ASTM D6363-98
(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
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.
General Information
Relations
Standards Content (Sample)
NOTICE: This standard has either been superceded and replaced by a new version or discontinued.
Contact ASTM International (www.astm.org) for the latest information.
Designation: D 6363 – 98
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 D 6363; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (e) indicates an editorial change since the last revision or reapproval.
1. Scope D 1356 Terminology Relating to Sampling and Analysis of
Atmospheres
1.1 This test method covers the determination of hydroper-
D 5012 Guide for Preparation of Materials Used for the
oxides, which include hydrogen peroxide (H O ) and com-
2 2
Collection and Preservation of Atmospheric Wet Deposi-
bined organic peroxides, in samples of atmospheric water by
tion
the method of horseradish peroxidase derivatization and fluo-
2,3
D 5085 Test Method for the Determination of Chloride,
rescence analysis of the derived dimer.
Nitrate, and Sulfate in Atmospheric Wet Deposition by
1.2 The range of applicable hydrogen peroxide concentra-
−6
Chemically Suppressed Ion Chromatography
tions was determined to be 0.6 - 176.0 3 10 M from
D 5111 Guide for Choosing Locations and Sampling Meth-
independent laboratory tests of the test method.
ods to Monitor Atmospheric Deposition at Non-Urban
1.3 The primary use of the test method is for hydrogen
Locations
peroxide, but it may also be used to quantitate organic
E 200 Standard Practice for Preparation, Standardization,
hydroperoxides. Determinations of organic hydroperoxide con-
−6
and Storage of Standard Solutions for Chemical Analysis
centration levels up to 30 3 10 M may be adequately
2,3
obtained by calibration with hydrogen peroxide. While
3. Terminology
organic hydroperoxides have not been detected at significant
3.1 Definitions—For definitions of terms used in this test
concentration levels in rain or cloud water, their presence may
method, refer to Terminologies D 1129 and D 1356 and Guide
be tested by operation of the test method with the addition of
3 D 5111.
catalase for destruction of H O .
2 2
3.2 Definitions of Terms Specific to This Standard:
1.4 Because of the instability of hydroperoxides in atmo-
3.2.1 atmospheric water, n—liquid or solid water sus-
spheric water samples, proper sample collection, at-collection
pended in the atmosphere or deposited from the atmosphere.
derivatization, and stringent quality control are essential as-
Forms of atmospheric water include rain, snow, fog, cloud
pects of the analytical process.
water, dew, and frost.
1.5 This standard does not purport to address all of the
3.2.2 derivatization, n—formation of the
safety concerns, if any, associated with its use. It is the
p-hydroxyphenylacetic acidic dimer by combination of
responsibility of the user of this standard to establish appro-
p-hydroxyphenylacetic acid, horseradish peroxidase reagent,
priate safety and health practices and determine the applica-
and hydroperoxide(s). Also the procedure of addition of the
bility of regulatory limitations prior to use.
derivatizing reagent to samples.
2. Referenced Documents 3.2.3 hydroperoxides, n—hydrogen peroxide and organic
peroxides dissolved in water.
2.1 ASTM Standards:
4 3.2.4 intrinsic hydroperoxides, n—hydroperoxides con-
D 1129 Terminology Relating to Water
tained in reagent water used for the method.
D 1193 Specification for Reagent Water
3.2.5 post-derivatization, n—addition of the derivatizing
reagent to the sample after collection.
This guide is under the jurisdiction of ASTM Committee D-22 on Sampling and
3.2.6 pre-derivatization, n—addition of the derivatizing re-
Analysis of Atmospheres and is the direct responsibility of Subcommittee D22.06
agent to the sample collection container prior to sample
on Atmospheric Deposition.
Current edition approved Dec. 10, 1998. Published February 1999.
collection.
Lazrus, A. L., Kok, G. L., Gitlin, S. N., and Lind, J. A., “Automated
3.2.7 systems blank, n—a field blank of reagent water that is
Fluorometric Method for Hydrogen Peroxide in Atmospheric Precipitation,” Anal.
subjected to a similar or identical environment and derivatiza-
Chem., 57, 1985, pp. 917–922.
Kok, G. L., Thompson, K., and Lazrus, A. L., “Derivatization Technique for the tion time as a collected atmospheric water sample.
Determination of Peroxides in Precipitation,” Anal. Chem., 58, 1986, pp.
1192–1194.
4 5
Annual Book of ASTM Standards, Vol 11.01. Annual Book of ASTM Standards, Vol 11.03.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.
NOTICE: This standard has either been superceded and replaced by a new version or discontinued.
Contact ASTM International (www.astm.org) for the latest information.
D 6363
3.2.8 systems standard, n—a H O calibration standard 7.1.6 Recorder.
2 2
solution subjected to a similar or identical environment and 7.2 Sample and Standards Containers—All containers used
derivatization time as a collected atmospheric water sample. for sample collection and sample transport, for storage and
analysis of samples and standards, and for reagents should be
4. Summary of Test Method
high density polyethylene, TFE-fluorocarbon, or borosilicate
4.1 The peroxidase enzyme fluorescence method is based on
glass, cleaned in accordance with procedures established for
the reaction of hydroperoxides, horseradish peroxidase, and
analyses of common inorganic ions (see Guide D 5012).
p-hydroxyphenylacetic (PHOPAA) acid, forming a fluorescent
7.3 Pipettes with Disposable Tips—Solution preparation
dimer of the latter. This dimer is detected using a fluorometric
and sample fixing operations are generally conducted using
technique, and the hydroperoxides are quantified by calibration
automatic pipettes. Solution volumes delivered by these de-
with hydrogen peroxide. The formation of the dimer (deriva-
vices should be verified to confirm consistent and accurate
tization) shall be accomplished soon after sample collection to
performance (see Test Method D 5085).
minimize H O decay. In addition, strict quality assurance
2 2
7.4 Reagent Bottles—All containers used for the prepara-
practices are part of the method, including use of systems
tion and storage of derivatizing and other reagent solutions
standards and systems blanks to estimate hydroperoxide loss
shall be dedicated for hydroperoxides. Containers for solutions
and to assess derivatizing solution effectiveness.
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) is a primary oxidizer of dissolved sulfur dioxide in
grade chemicals shall be used.
atmospheric water. Detection of H O in atmospheric water is
2 2
8.2 Purity of Water—Unless otherwise indicated, references
useful for inferring gas-phase H O concentrations and for
2 2
to water shall be understood to mean reagent water as defined
assessing the relative importance of various acidifying mecha-
by Type I of Specification D 1193, with the added stipulation
nisms under specific atmospheric conditions.
that the total organic carbon content be less than 20 μg/L. A
5.2 Hydroperoxides in samples to be analyzed are unstable
Type I water system equipped with an organic extraction
in water and can decay rapidly due to bacterial action or
cartridge and a 0.2 μm filter is an acceptable water source.
chemical reaction with other constituents. The test method
Water to be used for reagents, standard solutions, and analyti-
includes procedures for sample derivatization and methods for
cal rinsing should be stored in borosilicate glass.
estimating and correcting for hydroperoxide decay.
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
addition of the derivatizing reagent removes H O , but organic
effects of the following potentially interfering species. 2 2
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-
reaction with peroxides other than H O . Results of analyses of
press the negative interference of HMSA. In the absence of 2 2
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.
HMSA, displayed a fluorescence signal 5 % lower than that 2 2
8.3.1 Catalase, 1 + 49—Dilute 1 mL of catalase enzyme to
observed when HCHO was added to the derivatizing reagent.
a final volume of 50 mL with water. Before pipetting the
6.3 Trace Transition Metals and Common Ionic Compo-
concentrated solution, ensure that all the solid material is
nents of Atmospheric Water (Sodium, Ammonium, Hydrogen,
completely suspended by shaking or stirring the bottle of
Sulfate, Nitrate, Chloride, Formate)—Potential interference by
concentrate. Allow the dilute solution to stand at least 4 h
transition metals is overcome by the formation of ethylenedi-
before use. The solution can be stored for up to 48 h at 4°C.
aminetetraacetic acid (EDTA) complexes. Tests of simulated
8.4 Derivatizing Reagent, Concentrated—Dissolve 12.11 g
rain samples containing transition metals and common ionic
of Tris(hydroxymethyl)aminomethane, 0.38 g of EDTA, tetra-
components of precipitation have demonstrated both the gen-
sodium salt, 4.57 g of PHOPAA, 300 units of horseradish
eral applicability of this test method to samples containing
peroxidase, and 1 mL concentrated hydrochloric acid in water,
common contaminants and the stability of derivatized solutions
and dilute to 200 mL in a volumetric flask. The final pH of this
stored at 4°C for more than five days.
solution should be 9.0. If greater than 9.5 or less than 8.5,
7. Apparatus
7.1 Flow System, consisting of the following:
Reagent Chemicals, American Chemical Society Specifications, American
7.1.1 Automatic sampler or injection valve.
Chemical Society, Washington, DC. For suggestions on the testing of reagents not
7.1.2 Automated wet chemistry (peristaltic) pump.
listed by the American Chemical Society, see Analar Standards for Laboratory
7.1.3 Reagent manifold.
Chemicals, BDH Ltd., Poole, Dorset, U.K., and the United States Pharmacopeia
and National Formulary, U.S. Pharmacopeial Convention, Inc. (USPC), Rockville,
7.1.4 Mixing coil; 5-turn, 2-mm inner diameter.
MD.
7.1.5 Fluorometer; excitation at 320 nm and measurement
7 6
Catalase enzyme, 1.7 3 10 units/mL, has been found satisfactory for this
of the fluorescence signal at 400 nm; flow-through fluorescence
purpose. Available through Sigma Chemical Co., P.O. Box 14508, St. Louis, MO
cell. 63178.
NOTICE: This standard has either been superceded and replaced by a new version or discontinued.
Contact ASTM International (www.astm.org) for the latest information.
D 6363
remake. Prepare every four days and store at 4°C. Measure- D 5012 under inorganic ionic species (see 8.1 and 8.2 of Guide
ment of peroxides in aqueous atmospheric samples is based on D 5012).
the fluorescence of the PHOPAA dimer produced by reaction 9.3 Control procedures designed to ensure sample integrity
of hydroperoxides with PHOPAA. The fluorescence of samples in the field (see Section 10) are difficult to perform adequately
derivatized at the time of collection provide a measure of total if buckets or other high atmospheric-exposure collectors are
hydroperoxide (organic and H O ) content of the sample. used. Therefore, sampling for rain should be conducted using
2 2
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 The requirements for controlled derivatization of hydro-
Prepare daily as needed, and keep tightly sealed at 4°C. peroxides and timely analysis (see Section 10) dictate that
sampling for wet deposition be conducted on a daily or more
NOTE 1—The dilute derivatizing reagent is normally added to samples
frequent basis.
to be analyzed in the reagent:sample ratio of 1:1. Other concentrations of
dilute derivatizing reagent may be used as long as the final ratio entering
10. Derivatization
the analytical system is 1:1. Under special circumstances, other ratios may
10.1 The following procedures shall be in addition to those
be dictated by sampling conditions (see 10.6 and 10.7).
specified for preservation of inorganic anions and cations in
8.5 Hydrochloric Acid (HCl), (1 M)—Add 8.3 mL concen-
Guide D 5012 (see Table 1 of Guide D 5012).
trated HCl to water in a volumetric flask and dilute to 100 mL.
10.2 Hydroperoxides dissolved in atmospheric water solu-
8.6 Peroxide Solution, Standard Stock (1 %)—Dilute com-
tions are subject to decay at rates that are not predictable.
mercially available (pharmaceutical grade is acceptable) H O
2 2
Therefore, the derivatizing solution shall be added during
solution (30 % approximately1+29 with water in a volumet-
sample collection or within a known and controlled time after
ric flask. Add sodium stannate (Na SnO ) to a concentration of
2 3
sample collection.
10.65 mg/L and store at 4°C, and store in a borosilicate glass
10.3 The rate of decay of non-derivatized hydroperoxides
bottle. Determine the peroxide concentration by titration with
−1
may be quite fast: loss rates ranging from 1 to 28 % h were
standard permanganate solution (see 11.2) approximately 24 h
found for rain samples collected at Boulder, CO. The decay
after preparation. Update the concentration determination by
may be significant during the time of sampling, an especially
titration at one month intervals.
important consideration for sampling of precipitation. Thus,
8.6.1 Peroxide Solution, 1 + 199—Dilute 500 μL of the
addition of the derivatizing reagent to the collection container
standard stock (1 %) solution to 100 mL with water in a
prior to sampling (pre-derivatization) is the most desirable
volumetric flask. The approximate H O concentration of the
2 2
method. The pre-derivatized sample, however, is not suitable
resulting solution is 1500 μM (50 mg/L). Calibration standards
for analysis for other species, particularly hydrogen ion,
are prepared immediately
...
Questions, Comments and Discussion
Ask us and Technical Secretary will try to provide an answer. You can facilitate discussion about the standard in here.