. Samples with target analyte responses that exceed the 

highest standard require dilution and reanalysis (Sect. 11.4.4). 

12.3 Analyte identifications should be confirmed using the confirmation column specified in 

Table 3 (or Table 4) or another column that is dissimilar to the primary column. GC/MS 

confirmation is acceptable if the analyte concentrations are sufficient. 

12.4 Adjust the calculated concentrations of the detected analytes to reflect the initial sample 

volume and any dilutions performed. 

12.5 Analyte concentrations are reported in μg/L as the total free acid (usually to 2 significant 

figures); however, calculations should use all available digits of precision. 

13. METHOD PERFORMANCE 

13.1 PRECISION, ACCURACY, AND DETECTION LIMITS – Tables for these data are 

presented in Section 17. Detection Limits are presented in Table 5 and were calculated 

using the formula presented in Section 9.2.4. Single laboratory precision and accuracy 

data are presented in Tables 6-11. 

13.2 SAMPLE STORAGE STABILITY STUDIES – An analyte storage stability study was 

conducted by fortifying the analytes (10 μg/L of each analyte) into a chlorinated surface 

water that was collected, preserved, and stored as described in Section 8. The average of 

triplicate analyses, conducted on days 0, 3, 7, and 14 are presented in Table 12. 

13.3 EXTRACT STORAGE STABILITY STUDIES – Extract storage stability studies were 

conducted on TAME and MTBE extracts obtained from a chlorinated surface water 

fortified at 10 μg/L. The average of triplicate injections are reported in Tables 13 and 14. 

Extract storage stability can decline in the presence of peroxides. This is discussed in 

Section 8.4. 

552.3-31
14. POLLUTION PREVENTION 

14.1 This method utilizes liquid-liquid microextraction to extract analytes from water. It 

requires the use of very small volumes of organic solvent and very small quantities of 

pure analytes, thereby minimizing the potential hazards to both the analyst and the 

environment as compared to the use of large volumes of organic solvents in conventional 

liquid-liquid extractions. 

14.2 For information about pollution prevention that may be applicable to laboratory 

operations, consult “Less is Better: Laboratory Chemical Management for Waste 

Reduction” available from the American Chemical Society’s Department of Government 

Relations and Science Policy, 1155 16th Street N.W., Washington, D.C., 20036. 

15. WASTE MANAGEMENT 

15.1 The analytical procedures described in this method generate relatively small amounts of 

waste since only small amounts of reagents and solvents are used. The matrices of 

concern are finished drinking water or source water. However, the Agency requires that 

laboratory waste management practices be conducted consistent with all applicable rules 

and regulations, and that laboratories protect the air, water, and land by minimizing and 

controlling all releases from fume hoods and bench operations. Also, compliance is 

required with any sewage discharge permits and regulations, particularly the hazardous 

waste identification rules and land disposal restrictions. For further information on waste 

management, consult “The Waste Management Manual for Laboratory Personnel” also 

available from the American Chemical Society at the address in Section 14.2. 

16. REFERENCES 

1. Glaser, J.A., Foerst, D.L., McKee, G.D., Quave, S.A., and Budde, W.L., “Trace Analyses 

for Wastewaters,” Environ. Sci. Technol., 15 (1981) 1426-1435. 

2. Giam, C.S., Chan, H.S., and Nef, G.S., "Sensitive Method for Determination of Phthalate 

Ester Plasticizers in Open-Ocean Biota Samples," Analytical Chemistry, 47, 2225 (1975). 

552.3-32
3. Giam, C.S., and Chan, H.S., "Control of Blanks in the Analysis of Phthalates in Air and 

Ocean Biota Samples," U.S. National Bureau of Standards, Special Publication 442, 

701-708, (1976). 

4. "OSHA Safety and Health Standards, General Industry," (29CRF1910). Occupational 

Safety and Health Administration, OSHA 2206, (Revised, Jan. 1976). 

5. “Carcinogens-Working with Carcinogens,” Publication No. 77-206, Department of 

Health, Education, and Welfare, Public Health Service, Center for Disease Control, 

National Institute of Occupational Safety and Health, Atlanta, Georgia, August 1977. 

6. “Safety In Academic Chemistry Laboratories,” 3rd Edition, American Chemical Society 

Publication, Committee on Chemical Safety, Washington, D.C., (1979). 

7. ASTM Annual Book of Standards, Part II, Volume 11.01, D3370-82, "Standard Practice 

for Sampling Water," American Society for Testing and Materials, Philadelphia, PA, 

1986. 

8. Pepich, B.V., Domino, M.M., Munch, D.J. and Fair, P.S. “Validating Sample 

Preservation Techniques and Holding Times for the Approved Compliance Monitoring 

Methods for Haloacetic Acids Under the U.S. EPA’s Stage 1 D/DBP Rule,” Submitted to 

Wat. Res., Sept. 2002. 

9. Xie, Y., Rashid, I., Zhou, H., and Gammie, L. “Acidic Methanol Methylation for HAA 

Analysis: Limitations and Possible Solutions” JAWWA, 94:11, 115-122 (Nov. 2002). 

552.3-33
17. TABLES, DIAGRAMS, FLOWCHARTS, AND VALIDATION DATA 

TABLE 1: CHROMATOGRAPHIC CONDITIONS AND AVERAGE RETENTION TIME 

DATA FOR THE PRIMARY COLUMN (DB-1701) USING MTBE 

Compound 

Average Tr 

(min)* 

RSD 

(% ) 

Monochloroacetic acid (MCAA) 9.29 0.03 

Monobromoacetic acid (MBAA) 14.19 0.02 

Dalapon 14.69 0.02 

Dichloroacetic acid (DCAA) 15.07 0.02 

Bromoform** 17.12 0.01 

Trichloroacetic acid (TCAA) 18.89 0.01 

1,2,3 Trichloropropane (IS) 20.69 0.01 

Bromochloroacetic acid (BCAA) 21.22 0.01 

2-bromobutanoic acid (SUR) 21.63 0.01 

Bromodichloroacetic acid (BDCAA) 23.60 0.01 

Dibromoacetic acid (DBAA) 24.01 0.005 

Chlorodibromoacetic acid (CDBAA) 25.55 0.004 

Tribromoacetic acid (TBAA) 26.88 0.003 

*The average retention time represents the average of 8 injections of fortified reagent water extracts that had been fortified at

10 μg/L of each analyte. 

**Bromoform is not a target analyte. RT value is provided for information only. 

Primary Column: DB-1701, 30 m x 0.25 mm i.d., 0.25 μm film thickness.

Injector: Injector temperature, 210 oC; 2-mm straight quartz liner; injection volume 1 μL; splitless injection hold for 

45 sec then purge @ 30 mL/min. 

GC Program: (for MTBE) 40 oC initial held for 10 minutes, program at 2.5 oC/min to 65 oC, then 10 oC/min to 85 oC, then 

20 o

C/min to 205 o

C, hold for 7 min. 

Detector: Agilent Micro ECD (150 μL volume); detector temperature, 290 oC; detector make up gas 95% Ar/5% CH4 

at 20 mL/min. 

Carrier Gas: Helium (UHP), set at constant pressure. Initial carrier gas velocity (at 40 oC) 33 cm/sec. 

Data Collection: Agilent GC Chemstation with a digitization rate of 20 Hz. 

552.3-34
TABLE 2: CHROMATOGRAPHIC CONDITIONS AND AVERAGE RETENTION TIME

DATA FOR THE PRIMARY COLUMN (DB-1701) USING TAME

Compound 

Average Tr 

(min)* 

RSD 

(%) 

Monochloroacetic acid (MCAA) 5.94 0.08 

Monobromoacetic acid (MBAA) 8.76 0.04 

Dalapon 9.17 0.04 

Dichloroacetic acid (DCAA) 9.34 0.04 

Bromoform** 11.06 0.02 

Trichloroacetic acid (TCAA) 12.28 0.02 

1,2,3 Trichloropropane (IS) 13.56 0.01 

Bromochloroacetic acid (BCAA) 13.87 0.01 

2-bromobutanoic acid (SUR) 14.20 0.01 

Bromodichloroacetic acid (BDCAA) 15.78 0.004 

Dibromoacetic acid (DBAA) 16.14 0.003 

Chlorodibromoacetic acid (CDBAA) 17.58 0.002 

Tribromoacetic acid (TBAA) 18.89 0.02 

*The average retention time represents the average of 8 injections of fortified reagent water extracts that had been fortified at

10 μg/L of each analyte. 

**Bromoform is not a target analyte. RT value is provided for information only.

Primary Column: DB-1701, 30 m x 0.25 mm i.d., 0.25 μm film thickness.

Injector: Injector temperature, 210 o

C; 2-mm straight quartz liner; injection volume 1 μL; splitless injection hold for 

45 sec then purge @ 30 mL/min. 

GC Program: (for TAME) 55 oC initial hold 8 minutes, program at 2.5 oC/min to 65 oC, then 10 oC/min to 85 oC, then 20 o

C/min to 205 oC, hold for 7 min. 

Detector: Agilent Micro ECD (150 μL volume); detector temperature, 290 oC; detector make up gas 95% Ar/5% CH4 

at 20 mL/min. 

Carrier Gas: Helium (UHP), set at constant pressure. Initial carrier gas velocity (at 55 oC) 32 cm/sec. 

Data Collection: Agilent GC Chemstation with a digitization rate of 20 Hz. 

552.3-35
TABLE 3: CHROMATOGRAPHIC CONDITIONS AND AVERAGE RETENTION TIME 

DATA FOR THE CONFIRMATION COLUMN (DB-5.625) USING MTBE 

Compound 

Average Tr 

(min)* 

RSD 

(%) 

Monochloroacetic acid (MCAA) 5.28 0.04 

Monobromoacetic acid (MBAA) 8.31 0.01 

Dichloroacetic acid (DCAA) 9.05 0.01 

Dalapon 10.63 0.02 

Bromoform** 12.74 0.02 

Trichloroacetic acid (TCAA) 14.73 0.02 

Bromochloroacetic acid (BCAA) 15.25 0.02 

1,2,3 Trichloropropane (IS) 15.65 0.01 

2-bromobutanoic acid (SUR) 17.41 0.02 

Dibromoacetic acid (DBAA) 21.08 0.01 

Bromodichloroacetic acid (BDCAA) 21.39 0.01 

Chlorodibromoacetic acid (CDBAA) 24.35 0.002 

Tribromoacetic acid (TBAA) 25.94 0.000 

*The average retention time represents the average of 8 injections of fortified reagent water extracts that had been fortified at

10 μg/L of each analyte. 

**Bromoform is not a target analyte. RT value is provided for information only. 

Confirmation 

Column: DB-5.625, 30 m x 0.25 mm i.d., 0.25 μm film thickness.

Injector: Injector temperature, 210 o

C; 2-mm straight quartz liner; injection volume 1 μL ; splitless injection hold for 

45 sec then purge @ 30 mL/min. 

GC Program: (for MTBE) 40 oC initial held for 10 minutes, program at 2.5 oC/min to 65 oC, then 10 oC/min to 85 oC, then 

20 o

C/min to 205 o

C,. Post run 210 oC hold for 7 min. 

Detector: Agilent Micro ECD (150 μL volume); detector temperature, 290 oC; detector make up gas 95% Ar/5% CH4 

at 20 mL/min. 

Carrier Gas: Helium (UHP), set at constant pressure. Initial carrier gas velocity (at 40 oC) 32 cm/sec. 

Data Collection: Agilent GC Chemstation with a digitization rate of 20 Hz. 

552.3-36
TABLE 4: CHROMATOGRAPHIC CONDITIONS AND AVERAGE RETENTION TIME 

DATA FOR THE CONFIRMATION COLUMN (DB-5.625) USING TAME 

Compound 

Average Tr 

(min)* 

RSD 

(%) 

Monochloroacetic acid (MCAA) 4.07 0.26 

Monobromoacetic acid (MBAA) 5.47 0.14 

Dichloroacetic acid (DCAA) 5.81 0.12 

Dalapon 6.61 0.10 

Bromoform** 7.97 0.07 

Trichloroacetic acid (TCAA) 9.11 0.06 

Bromochloroacetic acid (BCAA) 9.44 0.05 

1,2,3 Trichloropropane (IS) 9.79 0.05 

2-bromobutanoic acid (SUR) 11.05 0.03 

Dibromoacetic acid (DBAA) 13.77 0.01 

Bromodichloroacetic acid (BDCAA) 14.01 0.01 

Chlorodibromoacetic acid (CDBAA) 16.45 0.008 

Tribromoacetic acid (TBAA) 17.96 0.004 

*The average retention time represents the average of 8 injections of fortified reagent water extracts that had been fortified at

10 μg/L of each analyte. 

**Bromoform is not a target analyte. RT value is provided for information only.

Confirmation 

Column: DB-5.625, 30 m x 0.25 mm i.d., 0.25 μm film thickness.

Injector: Injector temperature, 210 o

C; 2-mm straight quartz liner; injection volume 1 μL; splitless injection hold for 

45 sec then purge @ 30 mL/min. 

GC Program: (for TAME) 55 oC initial held for 8 minutes, program at 2.5 oC/min to 65 oC, then 10 oC/min to 85 oC, then 

20 o

C/min to 205 o

C,. Post run 210 oC hold for 7 min. 

Detector: Agilent Micro ECD (150 μL volume); detector temperature, 290 oC; detector make up gas 95% Ar/5% CH4 

at 20 mL/min. 

Carrier Gas: Helium (UHP), set at constant pressure. Initial carrier gas velocity (at 55 oC) 32 cm/sec. 

Data Collection: Agilent GC Chemstation with a digitization rate of 20 Hz. 

552.3-37
TABLE 5: DETECTION LIMITS IN REAGENT WATER USING THE MTBE AND TAME 

PROCEDURES 

Compound 

MTBE TAME 

Fortification 

Level 

(μg/L) 

Detection 

Limita 

(μg/L) 

Fortification 

Level 

(μg/L) 

Detection 

Limita 

(μg/L) 

Monochloroacetic acid (MCAA) 1.00 0.17 1.00 0.20 

Monobromoacetic acid (MBAA) 0.50 0.027 0.50 0.13 

Dalapon 0.50 0.024 0.50 0.14 

Dichloroacetic acid (DCAA) 0.50 0.020 0.50 0.084 

Trichloroacetic acid (TCAA) 0.50 0.019 0.50 0.024 

Bromochloroacetic acid (BCAA) 0.50 0.016 0.50 0.029 

Bromodichloroacetic acid (BDCAA) 0.50 0.034 0.50 0.031 

Dibromoacetic acid (DBAA) 0.50 0.012 0.50 0.021 

Chlorodibromoacetic acid (CDBAA) 0.50 0.054 0.50 0.035 

Tribromoacetic acid (TBAA) 0.50 0.11 0.50 0.097 

a

Fortified reagent waters were extracted and analyzed over 3 days for 7-9 replicates following the 

procedure outlined in Section 9. MTBE detection limits were determined with 9 replicates. TAME 

detection limits were determined with 7 replicates. 

552.3-38
TABLE 6: PRECISION AND ACCURACY IN REAGENT WATER FORTIFIED AT 1.0 μg/L 

Compound 

MTBE TAME 

Mean 

Recovery (%) 

RSD 

(%) 

(n=8) 

S/N 

Ratioa 

Mean 

Recovery 

(%) 

RSD 

(%) 

(n=8) 

S/N 

Ratioa 

Monochloroacetic acid (MCAA) 95.8 4.0 12 81.4 5.1 17 

Monobromoacetic acid (MBAA) 92.2 1.8 81 90.7 3.7 92 

Dalapon 97.5 1.6 110 92.8 2.1 150 

Dichloroacetic acid (DCAA) 93.8 1.6 190 97.8 2.2 190 

Trichloroacetic acid (TCAA) 105 0.52 580 107 0.90 520 

Bromochloroacetic acid (BCAA) 102 0.36 580 103 0.94 740 

Bromodichloroacetic acid (BDCAA) 117 1.2 1400 113 1.1 1800 

Dibromoacetic acid (DBAA) 105 0.63 1300 105 0.86 1800 

Chlorodibromoacetic Acid (CDBAA) 125 2.3 650 112 1.5 1600 

Tribromoacetic acid (TBAA) 128 4.1 970 109 1.8 1100 

a

Signal-to-noise ratios were calculated for each target compound peak by dividing the peak height for 

each compound by the peak-to-peak noise, which was determined for each component from the method 

blank over a period of time equal to the full peak width in the target analyte’s retention time window. 

552.3-39
TABLE 7: PRECISION AND ACCURACY IN REAGENT WATER FORTIFIED AT 10 μg/L 

Compound 

MTBE TAME 

Mean 

Recoverya 

(%) 

RSD 

(%) 

(n=8) 

Mean 

Recoverya 

(%) 

RSD 

(%) 

(n=8) 

Monochloroacetic acid (MCAA) 101 3.5 102 2.5 

Monobromoacetic acid (MBAA) 101 2.2 101 1.6 

Dalapon 99.8 0.88 99.9 0.50 

Dichloroacetic acid (DCAA) 98.2 0.73 100 0.33 

Trichloroacetic acid (TCAA) 102 1.2 101 1.1 

Bromochloroacetic acid (BCAA) 101 0.72 101 0.65 

Bromodichloroacetic acid (BDCAA) 107 2.4 103 1.2 

Dibromoacetic acid (DBAA) 102 0.52 101 0.59 

Chlorodibromoacetic acid (CDBAA) 111 3.8 104 1.6 

Tribromoacetic acid (TBAA) 113 4.7 104 2.0 

552.3-40
TABLE 8: PRECISION AND ACCURACY IN CHLORINATED SURFACE WATER 

FORTIFIED AT 1.0 μg/L 

Compound 

MTBE TAME 

Mean 

Recoverya 

(%) 

RSD 

(%) 

(n=8) 

Mean 

Recoveryb 

(%) 

RSD 

(%) 

(n=8) 

Monochloroacetic acid (MCAA) 112 6.2 131 6.2 

Monobromoacetic acid (MBAA) 99.5 2.3 98.6 4.2 

Dalapon 107 2.2 103 3.5 

Dichloroacetic acid (DCAA) 103 1.5 106 3.8 

Trichloroacetic acid (TCAA) 89.2 1.4 89.0 1.1 

Bromochloroacetic acid (BCAA) 99.5 1.6 102 2.2 

Bromodichloroacetic acid (BDCAA) 91.7 4.0 87.5 1.8 

Dibromoacetic acid (DBAA) 103 2.8 103 4.5 

Chlorodibromoacetic acid (CDBAA) 95.5 7.1 94.4 3.3 

Tribromoacetic acid (TBAA) 110 8.1 111 4.3 

a

Recoveries were corrected for haloacetic acid (HAA) concentrations in the unfortified matrix, based on 

an average value of three measurements. HAAs detected in the unfortified matrix included MBAA (0.14 

μg/L), DCAA (0.56 μg/L), TCAA (0.32 μg/L), BCAA (1.1 μg/L), BDCAA (0.68 μg/L), DBAA (2.2 

μg/L), CDBAA (0.91 μg/L), and TBAA (1.0 μg/L). 

b

Recoveries were corrected for haloacetic acid (HAA) concentrations in the unfortified matrix, based on 

an average value of three measurements. HAAs detected in the unfortified matrix included MBAA (0.22 

μg/L), DCAA (0.78 μg/L), TCAA (0.36 μg/L), BCAA (2.0 μg/L), BDCAA (0.62 μg/L), DBAA (4.0 

μg/L), CDBAA (0.82 μg/L), and TBAA (0.79 μg/L). 

552.3-41
TABLE 9: PRECISION AND ACCURACY IN CHLORINATED SURFACE WATER 

FORTIFIED AT 10 μg/L 

Compound 

MTBE TAME 

Mean 

Recoverya 

(%) 

RSD 

(%) 

(n=8) 

Mean 

Recoveryb 

(%) 

RSD 

(%) 

(n=8) 

Monochloroacetic acid (MCAA) 97.8 3.1 97.1 3.8 

Monobromoacetic acid (MBAA) 98.8 2.0 98.8 2.0 

Dalapon 99.6 0.90 97.7 0.42 

Dichloroacetic acid (DCAA) 98.0 1.1 97.6 0.42 

Trichloroacetic acid (TCAA) 99.8 0.80 100 1.4 

Bromochloroacetic acid (BCAA) 101 1.4 101 0.93 

Bromodichloroacetic acid (BDCAA) 103 4.3 104 2.3 

Dibromoacetic acid (DBAA) 102 1.4 101 0.93 

Chlorodibromoacetic acid (CDBAA) 105 5.5 105 3.2 

Tribromoacetic acid (TBAA) 108 6.3 106 3.4 

a

Recoveries were corrected for haloacetic acid (HAA) concentrations in the unfortified matrix, based on 

an average value of three measurements. HAAs detected in the unfortified matrix included MBAA (0.14 

μg/L), DCAA (0.56 μg/L), TCAA (0.32 μg/L), BCAA (1.1 μg/L), BDCAA (0.68 μg/L), DBAA (2.2 

μg/L), CDBAA (0.91 μg/L), and TBAA (1.0 μg/L). 

b

Recoveries were corrected for haloacetic acid (HAA) concentrations in the unfortified matrix, based on 

an average value of three measurements. HAAs detected in the unfortified matrix included MBAA (0.22 

μg/L), DCAA (0.78 μg/L), TCAA (0.36 μg/L), BCAA (2.0 μg/L), BDCAA (0.62 μg/L), DBAA (4.0 

μg/L), CDBAA (0.82 μg/L), and TBAA (0.79 μg/L). 

552.3-42
TABLE 10: PRECISION AND ACCURACY IN CHLORINATED GROUND WATER 

FORTIFIED AT 1.0 μg/L 

Compound 

MTBE TAME 

Mean 

Recoverya 

(%) 

RSD 

(%) 

(n=8) 

Mean 

Recoveryb 

(%) 

RSD 

(%) 

(n=8) 

Monochloroacetic acid (MCAA) 126 4.5 124 9.5 

Monobromoacetic acid (MBAA) 113 3.8 98.9 2.6 

Dalapon 112 1.1 87.3 2.7 

Dichloroacetic acid (DCAA) 102 3.7 107 3.2 

Trichloroacetic acid (TCAA) 92.2 1.2 103 2.1 

Bromochloroacetic acid (BCAA) 106 3.8 104 2.4 

Bromodichloroacetic acid (BDCAA) 105 6.1 106 4.0 

Dibromoacetic acid (DBAA) 111 5.3 105 1.7 

Chlorodibromoacetic acid (CDBAA) 103 8.8 103 4.5 

Tribromoacetic acid (TBAA) 99.2 7.3 103 3.2 

a

Recoveries were corrected for haloacetic acid (HAA) concentrations in the unfortified matrix, based on 

an average value of three measurements. HAAs detected in the unfortified matrix included MBAA (0.07 

μg/L), Dalapon (0.19 μg/L), DCAA (2.6 μg/L), TCAA (1.1 μg/L), BCAA (3.0 μg/L), BDCAA (1.8 

μg/L), DBAA (3.3 μg/L), CDBAA (1.8 μg/L), and TBAA (1.1 μg/L). 

b

Recoveries were corrected for haloacetic acid (HAA) concentrations in the unfortified matrix, based on 

an average value of three measurements. HAAs detected in the unfortified matrix included MBAA (0.28 

μg/L), Dalapon (0.35 μg/L) DCAA (2.7 μg/L), TCAA (1.6 μg/L), BCAA (3.2 μg/L), BDCAA (2.4 

μg/L), DBAA (2.5 μg/L), CDBAA (1.7 μg/L), and TBAA (0.54 μg/L). 

552.3-43
TABLE 11: PRECISION AND ACCURACY IN CHLORINATED GROUND WATER 

FORTIFIED AT 10 μg/L 

Compound 

MTBE TAME 

Mean 

Recoverya 

(%) 

RSD 

(%) 

(n=8) 

Mean 

Recoveryb 

(%) 

RSD 

(%) 

(n=8) 

Monochloroacetic acid (MCAA) 104 1.8 99.7 1.7 

Monobromoacetic acid (MBAA) 100 1.4 98.7 1.1 

Dalapon 97.8 1.1 97.9 0.63 

Dichloroacetic acid (DCAA) 95.9 1.0 97.1 0.83 

Trichloroacetic acid (TCAA) 98.2 0.81 101 1.4 

Bromochloroacetic acid (BCAA) 100 0.53 99.8 1.4 

Bromodichloroacetic acid (BDCAA) 106 1.8 106 2.3 

Dibromoacetic acid (DBAA) 101 0.89 101 1.6 

Chlorodibromoacetic acid (CDBAA) 110 2.8 106 2.8 

Tribromoacetic acid (TBAA) 116 3.4 105 2.6 

a

Recoveries were corrected for haloacetic acid (HAA) concentrations in the unfortified matrix, based on 

an average value of three measurements. HAAs detected in the unfortified matrix included MBAA (0.07 

μg/L), Dalapon (0.19 μg/L), DCAA (2.6 μg/L), TCAA (1.1 μg/L), BCAA (3.0 μg/L), BDCAA (1.8 

μg/L), DBAA (3.3 μg/L), CDBAA (1.8 μg/L), and TBAA (1.1 μg/L). 

b

Recoveries were corrected for haloacetic acid (HAA) concentrations in the unfortified matrix, based on 

an average value of three measurements. HAAs detected in the unfortified matrix included MBAA (0.28 

μg/L), Dalapon (0.35 μg/L) DCAA (2.7 μg/L), TCAA (1.6 μg/L), BCAA (3.2 μg/L), BDCAA (2.4 

μg/L), DBAA (2.5 μg/L), CDBAA (1.7 μg/L), and TBAA (0.54 μg/L). 

552.3-44
TABLE 12: SAMPLE HOLDING TIME DATA FOR SAMPLES FROM A CHLORINATED 

SURFACE WATER FORTIFIED WITH METHOD ANALYTESa 

Compound 

Day 0 

Rb 

(%) 

Day 3 

Rb 

(%) 

Day 7 

Rb 

(%) 

Day 14 

Rb 

(%) 

Monochloroacetic Acid (MCAA) 97.9 96.8 98.5 104 

Monobromoacetic Acid (MBAA) 107 99.6 105 109 

Dalapon 104 104 103 99.7 

Dichloroacetic Acid (DCAA) 101 99.0 102 98.8 

Trichloroacetic Acid (TCAA) 101 102 98.8 96.2 

Bromochloroacetic Acid (BCAA) 101 99.3 100 97.8 

Bromodichloroacetic Acid (BDCAA) 112 111 113 110 

Dibromoacetic Acid (DBAA) 102 99.1 100 96.9 

Chlorodibromoacetic Acid (CDBAA) 117 115 119 116 

Tribromoacetic Acid (TBAA) 123 118 124 121 

a

 All samples were fortified at 10 μg/L. All samples were stored at 10 o

C for 48 hours, and at 6 o

C 

thereafter. 

b

Recoveries were corrected for haloacetic acid (HAA) concentrations in the unfortified matrix, based on 

an average value of three measurements. HAAs detected in the unfortified matrix included MBAA (0.30 

μg/L), DCAA (0.73 μg/L), TCAA (0.33 μg/L), BCAA (2.0 μg/L), BDCAA (0.67 μg/L), DBAA (4.3 

μg/L), CDBAA (0.91 μg/L), and TBAA (1.1 μg/L). 

552.3-45
TABLE 13: MTBE EXTRACT HOLDING TIME DATA FOR SAMPLES FROM A 

CHLORINATED SURFACE WATER FORTIFIED WITH METHOD 

ANALYTESa 

Compound 

Initial 

R 

(%) 

Day 7 

R 

(%) 

Day 14 

R 

(%) 

Day 21 

R 

(%) 

Monochloroacetic Acid (MCAA) 99.0 98.6 100 99.3 

Monobromoacetic Acid (MBAA) 99.7 98.2 98.1 99.0 

Dalapon 99.0 96.7 98.1 97.8 

Dichloroacetic Acid (DCAA) 97.9 96.3 97.5 97.3 

Trichloroacetic Acid (TCAA) 99.3 99.6 98.5 99.1 

Bromochloroacetic Acid (BCAA) 100 101 100 101 

Bromodichloroacetic Acid (BDCAA) 99.3 106 102 103 

Dibromoacetic Acid (DBAA) 101 102 101 102 

Chlorodibromoacetic Acid (CDBAA) 99.7 107 102 103 

Tribromoacetic Acid (TBAA) 102 110 102 106 

a

Extract storage stability was conducted on three of the eight extracts from the precision and accuracy 

study conducted in chlorinated surface water (fortified at 10 μg/L) reported in Table 9. Sample storage 

stability is expressed as a recovery value (%) calculated as described in Section 9.8.2. All values have 

been corrected for background levels in the unfortified sample based on triplicate measurements as 

described in Table 9. Extracts were stored at < -10 °C. 

552.3-46
TABLE 14: TAME EXTRACT HOLDING TIME DATA FOR SAMPLES FROM A 

CHLORINATED SURFACE WATER FORTIFIED WITH METHOD 

ANALYTESa 

Compound 

Initial 

R 

(%) 

Day 7 

R 

(%) 

Day 14 

R 

(%) 

Day 21 

R 

(%) 

Day 28 

R 

(%) 

Monochloroacetic Acid (MCAA) 98.2 95.8 99.2 98.3 98.3 

Monobromoacetic Acid (MBAA) 98.9 98.7 99.3 98.8 98.8 

Dalapon 100 101 101 100 101 

Dichloroacetic Acid (DCAA) 97.5 96.4 97.3 97.4 97.0 

Trichloroacetic Acid (TCAA) 99.6 101 100 102 102 

Bromochloroacetic Acid (BCAA) 101 102 101 102 102 

Bromodichloroacetic Acid (BDCAA) 103 104 102 108 108 

Dibromoacetic Acid (DBAA) 100 101 100 102 102 

Chlorodibromoacetic Acid (CDBAA) 103 106 102 110 110 

Tribromoacetic Acid (TBAA) 104 108 103 110 111 

a

Extract storage stability was conducted on three of the eight extracts from the precision and accuracy 

study conducted in chlorinated surface water (fortified at 10 μg/L) reported in Table 9. Sample storage 

stability is expressed as a recovery value (%) calculated as described in Section 9.8.2. All values have 

been corrected for background levels in the unfortified sample based on triplicate measurements as 

described in Table 9. Extracts were stored at < -10 °C. 

552.3-47
TABLE 15: INITIAL DEMONSTRATION OF CAPABILITY (IDC) REQUIREMENTS

Method 

Reference 

Requirement Specification and 

Frequency 

Acceptance Criteria 

Section 

9.2.1 

Section 

9.2.2 

Section 

9.2.3 

Initial 

Demonstration of 

Low Method 

Background 

Initial 

Demonstration of 

Accuracy 

Initial 

Demonstration of 

Precision 

Analyze LRB prior to any 

other IDC steps 

Analyze QCS 

Analyze 4-7 replicate LFBs 

fortified at midrange 

concentration. 

Calculate standard deviation 

and RSD for replicates used 

in the Initial Demonstration 

of Accuracy (Sect. 9.2.2) 

Over a period of three days, 

prepare a minimum of 7 

replicate LFBs fortified at a 

concentration estimated to be 

near the Detection Limit. 

Analyze the replicates 

through all steps of the 

analysis. Calculate the 

Detection Limit using the 

equation in Section 9.2.4. 

Demonstrate that LRB is 

reasonably free of 

contamination. 

Recovery for analytes must be 

± 30% of expected value. 

Mean recovery must be within 

± 20% of expected value. 

RSD must be # 20%. 

Note: Data from DL replicates 

are not required to meet 

method precision and accuracy 

criteria. If the DL replicates 

are fortified at a low enough 

concentration, it is likely that 

they will not meet precision 

and accuracy criteria. 

Section 

9.2.4 

Detection Limit 

Determination 

Section 

9.2.5 

Minimum 

Reporting Limit 

Estimate after determining 

DL and prior to analysis of 

Field Samples 

At least 3 times the DL or at a 

concentration which yields a 

signal-to-noise ratio of at least 

5. 

552.3-48
TABLE 16: QUALITY CONTROL REQUIREMENTS (SUMMARY)

Method 

Reference 

Requirement Specification and 

Frequency 

Acceptance Criteria 

Section 8.3 Sample Shipment 

and Storage 

Section 8.4 Sample Holding 

Time 

Section 8.4 Extract Holding 

Time 

Sample results are valid only if 

extracts are properly stored and 

analyzed within extract hold 

time. 

Section 9.3 Laboratory 

Reagent Blank 

(LRB) 

With each extraction batch 

of up to 20 samples 

Demonstrate that all target 

analytes in LRB are below 1/3 

the MRL and that interferences 

do not prevent the identification 

and quantification of method 

analytes. 1/3 

the MRL, results for all 

problem analytes in extraction 

batch are invalid. 

If targets exceed 

Samples must not exceed 

10 °C within 48 hours after 

collection. 

in laboratory must not 

exceed 

6 °C. 

14 days with appropriate 

preservation and storage 

21 days at # -10 °C 

protected from light 

(MTBE) 

28 days at # -10 °C 

protected from light 

(TAME) 

Samples stored 

Sample results are valid only if 

samples are properly collected, 

preserved, and stored. 

Sample results are valid only if 

samples are extracted within 

sample hold time. 

552.3-49
TABLE 16: QUALITY CONTROL REQUIREMENTS (SUMMARY)

Method 

Reference 

Requirement Specification and 

Frequency 

Acceptance Criteria 

Sections 

9.4 and 

10.3 

Continuing 

Calibration 

Check (CCC) 

Verify initial calibration 

by analyzing a calibration 

standard at the beginning 

of each analysis batch, 

after every 10 samples, 

and after the last sample. 

Low CCC - # MRL 

Mid CCC - near midpoint 

in initial calibration curve 

High CCC - near highest 

calibration standard 

1) The result for each analyte must be 

70-130% of the expected value for all 

but the lowest standard. lowest 

standard must be 50-150% of the 

expected value. 

2) The peak area of internal standards 

must be 50-150% of the average peak 

area calculated during the initial 

calibration. 

Results for analytes that do not meet 

IS criteria or are not bracketed by 

acceptable CCCs are invalid. 

The

Section 9.6 Internal 

Standard 

(IS) 

Section 9.7 Surrogate 

Standard 

(SUR) 

Section 9.8 Laboratory 

Fortified 

Sample 

Matrix 

(LFSM) 

and 

Laboratory 

Fortified 

Matrix 

Duplicate 

(LFSMD) 

1,2,3 Trichloropropane is 

added to the extraction 

solvent. 

2-bromobutanoic acid 

[80-58-0] is added to all 

calibration standards and 

samples, including QC 

samples. 

Analyze one LFSM per 

analysis batch (20 

samples or less) fortified 

with method analytes at a 

concentration greater than 

the native concentration. 

LFSMD should be used in 

place of Field Duplicate if 

frequency of detects for 

target analytes is low. 

Peak area counts for the IS in all 

injections must be within 50-150% of 

the average peak area calculated 

during the initial calibration. 

does not meet criteria, corresponding 

target results are invalid. 

Surrogate recovery must be 70-130% 

of the expected value. If surrogate 

fails this criterion, report all results for 

sample as suspect/surrogate recovery. 

Recoveries at mid and high levels not 

within 70-130% or low-level 

recoveries not within 50-150% of the 

fortified amount may indicate a matrix 

effect. 

Target analyte RPDs for LFSMD not 

within ± 30% at mid and high levels 

of fortification and within ± 50% near 

MRL may indicate a matrix effect. 

If the IS 

552.3-50
Method 

Reference 

Requirement Specification and 

Frequency 

Acceptance Criteria 

Section 9.9 Field 

Duplicates 

(FD) 

Extract and analyze at 

least one FD with each 

extraction batch (20 

samples or less). A 

LFSMD may be 

substituted periodically 

for a FD when the 

frequency of detects for 

target analytes is low. 

Target analyte RPDs for FD should be 

within ± 30% at mid and high level 

concentrations and within ± 50% near 

MRL. 

Section 

9.10 

Quality 

Control 

Sample 

(QCS) 

Analyzed when new 

Primary Dilution 

Standards (PDS) are 

prepared, during the IDC, 

or quarterly. 

Results must be 70-130% of the 

expected values. 

Section 

10.2 

Initial 

Calibration 

Use internal standard 

calibration technique to 

generate a calibration 

curve. se at least 5 

standard concentrations. 

When each calibration standard is 

calculated as an unknown using the 

calibration curve, the result must be 

70-130% of the expected value for all 

except the lowest standard, which 

must be 50-150% of the expected 

value. 

U

552.3-51
FIGURE 1: CHROMATOGRAM OF THE HALOACETIC ACIDS ON A DB-1701 COLUMN USING THE 

CHROMATOGRAPHIC CONDITIONS GIVEN IN TABLE 1 USING THE MTBE PROCEDURE 

552.3-52
FIGURE 2: CHROMATOGRAM OF THE HALOACETIC ACIDS ON A DB-1701 COLUMN USING THE 

CHROMATOGRAPHIC CONDITIONS GIVEN IN TABLE 2 USING THE TAME PROCEDURE 

552.3-53
FIGURE 3: CHROMATOGRAM OF THE HALOACETIC ACIDS ON A DB-5.625 COLUMN USING THE 

CHROMATOGRAPHIC CONDITIONS GIVEN IN TABLE 3 USING THE MTBE PROCEDURE 

552.3-54
FIGURE 4: CHROMATOGRAM OF THE HALOACETIC ACIDS ON A DB-5.625 COLUMN USING THE 

CHROMATOGRAPHIC CONDITIONS GIVEN IN TABLE 4 USING THE TAME PROCEDURE 

552.3-55