Nuclear Instruments and Methods in Physics Research B 223–224 (2004) 313–317

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Technical progress in AMS microscale radiocarbon analysis

Masao Uchida a,b,*, Yasuyuki Shibata a, Minoru Yoneda a,
Toshiyuki Kobayashi a, Masatoshi Morita a
a
NIES-TERRA AMS facility, National Institute for Environmental Studies (NIES), Tsukuba 305-8506, Japan
b
Laboratory for Molecular Biogeochemistry and Chemical Paleoceanography, Mutsu Institute for Oceanography (MIO), Japan
Agency for Marine-Earth Science and Technology (JAMSTEC), Yokosuka 237-0061, Japan

Abstract

A graphitization technique for making precise and accurate radiocarbon measurements on small samples containing
less than 100 lg of carbon was developed. The development of a microscale radiocarbon analysis technique is very
important for advanced research in various fields of study, especially organic geochemistry. A detailed examination of
the sample preparation process for microscale radiocarbon analysis and data analysis using IAEA-C1, IAEA-C6 and
NIST HOXII standards show encouraging results. Small amounts of CO2 (10–50 lg C) from each standard were re-
duced to graphite over an iron catalyst (
1 mg) at 670 °C. In this study, a newly developed ultralow-background
graphite reactor was also used. Measured 14 C/12 C ratios of small samples were significantly decreased compared with
those of samples with
1 mg C probably because small amount of carbon induced isotopic fractionation effect. This
fractionation appears to be directly related to a decrease in the ion current generated by smaller sample sizes. We
confirmed that this fractionation could be compensated for by measuring the samples relative to standards of identical
size. We also achieved low-background measurement during the entire graphitization process with this newly estab-
lished system. Detailed results of sample preparation and data analysis are discussed.
Ó 2004 Elsevier B.V. All rights reserved.

Keywords: Microscale radiocarbon analysis; Compound-specific radiocarbon analysis (CSRA); Low-background graphitization;
PCGC

1. Introduction is a powerful tool for elucidation of the origin and

The recently developed compound-specific
radiocarbon analysis (CSRA) technique using
preparative capillary gas chromatography (PCGC)
*
Corresponding author. Address: Laboratory for Molecular
Biogeochemistry and Chemical Paleoceanography, Mutsu
Institute for Oceanography (MIO), Japan Agency for Marine-
Earth Science and Technology (JAMSTEC), Yokosuka 237-
0061, Japan. Tel.: +81-46-867-9491; fax: +81-46-867-9039.
E-mail address: uchidama@jamstec.go.jp (M. Uchida).
fate of organic carbon in the environment [1,2].
Some researchers in particular should find this
technique useful as an alternative tool for dating
carbonate fossils from marine sediment cores as
well as organic molecular tracers from various
sources in marine sediments [3–6]. By this tech-
nique, we are possible to measure 14 C in small
amount of carbon from marine sediments, aero-
sols, and some microfossils, which would make
breakthroughs in various fields such as geochem-
istry, paleoceanography, atmospheric chemistry,

0168-583X/$ - see front matter Ó 2004 Elsevier B.V. All rights reserved.
doi:10.1016/j.nimb.2004.04.062
314 M. Uchida et al. / Nucl. Instr. and Meth. in Phys. Res. B 223–224 (2004) 313–317
organic geochemistry and archaeology possible.
PCGC systems were installed in 1999 at NIES-
TERRA and in 2002 at MIO, JAMSTEC for use in
various environmental studies, especially paleoce-
anographic and biogeochemical studies. Moreover,
the system at NIES-TERRA was expanded to a
multi-dimensional PCGC in 2001. Concurrently
with the installation of the PCGC systems at the
two institutes, we began to pursue a reliable tech-
nique for expanding radiocarbon measurements to
ultrasmall samples containing less than 50 lg C.
When the PCGC system was first installed, a
large amount of sample, more than 100 lg C, was
required for AMS measurement. Thus far, some
AMS laboratories had already improved proce-
dural blanks and expanded their capability to
handle samples containing less than 100 lg C [7–
9]. By using modification of their techniques in
some levels, we studied compound-specific radio-
carbon analysis (CSRA) techniques, which were
successfully applied for CSRA such as fatty acids,
alkenones and n-alkanes from marine sediments
[4,5], fatty acids from aerosol samples [10,11]
and polycyclic aromatic hydrocarbons (PAHs) in
pond sediments from an urban reservoir [12].
In this paper, we demonstrate that it is possible
to measure the 14 C content of microgram carbon
targets for microscale radiocarbon analysis. Our
method for preparing graphite targets of less than
50 lg C, which includes a modified graphite reac-
tion system, is reported here. We also discuss the
evaluation of data along with the results of the
Fig. 1. Detailed diagram of the specially designed low-background graphite reactor vessel.
AMS analysis of the NIST oxalic acid II (HOxII)
standard. IAEA C-1 Carrara marble carbonate was
also used to test background during graphitization.
2. Material and methods
A batch of CO2 containing a few milligrams of
carbon was prepared from HOxII and IAEA C6
sucrose to evaluate 12 C3þ ion current trends relative
to mass sizes from 10 to 50 lg C and 14 C/12 C
relationships of similarly sized standards. A vac-
uum line was specially prepared for the microscale
14
C analysis. The vacuum line consists of two parts:
(1) a purification and quantification part and (2) a
graphite reactor part. This line was designed to
achieve low contamination by modern carbon
during graphitization of small-volume samples. To
facilitate the processing of samples containing
approximately 10–50 lg C for 14 C analysis, a new
4-port graphite reactor bank was constructed with
attached gas transfer/quantification devices and
including reactor vessels with minimized volumes
to improve the graphite-producing reaction. A
detailed diagram of the low-background graphite
reactor vessel (
1.2 ml), which was specially de-
signed to prevent contamination during graphiti-
zation, is shown in Fig. 1. In addition all of the
vessel connectors consist of a vacuum-backed
union with two Viton O-rings. For graphitiza-
tion of small amounts of carbon, we modified
the standard reduction procedure described by
 M. Uchida et al. / Nucl. Instr. and Meth. in Phys. Res. B 223–224 (2004) 313–317
Kitagawa et al. [13]. CO2 samples are introduced
into the ULB vessel, which has a vacuum-backed
union with a 40-mm quartz tube. A vertical, 50-mm
‘‘cold finger’’ made of Pyrex and containing glass
balls is kept at
)30 °C by a Peltier cell cooling
device. As pretreatment before graphitization, the
iron catalyst is reduced at 470 °C and 0.5 atm of H2
for 2 h. For graphitization, an excess of H2 (2.2–
5.0; H2 :CO2 , depending on sample size) is added to
the reactors, with a typical reaction lasting 2–4 h.
CO2 is reduced to graphite at 670 °C. The catalyst
used for graphitization was hydrogen-reduced
iron powder ()325 mesh, 97%; Aldrich Co. Ltd.,
USA). AMS measurement was conducted with the
MC-SNICS ion source for 40 solid samples and
Pelletron accelerator at the National Institute for
Environmental Studies [14–16].
3. Results and discussion
AMS performance was tested under the condi-
tions of reduced carbon ion beam intensity asso-
ciated with decreasing amounts of carbon in the
graphite targets. Measured 12 C3þ ion current values
for sample sizes below 50 lg C (Fig. 2) showed a
characteristic decrease with decreasing sample size.
Similar relationships between 12 C3þ currents and
sample size have been reported previously [8,9,17].
12 3þ
C currents associated with samples smaller than
20 lg C were apparently constant and had an
averaged value of
2.8 nA. Their 12 C3þ current
magnitudes decreased by approximately 1/20 of 1
mg C graphite. 12 C3þ ion currents are a reasonable
proxy for sample size, and the precision of the
measured isotope ratios, which result from mass-
dependant fractionation, is likely to be related
more directly to 12 C3þ ion current values than to
overall sample size (unpublished data). The insta-
bility of the 12 C3þ currents during the measurement
caused low precision of the isotope ratios. A mass-
dependent fractionation may limit the precision for
the smallest samples, especially those below 10 lg
C, to a few percent. These results are also consis-
tent with those of other studies [17,18]. The 12 C3þ
ion current is also found to be time-variant during
the AMS measurement, probably owing to condi-
tions of the graphite sample such as incomplete set-
315
y = -15.359 + 15.558log(x) R= 0.9415
20
ion current (nA)
15
10
12C3+
5
0
0 10 20 30 40 50 60
Sample size (µgC)
Fig. 2. Relationship between the measured 12 C3þ ion current
and the carbon weight of each graphite target made on Fe
powder.
up of graphite to AMS cathode holder. Small
HOxII standards below 100 lg C show a charac-
teristic decrease in 14 C/12 C values (data not shown),
a relationship that has also been observed previ-
ously by other groups [19,20].
The 14 C/12 C isotopic ratios for a series of stan-
dards of various masses measured by AMS show
decrease of 14 C/12 C if it is normalized to the cor-
responding values of a normal standard (1 mg C).
Although the data show some scattering due to the
different tuning conditions, deviation of the 14 C/
12
C ratio is likely to be correlated with deviation of
the corresponding 13 C/12 C ratio (data not shown).
Fig. 3 shows 14 C activity for small IAEA C6 su-
crose standards of 10–50 lg C calculated relative
to paired similarly sized standards of HOxII. The
corrected 14 C activity of the IAEA C6 standards
normalized to similarly sized samples of HOxII
shows overall agreement. This result shows that a
correction procedure for small samples that uses
similarly sized standards is effective, as has been
reported in some other studies [9,19]. Thus, the
contribution of mass fractionation can be cor-
rected by this procedure.
When we considered all measurements of car-
bon samples smaller than 50 lg C, we found that
316 M. Uchida et al. / Nucl. Instr. and Meth. in Phys. Res. B 223–224 (2004) 313–317
160
consensus value
150.61±0.11
155
14C(pMC)
150
145
140
10 15 20 25 30 35 40
Sample size (µgC)
Fig. 3. Results obtained for IAEA C6 sucrose small standards
of 10–50 lg C are calculated relative to paired similarly sized
standards of HOxII.
some data were somewhat less favorable because of
no sufficient ion current, which can probably be
attributed to insufficient graphitization rather than
to an AMS tuning problem such as a misalignment
of the Cs-beam focusing electrode in the sputter
source. It is usually difficult visualize any graphite
on the catalyst for samples smaller than 50 lg C.
To obtain successful graphitization, catalysts other
than iron should be tested. Some studies have al-
ready reported results of using a cobalt catalyst as
an alternative to an iron catalyst [9,21]. However,
to choose the best catalyst, it is necessary to take
into account purity and the possible formation of
molten beads during sputtering. Such problems
should be eliminated by future experiments. In
addition, after the reaction, it is important to re-
cover the small amount of graphite from the quartz
tube used as reactant after reaction, because a loss
of graphite occurs when it is transferred to the
aluminum AMS target holder. Consequently, we
expect that the consistent successful measurement
of 14 C in small carbon samples will be possible.
At NIES-TERRA, a typical series of IAEA C1
background samples containing about 1 mg C
gave us the lowest background level, which was a
14
C/12 C ratio of several times 1016 , corresponding
to 60,000 yrs [15]. Fig. 4 shows fraction-modern
0.01 -990
0.008 -992
0.1 µg "modern C"
0.006 -994
∆14C (ä)
fm
0.004 -996
0.002 -998
0 -1000
10 20 30 40 50 60
Sample mass (µgC)
45 50 Fig. 4. Calculated fm (fraction-modern) values from IAEA C-1
targets plotted against sample sizes between 10 and 50 lg C to
determine the graphite blank.
values (fm ) of the background versus mass for
normal samples (1 mg C) to small samples ranging
from 10 to 40 lg C. All of the results presented
here are given as the measured modern carbon (fm )
value. fm is defined as follows:
12
fm ¼ ð14 C= CÞs =ð14 C=12 CÞSTD : ð1Þ
The fm values for samples larger than
15 lg C
were almost constant at
0.002, corresponding to

50,000 yrs. In the smallest samples of 10 lg C, fm
values ranged from 0.005 to 0.007. The back-
ground is likely due to contamination during
sample treatment. In this study, we believe that
contamination during graphite preparation using
the newly designed graphite reactor, which pre-
vents atmospheric CO2 from entering the reactant
vessel, is extremely low. However, to evaluate the
cause of the contamination, it will be necessary in
a future study to investigate the main sources of
contamination, such as the machine source blank
and graphitization blank, including by using a
phosphoric acid treatment to produce CO2 from
IAEA C1.
4. Conclusion
The technical progress in microscale radiocar-
bon analysis at the NIES-TERRA AMS facility
greatly reduces carbon sample size requirements
 M. Uchida et al. / Nucl. Instr. and Meth. in Phys. Res. B 223–224 (2004) 313–317
compared to what was hitherto needed for AMS
analysis. The ability to use small carbon samples
of 10–50 lg C will allow scientists in many re-
search fields to take advantage of AMS analysis.
Preliminary results of graphitization of small
samples using a new vacuum line are presented.
Further modification of the vacuum line and the
newly designed reactor vessel should make it
possible for small amounts of CO2 to be reacted
reliably into high-quality graphite with low-back-
ground levels. Contamination by modern carbon
during graphitization of samples of approximately
20–50 lg was less than
0.1 lg C. We confirmed
that machine-induced fractionation, which causes
the 14 C/12 C ratio to decrease with decreasing
sample size, can be corrected for by measuring
similarly sized samples of the HOxII standard.
Moreover, substantial improvements in microscale
14
C analysis are required to decrease the uncer-
tainties of 14 C measurements at the level of a few
micrograms. This technique has advantages for
organic geochemical studies, especially for com-
pound-specific radiocarbon analysis (CSRA), and
will surely contribute greatly toward our under-
standing of the carbon cycle on the earth.
Acknowledgements
We gratefully appreciated C. Kobayashi, for
operating the AMS in NIES-TERRA. We also
appreciate J.M. Hayes, A.P. McNichol, K.L. Ga-
gnon for the technical support of micro-scale 14 C
analyses. A part of this study was financially
supported by the Budget for Nuclear Research of
the Ministry of Education, Culture, Sports, Sci-
ence and Technology, based on the screening and
counseling by the Atomic Energy Commission.
Part of this study also was financially supported
by MIO, JAMSTEC (MIO contribution #30).
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