Geochimica et Cosmochimica Acta 71 (2007) 655–669

www.elsevier.com/locate/gca

Carbonaceous cherts of the Barberton Greenstone Belt, South Africa:

Isotopic, chemical and structural characteristics
of individual microstructures
Mark A. van Zuilen *,1, Marc Chaussidon, Claire Rollion-Bard, Bernard Marty
Centre de Recherche Pétrographiques et Géochimiques RPG-CNRS, 15 rue Notre Dame des Pauvres, BP-20, 54501 Vandoeuvre les Nancy, France

Received 11 January 2006; accepted in revised form 25 September 2006

Abstract

Carbonaceous matter occurring in chert deposits of the 3.4–3.2 Ga old Barberton Greenstone Belt (BGB), South Africa, has experi-
enced low grade regional metamorphism and variable degrees of local hydrothermal alteration. Here a detailed study is presented of
in situ analysis of carbonaceous particles by LRS (laser Raman spectroscopy) and SIMS (secondary ion mass spectrometry), reporting
degree of structural disorder, carbon isotope ratio and nitrogen-to-carbon ratio. This combination of in situ analytical tools is used to
interpret the d13C values of only the best preserved carbonaceous remains, enabling the rejection of non-indigenous (unmetamorphosed)
material as well as the exclusion of strongly hydrothermally altered carbonaceous particles. Raman spectroscopy confirmed that all car-
bonaceous cherts studied here have experienced a regional sub- to lower-greenschist facies metamorphic event. Although this identifies
these organics as indigenous to the cherts, it is inferred from petrographic observations that hydrothermal alteration has caused small
scale migration and re-deposition of organics. This suggest that morphological interpretation of these carbonaceous particles, and in
general of putative microfossils or microlaminae in hydrothermally altered early Archean cherts, should be made with caution. A chert
in the Hooggenoeg Formation, which is older than and has been hydrothermally altered by a volcanic event 3445 Ma ago, contains
strongly altered carbonaceous particles with a uniform N/C-ratio of 0.001 and a range of d13C that is shifted from its original value.
Cherts of the Kromberg Formation post-date this volcanic event, and contain carbonaceous particles with a N/C-ratio between 0.002
and 0.006. Both the Buck Reef Chert and the Footbridge Chert of the Kromberg Formation have retained fairly well-preserved d13C
values, with ranges from 34& to 24& and 40& to 32 &, respectively. Abiologic reactions associated with hydrothermal serpent-
inization of ultramafic crust (such as Fischer–Tropsch synthesis) were an unlikely source for carbonaceous material in these cherts. The
carbonaceous matter in these cherts has all the characteristics of metamorphosed biologic material.
 2006 Elsevier Inc. All rights reserved.

1. Introduction Schopf et al., 2002; Garcia-Ruiz et al., 2003; Schopf and

Recent discussions regarding morphological identifica-
tion of microfossils (either by conventional microscopy or
by laser Raman imaging) in metamorphosed Archean ter-
rains, have highlighted the problems of recognizing a true
indigenous syndepositional biological structure (Brasier
et al., 2002, 2005; Pasteris and Wopenka, 2002, 2003;
*
Corresponding author.
E-mail address: vanzuilen@ipgp.jussieu.fr (M.A. van Zuilen).
1
Present address: Institut de Physique du Globe de Paris, Case 89,
4 Place Jussieu, 75252 Paris Cedex 05, France.
Kudryavtsev, 2005; Westall, 2005). Additional chemical
and isotopic characterization of individual carbonaceous
microstructures is therefore of crucial importance in studies
of early life. Carbon isotope ratios (d13C) have been used
extensively to trace back life over the geological record
(e.g. Schidlowski, 2001 and references therein), and the
d13C analysis of individual carbonaceous microstructures
by SIMS technique has been applied to Archean and Pro-
terozoic rock samples (Mojzsis et al., 1996; House et al.,
2000; Ueno and Isozaki, 2001; Ueno et al., 2002). Unfortu-
nately, metamorphism can shift the d13C of sedimentary
organic material to higher values, blurring the original

0016-7037/$ - see front matter  2006 Elsevier Inc. All rights reserved.
doi:10.1016/j.gca.2006.09.029
656 M.A. van Zuilen et al. 71 (2007) 655–669
biologic d13C signature. The wide range in d13C values that
is observed in metamorphosed Archean carbonaceous mat-
ter can therefore be interpreted in two ways; it either repre-
sents different degrees of metamorphic overprinting of a
uniform biologic source with a specific isotope ratio, or it
reflects the unaltered original variation in carbon fixation
pathways that were utilized by different types of organisms
(House et al., 2000, 2003; Grassineau et al., 2001). An addi-
tional complication is the fact that organic matter can form
abiologically during hydrothermal processes. The range of
carbon isotope ratios found in this type of carbonaceous
material overlaps with that of biologic material (McCollom
and Seewald, 2006). During prograde metamorphism car-
bonaceous matter also progressively looses heteroatoms
(H, O, N, S), obtains a higher degree of structural order
and ultimately transforms into crystalline graphite. There-
fore, the shifts in d13C due to metamorphism are correlated
to e.g. H/C-ratios of carbonaceous matter (Hayes et al.,
1983; Strauss et al., 1992a,b; Watanabe et al., 1997; Des
Marais, 2001), and N/C-ratios (Pinti et al., 2001). Further-
more, hydrothermal alteration and thermal cracking will
cause migration and re-deposition of organics. These pro-
cesses will lead to carbonaceous aggregates that can be mis-
taken for microfossil structures. The problems outlined
above indicate that it is of crucial importance to study
the detailed effects of metamorphism and hydrothermal
alteration before morphological, chemical, or isotopic
observations are interpreted as traces of early life.
Here a study is presented of a combined in situ analysis
(carbon isotopes, nitrogen/carbon-ratio, degree of disor-
der) of carbonaceous particles that occur in three hydro-
thermally altered cherts of the 3.4–3.2 Ga old Barberton
Greenstone Belt, South Africa. This study highlights the
complexity of different processes that have influenced the
characteristics of carbonaceous material in these rocks.
Previous studies presented morphological and/or bulk iso-
tope evidence for a biologic source of the carbonaceous
material (Hayes et al., 1983; Robert, 1988; Walsh, 1992;
Westall et al., 2001). Tice et al. (2004) provided a detailed
study of the thermal maturity and structural disorder of
carbonaceous particles in cherts of the BGB, using LRS.
In later studies Tice and Lowe (2004, 2006) provided geo-
logical, geochemical and isotopic evidence that strength-
ened the conclusion that much of this carbonaceous
matter represents the remains of anoxygenic photosynthe-
sizing bacteria. In the present study LRS and SIMS tech-
niques are used to determine the degree of structural
order, d13C, and N/C-ratio of individual carbonaceous
microstructures. The combination of these in situ tech-
niques is used to discard specific highly altered carbona-
ceous material, before carbon isotope ratios are
interpreted.
2. Geologic background and material
Cherts in the BGB have been interpreted as direct pre-
cipitates or diagenetic replacements of preexisting sedimen-
tary and pyroclastic deposits in a silica saturated Archean
ocean (Knauth and Lowe, 1978; Lowe and Byerly, 1986;
Knauth and Lowe, 2003). Alternatively, Paris et al.
(1985) have suggested that silica addition to both volcanic
units and overlying sediments is the result of convective
seawater circulation that acted directly on the oceanic crust
and overlying sediments. Higher heat flux and greater
availability of Mg, Fe-rich silicates (ultramafic/mafic bas-
alts) in the early Archean caused effective serpentinization
of oceanic crust that produced large quantities of silica-en-
riched hydrothermal fluids. Lateral migration of such fluids
caused silicification of the upper ocean floor basalts and
overlying sediments. Irrespective of the exact process of
formation of these cherts it is clear that hydrothermal pro-
cesses (either at an early or at a later stage) played an
important role in shaping these rocks, and could possibly
have caused abiologic synthesis of carbonaceous material.
Three carbonaceous chert samples from the Onverwacht
Group of the BGB, South Africa, are described here in
detail. The Onverwacht Group represents a predominantly
tholeiitic and komatiitic volcanic sequence in the lower part
of the Swaziland Supergroup. Throughout this paper the
stratigraphy (Fig. 1) as described by Lowe and Byerly
(1999) is used, but it should be emphasized that rigorously
different viewpoints exist for this specific interpretation
(de Wit et al., 1982; de Wit and Hart, 1993). Sample
BAR-4-1 was obtained from the Hooggenoeg Formation
(>3.445 Ga) and samples BAR-3-1 and BAR-3-3-B from
the overlying Kromberg Formation (<3.445 Ga). The
results for carbonaceous matter in these samples from
Barberton are compared with that of graphite found in a
talc-rich rock from the 3.8 Ga Isua Supracrustal Belt
(ISB) in southern West Greenland (sample IS-7-6-B2).
The graphite in this rock is thought to be derived from
high-temperature alteration of an ultramafic rock
(Naraoka et al., 1996), and therefore represents a clear
example of abiologic carbonaceous matter (Fig. 2).
2.1. Sample BAR-4-1
Location S 2556.200 , E 3050.200 . Hooggenoeg chert
H5c (Fig. 1) is a capping chert that shows characteristics
which are found in many other capping cherts in the
Hooggenoeg Formation (e.g. H1c, H3c). The underlying
unit H5v consists of a thick sequence of basaltic variolitic
pillows. As was pointed out by Lowe and Byerly (1999)
the continuity, and lithologic and sedimentological similar-
ity of most major cherts in the stratigraphy of the Hoogge-
noeg Formation point to an overall depositional and
environmental uniformity during the accumulation of this
sequence. Chert unit H5c consists of black massive to
laminated carbonaceous chert, alternating with thin layers
of silicified komatiitic ash. Near the top of the formation
also some thin, regionally traceable layers of accretionary
lapilli occur. This is similar to chert H3c that contains
alternations of black chert with silicified komatiitic ash.
In these cherts (H3c and H5c) the carbonaceous matter
 Archean carbonaceous cherts; isotopic, chemical, and structural characteristics 657

Fig. 1. Generalized stratigraphy of the Barberton Greenstone Belt (BGB), redrawn from Fig. 8, page 12 in Lowe and Byerly (2003). More details on this
general stratigraphy can be found in Lowe (1999). Sample BAR-4-1 is derived from carbonaceous capping chert unit H5c in the Hooggenoeg Formation,
BAR-3-1 from carbonaceous capping chert unit K1c1 and BAR-3-3-B from carbonaceous capping chert unit K3c in the Kromberg Formation. Thin
sections in plain polarized light show carbonaceous aggregates and hydrothermal veinlets in all three samples.

Fig. 2. (a) Map of the northeastern part of the 3.8 Ga Isua Supracrustal Belt (ISB), southern West Greenland. Location of sample 7.6B2 is shown by the
arrow. (b) Field appearance of talc–quartz–graphite rock, and sampling location directly below the hammer. (c) Thin section under plain polarized light
showing wavy patterns of graphite particles. (d) Thin section under plain polarized light showing how fluid-deposited graphite follows a preferred direction
of flow.

appears detrital (Lowe and Byerly, 1999, 2003), and con-
sists of sand-sized particles. In thin section gray to black
domains are seen that consist of finely dispersed carbona-
ceous matter in a fine-grained quartz matrix. Secondary
quartz veinlets cross-cut these features (Fig. 1).
2.2. Sample BAR-3-1
Location S 2601.650 , E 3059.370 . Buck Reef chert K1c1
occurs in the Kromberg Formation (Komati Gorge, East
limb of the Onverwacht anticline), and is different from
658 M.A. van Zuilen et al. 71 (2007) 655–669
K1 occurring in the west limb (Tice et al., 2004; Tice and
Lowe, 2004). In the east limb K1 overlies felsic igneous
rocks from the volcanic event at 3.445 Ga (unit H6 in
Fig. 1), which consists of dacitic conglomerate and deb-
ris-flow deposits. Near the top of this unit H6 dacitic turbi-
dites occur. The base of K1 consists of an ultramafic unit,
which includes peridotite near the base, a thick komatiitic
volcanic unit, and a heavily carbonated lapillistone/tuff
near the top. This is capped by chert unit K1c1. Several
cycles of basalt occur above K1c1, which are themselves
capped by K1c2 and K1c3. In the west limb, however,
K1 contains a lower detrital and volcaniclastic unit. In thin
section black domains of carbonaceous matter are
observed, that are dispersed in a fine-grained quartz matrix
(Fig. 1). This matrix is cross-cut by many secondary quartz
veinlets.
2.3. Sample BAR-3-3-B
Location S 2602.150 , E 3059.550 . Footbridge chert
K3c in the east limb of the Onverwacht anticline consists
of two chert layers that most likely represent a single unit
repeated by local faulting. The altered tholeiitic basalts
(unit K3v) directly below K3c have been interpreted as a
low-temperature flow-top alteration zone (Lowe and Byer-
ly, 2003). Lower in the section of K3v a sequence of car-
bonated mafic lapillistone and lapilli tuff occurs. The chert
unit K3c itself consists mainly of massive to laminated
black chert and black-and-white banded chert. A thin
dacitic tuff layer within the chert has been dated at
3.3 Ga (Byerly et al., 1986). Lowe (1999) reported the
absence of current structures, scour features, rip-up clasts,
or other evidence of current activity. Lowe (1999) recog-
nized local occurrences of laminated, highly carbonaceous
iron-rich rock representing partially silicified mixtures of
carbonaceous matter, iron formation, volcanic ash and
clay. This highly carbonaceous rock has been interpreted
by Lowe (1999) as a fine organic-rich mud deposited un-
der quiet, low-energy, fully sub-aqueous conditions. In
thin section an opaque, closely packed mixture of carbo-
naceous matter, magnetite, and rutile is observed in a fine
quartz matrix (Fig. 1). This quartz matrix is cross-cut by
many secondary quartz veinlets. Also, goethite and ana-
tase in small veinlets are evidence of post-metamorphic
fluid alteration.
2.4. Sample IS-7-6-B2
Location N 6510.950 , W 4947.560 . Rose et al. (1996)
describe many outcrops of talc-rich rocks in the ISB, and
associate these rocks with strongly altered ultramafic rocks.
A graphite-rich example (sample BAR-7-6-B2) of such a
rock occurs in the northeastern part of the ISB (Fig. 2a
and b). This schistose rock consists of cm-wide light
(quartz) and dark (quartz–talc–graphite) bands. Minor
mineral phases are apatite and xenotime. Petrographic
analysis of sample IS-7-6-B2 reveals that graphite particles
are concentrated in thin irregular bands and in fine hair-
thin black veinlets, surrounded by talc and quartz
(Fig. 2c and d). The straightforward conclusion can be
drawn that graphite formed epigenetically from a dehy-
drating CO2-rich fluid during silica-rich metasomatism of
an ultramafic protolith. This type of epigenetic graphite
has been observed by others; Naraoka et al. (1996) report
a rock from the northeastern part of the ISB (their sample
ILW-29) that exhibits a strong schistosity, comprises mostly
of talc, quartz and graphite. In this study it is suggested
that the observed graphite was formed either by reaction
of CH4 and CO2, or by direct reduction of CO2 in meta-
morphic fluids.
3. Methods
3.1. Sample preparation
Thin sections of rock samples were prepared for gen-
eral petrography and identification of carbonaceous
aggregates. Since thin sections contain glue within pore
space and grain boundaries, several polished rock sample
disks were specifically prepared for SIMS analysis. The
thin sections and rock sample disks do not represent
complementary sides of a common rock surface, but
are generally taken from the same rock slab of a sample.
Rock sample discs (diameter 2.54 cm) were cut to 3 mm
thickness and polished with silicon carbide and cerium
oxide, respectively. The discs were washed ultrasonically
using Millipore water, dried, and covered with a fine
layer of gold. Carbonaceous particles could be identified
in both thin sections and polished rock sample discs
using SEM-EDS and LRS.
3.2. LRS (laser Raman spectroscopy)
Raman spectra were obtained at G2R, Université Henri
Poincaré (Nancy, France) with a Labram (Horiba-Jobin
Yvon) Raman microprobe system. The instrument is con-
figured with an argon ion laser (spot size between 1 and
5 lm was used) which emits 514.5-nm visible light, an
1800-channel CCD array detector (30 C), and an Olym-
pus microscope that can be used in the transmitted- or
reflected-light mode. The degree of disorder in carbona-
ceous material can be enhanced during sample preparation.
Polishing or crushing will cause dislocations, and breakup
of the crystal structure, leading to a disordered Raman
spectrum (Pasteris, 1989) that could lead to erroneous iden-
tification of disorder. In some studies crushing or polishing
are therefore avoided by direct analysis of a cracked rock
surface (Tice et al., 2004), or by carefully pressing carbona-
ceous matter in a gold foil (Wopenka and Pasteris, 1993).
Alternatively, carbonaceous matter in thin sections can
be studied by Raman spectroscopy by focusing the laser
beam on the subsurface continuations of polished grains,
or focus below the polished surface on grains of interest
that are fully embedded in surrounding transparent miner-
 Archean carbonaceous cherts; isotopic, chemical, and structural characteristics
al phases (Pasteris, 1989). In this work, it is important to
identify carbonaceous matter in polished rock pellets that
is subsequently analyzed by ion microprobe. Laser Raman
spectroscopy was therefore carried out directly on the pol-
ished surface of the rock pellet that was prepared for ion
probe analysis. In order to study the effect of polishing,
laser Raman analysis was carried out on thin sections as
well, where a comparison was made between carbonaceous
matter that was affected by polishing and carbonaceous
matter that was fully enclosed in a quartz matrix. Finally,
Raman spectra were obtained in some ion probe pits from
SIMS analysis of the polished rock slabs, in order to study
the effect of the ion beam on carbonaceous matter. These
tests are important, since they provide a verification that
the actual point of interest (indigenous carbonaceous
matter, and not some contamination or artifact) was
analyzed by ion probe.
3.3. SIMS (secondary ion mass spectrometry)
Polished rock pellets were covered with a thin layer of
gold, and outgassed overnight in an offline vacuum cham-
ber. Using the IMS 1270 ion microprobe at CRPG-CNRS
(Nancy, France), a typical d13C analysis of individual car-
bonaceous particles was made with a 10–15 lm spot size,
and a mass resolution (M/DM) of 5000. No isobaric inter-
ferences of 1H12C over 13C peaks are present at this
resolution. Secondary 13C and 12C ions were simulta-
neously measured in multicollection mode. Since instru-
mental isotope fractionation is influenced by the
chamber vacuum, the chamber pressure was kept below
1 · 108 Torr during the entire session (Sangely et al.,
2005). Several organic carbon standards for SIMS d13C
analysis have been used here (described by Sangely
et al., 2005). These materials represent a range in H/C-,
O/C-, and N/C-ratio, and are therefore also suitable as
standards for SIMS N/C-ratio analysis. In order to con-
strain nitrogen concentration in these carbonaceous stan-
dard materials a correlation line is presented here of ion
probe CN/C versus real N/C-ratio (Fig. 3). This corre-
lation line is in accordance with the general correlation
line constructed by (Aléon et al., 2003) for N/C-ratio in
interplanetary dust particles. Repeated analyses of stan-
dard MB (pure graphite) during the different sessions con-
firmed that there is a reproducible fixed nitrogen
background contamination (Fig. 3). Since all measure-
ments on standards and samples have this additional
nitrogen, a blank correction was made by subtracting a
fixed average CN/Cratio (based on MB-standard).
Since the correlation line is linear, two standards (MB
graphite, Don anthracite) were used throughout the ana-
lytical sessions, before and after the sample analysis.
The procedures of drift correction, sample normalization
and error propagation are summarized in the Appendix
A. After SIMS analysis, ion beam pits on target particles
within the sample discs were analyzed by LRS, in order to
verify the carbonaceous character.
659
Fig. 3. Correlation between CN/C as measured by SIMS and the actual
bulk N/C-ratio as determined by conventional CHN-analysis. Standards
are described in Sangely et al. (2005).
4. Results
4.1. Petrography
Close inspection by SEM-EDS reveals that the carbona-
ceous matter present in the Barberton samples (BAR-3-1,
BAR-3-3-B, BAR-4-1) has migrated over small distances.
This is especially evident in sample BAR-3-3-B, in which
small carbonaceous veinlets can be observed (Fig. 4). The
large (>10 lm) carbonaceous particles that occur in all
three samples (Figs. 5–7), are interpreted here as large
aggregates of migrated organics. This material was most
likely mobilized during hydrothermal alteration of the chert
deposits. From these petrographic observations it is obvi-
ous that these carbonaceous particles do not represent indi-
vidual syndepositional microfossils. In fact carbonaceous
matter seems to have filled pore space, and has moved in
preferred direction of fluid flow. The shapes that are seen
in these SEM pictures are therefore of a secondary nature
and no morphological interpretation can be inferred. The
predominantly igneous units above and below the chert
units in question do not contain organic compounds. It is
therefore concluded that this material did not migrate from
other younger units, and is directly related to the bulk
organics that are present in these cherts. These individual
carbonaceous particles are therefore not exactly syngenetic,
but are indigenous to the rock and are related to the synge-
netic bulk of organics present in these rocks. The carbona-
ceous particles of the Isua sample (IS-7-6-B2) are shown in
Fig. 8. Most of these particles occur in a clear quartz matrix
and follow wavy patterns that likely represent the direction
of flow of CO2-bearing metamorphic fluids (Fig. 2c and d).
Individual carbonaceous particles consist of many individ-
ual graphitic flakes (Fig. 8c and d).
660 M.A. van Zuilen et al. 71 (2007) 655–669

Fig. 4. SEM backscatter electron image of a polished rock slab of sample BAR-3-3-B. (a) Within the quartz matrix there are small veinlets filled with
carbonaceous matter. (b) Void fillings of carbonaceous matter within a quartz matrix.

Fig. 5. SEM backscatter electron image of sample BAR-4-1 representing carbonaceous chert H5c, Hooggenoeg Formation. Four different regions within
the polished pellet are shown (a–d). Numbers correspond to point analyses of d13C and N/C-ratio by SIMS (see Table 1).

4.2. LRS results
The Raman spectra of altered (polished, ion probe) and
unaltered (below thin section surface) carbonaceous matter
are shown in Fig. 9. From the unaltered spectra it can be
concluded that the carbonaceous matter in the Barberton
chert samples has obtained a considerable degree of struc-
tural order; the D-peak (‘disorder’ peak at ca. 1355 cm1)
has a width (at half height) of ca. 60–70 cm1 and the
G-peak (‘graphite’ peak at ca. 1582 cm1) has a width of
ca. 50–65 cm1, the D/G intensity ratio is ca. 1.4–1.6,
and well-developed peaks occur in the second order spec-
trum. These characteristics have been observed before in
Raman spectra of carbonaceous matter in cherts from the
Barberton Greenstone Belt and are in agreement with ther-
mal alteration of at least 200 C (Tice et al., 2004). The car-
bonaceous particles in the sample from Isua (IS-7-6-B2)
display a high degree of structural order. Peak widths
(D-peak is ca. 40 cm1, G-peak is ca. 20 cm1), very well-
developed peaks in the second order spectrum, and a
D/G intensity ratio (ca. 0.14) are typical for graphite that
formed under amphibolite–facies metamorphic conditions
 Archean carbonaceous cherts; isotopic, chemical, and structural characteristics 661

Fig. 6. SEM backscatter electron image of sample BAR-3-3-B representing carbonaceous chert K3c, Kromberg Formation. Four different regions within
the polished pellet are shown (a–d). Numbers correspond to point analyses of d13C and N/C-ratio by SIMS (see Table 1).

Fig. 7. SEM backscatter electron image of sample BAR-3-1 representing carbonaceous chert K1c1, Kromberg Formation. Two different regions within
the polished pellet are shown (a and b). Numbers correspond to point analyses of d13C and N/C-ratio by SIMS (see Table 1).

(Wopenka and Pasteris, 1993; Beyssac et al., 2002) and has
been found elders in the Isua Supracrustal Belt (van Zuilen
et al., 2005). For the Barberton samples (BAR-3-1, BAR-3-
3-B, BAR-4-1), the effects of polishing and ion probe beam
are small, and approximately the same; in the first order
spectrum peak positions move to slightly lower wavenum-
ber and peak widths increase. The D/G intensity ratio
remains relatively unchanged. In contrast, for Isua graphite
(sample IS-7-6-B2) polishing causes a strong increase in
the intensity ratio of the D- and G-peaks, while the peak
positions and peak widths in the first order spectrum
remain relatively unchanged. The ion probe beam also
causes a strong increase in the intensity ratio of the
D- and G-peaks, as well as a sharp increase in peak widths
in the first order spectrum. The peak positions in the first or-
der spectrum, however, remain unchanged. From these
observations it can be concluded that polishing or ion beam
impact only affects rather highly crystalline graphitic mate-
rials (Pasteris, 1989), while finely dispersed carbonaceous
particles remain unchanged. Nevertheless, graphite that
has been altered by ion beam impact is still recognized by
peak positions in the Raman spectrum. A Raman spectrum
662 M.A. van Zuilen et al. 71 (2007) 655–669

Fig. 8. SEM backscatter electron image of sample IS-7-6-B2 representing a talc–quartz–graphite rock in the ISB. (a) Overview picture, showing four areas
of detail within the polished pellet (b–e). Numbers correspond to point analyses of d13C and N/C-ratio by SIMS (see Table 1).

of the exact ion probe pit on a sample provides proof that
the actual carbonaceous structure of interest was analyzed,
and not some artifact (e.g. carbon-bearing contamination).
4.3. SIMS results
The d13C and N/C-ratio of the individual carbonaceous
particles of samples BAR-3-1, BAR-3-3-B, BAR-4-1, and
IS-7-6-B2 are shown in Figs. 5–8, respectively. White cir-
cles represent the location and size of the ion probe pits.
The datapoints are listed in Table 1, and are plotted
(N/C-ratio versus d13C) in Fig. 10. The graphite sample
from Isua (sample IS-7-6-B2) has virtually no nitrogen
(which is reflected in the observed N/C-ratio). The average
bulk d13C value of 10.1& is in accord the expected
equilibrium isotope fractionation d13 CgrCO2 ¼ 6:4‰ at
550 C (Chacko et al., 2001) between graphite and mantle
type CO2 (d13C ca. 5&, Des Marais and Morre, 1984).
This graphite therefore has the typical isotopic characteris-
tics of epigenetic graphite that formed during alteration of
ultramafic rocks under amphibolite–facies metamorphic
conditions. The d13C values of individual graphite grains
within the quartz matrix of sample IS-7-6-B2 indeed scatter
around the bulk value, confirming that SIMS d13C analysis
is accurate and is not strongly influenced by matrix effects.
The chert samples from Barberton have experienced sub-
greenschist to lower-greenschist facies metamorphism. If
the carbonaceous matter in these cherts formed abiologi-
cally during metamorphism and in isotopic equilibrium
with mantle type CO2, then the d13C of this material could
 Archean carbonaceous cherts; isotopic, chemical, and structural characteristics 663

a b

c d

Fig. 9. Raman spectra of carbonaceous material in Barberton chert samples. The degree of order is proportional to the ratio of the D-peak (‘Disorder’, at
1355 cm1) and the G-peak (‘Graphite’, at 1582 cm1) (Wopenka and Pasteris, 1993; Beyssac et al., 2002, 2003; Tice et al., 2004). The three different
spectra for each sample represent resp. SIMS probe pit on a polished sample pellet (ion probe), the polished surface of a thin section (polished), the deeper
part of a thin section free of polishing (unaltered). The similarity among the spectra indicates the minimal effect of polishing, and confirms that in the
polished rock pellet actual indigenous carbonaceous material was analyzed by ion probe. (a) Sample BAR-4-1. Capping Chert H5c, Hooggenoeg
Formation. (b) Sample BAR-3-1. Buck Reef Chert, K1c1, Kromberg Formation. (c) Sample BAR-3-3-B. Footbridge Chert K3c, Kromberg Formation.
(d) Sample IS-7-6-B2. Raman spectra of graphite occurring in a talc schist from the eastern part of the ISB.

have obtained a d13C as low as 20& (assuming
d13 CgrCO2 ¼ 15‰ at 200 C, and mantle CO2 = 5&).
The carbonaceous particles in these Barberton cherts have
much lower d13C values than that.
5. Discussion
5.1. Alteration of carbonaceous matter
5.1.1. Migration of organics
Hydrothermal alteration played an important role dur-
ing metamorphic events that acted on the BGB. However,
such processes could also have played an important role
during the early stages of deposition causing direct serpent-
inization and silicification of seafloor crust and overlying
sediments (Paris et al., 1985). Under such complex circum-
stances it is to be expected that carbonaceous matter is dis-
placed and can accumulate along grain boundaries or
veinlets, or can form large aggregates in voids. This type
of small scale migration of hydrocarbons has been
described in detail before in studies of putative carbonaceous
microfossils occurring in the Apex chert, Pilbara, Western
Australia (Brasier et al., 2002, 2005). There it is argued that
carbonaceous microfossil artifacts are produced by the dis-
placement of abiologic amorphous carbon within a matrix
of recrystallizing silica. The carbonaceous material can
form filamentous reaction rims around individual crystals,
penetrates along microfissures to line or infill small vugs,
664 M.A. van Zuilen et al. 71 (2007) 655–669

Table 1
Results of d13C and CN/C by SIMS analysis of individual carbonaceous structures in Barberton and Isua samples
Sample d13C ±(2s) CN/C ±(2s) N/C (mol) ±(2s)
3.4–3.2 Ga Barberton Greenstone Belt, South Africa
1 4-1B-02 31.62 1.65
2 4-1B-02 28.12 1.65 4.41E04 6.71E03 2.06E05 3.14E04
3 4-1B-02 26.64 1.67 1.70E03 6.74E03 7.94E05 3.15E04
4 4-1B-02 31.30 1.65
5 4-1B-02 27.48 1.66
6 4-1B-02 28.51 1.65 3.05E03 6.81E03 1.43E04 3.19E04
7 4-1B-31 28.11 1.65 1.91E02 7.57E03 8.93E04 3.54E04
8 4-1B-31 20.09 1.70
9 4-1B-31 24.58 1.66 1.99E02 7.50E03 9.34E04 3.51E04
10 4-1B-27 22.72 1.65 1.42E02 7.08E03 6.63E04 3.32E04
11 4-1B-27 21.80 1.66
12 4-1B-22 27.20 1.65 2.66E02 8.12E03 1.25E03 3.80E04
13 4-1B-22 25.59 1.65 2.54E02 7.84E03 1.19E03 3.67E04
14 4-1B-22 23.94 1.65 1.52E02 7.23E03 7.12E04 3.39E04
15 3-3B-07 37.22 1.16 1.21E01 2.19E02 5.67E03 1.02E03
16 3-3B-07 32.81 1.04 1.13E01 2.08E02 5.31E03 9.72E04
17 3-3B-09 31.29 1.04 1.06E01 1.78E02 4.98E03 8.33E04
18 3-3B-15 32.44 1.08 9.17E02 1.78E02 4.29E03 8.33E04
19 3-3B-05 28.98 1.06
20 3-3B-39 33.29 1.16 9.47E02 1.55E02 4.43E03 7.27E04
21 3-3B-39 35.71 1.04 1.26E01 2.03E02 5.88E03 9.53E04
22 3-3B-39 33.93 1.12
23 3-1-22 20.59 1.83 9.25E02 1.12E02 4.33E03 5.25E04
24 3-1-22 30.92 1.83 6.29E02 7.78E03 2.95E03 3.64E04
25 3-1-22 22.35 1.83
26 3-1-09 31.12 1.83 4.41E02 7.68E03 2.06E03 3.60E04

3.8 Ga Isua Supracrustal Belt, southern West Greenland

27 IS-7B2 9.30 0.93 4.44E03 6.46E03 2.08E04 3.02E04
28 IS-7B2 5.40 0.94 4.44E03 6.46E03 2.08E04 3.02E04
29 IS-7B2 10.85 0.92 1.03E02 6.51E03 4.82E04 3.05E04
30 IS-7B2 14.12 0.94
31 IS-7B2 11.16 0.94 2.66E02 8.77E03 1.24E03 4.11E04
32 IS-7B2 8.47 0.92 2.66E02 8.77E03 1.24E03 4.11E04
33 IS-7B2 7.78 0.92
34 IS-7B2 8.92 0.95
35 IS-7B2 11.28 0.93
36 IS-7B2 5.75 0.94
Error propagation is based on 2s (see Appendix A for details). All individual points of SIMS analysis are shown in Figs. 5–8.

coats lithified clasts, and can accumulate as laminae or
soot-like dendrites within spherulitic fabrics. In case of
the Barberton carbonaceous cherts earlier studies reported
evidence for colonies of coccoid and sausage-shaped bac-
teria (paired carbon-lined spherules), bacterial biofilm
(smooth, ropy-textured carbonaceous laminaea) (Westall
et al., 2001), branching and twisting filamentous struc-
tures, carbonaceous grains, mat-like laminations (Walsh,
1992; Walsh and Lowe, 1999). Given the clear evidence
of migrated hydrocarbons in chert unit K3c (sample
BAR-3-3-B, Fig. 4), accumulation of large carbonaceous
aggregates in chert unit H5c (100 lm size particles in sam-
ple BAR-4-1, Fig. 5), it is concluded that the carbona-
ceous particles in this study represent secondary features.
In general, it is suggested here that interpretations about
primary microfossil structures should be made with
caution.
5.1.2. Effect of prograde metamorphism on carbon isotope
ratio
Thermal alteration during diagenesis and prograde
metamorphism will change the d13C of carbonaceous mat-
ter. Preferential breaking of the 12C–12C bond over the
12
C–13C bond leads to loss of isotopically light carbon
compounds and a 13C-enriched residue. As carbonaceous
matter is altered, it will also loose heteroatoms such as
H, N, O and S. Several studies have shown the inverse cor-
relation between 13C-enrichment and H/C-ratio (Strauss
et al., 1992a,b; Watanabe et al., 1997; Des Marais, 2001).
In general, it can be assumed that Archean carbonaceous
material that has experienced lower-greenschist metamor-
phism has obtained a d13C value that is enriched relative
to the original value by about 3&. Such material will also
have lost most of its H, and other heteroatoms such as
nitrogen (typical values for N/C-ratio found in early
 Archean carbonaceous cherts; isotopic, chemical, and structural characteristics
Fig. 10. Complete data set of SIMS analyses from Table 1. Lines are shown for d13C of mantle CO2 (5&, Des Marais and Morre, 1984) and graphite in
isotopic equilibrium at resp. 500 and 200 C with mantle CO2 (Chacko et al., 2001).
Archean carbonaceous cherts are in the range of 0.002–
0.02 (Watanabe et al., 1997; Beaumont and Robert,
1999)). Raman spectra of carbonaceous particles (Fig. 9)
in the different Barberton cherts are all identical, suggest-
ing that a uniform degree of structural order was attained
during a regional metamorphic event as late as 2.7 Ga,
when temperatures between 200 and 320 C were reached
(Weis and Wasserburg, 1987; Xie et al., 1997; Toulkeridis
et al., 1998; Tice et al., 2004). The observed N/C-ratio of
individual carbonaceous particles in the Barberton cherts
is in the order of 0.001–0.006, which is in line with typical
observed values in Archean carbonaceous cherts and is
significantly lower than that of unmetamorphosed biologic
material (N/C = 0.15, Redfield ratio C:N is 106:16). The
Raman spectra and N/C-ratios therefore identify all stud-
ied carbonaceous particles as indigenous to the samples,
and exclude any post-metamorphic artifacts. Furthermore,
the Raman spectra confirm that carbonaceous particles
did not experience alteration above lower-greenschist
facies metamorphism. Due to this uniform degree of low
regional peak metamorphism, it is concluded that thermal
cracking of 12C–12C bonds within the carbonaceous mat-
ter did not significantly change the original d13C value
by much more than 3&. Isotopic exchange with carbon-
ates did not significantly change the d13C values either,
since carbonates only form a minor phase in the samples
that have been studied here. Furthermore, isotope
exchange between carbonates and organic matter would
have been very sluggish at lower-greenschist facies
conditions.
665
5.1.3. Effect of hydrothermal alteration on carbon isotope
ratio
Despite the uniform degree of regional thermal alter-
ation and the absence of exchange reactions with carbon-
ates, there is variation in d13C values among the
carbonaceous particles in the different samples (Fig. 10).
The highest d13C values are observed in chert unit H5c of
the Hooggenoeg Formation (sample BAR-4-1). This chert
unit occurs stratigraphically below felsic igneous rocks
(unit H6, see Fig. 1) marking a volcanic event that occurred
3.445 Ga ago (Lowe and Byerly, 1999). Deformation and
magmatic intrusions associated with this event caused se-
vere alteration of the underlying formations, as is evident
from the coarse-grained sugary nature and low d18O (below
+16&) of cherts in the Hooggenoeg Formation relative to
the microcrystalline/amorphous nature and high d18O (as
high as +21&) of cherts in the overlying Kromberg For-
mation (Knauth and Lowe, 2003). A high water/rock ratio
during this event apparently caused recrystallization of
quartz, and could have influenced the carbon isotope ratio
of carbonaceous matter. It was shown before (Robert,
1988) that the d13C of carbonaceous matter inversely corre-
lates with d18O of the surrounding quartz matrix in pro-
gressively metamorphosed Archean cherts; the lowest
d13C values being found in cherts that were least altered
by water–rock interaction. The relatively high d13C values
in sample BAR-4-1 were therefore probably established
by isotopic exchange between carbonaceous matter and
dissolved CO2 in hydrothermal fluids. Samples BAR-3-1
and BAR-3-3-B are from cherts in the overlying Kromberg
666 M.A. van Zuilen et al. 71 (2007) 655–669
Formation, that was deposited after this volcanic event,
and the carbonaceous matter within these units has there-
fore been affected to a lesser degree by such hydrothermal
fluids. It is clear from Fig. 10 that such hydrothermal fluids
also affected the N/C-ratio; the values in sample BAR-4-1
are as low as those of abiologic graphite in sample IS-7-
6-B2 (N/C-ratio is 0.001). Based on geological constraints
and N/C-ratio it can be concluded that the original d13C
of sample BAR-4-1 cannot be adequately reconstructed.
Based on the geological constraints and the more elevated
N/C-ratios in the samples BAR-3-1 and BAR-3-3-B (N/C-
ratio between 0.002 and 0.006, Fig. 10), it can be concluded
that these carbonaceous particles have retained a d13C val-
ue that is closer to that of the original material (mostly
affected by thermal cracking). The original d13C of sample
BAR-3-1 probably was in the range of 34& to 24&,
and that of sample BAR-3-3-B in the range of 40& to
32&, if it is assumed that thermal cracking caused a max-
imum shift to higher values of about 3&. The measurement
of N/C-ratio can therefore be used as a tool for identifica-
tion of carbonaceous particles that have been least affected
by hydrothermal alteration. Together with analysis of
structural disorder and peak metamorphic grade by LRS,
this type of analysis can provide a basis for locating the
most well-preserved d13C value of carbonaceous remains
within samples of Archean deposits.
5.2. Abiologic origin of carbonaceous matter
Carbonaceous matter can form abiologically during
metamorphic processes; e.g. thermal disproportionation
of siderite (van Zuilen et al., 2002, 2003; McCollom,
2003), reactions between CO2 and CH4 or direct reduction
of CO2 in metamorphic fluids (Naraoka et al., 1996; Luque
et al., 1998). Organic compounds can also form by a
Fischer–Tropsch (FT-) synthesis during hydrothermal
alteration of ultramafic rocks (McCollom and Seewald,
2006). This latter process is of particular interest since
hydrocarbon production has been observed in modern
mid ocean ridge hydrothermal systems (Holm and Char-
lou, 2001), and FT-synthesis has been suggested as a source
for carbonaceous material in cherts of the 3.5 Ga Pilbara
Granitoid-Greenstone Belt in Western Australia (Brasier
et al., 2002, 2005; Lindsay et al., 2005). A recent study
has shown that hydrocarbons are formed during FT-syn-
thesis that have a low d13C (ca. 36& relative to a CO2-
source), which is in the same range as biologically pro-
duced organic compounds (McCollom and Seewald,
2006). Furthermore, it has been demonstrated that nitro-
gen can be incorporated in organic compounds during an
FT-reaction when the reactants CO2, H2, and NH4 are
present (Kung et al., 1979; Chang et al., 1983). FT-reac-
tions appear to generate organics that have many charac-
teristics that are typical for biologically produced
compounds. Since there are many units of altered ultramaf-
ic rocks in the stratigraphy of the Onverwacht Group of
the BGB, the possibility should be discussed that carbona-
ceous matter in these cherts is produced abiologically by
FT-synthesis. In such a scenario carbonaceous cherts
should be underlain by swarms of organic-rich black feeder
dikes and should only be found directly above hydrother-
mally altered ultramafic rocks. This is not the case. For in-
stance in the west limb of the Onverwacht Anticline the
Buck Reef Chert (formation K1) is a 250–400 m thick car-
bonaceous chert that overlies 1–2 km of felsic volcanic
units (Tice and Lowe, 2004). In all three carbonaceous
cherts that were studied here (H5c, K1c1, K3c) a clear divi-
sion is observed between the regionally traceable carbona-
ceous chert and underlying organics-free volcanic units. In
general there is no preferential development of carbona-
ceous cherts above units of serpentinized rocks in Barber-
ton. It can be concluded from geological observations
that, although hydrothermal alteration of mafic/ultramafic
basalts clearly occurred in Barberton, carbonaceous matter
in chert units did not form by FT-synthesis. It is possible,
however, that serpentinization of ultramafic crust generat-
ed hydrogen which formed a nutrient for metabolic pro-
cesses (Fig. 11).
5.3. Biologic origin of carbonaceous matter
From the discussion in Section 5.1 it was concluded that
the majority of carbonaceous particles that were studied in
samples BAR-3-1 and BAR-3-3-B represent an original
d13C between ca. 40& and 33&. If it is assumed that
mantle-derived CO2 at that time had a d13C similar to that
of today (5&, Des Marais and Morre, 1984), CO2-fixa-
tion by organisms should have produced a carbon isotope
fractionation between 35& and 28& relative to the
source. Photosynthesizing bacteria (both anoxygenic and
oxygenic) and chemolithoautotrophic bacteria (such as
methanogens) are capable of producing this degree of car-
bon isotope fractionation (House et al., 2003). An earlier
study of the Buck Reef Chert in in the Kromberg Forma-
tion has presented arguments for the dominant role of
anoxygenic photosynthesis in shallow marine depositional
settings (Tice and Lowe, 2004). Such organisms would have
required reduced species such as H2 and Fe2+ as electron
donors. Given the many ultramafic units within the stratig-
raphy of the Barberton Greenstone Belt, and the evidence
for episodes of hydrothermal alteration, it can be inferred
that these reduced chemical species were constantly being
generated by serpentinization of underlying crust and sub-
sequently transported to Archean ocean bottom waters.
High background levels of hydrogen in ocean bottom
water would be achieved by percolating gases along joints
and fractures within the oceanic crust. As was pointed
out by Paris et al. (1985) hydrothermal alteration could
have taken place directly after deposition of ultramafic
crust, leading to serpentinization (and hydrogen produc-
tion) and effective silicification of the ocean floor. It is pos-
sible that methanogens were abundant in such an
environment, using available H2 to reduce dissolved CO2
to methane. This latter scenario can be compared to a mod-
 Archean carbonaceous cherts; isotopic, chemical, and structural characteristics
Fig. 11. Generalized model of an Early Archean ocean environment.
Hydrothermal fluids circulate through ultramafic ocean floor crust causing
serpentinization and the production of hydrogen (Berndt et al., 1996):
Olivine (Fo88) + 1.34 H2O = 0.25 Serpentine + 0.26 Brucite + 0.08 Mag-
netite + 0.08 H2. Serpentinization could therefore have been an important
source of H2 in this early Archean seafloor setting, providing nutrients for
different types of organisms. In this model H2 would be generated on a
regional scale, percolating upward through joints and fractures within the
ocean floor. Possible groups of organisms that would benefit from
enhanced concentration of H2 in ocean water include anoxygenic
photosynthesizers in shallow marine settings (in the photic zone), and
methanogens in deep marine and seafloor hydrothermal settings. Possibly
these groups of organisms dominated in the Early Archean, although
other groups such as oxygenic photosynthesizers should certainly not be
excluded.
ern off-axis, peridotite-dominated low-temperature venting
system such as the Lost City hydrothermal field near the
Mid Atlantic Ridge. In this modern environment an entire
ecosystem has been observed that is characterized by meth-
ane-producing and methane-consuming organisms (Kelley
et al., 2001, 2005). A simplified model of the early Archean
ocean floor environment can therefore be constructed,
where H2, and to some extent Fe2+ are continuously being
produced by hydrothermal circulation through the oceanic
crust, providing a nutrient for methanogens and anoxygen-
ic photosynthesizers (Fig. 11). It should be noted, of
course, that such a model does not exclude other types of
metabolism, most notably oxygenic photosynthesis.
6. Conclusions
The Raman spectra, N/C-ratios, and geological con-
straints identify all studied carbonaceous particles as indig-
enous to the samples, and exclude any post-metamorphic
artifacts. Petrographic observations (pore spaces and small
veinlets filled by organics) suggest that within the chert
units carbonaceous matter has migrated over small distances.
This material was most likely mobilized during hydrother-
mal alteration of the chert deposits, and the observed
aggregates or particles of carbonaceous matter in thin sec-
tions or polished rock slabs are therefore of a secondary
nature. These observations suggest that morphological
interpretation of these carbonaceous structures, and in gen-
667
eral of putative microfossils or microlaminae in hydrother-
mally altered early Archean cherts, should be made with
care.
Thermal cracking or isotope exchange reactions with
carbonates did not significantly change the original d13C
value of carbonaceous matter by more than 3&. Isotope
exchange reactions with dissolved CO2 or CH4 in hydro-
thermal fluids, however, did significantly modify the car-
bon isotope ratio of carbonaceous material in one specific
highly altered chert. SIMS analysis of N/C-ratio and
LRS analysis of structural disorder have been used here
to identify and exclude such strongly altered carbonaceous
particles. This combination of in situ analytical tools there-
fore provides a basis for locating the most well-preserved
d13C values of carbonaceous remains within samples of
Archean chert deposits.
Carbonaceous particles in chert units of the Kromberg
Formation have retained a fairly pristine d13C value; carbo-
naceous particles in the Buck Reef Chert (chert unit K1c1)
have a range 34& to 24&, and those in the Footbridge
Chert (chert unit K3c) have a range of 40& to 32&.
Abiologic formation of carbonaceous matter by Fischer–
Tropsch synthesis during hydrothermal serpentinization of
ultramafic crust can be excluded. It is possible that serpent-
inization of ultramafic crust generated hydrogen which
formed a building block for metabolic processes higher up
in the water column. Likely candidates for such a hydro-
thermally influenced habitat include methanogens and, in
the photic zone, anoxygenic photosynthesizing bacteria.
Acknowledgments
Funding for this study has been provided by a Marie
Curie Individual Fellowship (M.v.Z.). We thank Denis
Mangin (SIMS analysis, CRPG, Nancy), Michel Champe-
nois (SIMS analysis, CRPG, Nancy), Alain Kohler (SEM
analysis, UHP, Nancy), Thérèse Lhomme (LRS analysis,
G2R-UHP, Nancy) for laboratory assistance. Samples for
this study were collected during the 2003 NASA Astrobiol-
ogy Institute Field Workshop ‘‘Archean Surface Processes’’,
in South Africa. M.v.Z. thanks the organizers Don Lowe
(Stanford University) and Gary Byerly (Louisiana State
University) for field guidance during this workshop. The
constructive comments of the reviewers (D.R. Lowe and
an anonymous reviewer) and the associate editor Juske
Horita were greatly appreciated.
Associate editor: Juske Horita
Appendix A. N/C-ratio analysis by SIMS
The analysis of nitrogen concentration by SIMS was
carried out by recording the count rates of CN ions,
and C ions. The error in count rates is based on a Poisson
distribution. The results are corrected for drift during the
analytical session, followed by a blank correction, and
finally a conversion to real N/C-(molar) ratio using several
668 M.A. van Zuilen et al. 71 (2007) 655–669

carbonaceous standards. The normalization and error Beyssac, O., Goffé, B., Chopin, C., Rouzaud, J.N., 2002. Raman spectra
propagation are outlined below: of carbonaceous material in metasediments: a new geothermometer. J.
Metamorph. Geol. 2002 (20), 859–871.
Beyssac, O., Goffé, B., Petitet, J., Froigneux, E., Moreau, M., Rouzaud,
A.1. Drift correction J.N., 2003. On the characterization of disordered and heterogeneous
carbonaceous materials by Raman spectroscopy. Spectrochim. Acta 59,
The drift is measured by comparing standards before 2267–2276.
and after analysis of a specific sample. Brasier, M., Green, O.R., Jephcoat, A.P., Kleppe, A., Van Kranendonk,
M.J., Lindsay, J.F., Steele, A., Grassineau, N.V., 2002. Questioning
ut = measured CN/C ratio of the sample at time t, du the evidence for Earth’s oldest fossils. Nature 416, 76–81.
is based on counting statistics. Brasier, M., Green, O.R., Lindsay, J.F., McLoughlin, N., Steele, A.,
xt = CN/C ratio of the standard at time t, inferred Stoakes, C., 2005. Critical testing of Earth’s oldest putative fossil
from least-squares fitting of xt1 and xt2 directly before assemblage from the 3.5 Ga Apex chert, Chinaman Creek, Western
and after sample measurement. Australia. Precambrian Res. 140, 55–102.
Byerly, G.R., Lowe, D.R., Walsh, M., 1986. Stromatolites from the 3300–
B = fixed CN/C ratio of standard (Don = 0.22, 3500-Myr Swaziland Supergroup, Barberton Mountain Land, South
Aléon et al., 2003) Africa. Nature 319, 489–491.
y = drift-corrected CN/C ratio of the sample Chacko, T., Cole, D.R., Horita, J., 2001. Equilibrium oxygen, hydrogen
sffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
 2  2 and carbon isotope fractionation factors applicable to geologic
Bu oy oy systems. In: Valley, J.W., Cole, D.R. (Eds.), Stable Isotope Geochem-
y¼ ; dy ¼ du þ dx istry, vol. 43. Mineralogical Society of America, pp. 1–81.
x ou ox Chang, S., Des Marais, D.J., Mack, R., Miller, S.L., Strathearn, G.E.,
s ffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
2  2 1983. Prebiotic organic syntheses and the origin of life. In: Schopf,
B Bu J.W. (Ed.), Earth’s Earliest Biosphere. Princeton University Press,
¼ du þ dx
x x2 Princeton, NJ, pp. 53–92.
de Wit, M.J., Hart, R., Martin, A., Abbott, P., 1982. Archean abiogenic
and probable biogenic structures associated with mineralized hydro-
A.2. Blank correction thermal vent systems and regional metasomatism, with implications
for greenstone belt studies. Econ. Geol. 77, 1783–1802.
de Wit, M.J., Hart, R.A., 1993. Earth’s earliest continental lithosphere,
A blank value for CN/C ratio was measured on graph- hydrothermal flux and crustal recycling. Lithos 30, 309–335.
ite standards. This drift-corrected value was subsequently Des Marais, D.J., 2001. Isotopic evolution of the biogeochemical carbon
subtracted from drift-corrected sample measurements. cycle during the Precambrian. In: Valley, J.W., Cole, D.R. (Eds.),
v = CN/C ratio of a blank graphite standard Stable Isotope Geochemistry, vol. 43. Mineralogical Society of Amer-
z = blank corrected CN/C ratio of the sample ica, pp. 555–578.
sffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi Des Marais, D.J., Morre, J.G., 1984. Carbon and its isotopes in mid-
 2  2 qffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi oceanic basaltic glasses. Earth Planet. Sci. Lett. 69, 43–47.
oz oz
z ¼ y  v; dz ¼ dy þ dv ¼ ðdy Þ2 þ ðdvÞ2 Garcia-Ruiz, J.M., Hyde, S.T., Carnerup, A.M., Van Kranendonk, M.J.,
oy ov Welham, N.J., 2003. Self-assembled silica-carbonate structures and
detection of ancient microfossils. Science 302, 1194–1197.
Grassineau, N.V., Nisbet, E.G., Bickle, M.J., Fowler, C.M.R., Lowry, D.,
Mattey, D.P., Abell, P.I., Martin, A., 2001. Antiquity of the biological
A.3. Conversion to real nitrogen concentration
sulphur cycle: evidence from sulphur and carbon isotopes in 2700
million-year-old rocks of the Belinwe Belt, Zimbabwe. Proc. Roy. Soc.
As is seen in Fig. 3, there is a good linear correlation London 268, 113–119.
between CN/C ratio and bulk molar N/C-ratio of differ- Hayes, J.M., Kaplan, I.R., Wedeking, W., 1983. Precambrian organic
ent standards. Using the Don standard (anthracite) as a geochemistry, preservation of the record. In: Schopf, J.W. (Ed.),
Earth’s Earliest Biosphere its Origin and Evolution. Princeton Univer-
reference, all CN/C data (Table 1, column 5) is convert-
sity Press, Princeton, NJ, pp. 93–134.
ed to molar N/C-ratios (Table 1, column 7): Holm, N.G., Charlou, J.L., 2001. Initial indicators of abiotic formation of
A = molar N/C-ratio of standard Don (N/C = 0.103, hydrocarbons in the rainbow ultramafic hydrothermal system. Earth
Sangely et al., 2005) Planet. Sci. Lett. 191, 1–8.
    House, C.H., Schopf, J.W., McKeegan, K.D., Coath, C.D., Harrison,
A A T.M., Stetter, K.O., 2000. Carbon isotopic composition of individual
r¼z ; dr ¼ dz
B B Precambrian microfossils. Geology 28, 707–710.
House, C.H., Schopf, J.W., Stetter, K.O., 2003. Carbon isotopic
fractionation by Archeans and other thermophilic prokaryotes. Org.
References Geochem. 34, 345–356.
Kelley, D.S., Karson, J.A., Blackman, D.K., Fruh-Green, G.L., Butter-
Aléon, J., Robert, F., Chaussidon, M., Marty, B., 2003. Nitrogen isotopic field, D.A., Lilley, M.D., Olson, E.J., Schrenk, M.O., Roe, K.K.,
composition of mantle macromolecular organic matter in interplane- Lebon, G.T., Rivizzigno, P., Party, A.-S., 2001. An off-axis hydro-
tary dust particles. Geochim. Cosmochim. Acta 67, 3773–3783. thermal vent field near the Mid-Atlantic Ridge at 30N. Nature 412,
Beaumont, V., Robert, F., 1999. Nitrogen isotope ratios of kerogens in 145–149.
Precambrian cherts: a record of the evolution of atmosphere chemistry. Kelley, D.S., Karson, J.A., Sylva, S.P., 2005. A serpentine-hosted
Precambrian Res. 96, 63–82. ecosystem: the lost city hydrothermal field. Science 307, 1428–
Berndt, M.E., Allen, D.E., Seyfried, W.E.J., 1996. Reduction of CO2 1434.
during serpentinization of olivine at 300 C and 500 bar. Geology 24 Knauth, L.P., Lowe, D.R., 1978. Oxygen isotope geochemistry of cherts
(4), 351–354. from the Onverwacht Group (3.4 billion years), Transvaal, South
 Archean carbonaceous cherts; isotopic, chemical, and structural characteristics
Africa, with implications for secular variations in the isotopic
composition of cherts. Earth Planet. Sci. Lett. 41, 209–222.
Knauth, L.P., Lowe, D.R., 2003. High Archean climatic temperature
inferred from oxygen isotope geochemistry of cherts in the 3.5 Ga
Swaziland Supergroup, South Africa. GSA Bull. 115, 566–580.
Kung, C., Hayatsu, R., Studier, M.H., Clayton, R.N., 1979. Nitrogen
isotope fractionations in the Fischer–Tropsch synthesis and in the
Miller-Urey reaction. Earth Planet. Sci. Lett. 46, 141–146.
Lindsay, J.F., Brasier, M.D., McLoughlin, N., Green, O.R., Fogel, M.,
Steele, A., Mertzman, S.A., 2005. The problem of deep carbon—An
Archean paradox. Precambrian Res. 143, 1–22.
Lowe, D.R., 1999. Petrology and sedimentology of cherts and related
silicified sedimentary rocks in the Swaziland Supergroup. In: Lowe,
D.R., Byerly, G.R. (Eds.), Geologic evolution of the Barberton Green-
stone Belt, South Africa, vol. 329. The Geological Society, pp. 83–114.
Lowe, D.R., Byerly, G.R., 1986. Archean flow-top alteration zones
formed initially in a low-temperature sulphate-rich environment.
Nature 324, 245–248.
Lowe, D.R., Byerly, G.R., 1999. Geologic Evolution of the Barberton
Greenstone Belt, South Africa. The Geological Society of America.
Lowe, D.R., Byerly, G.R. 2003. Field Guide to the Geology of the 3.5–
3.2 Ga Barberton Greenstone Belt, South Africa.
Luque, F.J., Pasteris, J.D., Wopenka, B., Rodas, M., Barrenechea, J.F.,
1998. Natural fluid-deposited graphite: mineralogical characteristics
and mechanisms of formation. Am. J. Sci. 298, 471–498.
McCollom, T.M., 2003. Formation of meteorite hydrocarbons from
thermal decomposition of siderite (FeCO3). Geochim. Cosmochim.
Acta 67, 311–317.
McCollom, T.M., Seewald, J.S., 2006. Carbon isotope composition of
organic compounds produced by abiotic synthesis under hydrothermal
conditions. Earth Planet. Sci. Lett. 243, 74–84.
Mojzsis, S.J., Arrhenius, G., McKeegan, K.D., Harrison, T.M., Nutman,
A.P., Friend, C.R.L., 1996. Evidence for life on Earth before 3800
million years ago. Nature 384, 55–59.
Naraoka, H., Ohtake, M., Maruyama, S., Ohmoto, H., 1996. Non-
biogenic graphite in 3.8-Ga metamorphic rocks from the Isua district,
Greenland. Chem. Geol. 133, 251–260.
Paris, I., Stanistreet, I.G., Hughes, M.J., 1985. Cherts of the Barberton
Greenstone Belt interpreted as products submarine exhalative activity.
J. Geol. 93, 111–129.
Pasteris, J.D., 1989. Insitu analysis in geological thin-sections by laser
Raman microprobe spectroscopy—a cautionary note. Appl. Spectrosc.
43, 567–570.
Pasteris, J.D., Wopenka, B., 2002. Images of the Earth’s earliest fossils?
Nature 420, 476–477.
Pasteris, J.D., Wopenka, B., 2003. Necessary, but not sufficient: Raman
identification of disordered carbon as a signature of ancient life.
Astrobiology 3, 727–738.
Pinti, D.L., Hashizume, K., Matsuda, J., 2001. Nitrogen and argon
signatures in 3.8 to 2.8 Ga metasediments: clues on the chemical state
of the Archean ocean and the deep biosphere. Geochim. Cosmochim.
Acta 65, 2301–2315.
Robert, F., 1988. Carbon and oxygen isotope variations in Precambrian
cherts. Geochim. Cosmochim. Acta 52, 1473–1478.
Rose, N.M., Rosing, M.T., Bridgwater, D., 1996. The origin of metacar-
bonate rocks in the Archaean Isua Supracrustal Belt, West Greenland.
Am. J. Sci. 296, 1004–1044.
Sangely, L., Chaussidon, M., Michels, R., Huault, V., 2005. Microanalysis
of carbon isotope composition in organic matter by secondary ion
mass spectrometry. Chem. Geol. 223, 179–195.
Schidlowski, M., 2001. Carbon isotopes as biogeochemical recorders of
life over 3.8 Ga of Earth history: evolution of a concept. Precambrian
Res. 106, 117–134.
Schopf, J.W., Kudryavtsev, A., 2005. Three-dimensional Raman imagery
of precambrian microscopic organisms. Geobiology 3, 1–12.
Schopf, J.W., Kudryavtsev, A., Agresti, D.G., Wdowiak, T.J., Czaja,
A.D., 2002. Laser-Raman imagery of Earth’s earliest fossils. Nature
416, 73–76.
669
Strauss, H., Des Marais, D.J., Summons, R.E., Hayes, J.M., 1992a. The
carbon isotopic record. In: Schopf, J.W., Klein, C. (Eds.), The
Proterozoic Biosphere. Cambridge University Press, Cambridge, pp.
117–127.
Strauss, H., Des Marais, D.J., Summons, R.E., Hayes, J.M., 1992b.
Concentrations of organic carbon and maturities and elemental
compositions of kerogens. In: Schopf, J.W., Klein, C. (Eds.), The
Proterozoic Biosphere. Cambridge University Press, Cambridge, pp.
95–100.
Tice, M.M., Bostick, B.C., Lowe, D.R., 2004. Thermal history of the 3.5–
3.2 Ga Onverwacht and Fig Tree Groups, Barberton greenstone belt,
South Africa, inferred by Raman microspectroscopy of carbonaceous
material. Geology 32, 37–40.
Tice, M.M., Lowe, D.R., 2004. Photosynthetic microbial mats in the 3416-
Myr-old ocean. Nature 431, 549–552.
Tice, M.M., Lowe, D.R., 2006. Hydrogen-based carbon fixation in the
earliest known photosynthetic organisms. Geology 34, 37–40.
Toulkeridis, T., Goldstein, S.L., Clauer, N., Kroner, A., Todt, W.,
Schidlowski, M., 1998. Sm–Nd, Rb–Sr and Pb–Pb dating of silicic
carbonates from the early Archaean Barberton Greenstone Belt, South
Africa. Evidence for post-depositional isotopic resetting at low
temperature. Precambrian Res. 92, 129–144.
Ueno, Y., Isozaki, Y., 2001. Carbon isotopic signatures of individual
Archean microfossils from Western Australia. Int. Geol. Rev. 43, 196–
212.
Ueno, Y., Yurimoto, H., Yoshioka, H., Komiya, T., Maruyama, S., 2002.
Ion microprobe analysis of graphite from ca. 3.8 Ga metasediments,
Isua supracrustal belt, West Greenland: relationship between meta-
morphism and carbon isotopic composition. Geochim. Cosmochim.
Acta 66, 1257–1268.
van Zuilen, M.A., Lepland, A., Arrhenius, G., 2002. Reassessing the
evidence for the earliest traces of life. Nature 418, 627–630.
van Zuilen, M.A., Lepland, A., Teranes, J.L., Finarelli, J., Wahlen, M.,
Arrhenius, A., 2003. Graphite and carbonates in the 3.8 Ga old Isua
Supracrustal Belt, southern West Greenland. Precambrian Res. 126,
331–348.
van Zuilen, M.A., Mathew, K., Wopenka, B., Lepland, A., Marti, K.,
Arrhenius, A., 2005. Nitrogen and argon isotopic signatures in
graphite from the 3.8-Ga-old Isua Supracrustal Belt, southern West
Greenland. Geochim. Cosmochim. Acta 69, 1241–1252.
Walsh, M., 1992. Microfossils and possible microfossils from the Early
Archean Onverwacht Group, Barberton Mountain Land, South
Africa. Precambrian Res. 54, 271–293.
Walsh, M.M., Lowe, D.R., 1999. Modes of accumulation of carbonaceous
matter in the Early Archean: a petrographic and geochemical study of
the carbonaceous cherts of the Swaziland Supergroup. In: Lowe, D.R.,
Byerly, G.R. (Eds.), Geologic Evolution of the Barberton Greenstone Belt,
South Africa, vol. 329. Geological Society of America, pp. 115–132.
Watanabe, Y., Naraoka, H., Wronkiewicz, D.J., Condie, K.C., Ohmoto,
H., 1997. Carbon, nitrogen, and sulfur geochemistry of Archean and
Proterozoic shales from the Kaapvaal Craton, South Africa. Geochim.
Cosmochim. Acta 61, 3441–3459.
Weis, D., Wasserburg, G.J., 1987. Rb–Sr and Sm–Nd isotope geochem-
istry and chronology of cherts from the Onverwacht Group (3.5 AE),
South Africa. Geochim. Cosmochim. Acta 51, 973–984.
Westall, F., 2005. Life on the early Earth: a sedimentary view. Science 308,
366–367.
Westall, F., de Wit, M.J., Dann, J., van der Gaast, S., de Ronde, C.E.J.,
Gerneke, D., 2001. Early Archean fossil bacteria and biofilms in
hydrothermally-influenced sediments from the Barberton greenstone
belt, South Africa. Precambrian Res. 106, 93–116.
Wopenka, B., Pasteris, J.D., 1993. Structural characteristics of kerogens to
granulite-facies graphite: applications of Raman microprobe spectros-
copy. Amer. Miner. 78, 533–557.
Xie, X., Byerly, G.R., Ferell Jr., R.E., 1997. IIb trioctahedral chlorite
from the Barberton greenstone belt: crystal structure and rock
composition constraints with implications to geothermometry. Contr.
Mineral. Petrol. 126, 275–291.