ASTM D6363-98(2003)e1
(Test Method)Standard Test Method for Determination of Hydrogen Peroxide and Combined Organic Peroxides in Atmospheric Water Samples by Peroxidase Enzyme Fluorescence Method
Standard Test Method for Determination of Hydrogen Peroxide and Combined Organic Peroxides in Atmospheric Water Samples by Peroxidase Enzyme Fluorescence Method
SIGNIFICANCE AND USE
Hydrogen peroxide (formed photochemically in the atmosphere) is a primary oxidizer of dissolved sulfur dioxide in atmospheric water. Detection of H2O2 in atmospheric water is useful for inferring gas-phase H2O2 concentrations and for assessing the relative importance of various acidifying mechanisms under specific atmospheric conditions.
Hydroperoxides in samples to be analyzed are unstable in water and can decay rapidly due to bacterial action or chemical reaction with other constituents. The test method includes procedures for sample derivatization and methods for estimating and correcting for hydroperoxide decay.
SCOPE
1.1 This test method covers the determination of hydroperoxides, which include hydrogen peroxide (H2O2) and combined organic peroxides, in samples of atmospheric water by the method of horseradish peroxidase derivatization and fluorescence analysis of the derived dimer.
1.2 The range of applicable hydrogen peroxide concentrations was determined to be 0.6-176.0 x 10-6 M from independent laboratory tests of the test method.
1.3 The primary use of the test method is for hydrogen peroxide, but it may also be used to quantitate organic hydroperoxides. Determinations of organic hydroperoxide concentration levels up to 30 x 10-6 M may be adequately obtained by calibration with hydrogen peroxide. While organic hydroperoxides have not been detected at significant concentration levels in rain or cloud water, their presence may be tested by operation of the test method with the addition of catalase for destruction of H2O2.
1.4 Because of the instability fo hydroperoxides in atmospheric water samples, proper sample collection, at-collection derivatization, and stringent quality control are essential aspects of the analytical process.
1.5 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.
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NOTICE: This standard has either been superseded and replaced by a new version or withdrawn.
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´1
Designation:D6363–98 (Reapproved 2003)
Standard Test Method for
Determination of Hydrogen Peroxide and Combined Organic
Peroxides in Atmospheric Water Samples by Peroxidase
Enzyme Fluorescence Method
This standard is issued under the fixed designation D6363; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision.Anumber in parentheses indicates the year of last reapproval.A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
´ NOTE—Editorial changes were made throughout this test method in August 2003.
1. Scope 2. Referenced Documents
1.1 This test method covers the determination of hydroper- 2.1 ASTM Standards:
oxides, which include hydrogen peroxide (H O ) and com- D1129 Terminology Relating to Water
2 2
bined organic peroxides, in samples of atmospheric water by D1193 Specification for Reagent Water
the method of horseradish peroxidase derivatization and fluo- D1356 Terminology Relating to Sampling and Analysis of
,
rescence analysis of the derived dimer. Atmospheres
1.2 The range of applicable hydrogen peroxide concentra- D5012 Guide for Preparation of Materials Used for the
−6
tions was determined to be 0.6 - 176.0 3 10 M from Collection and Preservation of Atmospheric Wet Deposi-
independent laboratory tests of the test method. tion
1.3 The primary use of the test method is for hydrogen D5111 Guide for Choosing Locations and Sampling Meth-
peroxide, but it may also be used to quantitate organic ods to Monitor Atmospheric Deposition at Non-Urban
hydroperoxides.Determinationsoforganichydroperoxidecon- Locations
−6
centration levels up to 30 3 10 M may be adequately E200 Practice for Preparation, Standardization, and Storage
2,3
obtained by calibration with hydrogen peroxide. While of Standard and Reagent Solutions for Chemical Analysis
organic hydroperoxides have not been detected at significant
3. Terminology
concentration levels in rain or cloud water, their presence may
be tested by operation of the test method with the addition of 3.1 Definitions—For definitions of terms used in this test
method, refer to Terminologies D1129 and D1356 and Guide
catalase for destruction of H O .
2 2
1.4 Because of the instability of hydroperoxides in atmo- D5111.
3.2 Definitions of Terms Specific to This Standard:
spheric water samples, proper sample collection, at-collection
derivatization, and stringent quality control are essential as- 3.2.1 atmospheric water, n—liquid or solid water sus-
pended in the atmosphere or deposited from the atmosphere.
pects of the analytical process.
1.5 This standard does not purport to address all of the Forms of atmospheric water include rain, snow, fog, cloud
water, dew, and frost.
safety concerns, if any, associated with its use. It is the
responsibility of the user of this standard to establish appro- 3.2.2 derivatization, n—formation of the
p-hydroxyphenylacetic acidic dimer by combination of
priate safety and health practices and determine the applica-
bility of regulatory limitations prior to use. p-hydroxyphenylacetic acid, horseradish peroxidase reagent,
and hydroperoxide(s). Also the procedure of addition of the
derivatizing reagent to samples.
This guide is under the jurisdiction of ASTM Committee D22 on Air Quality
3.2.3 hydroperoxides, n—hydrogen peroxide and organic
and is the direct responsibility of Subcommittee D22.03 on Ambient Atmospheres
and Source Emissions.
peroxides dissolved in water.
Current edition approved August 10, 2003. Published October 2003. Originally
3.2.4 intrinsic hydroperoxides, n—hydroperoxides con-
approved in 1998. Last previous edition approved in 1998 as D6363-98. DOI:
tained in reagent water used for the method.
10.1520/D6363-98R03E01.
Lazrus, A. L., Kok, G. L., Gitlin, S. N., and Lind, J. A., “Automated
Fluorometric Method for Hydrogen Peroxide in Atmospheric Precipitation,” Anal.
Chem., 57, 1985, pp. 917–922. For referenced ASTM standards, visit the ASTM website, www.astm.org, or
Kok,G.L.,Thompson,K.,andLazrus,A.L.,“DerivatizationTechniqueforthe contactASTM Customer Service at service@astm.org. ForAnnual Book ofASTM
Determination of Peroxides in Precipitation,” Anal. Chem., 58, 1986, pp. Standards volume information, refer to the standard’s Document Summary page on
1192–1194. the ASTM website.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.
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D6363–98 (2003)
3.2.5 post-derivatization, n—addition of the derivatizing 7. Apparatus
reagent to the sample after collection.
7.1 Flow System, consisting of the following:
3.2.6 pre-derivatization, n—addition of the derivatizing re-
7.1.1 Automatic sampler or injection valve.
agent to the sample collection container prior to sample
7.1.2 Automated wet chemistry (peristaltic) pump.
collection.
7.1.3 Reagent manifold.
3.2.7 systems blank, n—afieldblankofreagentwaterthatis
7.1.4 Mixing coil; 5-turn, 2-mm inner diameter.
subjected to a similar or identical environment and derivatiza-
7.1.5 Fluorometer; excitation at 320 nm and measurement
tion time as a collected atmospheric water sample.
ofthefluorescencesignalat400nm;flow-throughfluorescence
3.2.8 systems standard, n—a H O calibration standard
2 2
cell.
solution subjected to a similar or identical environment and
7.1.6 Recorder.
derivatization time as a collected atmospheric water sample.
7.2 Sample and Standards Containers—All containers used
for sample collection and sample transport, for storage and
4. Summary of Test Method
analysis of samples and standards, and for reagents should be
4.1 Theperoxidaseenzymefluorescencemethodisbasedon high density polyethylene, TFE-fluorocarbon, or borosilicate
the reaction of hydroperoxides, horseradish peroxidase, and glass, cleaned in accordance with procedures established for
analyses of common inorganic ions (see Guide D5012).
p-hydroxyphenylacetic (PHOPAA) acid, forming a fluorescent
dimer of the latter. This dimer is detected using a fluorometric 7.3 Pipettes with Disposable Tips—Solution preparation
and sample fixing operations are generally conducted using
technique,andthehydroperoxidesarequantifiedbycalibration
with hydrogen peroxide. The formation of the dimer (deriva- automatic pipettes. Solution volumes delivered by these de-
vices should be verified to confirm consistent and accurate
tization) shall be accomplished soon after sample collection to
minimize H O decay. In addition, strict quality assurance performance.
2 2
practices are part of the method, including use of systems 7.4 Reagent Bottles—All containers used for the prepara-
standards and systems blanks to estimate hydroperoxide loss tion and storage of derivatizing and other reagent solutions
and to assess derivatizing solution effectiveness. shallbededicatedforhydroperoxides.Containersforsolutions
of catalase shall not be used for non-catalase solutions.
5. Significance and Use
8. Reagents and Materials
5.1 Hydrogen peroxide (formed photochemically in the
8.1 Purity of Reagents—Unless otherwise noted, reagent
atmosphere)isaprimaryoxidizerofdissolvedsulfurdioxidein
atmospheric water. Detection of H O in atmospheric water is grade chemicals shall be used.
2 2
8.2 Purity of Water—Unlessotherwiseindicated,references
useful for inferring gas-phase H O concentrations and for
2 2
assessing the relative importance of various acidifying mecha- to water shall be understood to mean reagent water as defined
nisms under specific atmospheric conditions. by Type I of Specification D1193, with the added stipulation
5.2 Hydroperoxides in samples to be analyzed are unstable that the total organic carbon content be less than 20 µg/L. A
in water and can decay rapidly due to bacterial action or Type I water system equipped with an organic extraction
chemical reaction with other constituents. The test method cartridge and a 0.2 µm filter is an acceptable water source.
includes procedures for sample derivatization and methods for Water to be used for reagents, standard solutions, and analyti-
estimating and correcting for hydroperoxide decay. cal rinsing should be stored in borosilicate glass.
6 7
8.3 Catalase Enzyme (1.7 3 10 units/mL) —The enzyme
6. Interferences catalase may be used for the destruction of H O in atmo-
2 2
spheric water samples. Its addition to the sample before
6.1 The derivatizing reagent is formulated to counteract the
additionofthederivatizingreagentremovesH O ,butorganic
2 2
effects of the following potentially interfering species.
hydroperoxides are preserved. Subsequent addition of the
6.2 Hydroxymethane Sulfonate (HMSA)—The addition of
derivatizing reagent results in dimer formation by way of
formaldehyde (HCHO) to the derivatizing reagent will sup-
reactionwithperoxidesotherthanH O .Resultsofanalysesof
2 2
press the negative interference of HMSA. In the absence of
catalase-treated samples may be compared with the measure-
added HCHO, the PHOPAA dimer in a derivatized simulated
−5 −4
ment of peroxides in samples without catalase to determine
rain sample, containing 1.2 3 10 MH O and 1.0 3 10 M
2 2
H O by difference.
2 2
HMSA, displayed a fluorescence signal 5% lower than that
observed when HCHO was added to the derivatizing reagent.
6.3 Trace Transition Metals and Common Ionic Compo-
nents of Atmospheric Water (Sodium, Ammonium, Hydrogen,
Schwartz, L.M., “Calibration of Pipets: A Statistical View,” Analytical Chem-
istry, Vol. 61, 1989, pp. 1080–1083.
Sulfate, Nitrate, Chloride, Formate)—Potentialinterferenceby
Reagent Chemicals, American Chemical Society Specifications, American
transition metals is overcome by the formation of ethylenedi-
Chemical Society, Washington, DC. For suggestions on the testing of reagents not
aminetetraacetic acid (EDTA) complexes. Tests of simulated
listed by the American Chemical Society, see Analar Standards for Laboratory
rain samples containing transition metals and common ionic
Chemicals, BDH Ltd., Poole, Dorset, U.K., and the United States Pharmacopeia
and National Formulary, U.S. Pharmacopeial Convention, Inc. (USPC), Rockville,
components of precipitation have demonstrated both the gen-
MD.
eral applicability of this test method to samples containing
7 6
Catalase enzyme, 1.7 3 10 units/mL, has been found satisfactory for this
commoncontaminantsandthestabilityofderivatizedsolutions
purpose. Available through Sigma Chemical Co., P.O. Box 14508, St. Louis, MO
stored at 4°C for more than five days. 63178.
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D6363–98 (2003)
8.3.1 Catalase,1+49—Dilute 1 mLof catalase enzyme to 8.8 Sodium Hydroxide (NaOH) (0.1 M)—Dissolve 4.0 g of
a final volume of 50 mL with water. Before pipetting the sodium hydroxide in water and dilute to 1 L. Prepare weekly.
concentrated solution, ensure that all the solid material is 8.9 Sulfuric Acid (H SO ), 5 % (3.6 M)—Add 5 mL con-
2 4
completely suspended by shaking or stirring the bottle of centrated H SO to water in a volumetric flask, and dilute to
2 4
concentrate. Allow the dilute solution to stand at least 4 h 100 mL.
before use. The solution can be stored for up to 48 h at 4°C.
9. Sample Collection
8.4 Derivatizing Reagent, Concentrated—Dissolve 12.11 g
9.1 Select sampling locations and sampling methods in
of Tris(hydroxymethyl)aminomethane, 0.38 g of EDTA, tetra-
accordance with Guide D5111. Additional considerations spe-
sodium salt, 4.57 g of PHOPAA, 300 units of horseradish
cific to sampling for aqueous-phase hydrogen peroxide are
peroxidase, and 1 mLconcentrated hydrochloric acid in water,
provided in 9.3 and 9.4.
anddiluteto200mLinavolumetricflask.ThefinalpHofthis
9.2 Methods of preparation of sample containers for collec-
solution should be 9.0. If greater than 9.5 or less than 8.5,
tion, transport, and storage shall be those detailed in Guide
remake. Prepare every four days and store at 4°C. Measure-
D5012 under inorganic ionic species (see 8.1 and 8.2 of Guide
ment of peroxides in aqueous atmospheric samples is based on
D5012).
the fluorescence of the PHOPAA dimer produced by reaction
9.3 Control procedures designed to ensure sample integrity
ofhydroperoxideswithPHOPAA.Thefluorescenceofsamples
in the field (see Section 10) are difficult to perform adequately
derivatizedatthetimeofcollectionprovidesameasureoftotal
if buckets or other high atmospheric-exposure collectors are
hydroperoxide (organic and H O ) content of the sample.
2 2
used. Therefore, sampling for rain should be conducted using
8.4.1 Derivatizing Reagent,4+96—Dilute 4.0 mL of the
funnel-and-bottle type, or narrow-necked, collectors.
concentrated derivatizing reagent to 100 mL with water.
9.4 Therequirementsforcontrolledderivatizationofhydro-
Prepare daily as needed, and keep tightly sealed at 4°C.
peroxides and timely analysis (see Section 10) dictate that
NOTE 1—The dilute derivatizing reagent is normally added to samples
sampling for wet deposition be conducted on a daily or more
to be analyzed in the reagent:sample ratio of 1:1. Other concentrations of
frequent basis.
dilute derivatizing reagent may be used as long as the final ratio entering
theanalyticalsystemis1:1.Underspecialcircumstances,otherratiosmay
10. Derivatization
be dictated by sampling conditions (see 10.6 and 10.7).
10.1 The following procedures shall be in addition to those
8.5 Hydrochloric Acid (HCl), (1 M)—Add 8.3 mL concen-
specified for preservation of inorganic anions and cations in
trated HCl to water in a volumetric flask and dilute to 100 mL.
Guide D5012 (see Table 1 of Guide D5012).
8.6 Peroxide Solution, Standard Stock (1 %)—Dilute com-
10.2 Hydroperoxides dissolved in atmospheric water solu-
mercially available (pharmaceutical grade is acceptable) H O
2 2
tions are subject to decay at rates that are not predictable.
solution (30%) approximately1+29 with water in a volu-
Therefore, the derivatizing solution shall be added during
metric flask. Add sodium stannate (Na SnO ) to a concentra-
2 3
sample collection or within a known and controlled time after
tion of 10.65 mg/Land store at 4°C, and store in a borosilicate
sample collection.
glass bottle. Determine the peroxide concentration by titration
10.3 The rate of decay of non-derivatized hydroperoxides
−1
with standard permanganate solution (see 11.2) approximately
may be quite fast: loss rates ranging from 1 to 28% h were
24 h after preparation. Update the concentration determination
found for rain samples collected at Boulder, CO. The decay
by titration at one month intervals.
may be significant during the time of sampling, an especially
8.6.1 Peroxide Solution, 1 + 199—Dilute 500 µL of the
important consideration for sampling of precipitation. Thus,
standard stock (1%) solution to 100 mL with water in a
...
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