This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles
for the
Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.
Designation: D8473 − 22
Standard Test Method for
Determining the Biobased content of Liquid Hydrocarbon
Fuels Using Liquid Scintillation Counting with Spiked
Carbon-141
This standard is issued under the fixed designation D8473; 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
1.1 This test method covers quantitatively determining bio-
carbon content of liquid hydrocarbon fuels with a focus on
those produced in a typical petroleum refinery using liquid
scintillation counting (LSC). The method is designed to gen-
erate analogous results as Test Method D6866 Method C, for
low quench samples, without the need of benzene synthesis.
The purpose is to be able to use the produced data to report
biocarbon content of refinery products to regulatory agencies
and monitor refinery operation. The method does not address
regulatory reporting or fuel performance.
1.2 The method is needed to support refinery operations
when bio-feeds are co-processed with petroleum within a
reactor with a focus on samples with 100 % biocarbon or less
(not for 14C labeled species). It allows refineries to report the
biocarbon content of refinery products to regulatory agencies
such as the Environmental Protection Agency (EPA) or Cali-
fornia Air Resources Board (CARB) to comply with regulatory
statutes such as The Renewable Fuel Standard (RFS) or Low
Carbon Fuel Standard (LCFS).
1.3 This test method is applicable to any liquid fuel product,
petroleum based (pure hydrocarbon), biobased (such as renew-
able diesel or those that can contain oxygenates such as
ethanol), or blends, that contain 1 % to 100 % by mass
biocarbon where an instrument background can be experimen-
tally determined using a sample of similar matrix that contains
no measurable carbon-14.
1.4 This test method makes no attempt to teach the basic
principles of the instrumentation used although minimum
requirements for instrument selection are referenced in Refs
(1-11).2 However, the preparation of samples for the above test
methods is described. No details of instrument operation are
1
This test method is under the jurisdiction of ASTM Committee D02 on
Petroleum Products, Liquid Fuels, and Lubricants and is the direct responsibility of
Subcommittee D02.04.0F on Absorption Spectroscopic Methods.
Current edition approved Sept. 15, 2022. Published October 2022. DOI:
10.1520/D8473-22.
2
The boldface numbers in parentheses refer to a list of references at the end of
this standard.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
1
included here. These are best obtained from the manufacturer
of the specific instrument in use.
1.5 Pre-Requisite Requirements For Method Execution—
This test method uses artificial carbon-14 (14C) within the
method. Great care shall be taken to prevent laboratory
contamination of the elevated 14C. Once in the laboratory,
artificial 14C can contaminate a variety of laboratory surfaces
that can lead to artificially high sample biocarbon measure-
ments. If vigorous cleaning attempts to remove the artificial
14
C from a laboratory are unsuccessful, instrumentation and
sample preparation may have to be moved to a new laboratory
away from the contamination or the laboratory may have to
rely on outside third-party labs for analysis. Specific proce-
dural steps have been incorporated into this method that
minimize the risk of sample and lab contamination. Wipe tests
and quality assurance samples can validate absence of con-
tamination. In the event of contamination in the laboratory or
instrument, vigorous cleaning protocols shall be implemented,
and analysis cannot be resumed until the lab and instrument are
free of contamination. Accepted requirements are:
1.5.1 Working with the elevated 14C samples in a separate
and defined area away from the instrument and the preparation
of any non-spiked samples.
1.5.2 Using separate personnel to prepare the spiked
samples and non-spiked samples.
1.5.3 Using separate laboratory spaces with separate HVAC
systems for the handling of spiked and non-spiked samples.
The use of separate fume hoods that have separate exhaust
ventilation satisfies this requirement.
1.5.4 Weekly wipe tests of 14C sample handling area(s) to
detect lab contamination.
1.6 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 appro-
priate safety, health, and environmental practices and deter-
mine the applicability of regulatory limitations prior to use.
1.7 This international standard was developed in accor-
dance with internationally recognized principles on standard-
ization established in the Decision on Principles for the
 D8473 − 22
Development of International Standards, Guides and Recom-
mendations issued by the World Trade Organization Technical
Barriers to Trade (TBT) Committee.
2. Referenced Documents
2.1 ASTM Standards:3
D5291 Test Methods for Instrumental Determination of
Carbon, Hydrogen, and Nitrogen in Petroleum Products
and Lubricants
D6299 Practice for Applying Statistical Quality Assurance
and Control Charting Techniques to Evaluate Analytical
Measurement System Performance
D6300 Practice for Determination of Precision and Bias
Data for Use in Test Methods for Petroleum Products,
Liquid Fuels, and Lubricants
D6708 Practice for Statistical Assessment and Improvement
of Expected Agreement Between Two Test Methods that
Purport to Measure the Same Property of a Material
D6866 Test Methods for Determining the Biobased Content
of Solid, Liquid, and Gaseous Samples Using Radiocar-
bon Analysis
2.2 Other Standards:
DIN 51637 Liquid Petroleum Products – Determination of
the Bio-Based Hydrocarbon Content in Diesel Fuels and
Middle Distillates Using Liquid Scintillation Method4
CSN EN 16640 Bio-based products - Bio-based carbon
content - Determination of the Bio-Based Carbon Content
using the Radiocarbon Method5
3. Terminology
3.1 The definitions of terms used in this test method are
referenced in case the practitioner requires further information
regarding the practice of the art of isotope analysis and to
facilitate performance of these test methods.
3.2 Definitions:
3.2.1 background radiation, n—the radiation in the natural
environment; this includes cosmic radiation and radionucle-
otides present in the local environment, for example, materials
of construction, metals, glass, and concrete.
3.2.2 background sample, n—sample with no detectable 14C
(such as a carboniferous sample that should not contain any
measurable 14C because of its geologic age).
3.2.3 biobased, adj—containing organic carbon of renew-
able origin like agricultural, plant, animal, fungi,
microorganisms, marine, or forestry materials living in a
natural environment in equilibrium with the atmosphere.
3.2.4 biobased carbon content, n—the amount of biobased
carbon in the material or product as a percent of the total
organic carbon (TOC) in the product.
3
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 ASTM website.
4
Available from Deutsches Institut für Normung e.V. (DIN), Am DIN-Platz,
Burggrafenstrasse 6, 10787 Berlin, Germany, http://www.din.de.
5
Available from European Standards, Krimicka 134, 318 00 Pilsen, Czech
Republic, VAT: CZ09567909, https://www.en-standard.eu/.
2
3.2.5 biofeed, n—a feedstock sourced from a plant or
animal.
3.2.6 biogenic, adj—containing carbon (organic and inor-
ganic) of renewable origin like agricultural, plant, animal,
fungi, microorganisms, marine, or forestry materials.
3.2.7 biogenic carbon content, n—amount of biogenic car-
bon in the material or product as a percent of the total carbon
(TC) in the product.
3.2.8 carbon-14 (14C), n—naturally occurring radioactive
isotope of carbon that contains six protons and eight neutrons
with a true half-life of 5730 years.
3.2.9 cocktail, n—the solution in which samples are mixed
for measurement in an LSC.
3.2.10 coincidence circuit, n—a portion of the electronic
analysis system of an LSC which acts to reject pulses that are
not received from the two or three photomultiplier tubes (that
count the photons) within a given period of time and are
necessary to rule out background interference and required for
any LSC used in this test method.
3.2.11 coincidence threshold, n—the minimum decay en-
ergy required for an LSC to detect a radioactive event; the
ability to set that threshold is a requirement of any LSC used
in this test method.
3.2.12 coincidence time, n—the time period used by the
coincidence circuit that is used to determine if a detection event
is counted or rejected.
3.2.13 contemporary carbon, n—a direct indication of the
relative contributions of fossil carbon and “living” biospheric
carbon can be expressed as the fraction (or percentage) of
contemporary carbon, symbol fC; this is derived from “fraction
of modern” (fM) using the observed input function for atmo-
spheric 14C over recent decades, representing the combined
effects of fossil dilution of 14C (minor) and nuclear testing
enhancement (major); the relation between fC and fM is
necessarily a function of time; by 1985, when the particulate
sampling discussed in the cited reference was performed, the
fM ratio had decreased to approximately 1.2 (10, 11).
3.2.14 counting time, n—the total time used by the liquid
scintillation counter to count sample 14C decays.
3.2.15 counts per minute (cpm), n—the average number of
counts the liquid scintillation counter detections during analy-
sis; is used to derive dpm.
3.2.16 decay (radioactive), n—the spontaneous transforma-
tion of one nuclide into a different nuclide or into a different
energy state of the same nuclide; the process results in a
decrease, with time, of the number of original radioactive
atoms in a sample, according to the half-life of the radionu-
clide.
3.2.17 delay time, n—the time the instrument waits after
method is started before it starts counting; allows user to delay
the start of analysis to allow sample photoluminescence to stop
before counting is initiated.
3.2.18 discriminator, n—an electronic circuit which distin-
guishes signal pulses according to their pulse height or energy;
 D8473 − 22
used to exclude extraneous radiation, background radiation,
and extraneous noise from the desired signal.
3.2.19 disintegrations per minute (dpm), n—the quantity of
radioactivity; the measure dpm is derived from cpm or counts
per minute (dpm = (cpm – cpm of background) / counting
efficiency); there are 2.2 × 106 dpm / µCi.
3.2.20 effıciency, n—the ratio of measured observations or
counts compared to the number of decay events which oc-
curred during the measurement time; expressed as a percent-
age.
3.2.21 figures of merit, n—a term applied to a numerical
value used to characterize the performance of a system; in
liquid scintillation counting, specific formulas have been
derived for quantitatively comparing certain aspects of instru-
ment and cocktail performance and the term is frequently used
to compare efficiency and background measures (refer to Eq 1).
3.2.22 fluorescence, n—the emission of light resulting from
the absorption of incident radiation and persisting only as long
as the stimulation radiation is continued.
3.2.23 fossil carbon, n—carbon that contains no measurable
radiocarbon because its age is very much greater than the
5730-year true half-life of 14C.
3.2.24 half-life, n—the time in which one half the atoms of
a particular radioactive substance disintegrate to another
nuclear form; the true half-life of 14C is 5730 years.
3.2.25 intensity, n—the amount of energy, the number of
photons, or the numbers of particles of any radiation incident
upon a unit area per unit time.
3.2.26 internal standard, n—a known amount of radioactiv-
ity which is added to a sample to determine the counting
efficiency of that sample; the radionuclide used shall be the
same as that in the sample to be measured and have a certified
activity.
3.2.27 luminescence, n—scintillation flux of photons; can
produce coincidences if intense; causes—luminescent reac-
tions in sample, photoluminescence from bright, especially UV
excitation.
3.2.28 modern carbon, n—explicitly, 0.95 times the specific
activity of SRM 4990B (the original oxalic acid radiocarbon
standard), normalized to δ13C = −19 % (10); functionally, the
fraction of modern carbon equals 0.95 times the concentration
of 14C contemporaneous with 1950 wood (that is, pre-
atmospheric nuclear testing). To correct for the post 1950 bomb
14
C injection into the atmosphere, the fraction of modern
carbon is multiplied by a (REF) atmospheric adjustment value
representative of the excess 14C in the atmosphere at the time
of measurements.
3.2.29 noise pulse, n—a spurious signal arising from the
electronic and electrical supply of the instrument.
3.2.30 operational quality test (OQ), n—test done to verify
liquid scintillation counter is working properly.
3.2.31 photomultiplier tube (PMT, pmt), n—the device in
the LSC that counts the photons of light simultaneously at two
or three separate detectors.
3
3.2.32 pulse, n—the electrical signal result in when photos
are detected by the PMTs.
3.2.33 quenching, n—any material that interferes with the
(accurate) optimal conversion of decay energy to scintillation
photons captured by the PMT of the LSC (a significant
interference with this test method).
3.2.33.1 chemical quenching, n—a reduction in the scintil-
lation intensity seen by the PMT due to the materials present in
the sample that interfere with the processes leading to the
production of light.
3.2.33.2 color quenching, n—any material that absorbs
generated scintillation photons.
3.2.34 region, n—regions of interest, also called window or
channel, or both, regarding LSC; refers to an energy level or
subset specific to a particular isotope.
3.2.35 renewable, n—being readily replaced and of non-
fossil origin; specifically, not of petroleum origin.
3.2.36 scintillation, n—the sum of all photons produced by
a radioactive decay event; counters used to measure this as
described in this test method are Liquid Scintillation Counters
(LSC).
3.2.37 scintillation reagent, n—chemicals that absorbs de-
cay energy transferred from the solvent and emits light
(photons) proportional in intensity to the decay energy.
3.2.38 solvents and scintillators, n—chemicals that absorb
decay energy transferred from the solvent and emits light
(photons) proportional in intensity to the deposited energy.
3.2.39 solvent-in scintillation reagent, n—chemical(s)
which act as both a vehicle for dissolving the sample and
scintillator and the location of the initial kinetic energy transfer
from the decay products to the scintillator; that is, into
excitation energy that can be converted by the scintillator into
photons.
3.2.40 specific activity (SA), n—refers to the quantity of
radioactivity per mass unit of product, that is, decays per
minute per gram of carbon (dpm/gC).
3.2.41 standard count conditions (STDCT), n—LSC condi-
tions under which reference standards and samples are
counted.
3.2.42 triple to double count ratio (TDCR), n—the ratio of
counts detected by three detectors over the number of counts
that were detected by two detectors; requires use of a liquid
scintillation counter with three detectors.
3.2.43 wipe test, n—a test that is done to determine if a
surface has been contaminated with 14C or any other radioac-
tive isotope.
4. Summary of Test Method
4.1 Fuels or their component streams, or both, are mixed
with scintillation cocktail and placed into a liquid scintillation
counter with TDCR capabilities. The instrument is then used to
determine the number of 14C disintegrations per minute per
gram of carbon in the sample to calculate its biological carbon
content. 14C spiking using a separate aliquot of the same
 D8473 − 22
sample, and prepared at a different location, is used to
determine counting efficiencies of each sample.
5. Significance and Use
5.1 This test method provides accurate biobased/biogenic
carbon content results to materials whose carbon source was
directly in equilibrium with CO2 in the atmosphere at the time
of cessation of respiration or metabolism, such as the harvest-
ing of a crop or grass living in a field. Special considerations
are needed to apply the testing method to materials originating
from within artificial environments with non-natural levels of
14
C or if the biofeed was grown over the course of several
years such as trees and contains “bomb-carbon.” Application of
these test methods to materials derived from CO2 uptake within
artificial environments is beyond the present scope of this
standard.
5.2 This method uses LSC techniques to quantify the
biobased content of a liquid hydrocarbon fuels using sample
carbon that has been unmodified. It is designed to be able to
incorporate into a refinery laboratory to support biofeed and
petroleum coprocessing or blending operations to determine
the biocarbon content of the intermediate or finished products.
The test results can then be used for optimizing internal
parameters or reporting to regulatory agencies.
5.3 The use of this method requires that a pure petroleum-
based sample can be generated that has a similar matrix to each
product or stream to be analyzed. For example, gasoline and
diesel have very different matrices and will likely require the
use of different background measurements for each. Refer to
10.2 for how to determine if the same background sample can
be used for more than one product/stream.
6. Interferences
6.1 Sample Matrix—The sample matrix affects background
and quenching, so backgrounds (samples with no detectable
14
C) need to be measured for each stream or blend that is to be
analyzed.
6.2 Chemical/Color Quenching—Higher boiling samples
will likely contain higher levels of quenching molecules such
as conjugated aromatics. The presence of these species in high
concentrations can require sample dilutions or prevent use of
this method. Counting efficiency measurements can be con-
ducted to determine if the quench levels are too high. Counting
efficiency is required to be above 50.0 %.
6.3 Luminescence—In LSC context, unwanted non-
scintillation flux of photons. Consists of single photons and, if
moderate, does not markedly produce coincidences. If lumi-
nescence intensity increases, probability for two photons hit-
ting separate PMT’s within coincidence time increases and
random coincidences start to occur. Probability for triple
coincidences remains near negligible though. Common causes
of luminescence are luminescent reactions in sample (chemi-
luminescence) and photon emission after exposure to light,
especially UV illumination (photoluminescence). Precautions
to minimize luminescence include, avoid bright illumination
when preparing and handling samples, select triple coincidence
mode and/or set counting region (window) to exclude low-
4
amplitude luminescence pulses, and employ delay time before
counting starts, to allow photoluminescence to fade.
6.4 Sample and scintillation cocktail volumes need to be
consistent to obtain accurate efficiency measurements. Varia-
tions in the sample to cocktail ratio will change the dilution of
the quenching agents present in the sample resulting in an
increase or decrease in counting efficiency if too little sample
or too much sample is added in respect to the scintillation
cocktail, respectively. Measured volumes shall not fluctuate
more than 0.1 mL from the target volume to ensure data
accuracy.
6.5 Biological carbon contamination from outside sources,
sources of fossil or modern carbon such as corks, paper towels,
plant-based rope/string or dust shall be kept away from the
samples to prevent contamination of the sample.
6.6 14C Spike Standard Contamination in Samples—
Precaution is needed when preparing and handling the spike
solution and spiked samples to prevent sample and laboratory
contamination. Review 1.5 to make sure all necessary precau-
tions are being followed.
6.7 Sample Volatility—Addition of the sample to the vial
should always be the last step. When samples are weighed, a
cap needs to be tightly sealed when recording the final sample
mass measurement and the cap should not be reopened before
measuring. Background samples that are collected and stored
for extended periods of time shall be stored and sampled in a
manner to prevent loss of volatiles.
6.8 Potassium-40 (40K)—A radioactive isotope of potassium
that is present in glassware. 40K predominantly decays through
beta decay to 40Ca. The beta decay LSC spectrum of 40K
partially overlaps with that of 14C. Even low potassium glass
still has high enough levels of 40K to interfere with the results
produced via this method. For this reason, this method requires
the use of PTFE coated plastic vials and not glass vials to
prevent this interference.
6.9 Radon-222, see 14.4 for how to correct for radon
contamination.
6.10 Bomb Pulse:
6.10.1 The pMC can be greater than 100 % because of the
continuing but diminishing effects of the 1950s nuclear testing
programs, which resulted in a considerable enrichment of 14C
in the atmosphere. The decrease in 14C from the bomb testing
programs has been nonlinear in the past but has been linear
since at least 2004 to present. As of 2019 the 14C activity in the
atmosphere has reached the 1950 level of 13.56 dpm per gram
carbon that is defined as 100 pMC. Because all sample 14C
activities are referenced to a “prebomb” standard, and because
nearly all new biobased products are produced in a post-bomb
environment, all pMC values shall be adjusted by an atmo-
spheric correction factor (REF) to obtain the true biobased
content of the sample. The correction factor is based on the
excess or deficiency of 14C activity in the atmosphere at the
time the biological source was alive. A REF value of 102 pMC
was determined for 2015 based on the measurements of CO2 in
air in a rural area in the Netherlands (Lutjewad, Groningen).
The first version of Test Method D6866 – 04 in 2004 refer-
enced a value of 107.5 pMC and the Test Method D6866 – 10
 D8473 − 22
version (2010) cited 105 pMC. These data points equate to a
decline of 0.5 pMC per year. Therefore, on January 2 of each
year, the values in Table 1 are used as REF through 2019,
reflecting the same 0.5 pMC decrease per year until 2019 when
the pre bomb pulse level was reached. For all REF values after
2022, refer to the most recent version of Test Method D6866.
6.10.2 Atmospheric thermonuclear weapons testing was
extensive between 1952 and 1963. During this time period the
14
CO2 content in the air increased by 90 %. This means that a
plant living in 1965 would measure about 190 pMC. Since the
signing of the testing ban in 1963 this signature declined to
about 140 pMC by 1975, 120 pMC by 1985, and 101.5 pMC
by 2016. The consequence of this effect is error in biobased
content analysis relating to when the biobased material used in
the product was last actively part of a respiring/metabolizing
system. The error is predominant in products made from
forestry products. The rings within trees each represent the
previous growth season within which the previous year’s
14
CO2 signature was recorded. The center most ring of a tree
living today but planted in 1965 would be about 190 pMC
whereas the outermost ring/bark would be 100.0 pMC. If this
tree is harvested and used in manufacturing a biobased product,
the percent biobased carbon content of the product will be
dependent on where the carbon came from within the tree and
would be the average pMC of each of the rings and their size
in the portion of the tree used in the manufacturing process.
6.10.3 Bomb carbon is readily identified in a product when
the product’s pMC value is greater than 6 pMC above the
prescribed correction factor (REF). A high value can be
predicted based on the origin of the manufacturing compo-
nents. High values are typically observed in paper, cardboard,
forestry products, and forestry-derived chemicals. An exact
correction factor REF is not possible based strictly on the
measured pMC value of the product.
6.11 Variation in transmittance from vial to vial. Each PTFE
coated vial may have a slightly different transmittance through
it. As different vials are used to measure the background,
sample, and sample counting efficiency the variations in
transmittance from one vial to another will increase the error of
the calculated results.
7. Apparatus
7.1 Low level LSCs with active shielding that can produce
consistent background counts for a given sample matrix.
TABLE 1 Percent Modern Carbon (pMC) Reference
Year REF (pMC)
2015 102.0
2016 101.5
2017 101.0
2018 100.5
2019 100.0
2020 100.0
2021 100.0
2022 100.0
5
NOTE 1—Hidex 300 SL6 was used to conduct initial studies.
7.2 Anti-coincidence systems with at least three PMTs
(multidetector systems).
NOTE 2—Hidex brand LSC’s are currently the only known commercial
brand to have three detectors.6
7.3 Coincidence circuits.
7.4 Optimized counting regions to provide very low back-
ground counts while maintaining counting efficiency greater
than 50.0 %. The optimization of counting regions shall be
determined for each sample type to be analyzed. The level of
quench for a given sample type will change the optimal region
of interest.
NOTE 3—Initial work found that counting over channels 100 to 400 was
optimal for a diesel fuel sample.
NOTE 4—Test Method D6866 Method C recommends an efficiency of
60 % or higher. As that method is using a uniform and constant matrix
with little to no quenching present, this efficiency is more than adequate.
NOTE 5—This method analyzes the samples with no sample
preparation, and the analysis of samples with higher levels of quench may
be needed, but as the level of quench increases, it is more difficult to
distinguish low biocarbon containing samples from the noise. Depending
on the expected biocarbon concentration of the sample, it may be more
beneficial to dilute the sample to increase the counting efficiency as
sample dilution increases efficiency proportionally higher than it reduces
the amount of sample used, that is, a 25 % dilution will increase efficiency
by more than 25 % for high quench samples. Start running the sample with
a small dilution first and slowly increasing the dilution factor until the
measured efficiency is over 50.0 %. Some example dilutions ratios are
suggested here (mL sample+mL cocktail): 12+8, 10+10, 8+12, 5+15, and
2+18. 13.5 describes how to proceed if dilution is required.
7.5 No single LSC is specified for this method. However,
minimum counting efficiency and control of background inter-
ference is specified. Like all analytical instruments, LSCs
require study as to their specific components and counting
optimization. Currently Hidex6 is the only commercial source
of a three PMT LSC system that satisfies need stated in 7.2.
7.6 Standardization of sample preparation is required.
NOTE 6—Initial work used 15 mL of sample with 5 mL of scintillation
cocktail.
7.7 Unused 20 mL sample vials comprised of PTFE coated
plastic shall be used. Vials are not to be reused after sample
analysis.
NOTE 7—20 mL PTFE coated plastic vials were used to keep back-
ground levels as low as possible and allow for higher sample volumes to
decrease method detection limit.
7.8 The aforementioned optimizations shall be performed
using a suitable reference standard using the same reagents and
counting parameters as the samples. The reference standard
used for optimization shall be similar to the samples to be
6
The sole source of supply of the apparatus known to the committee at this time
is HIDEX, Lemminkäisenkatu 62 FIN-20520 Turku, Finland. If you are aware of
alternative suppliers, please provide this information to ASTM International
Headquarters. Your comments will receive careful consideration at a meeting of the
responsible technical committee,1 which you may attend.

analyzed. The standard should have low volatility and have a
similar matrix to that of the samples to be analyzed, that is, if
the samples are expected to contain oxygenates such as ethanol
the standard should also contain the same oxygenates in similar
concentrations.
NOTE 8—Initial work used diesel samples to determine optimized
conditions as diesel is the most colored sample with the most complex
matrix.
7.9 Counting interference concerns that shall be addressed
as part of specific instrument calibration and normalization
include luminance, chemical or color quench, static electricity,
random noise, temperature, and humidity variability.
7.10 Alternate regions of interest parameters may be used
based upon testing of 20, or more, 5 h counts of the same
reference standard that record the raw data throughout 14C
spectral region (channels 5 to 650 for Hidex6 instrumentation).
Optimal counting conditions shall be established by maximiz-
ing the Figures of Merit (see Eq 1) to obtain the highest count
efficiency and the lowest background. Counting efficiency of
less than 50.0 % is unacceptable and can be improved by LSC
instrument optimization and sample/reagent compatibility or
shielding improvements.
Figures of Merit 5 ~~ E 3 V ! 2 ⁄ bkg ! 3 k
where:
E = counting efficiency,
V = volume of sample,
bkg = CPM measured over a specific LSC channel range,
and
k = 1 CPM / (% × mL)2.
NOTE 9—Eq 1 is from DIN 51637.
8. Reagents and Materials
8.1 Unused 20 mL PTFE coated plastic vials. All vials are to
be single use to avoid sample carryover and reducing the risk
of lab contamination from artificial 14C present in the spiked
samples.
8.2 Unquenched standard kit used to verify instrument
performance.
8.3 Internal standard kit 14C, compatible with sample
matrix, used to determine counting efficiency of samples. (The
14
C labeled compound used to create the spike shall be a
non-volatile compound (vapor pressure below 1 Pa at 25 °C)
such as cholesterol to minimize potential of the compound
getting into the gas phase during sample preparation to reduce
risk of lab contamination.)
8.4 Scintillation cocktail, miscible with sample matrix and
not phase separate in the instrument.
8.5 Wipe test scintillation cocktail to dissolve wipes.
8.6 Wipe test strips designed to be dissolved in scintillation
cocktail.
9. Hazards
9.1 The specific safety and regulatory requirements associ-
ated with radioactivity, sample preparation, and instrument
operation are not addressed in this test method. It is the
D8473 − 22
responsibility of the user of this test method to establish
appropriate safety and health practices. It is also incumbent on
the user to conform to all the government legislation for their
location, especially those that relate to the use of open
radioactive source, in the performance of this test method.
Although 14C is one of the safest isotopes to work with, all
government regulations shall be followed in the performance
of this test method. As this method utilizes elevated levels of
14
C additional regulations and precautions may apply.
10. Sampling, Test Specimens, and Test Units
10.1 When sampling from large tanks avoid using compo-
nents that contain carbon-based material such as a cork or
non-synthetic rope or string. Metal or glass shall be used
wherever possible when safe to do so.
10.2 If this method is to be used to support coprocessing of
biofeeds with petroleum in a refinery or similar setting samples
shall be collected before coprocessing has started from each
stream or product, that will be analyzed after coprocessing has
started to allow for adequate backgrounds to be measured.
These samples serve as a background for the desired products/
streams and will allow for background corrections to be made.
(1) One background will have to be prepared and measured weekly
(all other backgrounds on a monthly basis, measurement
frequency outlined in more detail in 11.3.5) to verify laboratory
is not contaminated with artificial 14C and to account for any
instrumental drift, thus make sure enough sample is collected
as it may be needed for several years. More volatile samples,
such as gasoline boiling range material, shall be stored in a
cold area to prevent loss of volatile components over a long
period of time. The same background can be used for several
products/streams if the backgrounds of those products/streams
have been experimentally determined to be indistinguishable.
To determine if backgrounds can be considered the same or not
use a t-test after ten background measurements have been done
and use the t-value for a 95 % confidence interval.
10.3 Samples shall be thoroughly mixed before adding to
the scintillation vial.
11. Calibration, Standardization, and Quality Control
11.1 Apparatus—Operational quality of the instrument shall
be determined weekly to make sure the instrument is working
properly. This can be done with an unquenched standard of
known 14C activity. Some commercial instruments have a
designated procedure and kits to do this.
11.2 Quality Control (QC) Sample—A sample of known
biocarbon content shall be analyzed weekly. Preferred charac-
teristics of the QC sample are:
(1) Similar biocarbon content to samples,
(2) Similar matrix to samples, and
(3) Shelf stable with low volatility.
11.2.1 This QC shall be run following the same procedure
as samples as outlined in Section 13.
11.2.2 The measured biocarbon content of the QC shall be
control charted in accordance with Practice D6299 and moni-
tored for:
6
 D8473 − 22
11.2.2.1 Measurements trending away from the mean over
time. If this variation is observed analysis shall be halted until
reason for sustained variation has been determined and fixed.
11.2.2.2 Single points measured outside 63 sigma from the
mean. If this occurs rerun sample to verify result. If the rerun
is also outside 63 sigma from the mean, analysis shall be
halted until reason for variation has been determined and fixed.
If the rerun is inside 63 sigma from the mean, record this data
point in the control chart and discard the other. Keep a separate
record of when and the measured value for all points that are
outside of the 63 sigma limit for reference.
11.2.3 If laboratory 14C contamination or instrumental issue
has been identified, all data that has been collected since the
issue occurred shall be reprepared and reanalyzed once the
issue has been rectified. If an exact date of when the issue
started cannot be determined, all samples since the last good
QC measurement shall be reprepared and reanalyzed.
11.3 Background Sample—For each sample type to be
analyzed, a background needs to be measured to determine
proper background correction factor.
11.3.1 A background sample is a sample that contains no
measurable 14C.
11.3.2 As samples are analyzed directly, this is very impor-
tant as each sample type will likely have different types and
amounts of quenching species present. For example, different
gasoline blending components (such as reformate or aklylate)
need to have its own background unless experimentally deter-
mined to be identical. Refer to 10.2 for how to determine if
backgrounds are identical.
11.3.3 To establish a background for a sample, prepare the
same background sample five times using the exact same
reagents and conditions that will be used to analyze samples
such as sample vials, scintillation cocktail, sample to cocktail
ratio, counting time, and region of interest.
11.3.4 The measured CPM / gC for all five backgrounds is
averaged together and used as the background value for
samples of similar matrix.
11.3.5 A new background shall be measured monthly for all
products/streams. In addition to the monthly measurements,
one background sample shall be run weekly to help identify if
14
C lab contamination has occurred that was not detected via
the wipe test and to verify that no short-term instrument
fluctuations have occurred that would require the running of all
sample backgrounds. The background sample matrix shall
remain constant over the life to the sample, a sample with low
volatility and is stable at room temperature will simplify
sampling and storage requirements needed to maintain the
constant sample matrix.
11.3.6 The CPM / g C value of the new background is
averaged with the last nine values collected for that sample
matrix to maintain a rolling average to account for any minor
changes in background over time. If this is near to start of
analysis and nine previous backgrounds have not been taken
yet, average all backgrounds collected until a total of ten
backgrounds have been run.
7
11.3.7 A control chart shall be kept for all background
values measured (each background sample type should have
their own chart). Refer to 11.2.2 for control charting guide-
lines.
11.3.7.1 The background that is run on a weekly basis is to
aid in finding issues quickly so any issues can be addressed in
a timely manner.
11.3.8 If laboratory 14C contamination or an instrumental
issue has been identified, all data that has been collected since
the issue occurred shall be reprepared and reanalyzed once the
issue has been rectified. If an exact date of when the issue
started cannot be determined, all samples since the last good
background measurement shall be reprepared and reanalyzed.
11.4 Wipe Test—A weekly wipe test is to be performed in
any location that may potentially get contaminated with the
artificial 14C that is used during this analysis. The purpose of a
wipe test is to have a quick test to determine if 14C spike
solution or any spiked samples contaminated any part of the lab
so that cleanup can be done before the contamination is spread
too widely throughout the lab and/or contaminates any of the
samples. The most likely source of 14C contamination during
the use of this method is from a spill, if all method require-
ments are followed with regards to 14C spike sample materials
and handling.
11.4.1 Each location where the 14C spike solution, 14C spike
reagent, or the 14C spiked samples are handled shall be wipe
tested for possible contamination. Here is a list of example
locations: balance, area next to the balance, and hood where
14
C spiked samples are prepared, the knobs of any doors
between 14C spike sample preparation area and the instrument,
the instrument, the instruments autosampler tray, instrument
computer keyboard, etc. As the 14C spike solution and spiked
samples have a much higher concentration of 14C only a small
spill is easily detectable by a quick wipe test.
11.4.2 Example wipe test procedure.
11.4.2.1 Use dissolvable wipe test pads to wipe designated
area. Follow the wiping procedure designed specifically for
wipes chosen.
11.4.2.2 After wiping designated area, place wipe into a
20 mL liquid scintillation vial. Make sure the lid is labeled with
the location where the wipe was used. Place one wipe directly
into a vial without wiping any surface and label vial as blank.
11.4.2.3 Add 20 mL of scintillation cocktail designed to
dissolve chosen wipes.
11.4.2.4 Wait until wipe has fully dissolved and place it into
the liquid scintillation counter.
11.4.2.5 Set the instrument to count only the photons with
triple coincidence counts over the full 14C window range, range
designation may change depending on instrument used, for one
minute.
11.4.2.6 Control chart the results for each location and the
blank, as described in Practice D6299. As the runs are so short,
the standard deviation is quite high. If control charts indicate
that any of the wipe locations are trending upwards over the
course of several weeks, there is a potential that lab contami-
nation has occurred.
11.4.2.7 If the measured CPM for all samples is below 100
or 3× of the blank, whichever is lower, it is considered clean.
 D8473 − 22
If the CPM is above 100 or 3× of the blank, whichever is lower,
the location may be either dirty or contaminated. Clean surface
of any dirt or dust and repeat the wipe test in that area. Keeping
all lab surfaces free of dust and dirt will help reduce the
frequency of false positive measurements. If second run
measured CPM is also above 100 or 3× of the blank, there is
likely 14C contamination that needs to be cleaned before any
further analysis is performed.
11.4.2.8 Once contamination has been detected, cleanup
verification is required to be conducted by a 3rd party
laboratory.
11.4.2.9 If laboratory 14C contamination has been identified,
all data that has been collected since the issue occurred shall be
reprepared and reanalyzed once the contamination has been
cleaned. If an exact date of when the issue started cannot be
determined all samples since the last clean wipe test measure-
ment shall be reprepared and reanalyzed.
11.4.2.10 A wipe test shall be performed before any artifi-
cial 14C is handled within the laboratory to determine a
baseline level that is to be expected from the wipe tests.
12. Conditioning
12.1 All non-spiked samples, QC samples, and backgrounds
shall be allowed to sit in a dark environment after it has been
mixed with the scintillation cocktail for a minimum of 1 h
before analysis is started to reduce interference from sample
photoluminescence. If the sample is reintroduced to light, the
timer shall be reset to 1 h before sample is analyzed.
13. Procedure
13.1 Operational Quality Check:
13.1.1 A set of unquenched standards is to be run per
manufacturer’s instruction to verify that the instrument is
working properly.
13.2 Background Samples Preparation and Analysis:
13.2.1 Add optimized amount of scintillation cocktail to
vial. Use figures of merit to determine the optimal ratio of
sample to scintillation cocktail. (See Eq 1.)
NOTE 10—Initial work used 5 mL of scintillation cocktail.
13.2.2 Set vial onto analytical balance with cap on and tare
the vial.
13.2.3 Remove vial cap and add optimized amount of
background sample to vial, then recap the vial.
NOTE 11—Initial work used 15 mL of sample.
13.2.4 Record the mass of the sample added to the nearest
0.1 mg and label each vial cap with what sample was placed in
each vial (DO NOT write on the side of the vial as this will
interfere with the instrument detection).
13.2.5 Once sample is capped and properly labeled, place in
a dark place while other samples are being prepared.
13.2.6 Repeat 13.2.1 – 13.2.5 for each background sample
needing to be measured.
13.2.7 If this is the very first-time running backgrounds,
repeat 13.2.1 – 13.2.5 four more times to start a rolling
background average that will be continued each time back-
grounds are remade and run.
8
13.2.8 Make sure the outside of each sample vial is clean,
free of anything that could interfere with transmittance of light
out of the vial such as sample or dust, by wiping the outside
with a lint free cloth or similar before analysis.
13.2.9 Backgrounds will need to be remade and measured
weekly/monthly to make sure same background levels are
being measured or to account for any instrumental drift. See
11.3.5 for which frequency each background sample is to be
run.
13.2.10 Run the background samples using the optimized
LSC conditions. The following are some of the main param-
eters that need to be optimized before starting routine analysis:
analysis time, sample to cocktail ratio, coincidence time,
ionizer delay, and range of interest. Consult instrument manual
or instrument manufacturer if there are additional parameters
that must be optimized. When optimizing these parameters use
the same figures of merit question in Eq 1.
NOTE 12—Initial work used a 5 h analysis time as it was experimentally
determined to give a good balance of data variability while not requiring
a prohibitive amount of time. Shorter analysis time was found to greatly
increase data variability. A coincidence time of 35 ns was used. Ionizer
delay will be highly dependent on the instrument’s environment and the
personnel prepping the samples, start with default values and increase
them if static electricity interferences is found to be a problem. Range of
interest of channels 100 to 400 was determined using figures of merit
calculations on a diesel sample.
13.2.11 Record the measured CPM, to the nearest
0.01 CPM, for each of the backgrounds measured. Calculate
the CPM / gC and average this run with the previous runs of the
sample background sample to use when calculating biocarbon
content in the sample.
13.2.12 For sample types that are run infrequently that do
not justify running monthly background, instead of following
13.2.7 and 13.2.9, a single background run can be used but it
shall be run immediately before or after the sample and a new
background shall be prepared and analyzed each time that
sample type is run.
13.3 14C Spike Standard Preparation—Precaution is needed
when preparing and handling the spike solution to prevent
sample and laboratory contamination. Review 1.5 to make sure
all necessary precautions are being followed.
13.3.1 Place capsules from the Internal Standard Kit 14C
into one of the sample vials. (The 14C labeled compound used
to create the spike shall be a non-volatile compound (vapor
pressure below 1 Pa at 25 °C), such as cholesterol, to minimize
potential of the compound getting into the gas phase during
sample preparation to reduce risk of lab contamination.)
13.3.2 Mix with a solvent that is miscible with the samples
to be analyzed. Allow the solvent to dissolve the analyte for
30 min before proceeding. Prepare the sample to have a high
enough activity so that only 50 µL to 100 µL need to be spiked
in the sample for efficiency measurements. Should ideally
measure several thousand counts per minute (2000 cpm to
8000 cpm) when spiked into sample. The use of a solvent with
low volatility (vapor pressure below 1 Pa at 25 °C), such as the
scintillation cocktail, shall be used to help prevent any of the
artificial 14C molecules from being entrained into the gas phase
to reduce risk of lab contamination.
13.3.3 Store solution in a dry dark location.
 D8473 − 22
13.3.4 Make a new solution once depleted.
13.4 Sample Preparation and Analysis:
13.4.1 Sample CPM Measurement:
13.4.1.1 It is advised to prepare these samples before those
used to determine sample counting efficiency. Doing so allows
for these samples to sit in the dark while other samples are
prepared and analyzed.
13.4.1.2 Add optimized amount of scintillation cocktail to
vial, must be the same amount of scintillation cocktail used for
the background samples (13.2.1 and Note 10).
13.4.1.3 Set vial onto analytical balance with cap on and
tare the vial.
13.4.1.4 Remove cap and add optimized amount of sample
to vial then recap the vial, must be the same amount used for
background samples (13.2.3 and Note 11).
13.4.1.5 Record the mass of the sample added to the nearest
0.1 mg and label each vial cap with what sample was placed in
each vial (DO NOT write on the side of the vial as this will
interfere with the instrument detection).
13.4.1.6 Make sure vial cap is on tight and properly labeled,
place in a dark place while other samples are being prepared.
13.4.1.7 Repeat 13.4.1.2 – 13.4.1.6 for each sample.
13.4.1.8 Make sure the outside of all sample vials is clean,
free of anything that could interfere with transmittance of light
out of the vial such as sample or dust, by wiping the outside
with a lint free cloth or similar before analysis.
13.4.1.9 Once the samples have been allowed to sit in a dark
location for at least 1 h, run the samples using the same LSC
conditions that were used to measure the background samples.
It is best to do this after doing experiments to determine sample
counting efficiency which are described in the next two
sections.
13.4.1.10 Once the samples have been run, record the
measured CPM, to the nearest 0.01 CPM, for each sample.
13.4.2 Sample Counting Effıciency Measurement Part
1—Precaution is needed when preparing and handling the spike
solution to prevent sample and laboratory contamination.
Review 1.5 to make sure all necessary precautions are being
followed.
13.4.2.1 To an empty vial, add 50 µL to 100 µL of the
prepared 14C spike standard (13.3).
13.4.2.2 Make sure the spike is delivered to the bottom of
the vial to help facilitate it mixing completely with the
scintillation cocktail added in the next step.
13.4.2.3 Add optimized amount of scintillation cocktail to
vial, must be the same amount of scintillation cocktail used for
the background and sample analysis.
13.4.2.4 Cap and gently mix the spiked cocktail (must
ensure that cap is not wetted while mixing), once mixed, label
the top of the vial (DO NOT write on the side of the vial as this
will interfere with the instrument detection).
13.4.2.5 Run the spiked cocktail with the instrument set to
count all beta decays over the entire 14C range for 10 min to
30 min.
NOTE 13—Initial work used 10 min counting time.
13.4.2.6 The instrument needs to be set to collect counts that
are detected by two of the three detectors along with those
9
detected by all three detectors. Both numbers are used to
determine the TDCR of the spiked cocktail which is required
for efficiency calculation.
13.4.2.7 Record the measured CPM and TDCR, both to the
nearest 0.01, for each sample that was run.
13.4.3 Sample Counting Effıciency Measurement Part
2—Precaution is needed when preparing and handling the spike
solution to prevent sample and laboratory contamination.
Review 1.5 to make sure all necessary precautions are being
followed.
13.4.3.1 Once the 14C spiked cocktail from sample effi-
ciency measurement part 1 has been run, take the sample out of
the instrument.
13.4.3.2 Place vial with 14C spiked scintillation cocktail
onto the balance, with lid on, and tare the balance (this shall be
a different balance then the one used for non-spiked sample
preparation).
13.4.3.3 Remove cap and add optimized amount of sample
to the spiked cocktail and recap the vial, this shall be the same
amount that is used for the background and sample CPM
measurements (13.2.3 and Note 11). Record the mass of the
added sample to the nearest 0.1 mg.
13.4.3.4 Make sure vial cap is on tight and properly labeled.
13.4.3.5 Run the spiked sample using the same instrument
conditions that are used to run the samples except use the same
time frame that was used to run the spiked cocktail in sample
efficiency measurement part 1 (13.4.2.5 and Note 13).
13.4.3.6 Record the measured CPM for each sample to the
nearest 0.01 CPM.
13.5 Dilution—Determining if dilution is required and how
to proceed if it is.
13.5.1 Use the data that was recorded in 13.4.2.7 and
13.4.3.6 in Eq 3 (Assume CPMs = CPMb for this check).
13.5.1.1 If the calculated value is below 50.0 %, sample
dilution is required. Proceed to 13.5.2.
13.5.1.2 If calculated value is above 50.0 %, no additional
sample preparation is required.
13.5.2 In an unused vial, repeat 13.4.2 and 13.4.3 using
more cocktail and less sample. For example, if standard sample
preparation uses 5 mL of cocktail and 15 mL of sample, use
8 mL of cocktail and 12 mL of sample.
13.5.2.1 Dilute the sample as little as possible to achieve an
efficiency above 50.0 %.
13.5.2.2 If a sample dilution ratio of 80.0 % cocktail and
20.0 % sample or higher is required to achieve a counting
efficiency higher than 50.0 % the sample is not compatible with
this method and is required to be analyzed via a different
method such as those in Test Method D6866.
13.5.3 After the new, diluted sample has been run, repeat
13.5.1.
13.5.3.1 If calculated value is still below 50.0 % more
sample dilution is required and repeat 13.5.2.
13.5.3.2 If calculated value is above 50.0 % proceed to
13.5.4.
13.5.4 Prepare and analyze the sample as described in
13.4.1 but using the same cocktail to sample ratio that was
determined to have a counting efficacy above 50.0 % from
13.5.3.
 D8473 − 22
13.5.5 Prepare and analyze the appropriate background as
described in 13.2 for the sample that required dilution using the
same sample solvent to cocktail ratio from 13.5.4.
13.5.6 Once the efficiency, sample and background have all
been measured using the same scintillation cocktail to sample
ratio use the measured values to calculate biocarbon content
using equations in Section 14. Diluting a sample will result in
a proportionate increase in detection limit and limit of quanti-
tation as less sample is being analyzed. For example, if
normally 15 mL of sample is used and has a detection limit of
2 % by mass biocarbon, if the dilution requires the use of
10 mL of sample instead of the normal 15 mL the detection
limit for the diluted sample is 3 % by mass biocarbon.
14. Calculations
14.1 The counts shall be compared, directly to the currently
defined level of 100 % by mass modern carbon. Refer to Table
1 for REF correction factor for the last several years. The
inherent assumption is that all the organic components within
the analyzed material are either fossil or present day in origin.
14.2 The relative number of counts between the modern
reference and the sample is term “fraction of modern” (fM).
This is most commonly referred to as percent modern carbon
(pMC), the percent equivalent to fM (for example, fM =
pMC/100).
14.3 Use the following equations to convert the experimen-
tally obtained values to percent by mass biocarbon.
C m 5 S m 3 %C m⁄m (2)
where:
Cm = mass of carbon in the sample vial,
Sm = mass of sample in the sample vial, and
%Cm/m = percent carbon of the sample, to be determined
experimentally using methods such as CHN el-
emental analysis by D5291.
C e 5 ~ CPM ss 2 CPM s ! ⁄ ~ CPM SC ⁄ TDCR ! (3)
where:
Ce = counting efficiency,
CPMss = measured CPM of the spiked sample, 13.4.3
(sample efficiency measurement part 2),
CPMs = measured CPM of the sample, 13.4.1 (sample
CPM measurement),
CPMSC = measured CPM of spiked cocktail, 13.4.2 (sample
efficiency measurement part 1), and
TDCR = triple to double coincidence ratio, 13.4.2 (sample
efficiency measurement part 1).
DPM Cm 5 ~ CPM s ⁄ C m 2 ~ CPM ⁄ g C ! b ! ⁄C e (4)
where:
DPMCm = DPM of the sample,
10
CPMs = measured CPM of the sample, 13.4.1
(sample CPM measurement),
Cm = mass of carbon in the sample vial, Eq 2,
(CPM / gC)b = calculated CPM / gC of the sample
background, 13.2 (background samples
preparation and analysis), and
Ce = counting efficiency, Eq 3.
%Bio 5 DPM cm ⁄ ~ 13.56 3 ~ REF/100!! 3 100 (5)
where:
%Bio = percent biocarbon by mass expressed in units
of grams of modern carbon in sample relative
to total grams of carbon in the sample,
DPMcm = DPM of sample per gram of carbon in sample,
Eq 4,
13.56 = defined DPM per gram of carbon for a sample
with 100 % pMC, and
REF / 100 = correction factor for when the plant or animal
was alive.
14.3.1 This equation only applies for when the year(s) the
plant or animal was known to be living for a given biofeed and
the products produced from that feed. If the plant or animal
was alive for multiple years, the carbon uptake for each year
also needs to be known and averaged. If these are unknown, the
feed shall be analyzed to correct for the amount of 14C present
within. This will likely require the use of Test Method
D6866 Method B as most biofeeds are unable to be analyzed
via this method. The exception to this is if the plant or animal
was planted or born during or after 2019 as atmosphere 14C
concentrations have been constant since then.
14.3.1.1 Correction for use of biofeeds that accumulated
bomb carbon over the course of more than one year (more
detailed explanation in Test Method D6866).
(1) If feed pMC = 100 – 105, use Eq 5 as written.
(2) If feed pMC = 106 – 122, use REF = 112.
(3) If feed pMC = 122 or larger, use REF = 138.
14.4 Correcting for Radon-222 Contamination:
14.4.1 Crude oil, especially shale oil, has the potential to
contain small amounts of U-238. One of the decay products is
Rn-222. Rn-222 has the potential to make it through the
refining process and be retained in the finished products. The
decay of Rn-222 and its subsequent products can produce
interference in the 14C region of the LSC spectra. The half-life
of Rn-222 is short, 3.82 days. The short half-life of Rn-222
allows for calculation to be performed to correct for its
presence. Eq 6 and Eq 7 show how to perform this correction.
14.4.2 This correction only needs to be applied to samples
with Rn-222 contamination. The LSC full spectrum allows for
easy determination if the contamination is present. Fig. 1
shows LSC spectra with and without Rn-222 contamination.
CPM cor 5 Observed CPM C14 2 k ~ observed CPM 60021000
2 CPM bg60021000! (6)
 D8473 − 22

14
The orange line is the upper range of signal produced by C.

14 14
FIG. 1 LSC Spectra of a Sample with C and Rn-222 Contamination (top) and with Only C (bottom)

where: Observed CPM600 – 1000 = CPM measured over the counting

CPMcor = corrected CPM signal that can be window 600 – 1000 (If the win-
then input into equations in 14.3 dow chosen for CPMs measure-
for CPMs, ments extends above 600, this
Observed CPMC14 = CPM s for the contaminated window needs to be adjusted so it
sample over the specified region does not overlap with CPMs
of interest, window), and
k = sample type dependent correc-
tion constant (Eq 7),

11

CPMbg600 – 1000 = CPM measured for the contami-
nated samples background over
the counting window 600 – 1000
(this data is may already be
stored in the raw data collected
when the background samples
were run, if not, new back-
grounds need to be taken to mea-
sure over the 600 – 1000 channel
range so this equation can be
used).
k 5 ~ CPM s t1 2 CPM s t2 ! ⁄ ~ CPM 60021000 t1 2 CPM 60021000
where:
k = sample type dependent correction con-
stant (needed for Eq 6) (k is quench
dependent and needs to be calculated for
each sample type that contamination is
observed, that is, if the contamination is
observed in gasoline and jet fuel k would
need to be calculated for both sample
types),
CPMs t1 = CPMs initially measured,
CPMs t2 = CPMs measured again for exact same
sample (do not prepare a new one) after 3
to 6 days (Half-life of Rn-222 is 3.82
days, rerunning the sample after 3-6 days
helps determine the signal present in the
sample region that is from RN-222),
CPM600 – 1000 t1 = CPM measured over the counting win-
dow 600 – 1000 from the same run as
CPMs t1, and
CPM600 – 1000 t2 = CPM measured over the counting win-
dow 600 – 1000 from the same run as
CPMs t2.
15. Report
15.1 Final reports shall include the following:
15.1.1 Name of testing laboratory,
15.1.2 Date of testing,
15.1.3 Standard name and version,
15.1.4 REF value used, and
15.1.5 Percent by mass biobased carbon content (reported to
0.1 %, raw data shall be kept containing results to the 0.01 %
until full ILS is completed).
15.2 If regulatory reporting guidelines require the reporting
of results in units other than grams of modern carbon in sample
per total grams of carbon in the sample (percent by mass
biobased carbon content), then the conversion of the results
may be required. The required conversion will have to be
assessed on a case-by-case basis.
D8473 − 22
16. Precision and Bias
16.1 The repeatability and reproducibility will ultimately be
determined by an interlaboratory study in accordance with
ASTM methodology. This section reports interim precision
derived from a mini-interlaboratory study (ILS) participated by
three laboratories.
16.2 Precision—Interim repeatability was calculated over
the biocarbon ranges 0 % to 14 % by mass. The precision was
developed using 14, 12, and 12 fuel samples each of gasoline,
diesel, and jet standard, respectively, by either mixing renew-
able naphtha with the refinery produced gasoline or renewable
t2 ! diesel with the refinery produced diesel and jet fuel samples.
(7) Details of these samples can be found in Research Report
RR:D02-2038.7
16.2.1 Repeatability—The difference between two results
obtained by the same operator on the same apparatus under
constant operating conditions on identical test material would,
in the long run, in the normal and correct operation of the test
method, exceed the following values (see Tables 2 and 3) only
in one case in twenty (95 % confidence interval).
16.2.2 Reproducibility—The breadth of the initial ILS was
limited and not adequate for establishing reproducibility, com-
pliant with Practice D6300. A full ILS will be conducted within
five years of the method approval and publication.
16.2.3 Bias—No information can be presented on the bias of
the procedure for measuring percent biocarbon by mass be-
cause no material having an accepted reference value is
available at this time.
16.3 Relative Bias—A limited study to assess the relative
bias of this method versus Test Method D6866 – 20 using AMS
(accelerated mass spectrometry) for the determination of per-
cent biocarbon by mass was conducted involving one labora-
tory running the AMS method. The lack of participating AMS
labs prevents a reliable assessment of the relative bias in
accordance with Practice D6708. The AMS data collected are
documented in Research Report RR:D02-2038.
17. Keywords
17.1 biobased; bomb carbon; 14C (carbon-14); carbon dat-
ing; liquid scintillation counting; new carbon; old carbon;
percent modern carbon
7
Supporting data have been filed at ASTM International Headquarters and may
be obtained by requesting Research Report RR:D02-2038. Contact ASTM Customer
Service at service@astm.org.
TABLE 2 Interim Repeatability
Repeatability, r
Gasoline, jet, and diesel fuels 0.02723 (X + 22.6)A
A
X = percentage of biocarbon, between 0 % and 13.2 %.
12
 D8473 − 22
TABLE 3 Repeatability Calculation for Selected Biocarbon Concentrations
Biocarbon, % 0 2 4 6 8 10 12 14
Repeatability, 0.62 0.67 0.70 0.78 0.83 0.89 0.94 1.00
%

APPENDIX

(Nonmandatory Information)

X1. FIGURES

FIG. X1.1 Recommended Sample Analysis Workflow

13
 D8473 − 22
FIG. X1.2 List of Routine Analyses that Need to be Conducted, Not Including Normal Sample Analysis
REFERENCES
(1) Anderson, R., and Cook, G. T., “Scintillation Cocktail Optimization
for 14C Dating Using the Packard 2000 CA/LL and 2260 SL,”
Radiocarbon, Vol 33, 1991, pp. 1–7.
(2) Cook, G. T., Naysmith, P., Anderson, R., and Harkness, D. D.,
“Performance Optimization of the Packard 2000 CA/LL Liquid
Scintillation Counter for 14C Dating,” Nucl. Geophys., Vol 4, 1990a,
pp. 241–245.
(3) Koba, M., “Improved Results Using Higher Ratios of Scintillator
Solution to Benzene in Liquid Scintillation Spectrometry,”
Radiocarbon, Vol 42, No. 2, 2000, pp. 295–303.
(4) L’Annuniziata, M. F., “Effıciency Tracing DPM (ET-DPM) and
Direct-DPM-Instrument Performance Data. Counter Intel. Tri-Carb
LSC Application,” Packard BioScience Company, Meriden, CT, 1997.
(5) L’Annuniziata, M. F., Handbook of Radioactivity Analysis, Academic
Press, New York, 1998.
(6) Noakes, J. E., “Low-Level Liquid Scintillation Counter Array with
Computerized Data Acquisition and Age Calculation Capabilities for
14C Dating,” Radiocarbon, Vol 37, No. 2, 1995, pp. 773–779.
(7) Roessler, N., Valenta, R. J., and van Cauter, S., “Time-resolved Liquid
14
Scintillation Counting,” Liquid Scintillation Counting and Organic
Scintillators, Ross, H., Noakes, J. E., and Spaulding, J. D., Eds.,
Lewis Publishers, Chelsea, MI, 1991, pp. 501–511.
(8) Takiue, M., Natake, T., and Fujii, H., “Liquid Scintillation Radioassay
for Low-activity Beta Emitter Mixtures by the Method of Least
Squares,” J. Radioanal. Nucl. Chem. Lett., Vol 200, 1995, pp.
247–258.
(9) Thomson, J., and Burns, D. A., “LSC Sample Preparation by
Solubilization,” Counting Solutions CS-003, Packard Instrument
Company, Meriden, CT, 1996b.
(10) Currie, L. A., Stafford, T.W., Sheffield, A. E.,Wise, S. A., Fletcher, R.
A., Donahue, D. J., Jull, T. J. T., and Linick, T. W., “Microchemical
and Molecular Dating,” Radiocarbon, Vol 31, No. 3, 1989, pp.
448–463.
(11) Currie, Lloyd A., Klinedinst, Donna B., Burch, R., Feltham, N., and
Dorsch, R., “Authentication and Dating of Biomass Components of
Industrial Materials; Links to Sustainable Technology,” Nuclear
Instruments and Methods in Physics Research, Section B, Vol 172,
2000, pp. 281-287.
 D8473 − 22
ASTM International takes no position respecting the validity of any patent rights asserted in connection with any item mentioned
in this standard. Users of this standard are expressly advised that determination of the validity of any such patent rights, and the risk
of infringement of such rights, are entirely their own responsibility.

This standard is subject to revision at any time by the responsible technical committee and must be reviewed every five years and
if not revised, either reapproved or withdrawn. Your comments are invited either for revision of this standard or for additional standards
and should be addressed to ASTM International Headquarters. Your comments will receive careful consideration at a meeting of the
responsible technical committee, which you may attend. If you feel that your comments have not received a fair hearing you should
make your views known to the ASTM Committee on Standards, at the address shown below.

This standard is copyrighted by ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959,
United States. Individual reprints (single or multiple copies) of this standard may be obtained by contacting ASTM at the above
address or at 610-832-9585 (phone), 610-832-9555 (fax), or service@astm.org (e-mail); or through the ASTM website
(www.astm.org). Permission rights to photocopy the standard may also be secured from the Copyright Clearance Center, 222
Rosewood Drive, Danvers, MA 01923, Tel: (978) 646-2600; http://www.copyright.com/

15