Forensic Science International 163 (2006) 59–80

www.elsevier.com/locate/forsciint

Forensic comparison of soil samples: Assessment of small-scale

spatial variability in elemental composition, carbon and
nitrogen isotope ratios, colour, and
particle size distribution
Kenneth Pye a,*, Simon J. Blott a, Debra J. Croft a,b, James F. Carter c
a
Kenneth Pye Associates Ltd., Crowthorne Enterprise Centre, Crowthorne Business Estate,
Old Wokingham Road, Crowthorne, Berkshire RG45 6AW, UK
b
Croft Scientific and Technical, Blaen-y-Waun, Llanafan, Ceredigion SY23 4BD, UK
c
Mass Spec Analytical Ltd., Building O8P, Golf Course Lane, Filton, Bristol BS99 7AR, UK
Received 11 July 2005; received in revised form 3 November 2005; accepted 3 November 2005
Available online 13 December 2005

Abstract

Small-scale (<1 m2) spatial variability in soil properties was investigated at two locations in Berkshire, UK. At each site, nine
samples were collected sequentially in a regular grid pattern. The samples were compared in terms of four properties: major and
trace element composition of the <150 mm size fraction determined by inductively coupled plasma spectrometry (ICP-AES and
ICP-MS), stable carbon and nitrogen isotope ratios of the bulk <150 and >150 mm fractions determined by isotope ratio mass-
spectrometry (IRMS), colour of the <150 mm fractions determined by spectrophotometry, and particle size distribution determined
by laser granulometry. Results showed that analytical and within-site variations were much smaller than between-site variations,
and that the two sampling localities could be readily distinguished using any of the four soil properties. Significant within-site
variation in the elemental composition and nitrogen isotope ratio was found at both sites. One site also showed significant within-site
variability in particle size. Colour properties and carbon isotope ratios showed relatively low variability at both sites. Considering
the data as a whole, the two sites could be readily differentiated on the basis of a single sample from each site, using a minimum of
three comparison criteria. However, in order to adequately assess the potential variability at this scale it is recommended that a
minimum of three, and preferably five or more, samples should be taken to assess variation within a localised area of forensic
interest. Additional samples should also be taken from the wider surrounding area for purposes of comparison.
# 2005 Elsevier Ireland Ltd. All rights reserved.

Keywords: Soils; Major elements; Trace elements; Particle size distribution; Colour; Stable isotopes; Sampling; Principal components
analysis; Agglomerative hierarchical cluster analysis; ICP-AES; ICP-OES; ICP-MS; IRMS; Laser granulometry; Spatial variation

1. Introduction

A key task in forensic soil investigations is to compare a

questioned sample (e.g. from an item of clothing, footwear
* Corresponding author. Tel.: +44 1344 751610; or a digging implement) with samples taken from known
fax: +44 1344 751610. locations of interest to the enquiry (e.g. a crime scene) or
E-mail address: k.pye@kpal.co.uk (K. Pye). laboratory reference samples. There are three possible

0379-0738/$ – see front matter # 2005 Elsevier Ireland Ltd. All rights reserved.
doi:10.1016/j.forsciint.2005.11.008
60 K. Pye et al. / Forensic Science International 163 (2006) 59–80
conclusions which can be drawn from such comparisons:
(1) the questioned sample definitely did not come from the
location of interest (i.e. is excluded); (2) the questioned
sample could have come from the location of interest; and
(3) the questioned sample almost certainly did come from
the location of interest. The reliability of the conclusion
drawn is dependent on a number of factors, including (a) the
precision of the methods used to make the comparisons, (b)
the degree of variation which exists in the soil properties
considered at the location of interest, and (c) how common
the observed points of similarity or difference are in the
wider environment.
The precision with which a sample can be analysed
using any particular technique can be determined experi-
mentally. This involves determination of the instrumental
measurement precision (repeatability) and variation due
to sub-sampling and sample preparation in the laboratory
(method precision, or reproducibility). In all reputable
geochemistry, sedimentology, and soil laboratories repeat
analytical determinations are routinely made on individual
sub-sample preparations, and on replicate preparations.
Each analytical run normally includes several certified
reference materials (CRMs) and internal laboratory stan-
dards to provide both short- and long-term checks on the
quality of the results obtained. Issues of instrumental
measurement and method precision have been discussed
extensively elsewhere [1–4]. However, there is relatively
little published information regarding the magnitude of
small-scale (<1 m) spatial variability in soil and sediment
properties relative to instrumental measurement and
method precision. Furthermore, there is little published
guidance regarding the number of samples which should
be collected in order to adequately represent an area of
forensic interest such as a crime scene, how large the
samples should be, or what sampling pattern should be
adopted. Against this background, a study was undertaken
to assess small-scale spatial variability in four surface soil
properties at two locations in Berkshire, UK. The soil
properties selected were: (1) major and trace element
concentrations, (2) carbon and nitrogen isotope ratios,
(3) colour, and (4) particle size distribution.
2. Characteristics of the study sites
The study sites were at Simon’s Wood, located 2 km west
of Crowthorne (British National Grid reference SU 8133
6362), and Arborfield Bridge (SU 7449 6776), to the southeast
of Reading (Fig. 1). At Simon’s Wood, the soils are essentially
natural and are developed primarily on unconsolidated quartz-
rich, gravelly sands of the Camberley Sand Formation. The
‘soils’ at Arborfield Bridge are a mixture of locally derived
soil from the surrounding area, decayed leaf matter, and
materials derived from fly-tipping onto an asphalt and con-
crete surface. These sites represent actual crime scenes (relat-
ing to an armed robbery in the case of Simon’s Wood and a
murder in the case of Arborfield Bridge), and are representa-
tive of environments (woodland and waste ground) commonly
encountered in forensic case work.
The Simon’s Wood site is surrounded by mixed woodland
containing oak, birch, pine, and sweet chestnut, with Well-
ingtonia trees within 100 m. The understorey and ground
vegetation consists of saplings, bracken, brambles, grass
species, and moss. The soils consist of a 0–10 cm thick A0
horizon which in undisturbed profiles overlies an A1 horizon
(10–15 cm thick), a bleached A2 horizon (up to 50 cm thick)
and an iron-stained B horizon (up to 30 cm thick). The
underlying Camberley Sands (C horizon) are weakly cemen-
ted, especially near the surface, and consist largely of medium
to fine quartz sand grains with variable development of quartz
overgrowths and extensive silica dissolution features. Feld-
spars and glauconite grains are also present in low percen-
tages. The soil profile characteristics reflect podsolization
processes. In areas exposed to heavy pedestrian pressure
the A0 and A1 horizons are frequently eroded, and on steeper
slopes the soils are gullied, with water-transported sand
forming depositional aprons in lower lying areas.
The Arborfield Bridge site represents a hard-standing
area adjacent to the A327 road, and lies between the River
Loddon and a subsidiary stream channel. At the time of
sampling, the hard-standing was partly covered by ‘soil’ up
to 10 cm deep and contained abundant organic matter
derived from surrounding trees (mainly oak, poplar, haw-
thorn), bushes and brambles. There was no visible soil
profile development and considerable variation in the thick-
ness and visual appearance of the surface.
3. Methods
3.1. Sample collection
For the purposes of this study, a small area at each site was
selected for sampling. At Simon’s Wood, a sampling grid was
purposefully located on an ill-defined pathway through the
wood where footwear impressions were found following the
armed robbery. At Arborfield Bridge, a second sampling grid
was purposefully located in an area close to the location where
a burning body was discovered and where it is most likely that
an offender’s vehicle would have been parked.
Soil samples were collected at each of nine nodes within
each grid. Each sample was taken from an area of
10 cm  10 cm, to a depth of 1 cm, using a stainless steel
trowel. Samples were taken sequentially and numbered clock-
wise starting at the centre of each grid (Fig. 2). The samples
were placed in plastic pots and stored in a refrigerator at 3 8C
until required for further preparation and analysis.
3.2. Sub-sampling and sample preparation
Prior to sub-sampling, the bulk samples were homoge-
nized in the laboratory by manual mixing for 2 min using a
 K. Pye et al. / Forensic Science International 163 (2006) 59–80 61

Fig. 1. Locations and geological settings of the experimental sites.

stainless steel spatula and then split into two parts. Particle
size distribution analysis was carried out on one of the parts
after removal of leaves, twigs and other coarse debris using a
2 mm woven mesh sieve. Sub-samples weighing ca. 30 g
were then taken from the other part in ca. 2 g increments
using a stainless steel spatula. These were then separated into
<150 and >150 mm fractions by washing through ‘single
use’ plastic mesh [2]. The <150 mm fractions were oven
dried at 40 8C and then split into two parts, one of which was
ground to a fineness of ca. 10 mm using an agate pestle and
mortar. The >150 mm fractions were air dried for 48 h and
then ground using a pestle and mortar.
62 K. Pye et al. / Forensic Science International 163 (2006) 59–80
Fig. 2. Sequential sample numbering system used at both sites, and
different sample subsets used for statistical comparison.
3.3. Elemental analysis by ICP-AES and ICP-MS
Solutions were prepared from the ground <150 mm
fractions for ICP-AES and ICP-MS analysis at the Univer-
sity of Greenwich, UK, using the equipment and procedures
previously described by Pye et al. [2]. Analytical precision
for most elements determined by ICP-AES has previously
been shown to lie typically in the range 1–3%, and for
elements determined by ICP-MS typically in the range 2–5%
[5–7]. In this study, analytical and sub-sampling precision
were assessed using one soil sample from each grid (samples
A1 and B1), each of which was sub-sampled three times and
the solutions analysed in triplicate.
3.4. Light stable isotopic ratio analysis by isotope ratio
mass spectrometry (IRMS)
Three sub-samples of each bulk <150 mm soil fraction
were analysed for their carbon and nitrogen isotopic ratios,
together with two sub-samples of each of the >150 mm
fractions (insufficient >150 mm material was available to
allow triplicate analyses). Analysis was performed on the
bulk (combined organic and inorganic) material present in
both fractions. Although useful additional information can
often be gained by investigating the isotopic ratios of
separate organic and inorganic fractions [8–10], previous
work has shown that for preliminary forensic comparison
purposes it is often sufficient to consider the isotopic ratios
of bulk material [4].
Isotopic measurements were performed using a Thermo-
Finnigan Delta XP stable isotope ratio mass spectrometer
with a ConFlo III interface. Samples were crimped into tin
capsules (Elemental Microanalysis, UK) and introduced
using an AS200 auto-sampler. The oxidation reactor, com-
prising chromic oxide and silver/cobaltous oxide, was main-
tained at a temperature of 900 8C and the reduction reactor,
comprising electrolytic copper, was maintained at 680 8C.
The nitrogen and carbon dioxide formed by combustion/
reduction were separated using a gas chromatography col-
umn containing Porapak QS. The helium carrier gas was
maintained at 150 ml min 1 and a 5 s pulse of oxygen (BOC
Research Grade N5.5) introduced at a flow rate of
175 ml min 1. Water was removed by an anhydrous mag-
nesium perchlorate (supplier) trap. Isotopic values were
referenced against gaseous carbon dioxide (BOC CP Grade
N4.5), which was in turn calibrated against PEF1 polyethy-
lene foil (National Institute for Standards and Technology
(NIST) Reference Material RM8540) and gaseous nitrogen
(BOC Research Grade N5.5), which was in turn calibrated
against NBS19 USGS25 ammonium sulphate (NIST
RM8550). Data were acquired and processed using Thermo
Electron ISODAT NT software.
The isotopic ratio for carbon represents the ratio of the
two stable isotopes 13C and 12C measured in parts per
thousand (per mil, or %) relative to the international stan-
dard, Vienna PeeDee Belemnite (VPDB), and is expressed
by the notation d13C. The isotopic ratio for nitrogen repre-
sents the ratio between the two stable isotopes 15N and 14N
measured in parts per mil relative to atmospheric nitrogen,
and is expressed as d15N [11].
The precision of the measured values (which includes
both analytical and sub-sampling precision) was assessed by
comparing the results from the different sub-samples. Ana-
lytical precision, based on repeat analyses of standard
reference materials, was within 0.1% for both d13C and
d15N, which was comparable with results from previous
studies [4,12,13].
3.5. Colour analysis by spectrophotometry
Colour analysis was carried out on both the unground and
ground <150 mm solid powder fractions. A Minolta1
CM2002 spectrophotometer was used to obtain reflectance
data over the 400–700 nm wavelength range. The instrument
was calibrated negatively to a closed black box (zero reflec-
tance) and positively to an International Standard white tile
(100% reflectance) before each use. Samples were tested on
a clean, white ceramic tile background, under a standardised
illumination system of average daylight, including the ultra-
violet wavelength region, at a temperature of 6504 K. The
specular component was included in the analyses to reflect
the largely crystalline nature of many geological materials.
Two sets of colour indices were obtained from the analysis:
 K. Pye et al. / Forensic Science International 163 (2006) 59–80
Munsell indices of Hue, Value and Chroma, and CIE L*a*b*
indices, as well as the percentage of light reflected for each
10 nm interval of the wavelength from 400 to 700 nm [13].
Previous studies have demonstrated that analytical pre-
cision is <0.1% for repeat measurements made over periods
of several minutes, and repeat sub-sampling and measure-
ment of the same soil sample over several days has a
precision of 4–5% for the L*, a* and b* indices [13]. In
this study, sub-sampling precision was assessed by analysing
five sub-samples from one soil sample at each sampling
location. Measurement precision was quantified by five
determinations on each sub-sample.
3.6. Particle size distribution analysis by laser
granulometry
A CoulterTM LS230 laser granulometer, with a variable-
speed fluid module, was used to determine the particle size
distributions of the bulk size fractions in the size range 0.04–
2000 mm. Particle size distributions were calculated from the
diffraction data using the Fraunhofer model [3,14]. Several
statistical parameters were derived from the primary data,
including mean, median, mode, standard deviation, D10 (10th
percentile of the distribution), D90 (90th percentile of the
distribution) and the percentages of sand, silt, and clay. A
graphical output of volume percentage in 116 size classes
spanning the size range 0.04–2000 mm was also obtained for
each sample.
Previous experiments have demonstrated that measure-
ment precision for the Coulter LS230 laser granulometer is
better than 0.03% for the mean size of glass bead standard
samples. Typical values for combined instrumental measure-
ment and sub-sampling error are of the order of 0.1% for well
sorted coastal dune sands and 6.5% for poorly sorted silty soils
[14]. In this study, precision was quantified by taking five sub-
samples from one soil sample from each grid (samples A1 and
B1) and analyzing each sub-sample five times.
4. Results
4.1. Spatial variability in major and trace elemental
composition
A mean coefficient of variation for all elements of 2.2%
was obtained for the Simon’s Wood sample A1 and 1.4% for
the Arborfield Bridge sample B1, as discussed by Pye et al.
[2]. The variation due to sub-sampling was found to be
significantly larger, with a mean CV of 11.1% for the
Simon’s Wood samples and a mean CV of 3.9% for the
Arborfield Bridge samples. A number of elements showed
particularly high coefficients of variation, notably W, V, and
La in the Simon’s Wood sub-samples, and W and Pb in the
Arborfield Bridge samples.
Tables 1 and 2 summarise the within-grid variation in
elemental composition at each site. Variability within each
63
grid was quantified by calculating the mean, range, and
coefficient of variation (CV) for each element. The Simon’s
Wood grid was found to be more heterogeneous than the
Arborfield Bridge grid, with an average CV for all elements
of 27.9% and 20 elements showing a CV higher than 20%.
By contrast, the average CV for all elements at Arborfield
Bridge was 14.5%, with just five elements having a CV of
>20%. Although the variability for each element was not the
same for the two grids, several elements show similar trends,
for example W, Tl, Mo, V, Pb, Sn, and CaO showed con-
siderable variability in both grids, while Al2O3, SiO2, Na2O,
and Y were least variable at both sites.
The possibility of spatial trends within the grids, and the
effects of different sampling patterns, were investigated by
examining different subsets of samples from each grid and
comparing the observed means and ranges with those for all
nine samples. Eight sample subsets were defined, incorporat-
ing three, four, or five samples and representing different
spatial arrangements, as shown in Fig. 2. Tables 1 and 2 show
the mean and range of elemental values in each of the subsets,
expressed as a percentage of the total range. In general, at both
sites a higher percentage of the total range was found in
subsets containing larger numbers of samples. At the Arbor-
field Bridge site, the sample subset which included the corners
and centre of the grid (five samples) represented >50% of the
total variation in all of the elements, and 100% of the total
variation in more than half of the elements. At Simon’s Wood,
100% of the total variation was represented by 5 samples for
only 14 of the elements, but more than 50% of the total
variation range was recorded for all of the elements. No
consistent pattern was evident in terms of the order of ele-
ments for each sample subset or for each site.
The effect on the cumulative mean values of adding
sequential samples is illustrated in Fig. 3. A reasonable
approximation of the population mean (represented by nine
samples) was obtained by taking three samples, although in
most cases the approximation was improved by taking five
samples. Averaging based on six, seven or eight samples
only slightly improved the approximation compared with the
average based on five samples.
Distinct differences between the two sites are apparent on
the basis of their elemental composition. For example, the
Simon’s Wood samples contain markedly more SiO2 but less
Al2O3 and CaO than the Arborfield Bridge samples. One
statistical measure of similarity is provided by Pearson’s
product-moment correlation coefficient, a dimensionless
parameter which ranges in value from 1 (perfect negative
correlation) through 0 (no correlation) to +1 (perfect positive
correlation). Matrices of Pearson’s correlation coefficients
were constructed for the chemical data set as a whole and
also separately for the major, trace (excluding rare earths),
and 14 rare earth elements, as described by Pye et al. [15].
An example using the entire chemical data set is shown in
Table 3. The samples from within each site are clearly more
similar to each other (show higher correlation coefficients)
than to samples from the other site. Systematic variations
64 K. Pye et al. / Forensic Science International 163 (2006) 59–80

Table 1
Average and range of elemental concentrations in samples from the Simon’s Wood grid, together with the percentage of the total range recorded
in different sample subsets (see Fig. 2 for subset identification)
All samples (n = 9) Percentage of the total range recorded
Mean Maximum Minimum CV% Subset Subset Subset Subset Subset Subset Subset Subset
A (n = 3) B (n = 3) C (n = 3) D (n = 3) E (n = 4) F (n = 4) G (n = 5) H (n = 5)
ICP-AES elements
Al2O3 1.93 2.22 1.73 7.7 100.0 30.8 19.1 59.2 60.9 70.0 60.9 97.2
Ba 279.61 313.20 240.30 8.7 86.6 27.8 56.1 100.0 77.9 100.0 77.9 100.0
Be n/a <LoQ <LoQ n/a n/a n/a n/a n/a n/a n/a n/a n/a
CaO 0.88 1.48 0.34 43.6 78.5 55.2 45.0 100.0 74.2 100.0 74.2 100.0
Fe2O3 0.41 0.83 0.28 41.6 100.0 12.6 20.2 99.2 33.6 99.2 33.6 100.0
K2O 0.96 1.05 0.91 4.5 25.9 19.0 81.0 57.3 64.8 83.2 64.8 83.2
MgO 0.07 0.13 0.03 50.9 87.1 42.7 34.6 100.0 62.8 100.0 64.1 100.0
MnO 0.01 0.02 0.01 30.5 73.0 66.1 46.0 82.9 54.1 100.0 63.1 100.0
Na2O 0.09 0.10 0.08 8.1 69.8 85.9 34.4 79.7 99.5 66.1 100.0 80.2
Ni n/a <LoQ <LoQ n/a n/a n/a n/a n/a n/a n/a n/a n/a
P2O5 0.08 0.11 0.05 27.1 91.9 62.5 36.7 100.0 63.0 100.0 63.0 100.0
Sc <LLQ
SiO2 78.07 86.98 66.97 8.9 93.9 55.5 11.5 71.4 72.4 74.6 78.6 100.0
TiO2 0.33 0.39 0.27 11.3 38.4 78.6 35.6 78.4 64.4 78.4 64.4 78.4
Y 9.31 10.24 7.87 7.9 51.6 78.9 38.4 72.8 85.7 72.8 85.7 72.8
Zn 56.10 82.32 32.86 28.7 75.2 60.7 41.6 91.4 83.8 91.4 83.8 91.4
ICP-MS elements
V n/a 25.98 <LoQ n/a n/a n/a n/a n/a n/a n/a n/a n/a
Cr 38.47 52.23 24.81 23.7 21.9 100.0 36.5 61.0 82.6 78.3 82.6 78.3
Co 0.98 2.08 0.58 46.5 100.0 24.2 4.7 87.2 33.9 91.9 33.9 92.7
Cu 10.88 23.09 0.00 57.7 61.4 32.2 42.8 100.0 23.1 100.0 23.1 100.0
Ga 2.17 2.80 1.77 13.7 100.0 18.4 15.3 68.3 48.3 70.1 48.3 95.2
Rb 26.37 28.57 23.92 6.0 100.0 25.6 44.8 33.0 80.8 64.0 80.8 94.8
Sr 33.09 37.17 29.38 7.5 100.0 48.4 34.7 77.0 82.3 83.2 82.3 83.2
Zr 371.82 474.11 203.67 25.5 82.5 69.0 26.3 100.0 77.8 100.0 77.8 100.0
Nb 4.95 5.66 4.04 10.8 51.4 83.8 39.3 64.8 73.3 64.8 73.3 64.8
Mo 0.57 1.15 0.06 67.8 56.6 83.0 15.7 100.0 67.6 100.0 67.6 100.0
Sn 1.66 2.51 0.88 33.3 100.0 76.4 18.3 56.7 60.8 76.4 60.8 76.4
Cs 1.06 1.30 0.88 15.0 100.0 35.3 57.8 89.2 84.9 89.2 84.9 94.9
La 6.92 9.48 2.76 29.0 9.7 80.1 64.7 50.7 16.4 82.2 43.9 100.0
Ce 14.42 18.16 10.05 16.6 33.7 74.2 100.0 20.9 75.2 74.2 75.2 74.2
Pr 1.59 1.93 0.83 20.1 14.5 86.8 100.0 17.2 39.3 86.8 39.3 90.3
Nd 6.06 7.82 3.77 18.3 21.8 71.7 100.0 11.8 64.2 71.7 64.2 71.7
Sm 1.15 1.52 0.81 20.9 38.8 67.3 100.0 54.4 90.0 67.3 90.0 67.3
Eu 0.23 0.28 0.19 12.0 19.0 51.2 98.8 38.1 100.0 54.8 100.0 54.8
Gd 1.02 1.22 0.79 13.6 38.0 73.5 100.0 42.3 85.2 73.5 85.2 73.5
Tb 0.17 0.20 0.14 12.9 71.2 100.0 78.0 59.3 100.0 78.0 100.0 78.0
Dy 1.09 1.27 0.87 12.4 77.9 100.0 54.3 59.3 90.8 77.9 90.8 77.9
Ho 0.26 0.30 0.20 13.7 83.0 100.0 44.7 74.5 93.6 83.0 93.6 83.0
Er 0.79 0.89 0.60 13.2 73.3 95.0 50.2 78.3 97.5 78.3 97.5 78.3
Tm 0.13 0.15 0.09 15.4 74.5 90.9 41.8 83.6 92.7 83.6 92.7 83.6
Yb 0.92 1.11 0.65 17.1 78.4 89.4 38.9 77.1 100.0 78.4 100.0 78.4
Lu 0.15 0.17 0.10 17.0 80.9 94.1 36.8 86.8 100.0 83.8 100.0 83.8
Hf 8.60 10.63 4.71 25.4 85.7 77.7 28.1 100.0 84.2 100.0 84.2 100.0
Ta 0.40 0.48 0.32 12.9 50.3 100.0 34.0 61.4 69.9 80.4 69.9 80.4
W n/a 3.20 <LoQ n/a n/a n/a n/a n/a n/a n/a n/a n/a
Tl 0.16 0.37 0.07 56.8 22.8 50.5 29.7 86.1 80.2 50.5 93.1 50.5
Pb 18.40 25.82 8.74 34.6 97.9 98.5 63.0 26.0 79.2 100.0 79.2 100.0
Th 2.26 2.98 1.62 19.7 45.6 75.2 100.0 28.6 94.7 75.2 94.7 75.2
U 0.90 1.02 0.75 10.5 70.6 95.2 77.3 57.2 100.0 72.5 100.0 72.5
Mean 27.9 64.6 66.4 50.2 70.4 72.8 82.7 74.0 85.9
n/a indicates cases where element concentrations in one or more samples were below the limit of quantitation (LoQ).
 K. Pye et al. / Forensic Science International 163 (2006) 59–80 65

Table 2
Average and range of elemental concentrations in samples from the Arborfield Bridge grid, together with the percentage of the total range
recorded in different sample subsets (see Fig. 2 for subset identification)
All samples (n = 9) Percentage of the total range recorded
Mean Maximum Minimum CV% Subset A Subset B Subset C Subset D Subset E Subset F Subset G Subset H
(n = 3) (n = 3) (n = 3) (n = 3) (n = 4) (n = 4) (n = 5) (n = 5)
ICP-AES elements
Al2O3 5.52 5.98 5.12 5.8 47.5 88.3 57.7 85.7 100.0 87.3 100.0 87.3
Ba 479.18 597.50 427.30 12.0 89.6 68.8 9.8 100.0 62.7 100.0 62.7 100.0
Be 1.09 1.23 1.01 6.3 36.3 39.1 59.1 72.6 93.5 45.6 93.5 45.6
CaO 3.75 4.81 2.92 20.0 77.3 97.5 100.0 69.1 79.8 97.5 88.2 97.5
Fe2O3 3.72 3.98 3.45 5.4 63.3 34.2 47.9 100.0 76.9 81.4 79.3 81.4
K2O 1.30 1.36 1.26 2.4 27.7 75.2 100.0 34.7 40.6 100.0 40.6 100.0
MgO 1.29 1.56 1.00 15.5 67.5 80.5 100.0 77.8 69.6 97.9 73.4 97.9
MnO 0.08 0.10 0.07 13.0 48.5 50.0 74.1 96.2 47.5 100.0 47.5 100.0
Na2O 0.26 0.32 0.24 8.8 85.4 33.0 38.8 85.9 5.3 85.9 19.8 100.0
Ni 25.83 28.07 21.55 8.9 79.9 77.9 67.3 92.2 20.1 89.4 20.1 89.4
P2O5 0.50 0.56 0.44 8.0 53.7 62.4 56.2 71.7 43.8 36.3 43.8 88.4
Sc 5.09 5.61 4.58 7.3 56.8 38.6 44.0 82.8 82.6 58.6 100.0 68.2
SiO2 50.64 55.08 45.08 5.9 23.2 38.5 64.6 80.6 68.5 38.5 68.5 71.6
TiO2 0.45 0.48 0.41 4.8 61.8 76.8 34.4 65.6 6.1 100.0 34.0 100.0
Y 19.49 20.47 18.10 3.9 100.0 50.2 24.5 58.2 65.0 74.7 65.0 81.0
Zn 365.84 401.10 333.80 6.1 38.6 26.9 100.0 39.5 64.3 100.0 64.3 100.0
ICP-MS elements
V 61.43 83.59 43.25 23.9 100.0 97.0 53.2 57.2 80.4 76.8 80.4 76.8
Cr 74.15 86.41 60.30 14.2 73.5 100.0 18.4 84.3 99.4 84.8 99.4 84.8
Co 9.57 10.84 7.77 11.9 91.6 88.7 66.4 58.4 57.9 100.0 78.6 100.0
Cu 78.70 93.04 66.01 11.6 70.5 48.2 100.0 61.6 41.4 100.0 47.5 100.0
Ga 6.76 7.92 5.63 12.1 77.7 96.5 64.8 57.9 55.1 96.5 81.3 100.0
Rb 60.84 72.34 50.42 13.2 79.9 100.0 75.8 65.7 63.1 100.0 73.9 100.0
Sr 72.09 81.57 56.16 11.6 43.0 20.6 52.5 65.8 56.5 43.0 90.7 100.0
Zr 237.80 311.98 159.77 20.9 34.1 68.6 74.6 74.3 88.4 68.6 88.4 68.6
Nb 7.78 9.48 6.30 14.7 75.2 100.0 79.8 63.8 67.3 100.0 67.3 100.0
Mo 2.85 3.91 2.14 19.0 44.4 100.0 50.9 42.8 86.7 56.1 86.7 56.1
Sn 7.09 9.02 5.14 17.7 48.6 75.6 46.2 84.8 53.8 75.6 53.8 80.8
Cs 2.87 3.34 2.37 12.5 80.2 100.0 63.3 67.9 58.9 100.0 75.5 100.0
La 19.30 23.58 14.51 18.4 100.0 98.9 64.7 78.7 95.4 83.3 95.4 83.3
Ce 42.51 48.41 33.84 14.2 97.6 100.0 81.9 83.4 79.3 100.0 79.3 100.0
Pr 4.61 5.42 3.57 16.1 93.4 100.0 71.9 81.6 92.7 88.9 92.7 88.9
Nd 17.86 20.33 14.46 13.9 91.7 100.0 79.0 80.6 90.7 92.6 90.7 92.6
Sm 3.45 3.97 2.84 13.2 89.7 100.0 76.6 76.2 91.9 94.3 91.9 94.3
Eu 0.78 0.89 0.64 12.8 87.3 100.0 79.6 80.8 70.6 100.0 70.6 100.0
Gd 3.08 3.55 2.53 14.5 100.0 95.5 77.6 86.3 97.2 95.0 97.2 95.0
Tb 0.49 0.56 0.42 13.2 100.0 99.3 82.7 86.0 97.2 99.3 97.2 99.3
Dy 2.87 3.32 2.37 13.9 95.0 100.0 78.1 83.2 88.3 100.0 88.3 100.0
Ho 0.64 0.74 0.52 14.5 98.1 100.0 77.8 76.0 88.9 100.0 88.9 100.0
Er 1.77 2.05 1.46 14.6 96.4 100.0 78.9 77.0 90.0 100.0 91.1 100.0
Tm 0.26 0.31 0.22 15.3 86.7 100.0 86.3 75.6 96.7 100.0 96.7 100.0
Yb 1.71 2.11 1.37 15.4 74.9 100.0 78.4 60.3 65.4 100.0 65.4 100.0
Lu 0.25 0.30 0.21 15.5 81.3 93.8 81.5 81.3 91.7 93.8 91.7 93.8
Hf 5.95 7.86 4.01 22.0 43.0 73.6 78.0 76.0 87.6 73.6 87.6 73.6
Ta 0.61 0.76 0.49 16.7 64.2 95.6 89.9 65.3 63.8 95.6 73.4 100.0
W 0.89 1.74 0.44 45.5 53.1 100.0 80.3 40.9 50.1 100.0 50.1 100.0
Tl 0.22 0.42 0.08 53.6 60.0 100.0 78.9 57.4 44.3 100.0 48.4 100.0
Pb 30.42 40.37 20.14 19.8 52.8 55.5 67.0 89.5 49.0 67.0 58.0 67.0
Th 5.34 6.59 4.29 15.5 68.5 100.0 84.2 61.4 57.9 100.0 62.7 100.0
U 1.52 1.78 1.24 14.1 84.5 100.0 83.1 70.4 82.0 100.0 82.0 100.0

Mean 14.5 71.3 80.5 69.0 72.8 69.5 87.3 74.0 91.1
66 K. Pye et al. / Forensic Science International 163 (2006) 59–80

Fig. 3. Concentrations of selected major oxides and trace elements in soil samples A1–A9 at Simon’s Wood (a–d) and samples B1–B9 at
Arborfield Bridge (e–h). The solid lines show the cumulative averages.

within the Simon’s Wood grid are also apparent, with
samples from one side of the grid (A7, A8, and A9) showing
lower correlation coefficients with samples from other parts
of the grid. By comparison, the Arborfield grid appears more
homogeneous using this method of comparison. Examina-
tion of separate correlation matrices for the major, trace, and
rare earth elements showed that the within-grid differences
are attributable mainly to trace elements rather than to the
major elements or rare earths.
Fig. 4a compares the Simon’s Wood and Arborfield
Bridge samples in terms of a plot of SiO2/Al2O3 ratio against
SiO2/Fe2O3 ratio. The two sites are clearly differentiated but
Table 3
Pearson’s correlation values obtained by comparing <150 mm soil fractions from Simon’s Wood and Arborfield Bridge in terms of the concentrations of all 49 elements determined by ICP-AES
and ICP-MS

K. Pye et al. / Forensic Science International 163 (2006) 59–80

The highest correlation values are highlighted.

67
68 K. Pye et al. / Forensic Science International 163 (2006) 59–80

Fig. 4. Example bivariate plots comparing intra-grid variability at Simon’s Wood and Arborfield Bridge grids in terms of selected major element
ratios (a and b), trace elements (c–e) and trace element ratios (f).

the Arborfield Bridge site shows a much higher degree of
clustering (i.e. less intra-grid variability) than the Simon’s
Wood site. A similar pattern is shown by a plot of Al2O3/
Fe2O3 against Al2O3/K2O (Fig. 4b). Plots of several indivi-
dual trace element concentrations, such as Rb versus Sr, U
versus Th, and Nd versus Sm (Fig. 4c–e), also allow clear
discrimination between the two sites and show that values
for the Simon’s Wood grid are more tightly clustered than
those for the Arborfield Bridge grid. However, such differ-
ences are not necessarily apparent when trace element ratios
are considered (e.g. Fig. 4f).
It is well known that some element concentrations in soils
and sediments are positively correlated while others are
negatively correlated. The selection of the most suitable
elements for forensic discrimination purposes is to some
extent dependent on the nature of the soils or sediments in
question. However, experience has shown that a combination
of several different elements, and their ratios, which may show
either positive or negative correlation behaviour, can be
usefully employed. Plots of SiO2/Al2O3 versus SiO2/Fe2O3,
Al2O3/Fe2O3 versus Al2O3/K2O, Nd versus Sm, Ce versus La,
U versus Th, and Rb versus Sr have frequently proved valuable
in practice [15].
After removing elements which were below the limit
of quantitation, principal components analysis (PCA) was
performed on the elemental concentration data. Fig. 5a
 K. Pye et al. / Forensic Science International 163 (2006) 59–80 69

Fig. 5. Plots of the first two axes of the principal components analyses using data for the nine samples from each grid: (a) based on ICP-AES and
ICP-MS data for all elements except Be, Ni, Sc, V, and W which were below the limits of quantitation in at least one sample, and (b) based on
particle size parameters (mean, median, mode, D10, D90, % sand, % silt, and % clay).

shows that a plot of the first two components, which
together represent 95.5% of the total variance in the
dataset, clearly distinguishes between the two grids.
Variability within each grid is far less than the differences
between the grids, although samples A8 and A9 are
notably different from the other Simon’s Wood samples,
and the Arborfield Bridge samples cluster into two main
groups, B1–B4 and B5–B9. Fig. 6a shows the results of
agglomerative hierarchical cluster analysis (AHC) in the
form of a dendrogram, and also clearly demonstrates that
variability within each grid is far less than that between
the two grids. The figure also clearly shows that samples
A8 and A9 are clearly distinguishable from the other
Simon’s Wood samples, and that the Arborfield Bridge
samples fall into two sub-groups.
PCA was repeated for the eight subsets of samples at each
site shown in Fig. 2. In each case, a clear separation between
the two sites was found, and it could be concluded that the
samples were from different populations on the basis of just
three samples from each grid. Increasing numbers of sam-
ples did not significantly increase the discrimination
between the two sites, but did increase the confidence that
there are no overlapping outliers in the two groups.
The probability that the samples come from different
populations was also examined using Student’s t-test. A
statistically significant difference was found between the
two sites for all subsets of samples in terms of most elements
(Table 4). A few elements, especially Pb, Zr, and Hf, showed
significant variability, and it was not possible to distinguish
between the sites on the basis of only a small number of
determinations of these elements. For all elements, better
discrimination between the grids was achieved by increasing
the number of samples.
4.2. Spatial variability in stable carbon and nitrogen
isotope ratios
Table 5 summarises the technique precision based on
triplicate sub-samples of the <150 mm fractions from the
two sampling locations. The precision for d13C was found to
be very good (0.2–0.3%), but an order of magnitude lower
for d15N (5.0–16.4%).
Table 6 shows the d13C and d15N data for different
subsets of samples. Distinct differences between the two
grids are apparent, especially in terms of d15N. Variability
within the grids is significantly less than the differences
between the grids, d13C values at Simon’s Wood showing a
particularly low degree of variation. Bivariate plots of d13C
against d15N for the <150 and >150 mm fractions are shown
in Fig. 7. The Simon’s Wood data for both size fractions
cluster tightly, while the Arborfield Bridge data show a much
higher degree of scatter.
The effect on the cumulative mean of adding sequential
samples is shown in Fig. 8. A fairly good approximation of
the mean of all nine samples was obtained with just three
samples, although a slightly better approximation was
achieved using five samples, especially in the case of the
Arborfield Bridge grid.
The probability that samples from the two grids come
from different populations was examined using a two-tailed
Student’s t-test. A significant difference was found between
the two sites for all sample subsets in terms of d13C, and for
most subsets in terms of d15N (Table 7). Although a higher
degree of discrimination is possible for subsets containing a
larger number of samples, it is statistically possible to
distinguish between the sites in terms of stable isotope ratios
using five or even fewer samples.
70 K. Pye et al. / Forensic Science International 163 (2006) 59–80

Fig. 6. Agglomerative hierarchical cluster analysis dendrograms produced using data for the nine samples from each grid: (a) based on ICP data
for all elements excluding Be, Ni, Sc, V, and W, and (b) based on particle size parameters (mean, median, mode, D10, D90, % sand, % silt, and %
clay). Clustering calculated using Ward’s method, and dissimilarity expressed in terms of the Euclidean distance.

4.3. Spatial variability in soil colour
The instrumental measurement precision was better than
0.7% for all the colour parameters (Table 8). Sub-sampling
precision was also high, the maximum variation being 1.9%
for the L* parameter in the Arborfield Bridge sample.
Variation in the colour parameters for different sample
subsets is summarised in Table 9. In the case of the Simon’s
Wood unground <150 mm fractions, the largest variation
(CV = 16.7%) was found for the L* parameter. Variation was
generally much less (<8.0%) for the ground fractions. In the
case of the Arborfield Bridge samples, variation for all
parameters was in the range 5.2–8.2%, with similar varia-
bility for the ground and unground powder samples.
Bivariate plots of b* against a* provided good discrimi-
nation between the two sites for both unground and ground
<150 mm fractions (Fig. 9a and c), but a plot of a* against
L* did not provide clear discrimination between the two
groups (Fig. 9b and d). The best discrimination between the
sites was provided by the b* parameter. Within-grid varia-
bility in the a* and b* parameters was similar at both sites.
Variability in the L* parameter was greatest for the unground
Simon’s Wood samples.
Table 9 shows that a larger proportion of the total
variation in the grid (represented by nine samples) was
obtained with five samples in the subset than with three
or four samples. Subsets of five unground samples repre-
sented 71.9–97.9% of the total variation in the L*, a* and b*
parameters in the Simon’s Wood grid and 48.1–100% in the
Arborfield Bridge grid. At both sites, the corner samples plus
the middle sample (subset H) included the greatest percen-
tage of the total range.
The probability that samples from the two grids come
from different populations was examined using Student’s t-
test. The results (Table 10) confirmed that b* is a good
discriminator, and show that the grids could easily be
differentiated on the basis of only three samples. The L*
and a* parameters were found to be less discriminatory, even
when all nine samples from each grid were considered.
4.4. Spatial variability in particle size distribution
The instrumental precision of the laser granulometer was
generally high for all parameters, with a coefficient of
variation for the mean of 2.5–3.3% for Simon’s Wood
and 1.3–4.7% for Arborfield Bridge (Table 11). Sub-sam-
 K. Pye et al. / Forensic Science International 163 (2006) 59–80 71

Table 4
Probabilities resulting from two-tailed t-tests comparing the elemental concentrations in different subsets of <150 mm soil fractions from the two
sites
All samples Subset A Subset B Subset C Subset D Subset E Subset F Subset G Subset H
(n = 9) (%) (n = 3) (%) (n = 3) (%) (n = 3) (%) (n = 3) (%) (n = 4) (%) (n = 4) (%) (n = 5) (%) (n = 5) (%)
ICP-AES elements
Al2O3 0.0 0.0 0.3 0.1 0.1 0.0 0.0 0.0 0.0
Ba 0.0 2.9 3.1 0.3 3.8 0.1 1.1 0.0 0.2
Be n/a n/a n/a n/a n/a n/a n/a n/a n/a
CaO 0.0 0.7 3.4 3.2 0.5 0.1 0.4 0.0 0.1
Fe2O3 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0
K2O 0.0 0.0 0.2 0.2 0.3 0.0 0.0 0.0 0.0
MgO 0.0 0.5 1.0 2.1 0.9 0.0 0.2 0.0 0.0
MnO 0.0 0.2 0.1 0.9 0.9 0.0 0.1 0.0 0.0
Na2O 0.0 0.7 0.0 0.2 1.1 0.0 0.1 0.0 0.0
Ni n/a n/a n/a n/a n/a n/a n/a n/a n/a
P2O5 0.0 0.0 0.1 0.1 0.0 0.0 0.0 0.0 0.0
Sc n/a n/a n/a n/a n/a n/a n/a n/a n/a
SiO2 0.0 5.6 1.8 0.2 1.3 0.1 0.8 0.0 0.1
TiO2 0.0 0.4 2.7 0.5 5.1 0.5 1.1 0.0 0.2
Y 0.0 0.1 0.0 0.0 0.0 0.0 0.0 0.0 0.0
Zn 0.0 0.0 0.0 0.2 0.0 0.0 0.0 0.0 0.0
ICP-MS elements
V n/a 6.7 n/a n/a n/a 0.5 n/a 0.1 n/a
Cr 0.0 1.6 2.7 0.2 2.1 0.2 0.3 0.1 0.1
Co 0.0 0.4 0.9 0.4 0.0 0.0 0.0 0.0 0.0
Cu 0.0 0.1 0.1 0.6 0.1 0.0 0.0 0.0 0.0
Ga 0.0 0.4 1.8 0.7 0.2 0.0 0.1 0.0 0.0
Rb 0.0 1.8 3.0 1.5 1.4 0.1 0.4 0.0 0.1
Sr 0.0 0.1 0.0 0.7 1.0 0.1 0.0 0.1 0.1
Zr 0.3 23.9 39.9 1.7 20.7 6.5 13.4 1.7 4.4
Nb 0.0 4.1 5.8 5.0 3.3 0.6 2.0 0.2 0.8
Mo 0.0 0.3 3.5 0.8 0.6 0.2 0.1 0.0 0.0
Sn 0.0 0.3 1.4 0.5 2.8 0.1 0.1 0.0 0.0
Cs 0.0 0.8 2.3 0.4 0.2 0.0 0.2 0.0 0.0
La 0.0 4.9 1.9 0.3 1.8 1.2 0.1 0.2 0.0
Ce 0.0 2.1 1.4 0.5 1.7 0.1 0.1 0.0 0.0
Pr 0.0 3.2 1.4 0.4 2.3 0.7 0.1 0.1 0.0
Nd 0.0 2.0 1.0 0.4 1.6 0.2 0.1 0.0 0.0
Sm 0.0 1.2 1.1 0.3 0.6 0.1 0.1 0.0 0.0
Eu 0.0 1.4 1.4 0.6 1.0 0.1 0.2 0.0 0.0
Gd 0.0 1.8 1.4 0.5 1.4 0.3 0.1 0.0 0.0
Tb 0.0 1.1 1.0 0.7 0.9 0.2 0.1 0.0 0.0
Dy 0.0 1.3 1.4 1.0 1.1 0.2 0.2 0.0 0.0
Ho 0.0 1.8 1.7 1.3 0.9 0.2 0.2 0.0 0.0
Er 0.0 1.8 1.8 1.3 0.9 0.3 0.2 0.1 0.1
Tm 0.0 1.5 2.2 2.3 1.1 0.5 0.2 0.1 0.1
Yb 0.0 2.0 3.4 3.1 1.2 0.3 0.6 0.1 0.2
Lu 0.0 2.0 2.5 3.3 3.2 1.0 0.4 0.3 0.2
Hf 0.8 29.6 56.5 4.5 25.5 10.3 20.6 3.0 7.3
Ta 0.0 3.0 6.5 9.8 3.9 1.1 1.9 0.7 1.8
W n/a n/a n/a n/a n/a n/a n/a n/a n/a
Tl 30.6 25.8 37.9 32.9 98.5 99.0 21.6 98.1 24.9
Pb 0.1 6.6 5.9 6.6 51.6 7.6 3.4 1.5 0.8
Th 0.0 1.5 1.9 1.1 1.2 0.1 0.3 0.0 0.1
U 0.0 3.3 3.7 2.4 2.1 0.8 0.6 0.2 0.2
Values lower than 5% indicate a significant difference between the two grids at the 95% confidence level (see Fig. 2 for subset identification).
n/a indicates cases where elemental concentration for at least one sample was below limit of quantitation (LoQ).
72 K. Pye et al. / Forensic Science International 163 (2006) 59–80

Table 5
Repeatability of carbon and nitrogen stable isotope analyses for <150 and >150 mm fractions of samples A1 (Simon’s Wood) and B1 (Arborfield
Bridge)
Sample <150 mm fraction >150 mm fraction
13 15
d C (%) d N (%) d13C (%) d15N (%)
Simon’s Wood A1
Sub-sample 1 27.34 0.63 28.13 1.48
Sub-sample 2 27.42 0.54 28.38 1.77
Sub-sample 3 27.41 0.75 – –
Mean 27.39 0.64 28.26 1.62
Standard deviation 0.04 0.11 – –
CV% 0.2 16.4 – –
Arborfield Bridge B1
Sub-sample 1 24.81 3.51 26.70 4.06
Sub-sample 2 24.66 3.39 26.73 4.11
Sub-sample 3 24.69 3.74 – –
Mean 24.72 3.55 26.71 4.08
Standard deviation 0.08 0.18 – –
CV% 0.3 5.0 – –
Three sub-samples of the <150 mm fraction were analysed, but there was sufficient material for analysis of only two sub-samples of the
>150 mm fraction. Due to the destructive nature of the technique, instrumental precision can only be estimated from repeat analyses of standard
reference materials, which is reported to be ca. 0.1% for both d13C and d15N.

pling precision for the Simon’s Wood grid was better (CV of
2.7% for the mean and 0.8% for the median) than for the
Arborfield Bridge grid (CV of 4.6% for the mean and 4.3%
for the median). The most variable parameter for both sites
was the D10 value.
The results for different subsets of samples and the entire
sample population from the two grids are shown in Table 12.
Coefficients of variation for the parameters considered were
generally lower for the Simon’s Wood samples than for the
Arborfield samples. The latter are generally much coarser
and more poorly sorted than the Simon’s Wood samples. In
the Simon’s Wood grid 100% of the total variation in the
mode was recorded by every subset consisting of three
samples. In the case of the Arborfield Bridge samples,
24.3–75.7% of the total variation in the mode was repre-
sented by the subsets containing three samples. Arborfield

Table 6
Average and range of carbon and nitrogen isotope ratios in samples from the two sites, together with the percentage of the total range recorded in
different sample subsets (see Fig. 2 for subset identification)
All samples (n = 9) Percentage of the total range recorded
Mean Maximum Minimum CV% Subset A Subset B Subset C Subset D Subset E Subset F Subset G Subset H
(%) (%) (%) (n = 3) (n = 3) (n = 3) (n = 3) (n = 4) (n = 4) (n = 5) (n = 5)
Simon’s Wood site
<150 mm fraction
d15N 0.98 0.64 1.22 20.5 42.0 23.6 64.7 40.8 48.8 82.8 100.0 94.5
d13C 27.46 27.35 27.61 0.3 100.0 89.1 44.2 50.7 60.1 84.1 60.1 86.3
>150 mm fraction
d15N 1.90 1.51 2.35 18.5 26.9 25.0 98.7 68.9 100.0 90.3 100.0 90.3
d13C 27.96 27.74 28.26 0.5 33.1 40.2 21.0 59.1 22.0 60.7 70.4 100.0
Arborfield Bridge site
<150 mm fraction
d15N 4.21 4.80 3.55 10.7 46.5 82.9 58.7 75.3 87.5 70.7 100.0 95.4
d13C 25.07 23.92 26.36 3.2 40.8 65.7 41.7 100.0 83.4 82.3 83.4 82.3
>150 mm fraction
d15N 3.78 4.52 1.88 21.2 89.1 98.0 26.2 33.5 41.7 98.0 41.7 98.0
d13C 25.32 23.00 27.83 6.2 42.8 61.5 31.8 100.0 72.4 73.9 72.4 76.8
 K. Pye et al. / Forensic Science International 163 (2006) 59–80 73

Fig. 7. Bivariate plots of d13C (per mil) relative to VPDB against d15N (per mil) relative to atmospheric N2 for: (a) the <150 mm, and (b) the
>150 mm bulk fractions.

subsets G and H represented 75.7 and 44.4% of the total but also a clear discrimination between the two sites
variation in the mode, respectively. (Fig. 10).
Bivariate plots of selected particle size parameters The effect on the cumulative averages of adding sequential
showed greater scatter for the Arborfield Bridge samples samples is shown in Fig. 11. At both sites the average values

Fig. 8. d13C and d15N values for the bulk <150 mm fractions of sequential samples taken at Simon’s Wood (a and b) and Arborfield Bridge (c and
d). The solid lines show the cumulative averages. Note exaggeration of the vertical scale in the case Simon’s Wood samples to make intra-grid
variations more visible.
74 K. Pye et al. / Forensic Science International 163 (2006) 59–80

Table 7
Probabilities resulting from two-tailed t-tests comparing the carbon and nitrogen stable isotope ratios in subsets of samples from the two sites
All samples Subset A Subset B Subset C Subset D Subset E Subset F Subset G Subset H
(n = 9) (n = 3) (n = 3) (n = 3) (n = 3) (n = 4) (n = 4) (n = 5) (n = 5)
<150 mm fraction
d15N 0.0 0.0 0.3 0.0 0.2 0.0 0.0 0.0 0.0
d13C 0.0 0.8 3.3 2.7 10.5 1.3 1.5 0.2 0.3
>150 mm fraction
d15N 0.0 1.7 1.7 0.0 0.0 0.0 0.1 0.0 0.0
d13C 0.1 2.3 7.2 3.7 23.4 5.5 3.3 2.6 2.0
Values lower than 5% indicate a significant difference between the two grids at the 95% confidence level.

for the mean, median D90–D10 range and percentages of sand,
silt, and clay based on the first three samples were relatively
similar to those obtained by averaging the first five and all nine
samples, although the approximation was closer for the
Simon’s Wood samples than for the Arborfield samples.
PCA performed using the summary parameters provided
visual discrimination between the two sites (Fig. 5b),
although this was less clear than the discrimination based
on the elemental data (Fig. 5a). Greater clustering of the
Simon’s Wood samples is evident on the basis of the first two
components, which represent 96% of the variability. The
results of agglomerated hierarchical cluster analysis also
demonstrated clear discrimination between the two sites and
indicated that the Arborfield Bridge grid samples are less
clustered than the Simon’s Wood grid samples (Fig. 6b).
Comparison of the particle size parameters for different
subsets of samples using Student’s t-test demonstrated that
the degree of discrimination generally improved as the
number of samples was increased (Table 13). This was
especially evident for the percentages of sand, silt, and clay.
A high degree of statistically significant discrimination was
obtained using subsets with five samples (subsets G and H).
With one exception (subset C), relatively good statistical
discrimination in terms of most of the size parameters was
also achieved with subsets containing three or four samples.
5. Discussion
5.1. Comparison of spatial variability indicated by
different techniques
Using the coefficient of variation as a measure of within-
grid variability, the Simon’s Wood grid showed greatest

Table 8
Precision of colour parameters determined by spectrophotometry
Sample L* CV% (n = 5) a* CV% (n = 5) b* CV% (n = 5)
Simon’s Wood A1
Sub-sample 1 39.25 0.0 2.67 0.2 6.12 0.2
Sub-sample 2 39.47 0.0 2.71 0.6 6.17 0.2
Sub-sample 3 38.75 0.0 2.79 0.4 6.30 0.1
Sub-sample 4 38.78 0.0 2.71 0.3 6.16 0.2
Sub-sample 5 39.70 0.0 2.69 0.5 6.19 0.2
Mean (n = 25) 39.19 2.71 6.19
Standard deviation (n = 25) 0.38 0.04 0.06
CV% (n = 25) 1.0 1.6 1.0
Arborfield Bridge B1
Sub-sample 1 30.54 0.0 2.87 0.4 9.32 0.2
Sub-sample 2 30.21 0.0 2.85 0.7 9.28 0.2
Sub-sample 3 29.30 0.0 2.83 0.7 9.05 0.2
Sub-sample 4 29.93 0.0 2.88 0.3 9.28 0.2
Sub-sample 5 29.06 0.0 2.77 0.3 8.90 0.3
Mean (n = 25) 29.81 2.84 9.17
Standard deviation (n = 25) 0.57 0.04 0.17
CV% (n = 25) 1.9 1.5 1.8
Sub-sampling precision was assessed by taking five different ground sub-samples from sample A1 (Simon’s Wood) and sample B1 (Arborfield
Bridge). Instrumental measurement precision was assessed by five repeat determinations of each sub-sample. The overall average, standard
deviation and coefficient of variation represent the instrumental and sub-sampling precision across all 25 determinations.
 K. Pye et al. / Forensic Science International 163 (2006) 59–80 75

Table 9
Average and range of colour L*, a* and b* indices in the <150 mm fraction of soil samples from the two sites, together with the percentage of the
total range recorded in different sample subsets (see Fig. 2 for subset identification)
All samples (n = 9) Percentage of the total range recorded
Mean Maximum Minimum CV% Subset A Subset Subset C Subset D Subset E Subset F Subset G Subset H
(n = 3) B (n = 3) (n = 3) (n = 3) (n = 4) (n = 4) (n = 5) (n = 5)
Simon’s Wood site
Ground <150 mm fraction
L* 30.21 33.94 27.32 8.0 36.7 16.8 87.1 70.6 65.0 87.1 95.7 100.0
a* 2.78 3.08 2.53 7.0 53.0 41.6 100.0 63.2 73.7 68.0 73.7 97.5
b* 6.10 6.63 5.51 6.1 61.1 43.2 91.8 49.3 71.1 75.8 71.1 75.8
Unground <150 mm fraction
L* 33.15 41.55 25.22 16.7 74.7 51.8 20.9 73.6 97.9 65.7 97.9 71.9
a* 2.17 2.41 1.92 7.9 13.8 89.3 93.3 100.0 96.0 93.3 96.0 93.3
b* 4.89 5.26 4.47 5.4 100.0 41.7 51.4 53.0 80.3 91.5 80.3 91.5
Arborfield Bridge site
Ground <150 mm fraction
L* 27.05 29.87 22.96 8.2 37.1 15.5 50.0 50.0 92.5 52.6 92.5 68.4
a* 2.95 3.36 2.70 7.1 35.3 52.5 87.2 52.9 35.7 84.0 47.5 100.0
b* 8.97 10.27 7.89 7.9 31.6 37.6 73.5 48.3 71.1 69.2 71.1 76.9
Unground <150 mm fraction
L* 26.02 27.81 24.31 5.2 79.3 56.6 48.3 88.6 50.9 44.8 51.7 100.0
a* 2.51 2.72 2.20 7.9 70.4 96.2 42.5 74.2 100.0 70.4 100.0 80.0
b* 7.80 8.66 7.20 6.4 82.4 80.8 52.7 22.1 48.1 53.0 48.1 98.9

Fig. 9. Bivariate plots comparing the a*, b* and L* colour indices in unground (a and b) and ground (c and d) <150 mm soil fractions.
76 K. Pye et al. / Forensic Science International 163 (2006) 59–80

Table 10
Probabilities resulting from two-tailed t-tests comparing the colour L*, a* and b* indices in different subsets of samples from the two sites
All samples Subset A Subset B Subset C Subset D Subset E Subset F Subset G Subset H
(n = 9) (n = 3) (n = 3) (n = 3) (n = 3) (n = 4) (n = 4) (n = 5) (n = 5)
Ground samples
L* 1.1 24.8 2.7 26.9 3.2 9.2 38.5 1.9 11.3
a* 8.6 39.7 6.9 24.5 81.1 19.6 23.0 15.2 25.0
b* 0.0 0.1 0.1 0.9 1.2 0.2 0.1 0.0 0.0
Unground samples
L* 0.5 36.0 72.7 0.7 10.0 7.8 20.1 2.2 5.9
a* 0.1 11.0 19.4 5.7 10.7 6.4 2.4 3.6 2.1
b* 0.0 0.4 0.7 0.2 0.0 0.0 0.0 0.0 0.0
Values lower than 5% indicate a significant difference between the two grids at the 95% confidence level (see Fig. 2 for subset identification).

internal variation in elemental composition (27.9%), fol-
lowed by nitrogen isotope ratio (20.5%), the L* colour
parameter (16.7%), mean particle size (8.5%), the colour
a* parameter (7.0%), the colour b* parameter (6.1%),
median particle size (5.3%), carbon isotope ratio (0.3%)
and modal particle size (0.0%). The Arborfield Bridge site
showed largest within grid variation in terms of modal
particle size (25.4%), median particle size (24.9%), mean
particle size (19.4%), elemental composition (14.5%), nitro-
gen isotope ratio (10.7%), the a* parameter (7.9%), b*
parameter (6.4%), L* parameter (5.2%), and carbon isotope
ratio (3.2%).
However, the within-grid variation was found to be much
smaller than the between-site variation, and the two sites
could be easily differentiated on the basis of any single
attribute considered, and especially so when the results of the
four methods are considered in combination.
The relatively higher degree of elemental variability at
the Simon’s Wood site arises primarily because three sam-
ples on one side of the grid were compositionally different to
the others, which showed a high degree of homogeneity. The
former samples showed lower SiO2/Al2O3, SiO2/Fe2O3 and
Al2O3/Fe2O3 ratios but higher Al2O3/K2O ratio and rela-
tively higher contents of some trace metals (Co, Cu, Pb, V,

Table 11
Precision of particle size parameters determined by laser granulometry
Sample Mean CV% Mode CV% D10 CV% D50 CV% D90 CV% Sand % CV% Silt % CV% Clay % CV%
(mm) (n = 5) (mm) (n = 5) (mm) (n = 5) (mm) (n = 5) (mm) (n = 5) (n = 5) (n = 5) (n = 5)
Simon’s Wood A1
Sub-sample 1 179.8 3.3 188.9 4.3 26.38 5.5 204.0 1.6 1131.0 3.2 82.8 0.6 15.7 3.2 1.5 2.5
Sub-sample 2 172.9 2.9 203.5 0.0 24.07 4.7 200.9 1.3 1124.8 5.5 82.0 0.5 16.4 2.5 1.5 2.1
Sub-sample 3 185.5 3.2 203.5 0.0 31.19 5.5 205.2 1.3 1143.4 7.1 84.2 0.5 14.4 2.7 1.4 2.7
Sub-sample 4 182.8 3.2 185.3 0.0 31.66 7.1 202.2 1.3 1091.6 3.7 84.2 0.7 14.5 3.9 1.3 3.4
Sub-sample 5 177.7 2.5 185.3 0.0 25.43 4.7 203.1 1.1 1136.4 2.7 82.5 0.5 16.0 2.5 1.5 2.4
Mean (n = 25) 179.7 193.3 27.7 203.1 1125.4 83.2 15.4 1.4
Standard deviation 4.8 9.4 3.5 1.6 20.1 1.0 0.9 0.1
(n = 25)
CV% (n = 25) 2.7 4.9 12.5 0.8 1.8 1.2 5.9 6.7
Arborfield Bridge B1
Sub-sample 1 341.9 2.3 1192.8 19.4 51.33 4.0 482.4 2.3 1517.4 1.6 88.3 0.3 10.9 2.4 0.8 3.6
sub-sample 2 343.7 4.6 1180.5 9.8 51.38 7.1 485.9 4.9 1499.6 2.9 88.3 0.6 10.8 4.6 0.8 6.4
Sub-sample 3 326.4 4.7 1032.5 5.2 49.32 7.2 450.9 4.7 1450.0 2.9 87.9 0.7 11.2 4.7 0.9 5.9
Sub-sample 4 311.6 3.0 1013.0 4.3 43.23 5.5 438.9 3.1 1453.0 1.4 86.7 0.5 12.4 3.5 0.9 4.0
Sub-sample 5 349.1 1.3 993.5 0.0 59.43 3.8 468.1 0.9 1468.8 0.5 89.5 0.4 9.8 3.2 0.7 3.8
Mean (n = 25) 334.6 1082.5 50.9 465.2 1477.8 88.1 11.0 0.8
Standard deviation 15.3 96.2 5.8 20.2 29.6 1.0 0.9 0.1
(n = 25)
CV% (n = 25) 4.6 8.9 11.4 4.3 2.0 1.2 8.5 9.4

Sub-sampling precision was assessed by taking five different sub-samples from sample A1 (Simon’s Wood) and sample B1 (Arborfield Bridge).
Instrumental measurement precision was assessed by five repeat determinations of each sub-sample. The overall average, standard deviation and
coefficient of variation represent the instrumental and sub-sampling precision across all 25 determinations.
 K. Pye et al. / Forensic Science International 163 (2006) 59–80 77

Table 12
Average and range of particle size parameters for samples from the two sites, together with the percentage of the total range recorded in different
sample subsets (see Fig. 2 for subset identification)
All samples (n = 9) Percentage of the total range recorded
Mean Maximum Minimum CV% Subset Subset Subset C Subset D Subset E Subset F Subset G Subset H
A (n = 3) B (n = 3) (n = 3) (n = 3) (n = 4) (n = 4) (n = 5) (n = 5)
Simon’s Wood site
Mean 111.8 127.1 97.5 8.5 60.8 59.8 90.5 36.8 100.0 51.4 100.0 58.4
Mode 185.3 185.3 185.3 0.0 100.0 100.0 100.0 100.0 100.0 100.0 100.0 100.0
D10 14.1 16.4 11.9 11.9 90.6 62.9 100.0 67.3 62.4 95.1 62.4 95.1
D50 160.4 177.2 149.8 5.3 86.5 58.0 71.5 20.4 100.0 58.0 100.0 58.0
D90 599.5 753.7 389.0 24.0 13.5 43.9 89.7 90.7 86.5 32.5 86.5 74.7
% Sand 72.8 76.4 70.6 2.8 98.3 84.5 72.4 41.4 100.0 82.8 100.0 82.8
% Silt 25.1 27.4 21.7 8.1 96.0 84.8 68.1 39.8 100.0 80.7 100.0 80.7
% Clay 2.1 2.3 1.9 7.1 71.4 40.5 73.8 45.2 81.0 71.4 81.0 92.9
Arborfield Bridge site
Mean 206.3 293.7 160.7 19.4 28.4 44.1 100.0 58.5 41.5 86.4 41.5 86.4
Mode 897.2 1198.0 356.1 25.4 10.5 75.7 34.8 24.3 75.7 34.8 75.7 44.4
D10 22.7 33.0 16.8 23.0 43.9 39.6 100.0 88.8 31.6 97.7 66.2 97.7
D50 287.1 442.7 211.3 24.9 21.9 43.3 99.2 61.0 51.6 82.5 51.6 82.5
D90 1234.4 1483.0 951.4 12.8 12.6 78.7 70.5 37.6 72.7 50.2 72.7 65.7
% Sand 78.4 84.4 73.5 4.3 47.7 49.5 100.0 61.5 41.3 90.8 71.6 90.8
% Silt 20.1 24.6 14.5 15.5 47.7 49.9 100.0 62.0 39.9 91.5 70.5 91.5
% Clay 1.5 1.9 1.1 16.9 47.5 45.0 100.0 55.0 58.8 82.5 85.0 82.5

and Zn) than other samples within the grid. Other character-
istics considered (C and N isotope ratios, colour, and particle
size distribution) in these samples showed less degree of
difference when compared with other samples in the grid.
This reflects the sensitivity of elemental concentrations
(especially of trace elements) to small variations in soil
and sediment mineralogical composition.
The Arborfield Bridge grid showed greater variability
than the Simon’s Wood grid in terms of particle size and
carbon isotope ratio, but a broadly similar level of variability
in terms of nitrogen isotope ratio and colour. The within-grid
variability in elemental composition was also relatively
high, although less than at Simon’s Wood. Given the nature
of the Arborfield site, the degree on within-grid variability in
elemental composition is less than might have been
expected.
In assessing the discriminatory power of each of the
techniques used, two aspects are important: (a) the
amount of information provided by each technique (i.e.
the number of quantitative parameters which can be used
for comparison) and (b) the degree to which the para-
meters vary in the soil samples under investigation. In

Fig. 10. Bivariate plots of (a) median against mean particle size and (b) the D90 against the D10 size parameters.
78 K. Pye et al. / Forensic Science International 163 (2006) 59–80

Fig. 11. Values of the mean, median, D90–D10 range and percentages of sand, silt, and clay for sequential samples in the grids at Simon’s Wood
(a–d) and Arborfield Bridge (e–h), plotted on the same vertical scale. The solid lines in a–c, e–g show the cumulative average values.

terms of the number of parameters generated, the laser
diffraction particle size analysis method used in this study
provides the largest number (volume frequency data in
116 size classes, plus 8 summary parameters used in this
study), followed by ICP-AES and ICP-MS (concentration
data for 49 elements analysed), colour (reflectance mea-
sured at 31 different wavelengths, plus 3 summary para-
meters used in this paper) and stable isotopes (one isotope
ratio for each of the two elements considered in this
study).
The degree of variability in soil parameters is closely
dependent on spatial scale. For a technique to be of use
 K. Pye et al. / Forensic Science International 163 (2006) 59–80 79

Table 13
Probabilities resulting from two-tailed t-tests comparing the particle size parameters of different subsets of soil samples from the two sites
All samples Subset A Subset B Subset C Subset D Subset E Subset F Subset G Subset H
(n = 9) (n = 3) (n = 3) (n = 3) (n = 3) (n = 4) (n = 4) (n = 5) (n = 5)
Mean 0.0 0.9 1.5 9.2 3.4 0.5 1.7 0.1 0.4
Mode 0.0 0.2 0.1 1.0 0.6 2.6 0.1 0.6 0.0
D10 0.1 6.4 9.3 16.0 10.8 0.9 5.6 1.1 1.2
D50 0.1 1.3 1.5 13.5 4.4 4.3 2.6 1.1 1.0
D90 0.0 0.0 0.1 0.8 0.5 0.1 0.0 0.0 0.0
% Sand 0.1 15.0 7.9 19.1 3.8 6.9 3.3 2.4 0.8
% Silt 0.1 17.4 8.7 20.1 4.1 9.0 3.5 3.2 0.9
% Clay 0.0 3.9 5.2 10.2 1.5 0.9 1.5 0.3 0.1
Values lower than 5% indicate a significant difference between the two grids at the 95% confidence level (see Fig. 2 for subset identification).

forensically, the measured parameters must vary sufficiently
between geographically separate sampling sites for them to
be discriminatory, but should show relatively low variability
at small spatial scales. The degree to which the soil para-
meters under consideration vary will depend on a variety of
factors, including the type of soils or sediments in any area,
the parent material, and land use history.
The two experimental sites in this study, which were
8 km apart, could be easily distinguished on the basis of any
one of the four properties investigated (elemental composi-
tion, nitrogen isotope ratio, colour, and particle size dis-
tribution). These attributes were found to show relatively
low variability on a small (<1 m grid) scale, although subtle
within-grid differences could be identified. Major and trace
element analysis was found to be particularly useful in
providing discrimination between the sites, whilst also being
sensitive to systematic within-grid spatial variation. In the
grids sampled, colour showed the least variation, but there
are many instances where major variations in the colour of
soils and sediment can be observed over distances of a few
metres, and also over time, as a consequence of variations in
redox conditions and other factors. Previous studies have
also shown that particle size can vary significantly on a
small-scale, especially in poorly sorted soils and sediments,
and care must be taken to base comparisons on the least
variable (at a local scale) parameters, such as the modal size,
which may provide a better basis for comparison than
measures such as the mean size.
In the case of 1 m  1 m soil grids investigated, a total of
100 samples, each measuring 0.1 m  0.1 m  0.01 m
would have been required to fully represent the composi-
tional variation in the surface soil of each grid, but such
detailed sampling can rarely, if ever, be justified in forensic
casework. In this study, five samples was found to be
sufficient to characterise a high percentage (ca. 90%) of
the total variability in the grid represented by nine samples,
while on average three samples represented ca. 70% of the
total variability. With hindsight, a single sample would have
been sufficient to discriminate between the two sites in
question, but this could not be known in advance and the
degree to which a single sample is representative is always
open to some uncertainty. It is therefore recommended that
in routine casework a minimum of three, and preferably five
or more, samples should be taken to represent a small area of
investigative interest such as a crime scene. Additional
samples should also been be taken for purposes of compar-
ison from the wider area and any other specific locations of
possible interest identified in the enquiry.
Acknowledgements
We gratefully acknowledge Samantha Witton, David
Wray and Lorna Dyer for assistance with sample preparation
and analysis, and two anonymous referees for helpful com-
ments on an earlier version of the paper.

6. Conclusions References

In this study, the relative between-site discriminatory
capacity of the four techniques investigated was as follows:
(1) multi-element analysis by ICP-AES and ICP-MS, (2)
stable C and N isotope ratio analysis, (3) colour analysis, and
(4) particle size analysis. However, under different circum-
stances this order may change, and it is therefore recom-
mended that studies should always utilise a multi-technique
approach which may include methods other than those
considered in this paper.
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