ASTM D6363-98(2009)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 × 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 × 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 of 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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´1
Designation: D6363 − 98(Reapproved 2009)
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—Reapproved with an editorial change to Section 14 in October 2009.
1. Scope 2. Referenced Documents
1.1 This test method covers the determination of 2.1 ASTM Standards:
hydroperoxides, which include hydrogen peroxide (H O ) and D1129Terminology Relating to Water
2 2
combined organic peroxides, in samples of atmospheric water D1193Specification for Reagent Water
by the method of horseradish peroxidase derivatization and D1356Terminology Relating to Sampling and Analysis of
2,3
fluorescence analysis of the derived dimer. Atmospheres
D5012Guide for Preparation of Materials Used for the
1.2 The range of applicable hydrogen peroxide concentra-
−6 Collection and Preservation of Atmospheric Wet Deposi-
tions was determined to be 0.6 - 176.0 × 10 M from
tion
independent laboratory tests of the test method.
D5111Guide for Choosing Locations and Sampling Meth-
1.3 The primary use of the test method is for hydrogen
ods to Monitor Atmospheric Deposition at Non-Urban
peroxide, but it may also be used to quantitate organic
Locations
hydroperoxides.Determinationsoforganichydroperoxidecon-
E200Practice for Preparation, Standardization, and Storage
−6
centration levels up to 30 × 10 M may be adequately
of Standard and Reagent Solutions for ChemicalAnalysis
2,3
obtained by calibration with hydrogen peroxide. While
organic hydroperoxides have not been detected at significant
3. Terminology
concentration levels in rain or cloud water, their presence may
3.1 Definitions—For definitions of terms used in this test
be tested by operation of the test method with the addition of
method, refer to Terminologies D1129 and D1356 and Guide
catalase for destruction of H O .
2 2
D5111.
1.4 Because of the instability of hydroperoxides in atmo-
3.2 Definitions of Terms Specific to This Standard:
spheric water samples, proper sample collection, at-collection
3.2.1 atmospheric water, n—liquidorsolidwatersuspended
derivatization, and stringent quality control are essential as-
in the atmosphere or deposited from the atmosphere. Forms of
pects of the analytical process.
atmospheric water include rain, snow, fog, cloud water, dew,
1.5 This standard does not purport to address all of the
and frost.
safety concerns, if any, associated with its use. It is the
3.2.2 derivatization, n—formation of the
responsibility of the user of this standard to establish appro-
p-hydroxyphenylacetic acidic dimer by combination of
priate safety and health practices and determine the applica-
p-hydroxyphenylacetic acid, horseradish peroxidase reagent,
bility of regulatory limitations prior to use.
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
peroxides dissolved in water.
and Source Emissions.
Current edition approved Oct. 1, 2009. Published December 2009. Originally
3.2.4 intrinsic hydroperoxides, n—hydroperoxides con-
ϵ1
approved in 1998. Last previous edition approved in 2003 as D6363-98(03) .
tained in reagent water used for the method.
DOI: 10.1520/D6363-98R09E01.
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
´1
D6363 − 98 (2009)
3.2.5 post-derivatization, n—addition of the derivatizing commoncontaminantsandthestabilityofderivatizedsolutions
reagent to the sample after collection. stored at 4°C for more than five days.
3.2.6 pre-derivatization, n—addition of the derivatizing re-
7. Apparatus
agent to the sample collection container prior to sample
collection. 7.1 Flow System, consisting of the following:
7.1.1 Automatic sampler or injection valve.
3.2.7 systems blank, n—afieldblankofreagentwaterthatis
7.1.2 Automated wet chemistry (peristaltic) pump.
subjected to a similar or identical environment and derivatiza-
7.1.3 Reagent manifold.
tion time as a collected atmospheric water sample.
7.1.4 Mixing coil; 5-turn, 2-mm inner diameter.
3.2.8 systems standard, n—aH O calibration standard
2 2
7.1.5 Fluorometer; excitation at 320 nm and measurement
solution subjected to a similar or identical environment and
ofthefluorescencesignalat400nm;flow-throughfluorescence
derivatization time as a collected atmospheric water sample.
cell.
7.1.6 Recorder.
4. Summary of Test Method
7.2 Sample and Standards Containers—All containers used
4.1 Theperoxidaseenzymefluorescencemethodisbasedon
for sample collection and sample transport, for storage and
the reaction of hydroperoxides, horseradish peroxidase, and
analysis of samples and standards, and for reagents should be
p-hydroxyphenylacetic (PHOPAA) acid, forming a fluorescent
high density polyethylene, TFE-fluorocarbon, or borosilicate
dimer of the latter. This dimer is detected using a fluorometric
glass, cleaned in accordance with procedures established for
technique,andthehydroperoxidesarequantifiedbycalibration
analyses of common inorganic ions (see Guide D5012).
with hydrogen peroxide. The formation of the dimer (deriva-
7.3 PipetteswithDisposableTips—Solutionpreparationand
tization) shall be accomplished soon after sample collection to
sample fixing operations are generally conducted using auto-
minimize H O decay. In addition, strict quality assurance
2 2
matic pipettes. Solution volumes delivered by these devices
practices are part of the method, including use of systems
should be verified to confirm consistent and accurate perfor-
standards and systems blanks to estimate hydroperoxide loss
mance.
and to assess derivatizing solution effectiveness.
7.4 Reagent Bottles—Allcontainersusedforthepreparation
5. Significance and Use
and storage of derivatizing and other reagent solutions shall be
dedicated for hydroperoxides. Containers for solutions of
5.1 Hydrogen peroxide (formed photochemically in the
catalase shall not be used for non-catalase solutions.
atmosphere)isaprimaryoxidizerofdissolvedsulfurdioxidein
atmospheric water. Detection of H O in atmospheric water is
2 2
8. Reagents and Materials
useful for inferring gas-phase H O concentrations and for
2 2
assessing the relative importance of various acidifying mecha- 8.1 Purity of Reagents—Unless otherwise noted, reagent
nisms under specific atmospheric conditions. grade chemicals shall be used.
5.2 Hydroperoxides in samples to be analyzed are unstable 8.2 Purity of Water—Unless otherwise indicated, references
in water and can decay rapidly due to bacterial action or to water shall be understood to mean reagent water as defined
chemical reaction with other constituents. The test method by Type I of Specification D1193, with the added stipulation
includes procedures for sample derivatization and methods for that the total organic carbon content be less than 20 µg/L. A
estimating and correcting for hydroperoxide decay. Type I water system equipped with an organic extraction
cartridge and a 0.2 µm filter is an acceptable water source.
6. Interferences Water to be used for reagents, standard solutions, and analyti-
cal rinsing should be stored in borosilicate glass.
6.1 The derivatizing reagent is formulated to counteract the
6 7
effects of the following potentially interfering species. 8.3 Catalase Enzyme (1.7 × 10 units/mL) —The enzyme
catalase may be used for the destruction of H O in atmo-
2 2
6.2 Hydroxymethane Sulfonate (HMSA)—The addition of
spheric water samples. Its addition to the sample before
formaldehyde (HCHO) to the derivatizing reagent will sup-
additionofthederivatizingreagentremovesH O ,butorganic
2 2
press the negative interference of HMSA. In the absence of
hydroperoxides are preserved. Subsequent addition of the
added HCHO, the PHOPAA dimer in a derivatized simulated
−5 −4 derivatizing reagent results in dimer formation by way of
rain sample, containing 1.2 × 10 MH O and 1.0 × 10 M
2 2
HMSA, displayed a fluorescence signal 5% lower than that
observed when HCHO was added to the derivatizing reagent.
Schwartz, L.M., “Calibration of Pipets: A Statistical View,” Analytical
Chemistry, Vol. 61, 1989, pp. 1080–1083.
6.3 Trace Transition Metals and Common Ionic Compo-
Reagent Chemicals, American Chemical Society Specifications, American
nents of Atmospheric Water (Sodium, Ammonium, Hydrogen,
Chemical Society, Washington, DC. For suggestions on the testing of reagents not
Sulfate, Nitrate, Chloride, Formate)—Potentialinterferenceby
listed by the American Chemical Society, see Analar Standards for Laboratory
transition metals is overcome by the formation of ethylenedi-
Chemicals, BDH Ltd., Poole, Dorset, U.K., and the United States Pharmacopeia
and National Formulary, U.S. Pharmacopeial Convention, Inc. (USPC), Rockville,
aminetetraacetic acid (EDTA) complexes. Tests of simulated
MD.
rain samples containing transition metals and common ionic
7 6
Catalase enzyme, 1.7 × 10 units/mL, has been found satisfactory for this
components of precipitation have demonstrated both the gen-
purpose. Available through Sigma Chemical Co., P.O. Box 14508, St. Louis, MO
eral applicability of this test method to samples containing 63178.
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D6363 − 98 (2009)
reactionwithperoxidesotherthanH O .Resultsofanalysesof the solution with water to 1 L. Seal tightly, and store in an
2 2
catalase-treated samples may be compared with the measure- amberborosilicateglassbottleinthedark.Standardizefollow-
ment of peroxides in samples without catalase to determine ing the procedure in Practice E200, Sections 64–68; adjust
H O by difference. chemical proportions according to 9.1 of that Practice.
2 2
8.3.1 Catalase,1+49—Dilute 1 mL of 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.
8.4 Derivatizing Reagent, Concentrated—Dissolve 12.11 g
9. Sample Collection
of Tris(hydroxymethyl)aminomethane, 0.38 g of EDTA, tetra-
9.1 Select sampling locations and sampling methods in
sodium salt, 4.57 g of PHOPAA, 300 units of horseradish
accordance with Guide D5111. Additional considerations spe-
peroxidase, and 1 mLconcentrated hydrochloric acid in water,
cific to sampling for aqueous-phase hydrogen peroxide are
anddiluteto200mLinavolumetricflask.ThefinalpHofthis
provided in 9.3 and 9.4.
solution should be 9.0. If greater than 9.5 or less than 8.5,
remake. Prepare every four days and store at 4°C. Measure-
9.2 Methods of preparation of sample containers for
ment of peroxides in aqueous atmospheric samples is based on
collection, transport, and storage shall be those detailed in
the fluorescence of the PHOPAA dimer produced by reaction
Guide D5012 under inorganic ionic species (see 8.1 and 8.2 of
ofhydroperoxideswithPHOPAA.Thefluorescenceofsamples
Guide D5012).
derivatizedatthetimeofcollectionprovidesameasureoftotal
9.3 Control procedures designed to ensure sample integrity
hydroperoxide (organic and H O ) content of the sample.
2 2
in the field (see Section 10) are difficult to perform adequately
8.4.1 Derivatizing Reagent, 4 + 96—Dilute 4.0 mL of the
if buckets or other high atmospheric-exposure collectors are
concentrated derivatizing reagent to 100 mL with water.
used. Therefore, sampling for rain should be conducted using
Prepare daily as needed, and keep tightly sealed at 4°C.
funnel-and-bottle type, or narrow-necked, collectors.
NOTE 1—The dilute derivatizing reagent is normally added to samples
9.4 The requirements for controlled derivatization of hy-
to be analyzed in the reagent:sample ratio of 1:1. Other concentrations of
droperoxides and timely analysis (see Section 10) dictate that
dilute derivatizing reagent may be used as long as the final ratio entering
theanalyticalsystemis1:1.Underspecialcircumstances,otherratiosmay
sampling for wet deposition be conducted on a daily or more
be dictated by sampling conditions (see 10.6 and 10.7).
frequent basis.
8.5 Hydrochloric Acid (HCl), (1 M)—Add 8.3 mL concen-
trated HCl to water in a volumetric flask and dilute to 100 mL.
10. Derivatization
8.6 Peroxide Solution, Standard Stock (1%)—Dilute com-
10.1 The following procedures shall be in addition to those
mercially available (pharmaceutical grade is acceptable) H O
2 2 specified for preservation of inorganic anions and cations in
solution (30%) approximately1+29 with water in a volu-
Guide D5012 (see Table 1 of Guide D5012).
metric flask. Add sodium stannate (Na SnO ) to a concentra-
2 3
10.2 Hydroperoxides dissolved in atmospheric water solu-
tion of 10.65 mg/Land store at 4°C, and store in a borosilicate
tions are subject to decay at rates that are not predictable.
glass bottle. Determine the peroxide concentration by titration
Therefore, the derivatizing solution shall be added during
with standard permanganate solution (see 11.2) approximately
sample collection or within a known and controlled time after
24 h after preparation. Update the concentration determination
sample collection.
by titration at one month intervals.
10.3 The rate of decay of non-derivatized hydroperoxides
8.6.1 Peroxide Solution, 1 + 199—Dilute 500 µL of the
−1
standard stock (1%) solution to 100 mL with water in a may be quite fast:
...
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