ASTM D5412-93(2011)
(Test Method)Standard Test Method for Quantification of Complex Polycyclic Aromatic Hydrocarbon Mixtures or Petroleum Oils in Water
Standard Test Method for Quantification of Complex Polycyclic Aromatic Hydrocarbon Mixtures or Petroleum Oils in Water
SIGNIFICANCE AND USE
This test method is useful for characterization and rapid quantification of PAH mixtures including petroleum oils, fuels, creosotes, and industrial organic mixtures, either waterborne or obtained from tanks.
The unknown PAH mixture is first characterized by its fluorescence emission and synchronous scanning spectra. Then a suitable site-specific calibration standard with similar spectral characteristics is selected as described in Annex A1. This calibration standard may also be well-characterized by other independent methods such as gas chromatography (GC), GC-mass spectrometry (GC-MS), or high performance liquid chromatography (HPLC). Some suggested independent analytical methods are included in References (1–7) and Test Method D4657. Other analytical methods can be substituted by an experienced analyst depending on the intended data quality objectives. Peak maxima intensities of appropriate fluorescence emission spectra are then used to set up suitable calibration curves as a function of concentration. Further discussion of fluorescence techniques as applied to the characterization and quantification of PAHs and petroleum oils can be found in References (8–18).
For the purpose of the present test method polynuclear aromatic hydrocarbons are defined to include substituted polycyclic aromatic hydrocarbons with functional groups such as carboxyl acid, hydroxy, carbonyl and amino groups, and heterocycles giving similar fluorescence responses to PAHs of similar molecular weight ranges. If PAHs in the more classic definition, that is, unsubstituted PAHs, are desired, chemical reactions, extractions, or chromatographic procedures may be required to eliminate these other components. Fortunately, for the most commonly expected PAH mixtures, such substituted PAHs and heterocycles are not major components of the mixtures and do not cause serious errors.
SCOPE
1.1 This test method covers a means for quantifying or characterizing total polycyclic aromatic hydrocarbons (PAHs) by fluorescence spectroscopy (Fl) for waterborne samples. The characterization step is for the purpose of finding an appropriate calibration standard with similiar emission and synchronous fluorescence spectra.
1.2 This test method is applicable to PAHs resulting from petroleum oils, fuel oils, creosotes, or industrial organic mixtures. Samples can be weathered or unweathered, but either the same material or appropriately characterized site-specific PAH or petroleum oil calibration standards with similar fluorescence spectra should be chosen. The degree of spectral similarity needed will depend on the desired level of quantification and on the required data quality objectives.
1.3 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.
1.4 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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Standards Content (Sample)
NOTICE: This standard has either been superseded and replaced by a new version or withdrawn.
Contact ASTM International (www.astm.org) for the latest information
Designation:D5412 −93(Reapproved2011)
StandardTest Method for
Quantification of Complex Polycyclic Aromatic Hydrocarbon
Mixtures or Petroleum Oils in Water
This standard is issued under the fixed designation D5412; 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 (´) indicates an editorial change since the last revision or reapproval.
1. Scope D3326 Practice for Preparation of Samples for Identification
of Waterborne Oils
1.1 This test method covers a means for quantifying or
D3415 Practice for Identification of Waterborne Oils
characterizing total polycyclic aromatic hydrocarbons (PAHs)
D3650 Test Method for Comparison of Waterborne Petro-
by fluorescence spectroscopy (Fl) for waterborne samples.The
leum Oils By Fluorescence Analysis
characterization step is for the purpose of finding an appropri-
D4489 Practices for Sampling of Waterborne Oils
ate calibration standard with similiar emission and synchro-
D4657 TestMethodforPolynuclearAromaticHydrocarbons
nous fluorescence spectra.
in Water (Withdrawn 2005)
1.2 This test method is applicable to PAHs resulting from
E131 Terminology Relating to Molecular Spectroscopy
petroleum oils, fuel oils, creosotes, or industrial organic
E169 PracticesforGeneralTechniquesofUltraviolet-Visible
mixtures.Samplescanbeweatheredorunweathered,buteither
Quantitative Analysis
the same material or appropriately characterized site-specific
E275 PracticeforDescribingandMeasuringPerformanceof
PAH or petroleum oil calibration standards with similar fluo-
Ultraviolet and Visible Spectrophotometers
rescence spectra should be chosen. The degree of spectral
E388 Test Method for Wavelength Accuracy and Spectral
similarity needed will depend on the desired level of quantifi-
Bandwidth of Fluorescence Spectrometers
cation and on the required data quality objectives.
E578 Test Method for Linearity of Fluorescence Measuring
1.3 The values stated in SI units are to be regarded as Systems
E579 Test Method for Limit of Detection of Fluorescence of
standard. No other units of measurement are included in this
standard. Quinine Sulfate in Solution
1.4 This standard does not purport to address all of the
3. Terminology
safety concerns, if any, associated with its use. It is the
responsibility of the user of this standard to establish appro-
3.1 Definitions—For definitions of terms used in this test
priate safety and health practices and determine the applica-
method, refer to Terminology D1129, Terminology E131, and
bility of regulatory limitations prior to use.
Practice D3415.
2. Referenced Documents
4. Summary of Test Method
2.1 ASTM Standards:
4.1 This test method consists of fluorescence analysis of
D1129 Terminology Relating to Water
dilute solutions of PAHs or petroleum oils in appropriate
D1193 Specification for Reagent Water
solvents (spectroquality solvents such as cyclohexane or other
D2777 Practice for Determination of Precision and Bias of
appropriate solvents, for example, ethanol, depending on
Applicable Test Methods of Committee D19 on Water
polarity considerations of the sample). The test method re-
D3325 Practice for Preservation of Waterborne Oil Samples
quiresaninitialqualitativecharacterizationstepinvolvingboth
fluorescence emission and synchronous spectroscopy in order
to select appropriate calibration standards with similar fluores-
cence spectra as compared to the samples (see Annex A1 for
This test method is under the jurisdiction of ASTM Committee D19 on Water
andisthedirectresponsibilityofSubcommitteeD19.06onMethodsforAnalysisfor
the definition of spectral similarity). Intensities of peak
Organic Substances in Water.
maxima of suitable emission spectra are then used to develop
Current edition approved May 1, 2011. Published June 2011. Originally
calibration curves for quantification.
approved in 1993. Last previous edition approved in 2005 as D5412 – 93 (2005).
DOI: 10.1520/D5412-93R11.
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
Standards volume information, refer to the standard’s Document Summary page on The last approved version of this historical standard is referenced on
the ASTM website. www.astm.org.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
D5412−93(Reapproved2011)
NOTE1—Althoughsomesectionsofthecharacterizationpartofthistest NOTE 4—This test method is normally used without a matrix spike due
method are similar to Test Method D3650, there are also significant to possible fluorescence interference by the spike. If a spike is to be used,
differences (See Annex A1). Since the purpose and intent of the two test it must fluoresce in a spectral region where it will not interfere with the
methods are different, one should not be substituted for the other. quantification process. Compounds that could be used are dyes that
fluoresce at longer wavelengths than the emission of the PAH mixture.
5. Significance and Use
6.2 If the PAH mixture to be analyzed is a complex mixture
5.1 This test method is useful for characterization and rapid
such as an oil or creosote, it is assumed that a well-
quantification of PAH mixtures including petroleum oils, fuels,
characterized sample of the same or similar material is avail-
creosotes,andindustrialorganicmixtures,eitherwaterborneor
able as a calibration standard so the fluorescent fraction of the
obtained from tanks.
mixture can be ratioed against the total mixture. Otherwise,
since the samples and standards are weighed, the nonfluores-
5.2 The unknown PAH mixture is first characterized by its
cent portion of the mixture would bias the quantification
fluorescence emission and synchronous scanning spectra.Then
although the characterization portion of the test method for
asuitablesite-specificcalibrationstandardwithsimilarspectral
PAHs given in Annex A1 would be unaffected.
characteristics is selected as described in Annex A1. This
calibration standard may also be well-characterized by other
7. Apparatus
independent methods such as gas chromatography (GC), GC-
7.1 Fluorescence Spectrometer—An instrument recording
mass spectrometry (GC-MS), or high performance liquid
in the spectral range of 250 nm to at least 600 nm for both
chromatography (HPLC). Some suggested independent ana-
excitation and emission responses and capable of scanning
lytical methods are included in References (1–7) and Test
both monochromators simultaneously at a constant speed with
MethodD4657.Otheranalyticalmethodscanbesubstitutedby
a constant wavelength offset between them for synchronous
an experienced analyst depending on the intended data quality
scanning. The instrument should meet the specifications in
objectives. Peak maxima intensities of appropriate fluores-
Table 1. (Also known as spectrofluorometer or fluorescence
cence emission spectra are then used to set up suitable
spectrophotometer). Consult manufacturer’s instrument manu-
calibration curves as a function of concentration. Further
als for specific operating instructions.
discussion of fluorescence techniques as applied to the char-
acterization and quantification of PAHs and petroleum oils can
NOTE 5—Although the characterization section of this test method
be found in References (8–18).
(given in Annex A1) is similar to Test Method D3650 in many respects,
there are differences in the purpose and intents of the two test methods.
5.3 For the purpose of the present test method polynuclear
The purpose of the characterization step of this test method is to find an
aromatic hydrocarbons are defined to include substituted poly-
oil with similar fluorescence properties as the sample in order to serve as
cyclic aromatic hydrocarbons with functional groups such as
an appropriate calibration standard for quantification. Other differences
between the test methods are instrumentation requirements and the use of
carboxyl acid, hydroxy, carbonyl and amino groups, and
synchronous spectra as well as emission spectra for this test method.
heterocycles giving similar fluorescence responses to PAHs of
7.2 Excitation Source—A high-pressure xenon lamp (a
similar molecular weight ranges. If PAHs in the more classic
definition, that is, unsubstituted PAHs, are desired, chemical 150-W continuous xenon lamp or a 10-W pulsed xenon lamp
reactions, extractions, or chromatographic procedures may be has been proven acceptable). Other continuum sources (either
required to eliminate these other components. Fortunately, for continuousorpulsed)havingsufficientintensitythroughoutthe
the most commonly expected PAH mixtures, such substituted ultraviolet and visible regions may also be used.
PAHs and heterocycles are not major components of the
7.3 Fluorescence Cells—Standard cells made from
mixtures and do not cause serious errors.
fluorescence-free fused silica with a path length of 10 mm and
a height of at least 45 mm. Stoppered cells may be preferred to
6. Interferences
prevent sample evaporation and contamination.
6.1 The fluorescence spectra may be distorted or quantifi-
7.4 Data Recording System—Preferably the instrument
cation may be affected if the sample is contaminated with an
should be interfaced to a suitable computer system compatible
appreciable amount of other fluorescent chemicals that are
with the instrument and with suitable software for spectral data
excited and which fluoresce in the same spectral regions with
relatively high fluorescence yields. Usually the fluorescence
spectra would be distorted at levels greater than 1 to 2 % of
TABLE 1 Specifications for Fluorescence Spectrometers
such impurities before the quantification would be seriously
Wavelength Reproducibility
affected. Excitation monochromator ±2 nm or better
Emission monochromator ±2 nm or better
NOTE 2—Caution: Storage of samples in improper containers (for Gratings (Typical Values)
example, plastics other than TFE-fluorocarbon) may result in contamina- Excitation monochromator minimum of 600 lines/mm
blazed at 300 nm
tion.
Emission monochromator minimum of 600 lines/mm
NOTE 3—Spectroquality solvents may not have low enough fluores-
blazed at 300 nm or 500 nm
cence background to be used as solvent blanks. Solvent lots vary in the
Photomultiplier Tube
content of fluorescent impurities that may increase with storage time even
S-20 or S-5 response or equivalent
for unopened bottles.
Spectral Resolutions
Excitation monochromator spectral bandpass of 2.5 nm or less
Emission monochromator spectral bandpass 2.5 nm or less
The boldface numbers in parentheses refer to the list of references at the end of Maximum bandpasses for both monochromators at least 10 nm
this standard.
D5412−93(Reapproved2011)
manipulation. Use of a strip chart or X-Y recorder with a a problem only at PAH levels in the low µg/Lrange. Extraction
response time of less than 1 s for full-scale deflection is methods (or separation by column chromatography) are listed
acceptable. in Practice D3326.
9.4.1 An extraction method that proved satisfactory for the
7.5 Micropipet, glass, 10 to 50-µL capacity.
collaborative test is as follows:
7.6 Weighing Pans,5to7-mmdiameter,18-mmthick,made
9.4.1.1 Pour50.0mLofthesampleintoaseparatoryfunnel,
of aluminum or equivalent. Check pans for contamination.
add 5.0 mL of cyclohexane and shake for 2 min. Vent the
separatory funnel occasionally. Withdraw the aqueous layer
8. Reagents and Materials
(keep this for a second extraction). Collect the cyclohexane
extract in a 10-mL volumetric flask. Add 5.0 mL of cyclo-
8.1 Purity of Reagents—Use spectroquality grade reagents
hexane to the aqueous layer and perform a second extraction.
in all instances unless otherwise stated. Since the goal is to
Combine the two extracts and dilute to 10.0 mL with cyclo-
have as low a fluorescence blank as possible, and since
hexane.
different brands and lots of spectroquality solvent may vary,
9.4.1.2 For field use, it has proven satisfactory to use a
check reagents frequently.
reagent bottle instead of a separatory funnel. Pour 50.0 mL of
8.2 Purity of Water—References to water mean Type IV
the sample in the bottle and add 5.0 mLof cyclohexane, shake
water conforming to Specification D1193. Since fluorescent
for 2 min and collect most of the top layer with a Pasteur pipet.
organic impurities in the water may introduce an interference,
It is important to collect most of the top layer to maximize
check the purity of the water by analyzing a water blank using
percentrecovery(tilttheflasktoseetheseparationbetweenthe
the same instrumental conditions as for the solvent blank.
two layers more easily). Add 5.0 mL of cyclohexane to the
8.3 Acetone, spectroquality, (CH COCH ). aqueous layer and perform a second extraction. Combine the
3 3
two cyclohexane extracts and dilute to 10.0 mL with cyclo-
8.4 Cyclohexane, spectroquality or HPLC grade. The fluo-
hexane.
rescencesolventblankmustbeaslowaspossibleandlessthan
9.4.1.3 See 12.6 to check extraction recoveries. Other ex-
5 % of the intensity of the maximum emission peak for the
tractionmethodscanbeusedatthediscretionoftheanalyst,by
lowest concentration of PAHs analyzed. Dispense cyclohexane
adding an appropriate solvent exchange step to cyclohexane
during the procedure from either a TFE-fluorocarbon or glass
and by checking for recoveries and interferences.As is always
wash bottle, but, for prolonged storage, store cyclohexane only
the case, the analyst shall demonstrate method performance
in glass.
when changing the method. At the mg/L level or above, the
8.5 Nitric Acid(1+1)—Carefully add one volume of con-
PAH mixture might not be totally in solution. If the PAH
centrated HNO (sp gr 1.42) to one volume of water.
3 mixture is emulsified in water, is sparingly soluble in water, or
if the concentration of the unknown must be known more
8.6 TFE-Fluorocarbon Strips, 25 mm by 75 mm, 0.25-mm
accurately, it may be necessary to evaporate the solution to
thickness. Use TFE strips when sampling neat PAH films on
dryness or to extract the PAH mixture into a suitable solvent,
water as described in Practice D4489.
followed by evaporation, weighing, and redissolving in cyclo-
hexane.
9. Sampling and Sample Preparation
9.4.1.4 At the mg/L level or above, the PAH mixture in
9.1 Collect a representative sample (see Practice D4489 for
water might not be totally in solution.
water samples).
9.5 Sample bottles must be made of glass, precleaned with
9.2 Preserve samples in containers as specified in Practice
dilute nitric acid (1 + 1) and sealed with plastic screw caps
D3325. Do not cool sa
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