Anal. Chem.

2003, 75, 5037-5045

Direct Dating of Archaeological Pottery by

Compound-Specific 14C Analysis of Preserved
Lipids
Andrew W. Stott, Robert Berstan, and Richard P. Evershed*

Organic Geochemistry Unit, Biogeochemistry Research Centre, School of Chemistry, University of Bristol,
Cantocks Close, Bristol, BS8 1TS, U.K.
Christopher Bronk-Ramsey, Robert E. M. Hedges, and Martin J. Humm

Oxford Radiocarbon Accelerator Unit, Research Laboratory for Archaeology and the History of Art, Oxford University,
6 Keble Road, Oxford, OX1 3QJ, U.K.

A methodology is described demonstrating the utility of

the compound-specific 14C technique as a direct means
of dating archaeological pottery. The method uses automated preparative capillary gas chromatography employing wide-bore capillary columns to isolate individual
compounds from lipid extracts of archaeological potsherds in high purity (>95%) and amounts (>200 g)
sufficient for radiocarbon dating using accelerator mass
spectrometry (AMS). A protocol was developed and tested
on n-alkanes and n-carboxylic acids possessing a broad
range of 14C ages. Analytical blanks and controls allowed
background 14C measurements to be assessed and potential sources of errors to be detected, i.e., contamination
with modern or dead 14C, isotopic fraction effects, etc. A
Russian doll method was developed to transfer isolated
target compounds onto tin powder/capsules prior to
combustion and AMS analyses. The major advantage of
the compound-specific technique is that 14C dates obtained for individual compounds can be directly linked
to the commodities processed in the vessels during their
use, e.g., animal fats. The compound-specific 14C dating
protocol was validated on a suite of ancient pottery whose
predicted ages spanned a 5000-year date range. Initial
results indicate that meaningful correlations can be
obtained between the predicted date of pottery and that
of the preserved lipids. These findings constitute an
important step forward to the direct dating of archaeological pottery.
Radiocarbon dating is an important analytical technique in
archaeology and is routinely used to date a wide variety of biogenic
materials, e.g., wood, bone, and plant remains. However, many
14C analyses of natural materials provide ages that only reflect
the bulk organic matter, which may be misleading in the case of
chemically complex heterogeneous samples. An example of this
in buried artifacts or ecofacts arises through contamination from
* Corresponding author. E-mail: r.p.evershed@bris.ac.uk.
Present address: NERC 15N Stable Isotope Facility, CEH-Merlewood,
Grange-over-Sands, Cumbria, LA11 6JU.
10.1021/ac020743y CCC: $25.00
Published on Web 09/03/2003

2003 American Chemical Society

migration of older or younger carbon-containing organic matter,

e.g., humic substances, thereby complicating 14C age-based
determinations. This problem of sample/age homogeneity can
be partially resolved through purification of the organic matter
into characteristic subfractions that directly equate to the original
sample, e.g., collagen from bone or cellulose from wood. However,
contamination from the burial environment may still present a
problem even though rigorous preparation procedures have been
employed. The ideal approach would be to date representative
compounds whose structures establish an unequivocal link to their
source in antiquity. Previous attempts at molecular 14C dating have
included individual amino acids1 and peptides;2 however, these
are not regarded as routine analytical procedures. More recently,
a compound-specific 14C dating approach has been demonstrated
in the dating of sedimentary and petroleum-derived organic
matter.3,4 These researchers demonstrated the feasibility of using
preparative capillary gas chromatography (PCGC) to isolate lipids
from complex mixtures in sufficient concentrations to allow 14C
analysis using high-precision accelerator mass spectrometry
(AMS).
Relative dating of archaeological pottery has focused extensively on empirically derived sequences based upon changes in
pottery characteristics with time, i.e., seriation. These chronological sequences form a routine part of establishing correlations and
differences between areas and phases of archaeological sites.
Absolute dates within these chronological sequences are often
fixed using 14C dating of associated organic artifacts, e.g., bone
or seed, or by dendrochronology. However, these chronologies
may become vague if, for example, (i) unconformities arise in the
archaeological record, (ii) there is a lack of associated datable
organic matter, or (iii) misinterpretation of typologically based
ceramic chronologies occurs.
(1) Bada, J. L.; Gillespie, R.; Gowlett, J. A. J.; Hedges, R. E. M. Nature 1984,
312, 442.
(2) van Klinken H. J.; Hedges, R. E. M. Radiocarbon 1992, 34, 292.
(3) Eglinton, T. I.; Aluwihare, L. I.; Bauer, J. E.; Druffel, E. R. M.; McNichol, A.
P. Anal. Chem. 1996, 68, 904.
(4) Eglinton, T. I.; Benitez-Nelson, B. C.; Pearson, A.; McNichol, A. P.; McNichol,
A. P.; Bauer, J. E.; Druffel, E. R. M. Science 1997, 277, 796.

Analytical Chemistry, Vol. 75, No. 19, October 1, 2003 5037

Potentially, one of the best methods of dating ceramics would

be to date organic matter directly associated with the pottery
(either surface organic residues or organic matter absorbed within
the fabric). Hedges et al.5 attempted to 14C date several organic
carbon-containing fractions from pottery with varying degrees of
success. The variability in dates could be assigned to fraction
impurity and the presence of contaminants. The dating of
archaeological ceramics thus presents a challenging goal within
the archaeological discipline, specifically with regard to dating
organic matter directly associated with the pottery itself.
Organic residues in archaeological ceramics are the focus of
much ongoing research with the results obtained to date clearly
demonstrating that lipids can be consistently used as carriers of
important archaeological information relating to the following: (i)
the nature of the commodities utilized in the vessels (Evershed
et al. and references therein)6 and (ii) determining the mode of
vessel use.7 During this latter work, it was recognized that lipids
are preserved in ceramics in sufficient quantities to serve as a
source of carbon for radiocarbon dating. Additional properties of
lipids favor their use in 14C dating, namely: (i) their structures,
distributions, and stable isotope values at natural abundance
can be confirmed by gas chromatography/mass spectrometry
(GC/MS) and compound-specific stable isotope techniques, which
unambiguously define their origins and identify potential contaminants, (ii) that lipids have fast metabolic turnovers and thus
young ages at the time of deposition, and (iii) they are likely to
be largely indigenous to the ancient pottery vessels due to their
relative immobility and hydrophobicity in the burial environment
(soil). Up to now, radiocarbon measurements have been reported
for two individual lipids isolated from an archaeological sample,
namely, an oil from ancient Egypt. The compound-specific 14C
dates measured for the two compounds recovered from the
Egyptian oil agreed reasonably well with the historic date of the
find spot from which the oil derived.3
This paper describes the results of a systematic study aimed
at developing and testing an analytical protocol for the dating of
individual lipids preserved in archaeological ceramics using a
compound-specific 14C approach. n-Hexadecanoic (C16:0) and
n-octadecanoic (C18:0) acids were targeted for analysis since these
are the most widely occurring lipids preserved in archaeological
pottery. These compounds also persist in sufficient concentrations
(10-4-10-3 g g-1 of potsherd) to make them realistic candidates
for radiocarbon dating. PCGC was used to isolate individual fatty
acids from lipid extracts of archaeological pottery in high purity
and amounts sufficient for radiocarbon dating using AMS.
EXPERIMENTAL SECTION
Test Reagents and Samples. All archaeological samples are
listed in Table 1. High-purity solvents, glass distilled and HPLC
grade (Rathburns Chemicals Ltd.), were used during all experimental procedures and subsequent aliquots taken from the same
bulk solvent. Authentic reference compounds, n-octadecane and
n-heneicosanoic acids, were purchased from Sigma Chemicals Co.
(5) Hedges R. E. M.; Tiemei, C.; Housley R. A. Radiocarbon 1992, 34, 906.
(6) Evershed, R. P.; Dudd, S. N.; Charters, S.; Mottram, H.; Stott, A. W.; Raven,
A.; van Bergen, P. F.; Bland, H. A. Philos. Trans. R. Soc. London B 1999,
354, 19.
(7) Mottram, H. R.; Dudd, S. N.; Lawrence, G. J.; Stott, A. W.; Evershed, R. P.
J. Chromatogr., A 1999, 833 (2), 209.

5038

Analytical Chemistry, Vol. 75, No. 19, October 1, 2003

Ltd. Glassware was either washed in Micro-90 (International

Products Corp.) and rinsed with double-distilled water, acetone,
and dichloromethane or heated in a furnace at 600 C for 12 h
prior to use.
Sample Preparation. Reference compounds, i.e., n-octadecane and n-heneicosanoic acids, were dissolved in dichloromethane (1 mg mL-1 solution) prior to use. Fatty acids were
converted to methyl esters using dry methanol (anhydrous Na2SO4) acidified with concentrated H2SO4, heated at 70 C for 1 h.
Fatty acid methyl esters (FAMES) were then recovered by the
addition of 2 mL of double-distilled water and diethyl ether.
Following removal of the diethyl ether and evaporation under
nitrogen, the methyl esters were eluted through a short column
of anhydrous Na2SO4 for water removal, evaporated, and redissolved in hexane.
Compositional analyses of lipids in archaeological potsherds
largely followed our previously published methodologies.7,8 However, for the purposes of compound-specific 14C dating, Soxhlet
extraction [(dichloromethane/methanol (2:1 v/v)] of powdered
sherds weighing >8 g was employed to recover larger amounts
of total lipid extract (TLE) from which target compounds could
eventually be isolated. An aliquot of the TLE was derivatized with
N,O-bis(trimethylsilyl)trifluoroacetamide containing 1% trichlorosilane (70 C, 1 h) prior to screening by high-temperature GC
(HT-GC) and HT-GC/MS. The remainder of the TLE was
hydrolyzed with methanolic NaOH (0.5 M, 70 C, 1 h), acidified
to pH 3 (HCl, 1 M), and extracted into hexane to yield a fatty
acid fraction. An aliquot of this fraction was analyzed by GCcombustion-isotope ratio MS (GC-C-IRMS) to provide the 13C
values of the individual FAMES before PCGC. Potsherd FAMES
were then submitted to PCGC as described below.
Instrumental Analyses. HT-GC, HT-GC/MS, and automated
GC-C-IRMS analyses were performed as described previously.8,9
Fully automated PCGC was carried out using a Hewlett-Packard
7673 autosampler coupled to a Hewlett-Packard 5890 series II GC
fitted with a Megabore fused-silica capillary column (30 m 0.53
mm i.d.) coated with a dimethyl polysiloxane stationary phase
(DB-1, 0.5-m film thickness). The GC was interfaced to a Gerstal
preparative fraction collector via a zero dead volume effluent
splitter connected to the flame ionization detector (FID) and a
temperature regulated (200-300 C) transfer capillary line interfaced to the eight-port fraction collector. The zero dead volume
preparative fraction collector was installed in a temperaturecontrolled oven (250-300 C) and consists of seven separate
capillary lines linked to individual glass U-tube traps (one waste,
six sample traps). The autosampler, GC, and preparative fraction
collector are fully controlled by a microprocessor, which allows
programmable operation throughout the PCGC trapping sequences. The GC temperature program was from 50 (2 min) to
210 C at a rate of 10 C min-1 and then to 260 C at 4 C min-1
and finally to 300 C at 10 C min-1. Data were continuously
monitored during PCGC run sequences via a Hewlett-Packard
3396 series II integrator.
Samples were combusted at the Oxford Radiocarbon Accelerator Unit using a continuous-flow CHN analyzer (Europa-ANCA)
(8) Evershed, R. P.; Heron, C.; Goad, L. J. Analyst 1990, 115, 1339.
(9) Copley, M. S.; Berstan, R.; Dudd, S. N.; Docherty, G.; Mukherjee, A. J.;
Straker, V.; Payne, S.; Evershed, R. P. Proc. Natl. Acad. Sci. U.S.A. 2003,
100, 1524.

Analytical Chemistry, Vol. 75, No. 19, October 1, 2003

5039

849-10
806-10
806-11

C18:0

C16:0
C18:0

850-13
850-14

450
1000

511
794

442
365

350

599
769
204
104
242
280

27
293
245
79
416
157
614
489
552
871
558
615
326

371

29

yield CO2
(g of C)
combusted

-28.6
-28.5
-30.8
-26.7
-27.2
-29.5
-27.5
-27.4
-31.3
-28.0
-31.2
-27.9
-31.3
-33.4
-28.1
-30.0
-29.5
-30.0
-32.5
-35.2
-31.3
28.0
-33.1

-27.6
-30.2

-28.5
-33.6

-29.6
-29.6
-31.7
-27.8
-28.4
-30.5
-28.6
-28.6
-32.2
-29.2
-32.1
-29.1
-32.2
-34.2
-29.3
-30.9
-30.5
-31.1
-33.3
-35.9
-32.2
-29.2
-33.9

-28.8
-31.1

-29.6
-34.4

51.9
51.8

52.0
51.9

55.3
53.8

53.9

55.7
56.3
56.9
57.1
52.6
53.6

80.9
86.7
86.4
84.9
83.7
86.3
78.2
76.7
79.2
77.5
77.9
76.2
55.5

83.1

94.0

%
mod.obs

0.4
0.4

0.5
0.4

0.4
0.6

0.6

0.5
0.5
0.8
0.8
0.5
0.9

0.6
0.6
0.7
0.5
0.6
0.7
0.5
0.5
0.5
0.4
0.7
0.5
0.8

0.5

1.8

% mod.
error (()

55.1
54.6

55.1
54.7

58.7
56.7

56.8

59.1
59.4
60.0
60.6
55.5
56.5

85.9
91.4
91.2
90.0
88.3
91.0
83.0
80.9
84.0
81.7
82.7
80.4
58.5

87.6

99.8

corrected
% mod.
14Ca

4790 ( 60
4860 ( 60

4790 ( 80
4840 ( 60

4280 ( 60
4550 ( 90

4540 ( 80

4220 ( 70
4190 ( 70
4100 ( 110
4020 ( 110
4730 ( 80
4580 ( 130

1230 ( 60
720 ( 60
740 ( 60
850 ( 50
1000 ( 60
760 ( 60
1500 ( 50
1700 ( 50
1400 ( 50
1620 ( 40
1530 ( 70
1760 ( 50
4300 ( 110

1060 ( 50

20 ( 150

date (
errora
(years BP)

date of associated material; D, dendrochronology.

-28.3

-29.5

14C

13C FA
()

13C
FAME
()

Corrected for contribution from methanol (derivatization reagent). bS, stylistic; A,

C16:0
C18:0

7589
7588
7590
7358
7356
849-08

C16:0
C18:0
C18:1
C16:0
C18:0
C18:0

Sweet Track carinated

bowl SW 2

7586
7587
7585
7582
7583
7584
7591
7592
807-06
807-07
807-08
807-09
806-07

C16:0
C18:0
C18:1
C16:0
C18:0
C18:1
C16:0
C18:0
C16:0
C18:0
C16:0
C18:0
C18:0

807-12
807-13

7581

C18:0

C16:0
C18:0

7355

OxA/OxA-X
sample codes

Data Measured on Fatty Acids Extracted from Medieval to Neolithic Potsherds

C16:0

Hambledon Hill
carinated bowl ST-81-96
(Stepleton enclosure)

Eton Rowing Lake

plain bowl DBC 1
Eton Rowing Lake
carinated bowl
DBC-E
Hambledon Hill plain
bowl HH77-1924
Hambledon Hill plain
bowl ST-81-938
(Stepleton enclosure)

Stanwick Mortaria
ST-250
Stanwick Mortaria
ST-250
Stanwick Mortaria
ST-250
Yarnton Mortlake
ware R 31
Yarnton Fengate
ware R 5-9

West Cotton Shelly

ware jar RP78

West Cotton Lyveden A

ware jar RP22
West Cotton St. Neots
ware bowl RP73
West Cotton Lyveden A
ware jar RP25

sherd site/type

14C

target
fatty
acids

Table 1. Compound-Specific

3670-3370 BC
3780-3510 BC

3700-3370 BC
3370-3380 BC

3090-2670 BC
3550-2900 BC

3550-2900 BC

3020-2570 BC
2910-2570 BC
2950-2350 BC
2900-2200 BC
3650-3360 BC
3650-2900 BC

670-960 AD
1210-1400 AD
1160-1400 AD
1040-1280 AD
890-1190 AD
1150-1400 AD
430-650 AD
210-440 AD
540-770 AD
260-540 AD
390-660 AD
130-410 AD
3350-2550 BC

880-1040 AD

modern

corrected
datea (2)

4870 ( 35 BP; 3710-3540 BC

(A)
sample later than UB-4138
(4648 ( 21 BP), OxA-7042
(4735 ( 60 BP), earlier
than OXA-7041 (4485+/
-60 BP), OxA-7048
(4670 ( 40 ( BP) (A)
sample later than UB-4138
(4648 ( 21 BP), OxA-7042
(4735 ( 60 BP), earlier
than OXA-7041 (4485
( 60 BP), OxA-7048 (4670
( 40 BP) (A)
3807/6 BC (D)

4000-3600 BC (S)

3700-3300 BC (S)

3400-2900 BC (S)

3400-2900 BC (S)

150-250 AD (S)

150-250 AD (S)

150-250 AD (S)

1150-1225 AD (S)

1225-1250 AD (S)

1250-1300 AD (S)

1250-1300 AD (S)

date by
associationb

fitted with a CO2 collection facility. The CHN analyzer uses GC

to separate the CO2 from the other gases formed through
combustion and collects the resultant CO2 as the target material
for the gas source10 of the AMS system.
RESULTS AND DISCUSSION
There were two main phases in the development of the
analytical protocol. The first phase focused on the implementation
and testing of the analytical techniques required for the isolation
of individual components from archaeological pottery. The second
phase, i.e., the AMS dating of individual lipids from archaeological
pottery, depended upon whether acceptable controls and blanks
were obtained. Validation of the method was achieved by
radiocarbon dating individual lipids isolated from archaeological
pottery of known date by correlation with materials from the same
archaeological contexts and related sequences dated by conventional methods.
Method Development. Analytical Blanks and Controls.
Particular consideration was given to the suitability of the PCGC
capillary column used for the separation and isolation of individual
lipids. The three key factors are (i) column dimensions, (ii)
durability of the stationary phase, and (iii) reproducibility of
compound retention times during repetitive trapping sequences
typically of >100 h. The GC was fitted with a Megabore fusedsilica capillary column (30 m 0.53 mm i.d.) coated with a
dimethyl polysiloxane stationary phase (DB-1, 0.5-m film thickness) as the use of such columns had been previously shown to
result in minimal amounts of methylsilicone degradation products
accumulating in the traps during a sequence.3 To assess the
contribution of carbon from the stationary phase, i.e., column
bleed, a chromatographic blank was trapped, i.e., collection of
the entire chromatographic eluent from a sequence of 120 GC
runs (temperature programmed to 300 C), and submitted for 14C
analysis. Upon combustion, the amount of carbon (recovered as
CO2) from the chromatographic blank was only 0.9 g. As the
chromatographic blank was trapped at temperatures well in
excess of the elution temperatures of the target compounds (<230
C) and each blank GC run was 40 times longer (34.5 min)
than a typical target compound trapping window (0.8 min), it
was concluded that the amount of stationary phase (column
bleed)-derived carbon would be minimal in any compoundspecific AMS analyses. This analysis also showed that contamination from the instrument and carrier gas supply was negligible.
Obtaining reproducible retention times for each compound is
essential since in excess of 80 GC runs are normally required for
a single trapping sequence. The drift in retention times during
several >100 GC run automated sequences was observed to vary
by only 5 s. This confirmed the excellent reproducibility of the
automated instrument and thus enabled the precise setting of trap
opening times during subsequent trapping sequences.
Our initial approach involved the analysis of appropriate
analytical controls in order to assess potential sources of contamination during manipulation and isolation of the individual
compounds. Four petroleum-derived solvents (dichloromethane,
diethyl ether, hexane, methanol) were analyzed by AMS. These
solvents were an essential component of the analytical protocol,
(10) Bronk-Ramsey, C.; Hedges, R. E. M. Nucl. Instrum. Methods B 1987, 29,
45.

5040

Analytical Chemistry, Vol. 75, No. 19, October 1, 2003

being required for the extraction of lipids from the archaeological

pottery and subsequent derivatization and transfer procedures
prior to AMS dating. The percentage of modern 14C in these
solvents was <0.9%, and on this basis, all four solvents were
classed as 14C dead.
Reference compounds including a C21 fatty acid (60% modern
14C), modern cocoa butter, and a petroleum-derived n-C alkane
18
were chosen for development work because of their differing ages
and GC properties similar to the compounds ultimately to be
isolated from archaeological pottery. Analysis of these standards
enabled the assessment of whether extraneous 14C was being
introduced. The quantity of modern carbon added in the method
was less than 4 g, with the amount of infinite age carbon added
less than 5 g. The correction applied for date calculation was
2.9 ( 0.3 g of modern carbon based on the comparison of AMS
measurements made on the bulk petroleum-derived n-C18 alkane
with measurements on smaller samples (down to 18 g of C)
combusted in the same way as the archaeological samples.
A concern at this stage was the potential isotopic fractionation
occurring during PCGC sequences. This was tested by determination of the 13C values of authentic compounds before and after
PCGC isolation. The n-C18 alkane was isolated after 100 PCGC
runs (72-h run time) and its 13C value measured. Comparison of
the 13C values before (-33.3) and after (-34.8) trapping
suggested a fractionation of 1.5, similar to that observed by
Eglinton and co-workers,3 i.e., <5. Similarly, the C21 fatty acid
standard showed a fractionation of 1.1% after 80 PCGC runs, i.e.
-28.2 before and -29.3 after isolation. These data indicate
that fractionation does indeed occur to a small extent during
isolation, although the effect is minimized provided that (i) the
PCGC instrument is carefully checked for leaks at the splitter,
traps, and fraction collector and (ii) the trapping window is
sufficiently wide to span the whole elution time range of the target
compound.
Trapping Recoveries and Sample Transfer to AMS. A split
ratio of 98:2 was set in the effluent splitter using a separate control
capillary installed below the jet of the FID. This directed 98% of
the column effluent via a heated transfer line to the preparative
fraction collector and 2% to the FID to enable continuous
monitoring during the PCGC sequence. Trapping recovery experiments were carried out using a stock solution of C16:0 and C18:0
fatty acids of known concentrations combined with a n-C18 alkane
internal standard. By comparing the peak areas of the target
components with the internal standard before and after trapping,
the percentage recovery of each compound was calculated.
Trapping efficiencies for the C16:0 and C18:0 fatty acids were found
to be >90%.
An important logistical component of the project was the
transfer of the isolated individual compounds from the PCGC traps
to the tin capsules for combustion to CO2. A method of depositing
the trapped target components onto tin capsules containing a small
quantity of tin powder was developed. The feasibility of the method
was determined by injecting solutions of fatty acids of known
concentrations (50, 100, 150, and 200 g) onto tin capsules
containing tin powder. By comparing the GC peak area responses
obtained from solvent washings with an internal standard, the
recoveries of the fatty acids (C16:0 and C18:0) from the tin were
calculated to be >95%. Solvent creep (hence, sample loss) was

eliminated by employing a Russian doll method in which a tin

capsule was partially filled with tin powder and then inserted inside
a larger diameter tin capsule prior to spiking with the target
compound. A key factor in the transfer method was the rapid
solvent evaporation (aided by heating at 40 C) under a gentle
stream of nitrogen during the spiking procedure.
Correction Factors and Dating. Additional corrections need
to be applied to the Oxford AMS %modOBS value to account for
the effect on the measured radiocarbon age by the methyl carbon
atom added through derivatization. These corrections are achieved
as follows:
A simple mass balance calculation is used to correct for the
infinite age carbon added to the fatty acid from the methanol:

% mod(corrected for 14C content of

MeOH)

no. of CFAME % modOBS

no. of CFA

where no. of CFAME is the total number of carbon atoms in

derivatized fatty acid, e.g., 19 for a C18 fatty acid, % modOBS is the
percent modern 14C of the derivatized fatty acid (corrected to 13C
value, -25), and no. of CFA is the total number of carbon atoms
in underivatized fatty acid, e.g., 18 for a C18 fatty acid.
To account for isotopic fractionation effects, each of the
radiocarbon measurements is corrected to the accepted convention 13C value of -25. Since the AMS at Oxford measures the
14C/13C ratio rather than conventional 14C/12C ratio measured by
other laboratories, the isotopic fractionation effects on the
measured values are smaller by a factor of 2 (i.e., 1 variation
equates to 8.2 radiocarbon years). The above isotopic fractionation
correction is already incorporated into the % modOBS values;
however, as the 13C value of the original fatty acid is altered
during the derivatization procedure, a further correction is
necessary. The 13C value of the fatty acid (13CFA) is required
and determined by a simple mass balance calculation:

13CFA )

(no. of CFAME 13CFAME) - 13CMeOH

no. of CFA

where no. of CFAME is the total number of carbon atoms in

derivatized molecule; 13CFAME ) 13C value of fatty acid methyl
ester; 13CMeOH ) 13C value of methanol (-48.0); and no. of
CFA is the total number of carbon atoms in the original fatty acid.
By combining the correction factors due to the methanol (14C
content and 13C alteration), an expression for the corrected
percent modern 14C (%modCORR) can be derived:
% modCORR )

)(

no. of CFAME % modOBS

no. of CFA

1 + [(-25 + 13CFAME)/1000]
1 + [(-25 + 13CFA)/1000]

This is then expressed as a radiocarbon age (in years BP)

according to the following equation:

radiocarbon years (BP) ) -8033 ln(% modCORR/100)

A calibrated calendar range was obtained using OxCal
with the Stuiver et al. calibration curve.12

v3.311

The previously described PCGC conditions were optimized to

enable complete isolation of individual compounds in high purity
and concentration. Trapped components were therefore guaranteed to be free from any major 14C contamination, which potentially
may have been introduced at any stage of the workup procedures
during trapping and subsequent AMS analysis.
Validation of the Protocol Using Archaeological Pottery.
The method was validated by dating individual lipids recovered
from archaeological pottery ranging in age from Early Neolithic
to Medieval, a range spanning 5000 years. Sherds were made
available for analysis from a variety of archaeological sites
including the Raunds Area Project [West Cotton assemblage (Late
Saxon/Early Medieval) and Stanwick (Roman)], Sweet Track
(Early Neolithic), Eton Rowing Lake (Early Neolithic), Hambledon
Hill (Early Neolithic), and Yarnton (Mid/Late Neolithic). Sherd
TLEs, derived from the processing or cooking of foodstuffs and
commodities, were initially screened by GC and quantified, and
those containing > 300 g g-1 of lipid were selected as candidates
for compound-specific 14C dating. The C16:0 and C18:0 fatty acids
(mainly animal fat derived, based on the high relative abundance
of the latter component) were the most dominant components in
all the selected TLEs. The 13C values of these fatty acids were
determined before and after trapping, from which it was evident
that isotopic fractionation occurred to a small extent, with the 13C
values always agreeing to within 2 and in the majority of cases
within the analytical precision of the GC-C-IRMS instrument
((0.3).
Medieval Vessels. Four samples (RP22, RP25, RP73, RP78)
were selected from the West Cotton assemblage, Northamptonshire (Northampton Archaeology Unit) ranging in date from 1150
to 1300 AD (dates assigned by typology and stylistic criteria;
Blinkhorn, personal communication). Table 1 shows the dates of
each of the sherds based on these criteria. Sherds from this
assemblage had previously been analyzed in our laboratory as
part of a larger study and were initially chosen because of their
high content of preserved lipids: RP25, 2031 g g-1, RP73, 2821
g g-1, and RP78 4840 g g-1. Sherd RP22 was chosen as the
exception as it yielded only 200 g g-1 of lipid and served as a
guide in defining the lower limits of lipid yield that could be
successfully used for PCGC. Determinations yielded target
compounds ranging between 29 and 416 g of combustible carbon
(as CO2). The fatty acids from sherd RP22 were among the first
to be isolated and dated using the developed protocol. However,
the low amounts of CO2 produced from combustion indicated that
200 g is the lower limit of sample size for valid compoundspecific dating. Figure 1 shows the GC traces of trapped products
collected from 95 preparative GC runs from sherd RP25. Each
compound has been isolated in the high purity essential for
compound-specific 14C analyses. Table 1 shows the 14C data
obtained for the most abundant fatty acids purified from the lipid
extracts of the four Medieval archaeological potsherds by PCGC.
The very substantial pottery assemblage from West Cotton
(>105 potsherds) has allowed typological chronologies to be
ascertained. A number of timber posts were also recovered from
(11) Bronk,-Ramsey, C. Oxcal v3.3, 1999.
(12) Stuiver, M.; Reimer, P. J.; Bard, E.; Beck, J. W.; Burr, G. S.; Hughen, K. A.;
Kromer, B.; McCormac, G.; van der Plicht, J.; Spurk, M. Radiocarbon 1998,
40 (3), 1041.

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5041

Figure 1. Partial gas chromatograms of products collected from

95 preparative GC runs in which the C16:0, C18:0, and C18:1 fatty acids
were targeted from a West Cotton Medieval sherd lipid extract. The
upper chromatogram corresponds to the total lipid extract while those
below are analyses of the contents of four traps after the sequence
was completed.

a sequence of water mills at the site and radiocarbon dated

independently. Sherd RP78 was recovered from a clay bank
overlying the infilled leat of the final watermill, which on the basis
of the pottery would date the construction of the bank to about
cal. 1150 AD (personal communication, A. Chapman, Northamptonshire Archaeology Unit). Timber posts (941 ( 53 BP; 88.9%
mod. 14C, 1014 ( 51 BP; 88.1% mod.14C) from the surviving
structure of the mill yielded radiocarbon dates suggesting that
the mill was in operation for about a century between 1025 and
1125 AD. The Shelly Ware pottery associated with this construction has been dated typologically to 1150-1225 AD. Compoundspecific 14C dates on the C16:0, C18:0, and C18:1 fatty acids from RP78
yielded dates of cal. 850 ( 50 BP; 90.0% mod. 14C (cal. 10401280 AD), 1000 ( 60 BP; 83.3% mod. 14C (cal. 890-1190 AD) and
760 ( 60 BP; 91.0% mod. 14C (cal. 1150-1400 AD), respectively,
which are in good agreement with the predicted age (Table 1).
Roman Vessels. Two fatty acids (C16:0 and C18:0) were isolated
from the total lipid extract of a Roman Mortaria (ST-250, dated
typologically at 150-250 AD) by PCGC. Upon combustion, yields
of CO2 were 614 and 489 g, respectively, with AMS analysis
producing dates of 1500 ( 50 BP; 83.0% mod. 14C (cal. 430-650
AD) and 1700 ( 50 BP; 80.9% mod. 14C (cal. 210-440 AD) for the
C16:0 and C18:0 fatty acids, respectively. The date for the C18:0 acid
correlates well with the expected age of the vessel, i.e., 79.5% mod.
14C. However, the C
16:0 fatty acid yielded a slightly later date (>%
mod. 14C). To investigate the reproducibility of this result, two
further extractions, PCGC isolations and AMS analyses, were
performed on the same sherd. The radiocarbon dates for the
duplicate C16:0 fatty acids were comparable with those obtained
5042 Analytical Chemistry, Vol. 75, No. 19, October 1, 2003

previously, i.e., later than second-third century Roman, and the

C18:0 fatty acids again correlated well with the expected age of
the vessel. The calibrated calendar date ranges for the fatty acids
are shown in the histograms displayed in Figure 2. The yields of
combustible carbon for the replicate analyses exceeded 550 g
in all cases (Table 1); therefore, the difference in age cannot be
explained on the basis of sample size.
Neolithic Vessels. Three fatty acids (C16:0, C18:0, and C18:1)
isolated in high purity by preparative GC (95 runs) from Yarnton
Mid-Late Neolithic Fengate Ware. R5-9 yielded very consistent
radiocarbon ages ranging between 59.1 and 60.0% mod. 14C (cal.
3020 and 2350 BC); cf. Table 1. Two of the three compounds
yielded larger amounts of CO2 than had previously been recovered
from other vessels, i.e., 599 and 769 g, which correlated well
with the predicted amounts (550 and 685 g) calculated from
an external C21 fatty acid standard of known concentration run
after the PCGC sequence. The C18:0 fatty acid isolated from a
second sherd (R31, Mid-Late Neolithic Mortlake Ware) yielded a
radiocarbon age of 4300 ( 110 BP; 58.5% mod. 14C (cal. 33502550 BC). All compound-specific 14C dates obtained for the
Yarnton pottery show that the individual lipids correlated with
the expected age of the vessels (Table 1).
Fatty acids (C16:0 and C18:0) were isolated in high purity from
a plain bowl sherd (DBC-1) and a carinated bowl sherd (DBC-E)
recovered from the Early Neolithic site at Eton Rowing Lake,
Oxfordshire (Table 1), yielding, upon combustion 104, 242
(DBC-1, C16:0, C18:0), and 280 g (DBC-E, C18:0) of carbon,
respectively. The compound-specific radiocarbon dates obtained
closely correlate with the expected range of the Early Neolithic
Eton pottery, particularly those of the C18:0 fatty acids, i.e., 4730
( 80 BP; 55.5% mod. 14C (cal. 3650-3360 BC) and 4580 ( 130
BP; 56.5% mod. 14C (cal. 3650-2900 BC), respectively.
Early Neolithic pottery from the Hambledon Hill prehistoric
monument was sampled (Dorset County Museum) and screened
for lipids. These sherds were chosen specifically because they
originated from contexts that contained bone or charcoal fragments that had been independently radiocarbon dated. Of the
20 sherds initially screened, 40% yielded lipid, of which three
(ST-81/938, ST-81/96, HH77-1924) were selected for compoundspecific 14C analysis on account of their high lipid content.
HH77-1924 was recovered together with other datable materials,
which could be classed as directly associated with the sherd.
ST-81/938 and ST-81/96 were recovered in secondary silts from
segments 14 L3B and 15 L3B, respectively, if the sequence in other
segments were contemporary then the potsherds should be later
in date than the following artifacts: (a) UB-4138 (articulated dog
skeleton, 4648 ( 21 BP), (b) UB-4151 (red deer antler pick, 4772
( 19 BP), (c) UB-4152 (red deer antler pick, 4792 ( 20 BP), (d)
UB-4153 (red deer antler rake, 4740 ( 19 BP), and (e) OxA-7042
(antler, 4735 ( 60 BP). The potsherds should be earlier than the
following: (a) OxA-7048 (Maloideae charcoal, 4670 ( 40 BP), (b)
OxA-7050 (Maloideae charcoal, 4730 ( 40 BP), and (c) OxA-7814
(adult female humerus, 4680 (30 BP).
Compound-specific AMS dates measured on the target C16:0
and C18:0 acids resulted in values of 4790 ( 80 BP; 55.2% mod. 14C
(cal. 3710-3370 BC) and 4840 ( 60 BP; 54.7% mod. 14C (cal. 37703380 BC) for sherd ST81/96 and 4280 ( 60 BP; 58.7% mod. 14C

Figure 2. Calibrated calendar ranges from the analyses of the C16:0 and C18:0 fatty acids extracted from a Roman Mortaria sherd (ST-250)
from the site of Stanwick. The C16:0 fatty acids (top three histograms) and the C18:0 fatty acids (bottom three histograms) were measured in
triplicate and compared with the typologically expected date range for the sherd (lower panel).

(cal. 3090-2670 BC) and 4550 ( 90 BP, 56.7% mod.14C (cal. 35502900 BC) for sherd ST-81/938. As mentioned previously, sherd
HH77-1924 was found directly associated with OxA-8845 (red
deer antler crown, 4870 ( 35 BP), i.e., from the bottom of the
same ditch. Compound-specific AMS dates measured on the C18:0
acid resulted in a value of 4540 ( 80 BP; 56.8% mod. 14C (cal.
3550-2900 BC). The overall compound-specific results on the
Early Neolithic pottery from Hambledon Hill correlated well with
the ages of the sherds; however, it is difficult to know the true
age of the contexts where the availability of associated precisely
datable materials is limited.
Lipids were extracted from an Early Neolithic carinated bowl
(SW2) from the Somerset levels in south west England. The
potsherd was discovered adjacent to an elevated wooden walkway,
known as the Sweet Track, that spanned 2 km of the swamp land.
This manmade structure has been accurately dated by dendrochronolgy to 3807/6 BC and was believed to be only used for
10 years.13 The C16:0 and C18:0 fatty acids were isolated in large
amounts from the sherd, producing yields of carbon upon
combustion in excess of 400 g. Compound-specific radiocarbon
ages for the C16:0 and C18:0 fatty acids were 4790 ( 60 BP; 55.1%
mod. 14C (cal. 3670-3370 BC) and 4860 ( 60 BP; 54.6% mod. 14C
(cal. 3780-3510 BC), respectively. The calibrated calendar ranges
for both the isolated fatty acids, particularly the C18:0 fatty acid,
compare very favorably with the associated dendrochronological
date for the Sweet track (Table 1 and Figure 3).

Figure 3. Radiocarbon age determination and calibrated calendar

range for the C18:0 fatty acid isolated from a carinated bowl sherd
(Sweet Track, SW2). The calibrated calendar range is compared with
the associated dendrochronological date (3807/6 BC).

CONCLUSIONS
The experiments and determinations reported herein confirm
that PCGC is capable of isolating individual compounds from lipid
extracts of potsherds via repeated injections, in sufficient amounts
for AMS dating (>200 g of C). Through AMS analyses of
appropriate blanks and controls it has been established that
introduction of background 14C contamination was minimal. AMS
measurements on individual fatty acids isolated from potsherds
ranging in date from Early Neolithic to Medieval yielded percent
modern 14C values, which were in good agreement with the sample
history.

In general, the radiocarbon ages obtained for the C18:0 fatty

acids correlated best with the associated ages, while a trend
toward a slightly more modern date was observed for several of
the trapped C16:0 fatty acids. Interestingly, the triplicate measurements made on the Roman Mortaria vessel (ST-250) showed little
variation, suggesting the age offset between the fatty acids does
not derive from laboratory methodologies. Environmental contamination, involving migration of the C16:0 fatty acid (the most
dominant fatty acid in soil organic matter in this carbon number
range) into the sherd during burial, could explain this apparent
discrepancy. An enrichment of 14C has previously been observed
in the bulk soil carboxylic acid fraction with increasing depth
presumed to result from the downward mobility of free fatty
acids originating from modern vegetation, fauna, and microorganisms, during groundwater leaching.14 The somewhat enhanced
migration/leaching predicted for the more soluble shorter chain
C16:0 fatty acid is consistent with the slightly younger values
observed for this component.

(13) Hillam, J.; Groves, C. M.; Brown, D. M.; Baillie, M. G. L.; Coles, J. M.; Coles,
B. J. Antiquity 1990, 210.

(14) Bol, R.; Huang Y.; Meredith, J. A.; Eglinton, G.; Harkness, D. D.; Ineson, P.
Eur. J. Soil Sci. 1996, 47, 215.

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Figure 4. Difference between the 14C ages of the individual fatty acids and the associated ages of each sample: (a) C16:0 fatty acids and (b)
C18:0 fatty acids. Gray rectangles represent the associated age range, and the black bars correspond to the offset from the associated age.

The average age difference between the C16:0 and C18:0

components (in samples that produced >100 g of C) was 216
radiocarbon years. If the average age of the exogenous C16:0 fatty
acid was half that of the endogenous component, the degree of
addition to the potsherd would have to be as high as 5-15% of
the exogenous soil C16:0 fatty acid. Considering the fatty acid
concentration difference between the potsherd and the soil is great
(potsherd concentrations at least 10 times greater15), it is difficult
to envisage that the addition of C16:0 fatty acid from the soil could
occur to the required extent.
The complete data set presented above reveals disparities
between some of the dates of the lipid from the potsherds and
(15) Simpson, I. A.; van Gerben, V.; Perret, V. P.; Elhmmali, M. M.; Roberts, D.
J.; Evershed, R. P. Holocene 1999, 9(2), 223.

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Analytical Chemistry, Vol. 75, No. 19, October 1, 2003

those from the associated finds (Figure 4). When assessing the
accuracy of the compound-specific dates relative to the associated
datable artifacts, it is important to note that the reliability of that
association is dependent upon a number of variables affecting the
formation of deposits at the archaeological sites, e.g., primary
versus secondary deposits, bioturbation, etc. As a result, caution
should be exercised when comparing the compound-specific dates
with some of the associated finds. Possibly the best associated
date related to SW2, which was discovered adjacent to the Sweet
Track. The compound-specific 14C dates for the isolated fatty acids
showed excellent agreement with the age of the Sweet Track.
Using 50 years as a more conservative estimate for the use
lifetime of the Sweet Track and decalibrating its calendrical
date range into radiocarbon years, the midpoint of its range was

found to be about 3 (160 14C years) and 5% (230 14C years) older
than the date range midpoints that corresponded to the C18:0 and
C16:0 fatty acids, respectively. These results are extremely encouraging considering the Sweet Track was constructed over 5800
years ago during the beginning of the Neolithic period of Britain.
As we proceed toward routine compound-specific dating
studies of individual lipids associated with potsherds it may be
that we will come to recommend that only the C18 component
need be considered. Clearly, these findings constitute an important
step forward in the direct dating of archaeological ceramics,
targeting for the first time pure organic compounds derived from
organic residues of commodities processed in the vessels during
their use.

ACKNOWLEDGMENT
The research was supported by a Natural Environment
Research Council (NERC) grant to R.P.E. and R.E.M.H. (GR3/
10641). Jim Carter and Andy Gledhill are thanked for assistance
with GC/MS and GC-C-IRMS. NERC are also thanked for organic,
stable isotope and accelerator MS facilities. We thank Andy
Chapman, Rob Perrin, Alistair Barclay, Tim Allen, Gill Hey,
Frances Healey, Roger Mercer, Peter Woodward, and Stephen
Minnit for provision of pottery and advice on aspects of the
archaeology.
Received for review December 6, 2002. Accepted April 17,
2003.
AC020743Y

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