CHAPTER 

23

Utility of stable isotope ratios of tap

water and human hair in determining
region of origin in Central and Southern
Mexico: Modeling relationships
between δ2H and δ18O isotope inputs
in modern Mexican hair
Chelsey Juarez1, Robin Ramey2, David T. Flaherty3 and Belinda S. Akpa3
1
 Department of Anthropology, California State University, Fresno, California, USA
2
 Fairfax County, Virginia, USA
3
 Department of Molecular Biomedical Sciences, North Carolina State University, Raleigh, North,
Carolina, USA

23.1 Introduction

In the last 15 years, there has been growing interest in the application of water
isotope analysis to provenance and migratory behavior for populations of forensic
significance (Thompson et  al., 2010; Bowen et  al., 2005, 2009; Bowen, 2010;
Ehleringer et al., 2008; Podlesak et al., 2008; Meier‐Augenstein and Kemp, 2010;
Meier‐Augenstein and Fraser, 2008; O’Brien and Wooller 2007; Warner et  al.,
2018). More recently, studies have focused on the development and testing of
semi‐mechanistic models to predict region of origin based on the O and H compo-
sition of scalp hair (Thompson et al., 2010; Bowen, 2010; Podlesak et al., 2008).
These models, constructed with assumptions based on modern American popula-
tions, demonstrated that the non‐exchangeable O and H isotopic composition of
hair is dictated in large part by tap water. Other significant contributions come
from non‐locally‐derived foods and locally produced foods, the latter of which is
presumed to be isotopically related to local water resources (Hobson et al., 1999).
Versions of these models with additional adjustable parameters were tested on
historic and modern international populations (Thompson et al., 2010; Bowen,
2010). Successful application of the Bowen model to non‐American populations
indicates that the model may be transferred to other regions.

Forensic Science and Humanitarian Action: Interacting with the Dead and the Living, First Edition.
Edited by Roberto C. Parra, Sara C. Zapico and Douglas H. Ubelaker.
© 2020 John Wiley & Sons Ltd. Published 2020 by John Wiley & Sons Ltd.

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 346    Forensic science and humanitarian action

The deaths of undocumented immigrants from Latin America in the United

States and Mexico present an opportunity where discriminating region‐of‐origin
information would be helpful in initiating identifications (Kimmerle et al., 2010;
Ross et al., 2016). Hair isotopes acclimate to new locations within weeks and record
region‐of‐origin information, as well as migratory data (Podlesak et  al., 2008).
Thus, hair isotopes may offer particularly valuable discriminating information in
the case of long‐distance migrants. Beyond the traditional difficulties with
identification in border death cases, deceased undocumented border‐crossers from
Latin America offer unique challenges for region‐of‐origin prediction using water
isotopes (Kimmerle et al., 2010). These challenges include drinking water insecu-
rity, high bottled water dependency, a lack of continuity between drinking water
and food water, and dramatic differences in socioeconomic status (SES) in these
regions (Sosa‐Rodriguez, 2012; Fernando and Cervantes, 2016; Martinez et  al.,
2015; Joshua, 2018; National Water Commission (CONAGUA), 2015).
This study explores the relationships between tap water, bottled water, and hair
samples of known Mexican origin from Central and Southern Mexico. The
purpose of this study was twofold. First, we wanted to quantify the relationship
between Mexican hair and tap water values; secondly, we wanted to investigate
whether the semi‐mechanistic model modified by Bowen was an appropriate
choice for Mexican populations, given the established challenges.

23.2  Water stress in Mexico

In Mexico, it is often the case that drinking water, cooking water, and bathing
water come from different sources (Fernando and Cervantes, 2016; Rodríguez‐
Tapia et  al., 2017; Rowles III et  al., 2017). Water stress is driven by uneven
development, poor water sanitation, epidemics, and municipal deficiencies (Sosa‐
Rodriguez, 2012; Rodríguez‐Tapia et al., 2017; Ayala and O’Rourke, 1989; Borroto
and Martinez‐Piedra, 2000; Davis, 2005; Armienta and Segovia, 2008; Bundschuh
et al., 2012; Espinosa‐García et al., 2015; Arcega‐Cabrera et al., 2017). In the sec-
tion below, we outline how the populations in seven Mexican regions at the focus
on this study are impacted by water stress, and how these nuanced experiences
impact drinking water choices and water isotope mapping. We also investigate
differences in the sources of agricultural water and drinking water in Mexico, and
the impact these differences have on modeling assumptions.
In Mexico, municipal water access is managed by hydrological administrative
regions (HAR), and 60% of municipal water is derived from groundwater sources
(CONAGUA, 2015). Michoacán is classified as a region of high water stress
(CONAGUA, 2015). The region has been negatively impacted by seismic activity,
and water stress comes from leaks in the municipal water system, drought, over‐
extraction of surface waters and industrial pollution (Ayala and O’Rourke, 1989;
Davis, 2005; Chávez‐García and Bard, 1994). Municipal water in Michoacán

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 Stable isotope ratios of tap water and human hair in Mexico    347

comes mainly from surface waters, and water quality in the Rio Lerma river basin
is not fit for drinking, cooking or bathing (Sedeño‐Díaz and López‐López, 2007).
Mexico State and Mexico City are the most water‐stressed regions in Mexico
(Martinez et al., 2015; CONAGUA, 2015; Kimmelman, 2017). Ground and sur-
face water in the region suffer from the second highest levels of pollution in the
country (Rodríguez‐Tapia et al., 2017; Espinosa‐García et al., 2015; Cifuentes and
Rodriguez, 2005). Consequently, as many as 87% of homes in the region rely
completely on bottled water for drinking and cooking (Cifuentes and Rodriguez,
2005). Morelos is classified as having a high level of water stress due to industrial
and sewage pollution. For Veracruz, water scarcity is a concern due to the unequal
distribution of treated drinking water (Martínez‐Mier et  al., 2005; CONEVAL,
2013; Mokondoko et al. 2016). Only 68.1% of rural inhabitants have access to
treated tap water compared with 91.1% of urban populations, which leads rural
populations to rely unsustainably on rivers and streams for water (Pacheco‐Vega,
2015; Návar, 2011). Similarly, in Chiapas, only 78% of the total population have
access to municipal water, and 32% of rural populations have no access to
municipal water (CONAGUA, 2015). In Chiapas, soda is cheaper than water and
the region consumes as much as half a gallon of soda per person per day (Lopez
and Jacobs, 2018). Over 30% of Oaxacans lack municipal water or sanitation ser-
vices, and almost 50% live in poverty (Rowles III et al., 2017). As a result, both
municipal supplied water, well water, and unimproved water sources (stream or
river water) all suffer from unsafe microbial levels. Bottled water usage for
drinking water is as high as 100% in many regions of the state (Rowles III et al.,
2017; Vertiz, 2011; McCulligh and Tetreault, 2017).
Depending on their state of residence, between 80 and 100% of Mexicans
report using bottled water as their only source of drinking water (Joshua, 2018;
Espinosa‐García et  al., 2015; Kimmelman, 2017; Guardiola et  al., 2013). As a
consequence, expenditure on bottled water can be more than 14% of monthly
income per household (Gutieŕrez et  al., 2012). Bottled water is primarily pro-
duced by four major companies in the Mexican industry: Coca‐Cola/FEMSA,
Nestle, PepsiCo and Danone. Together, these companies hold 80% of the bottled
water market share, with more than 6000 small companies constituting the
remaining 20% of market share (Greene, 2014). The large companies have
­several bottling plants in the country that draw water from local sources but dis-
tribute their products nationally (Franco et al., 2015). For example, water from
the Chiapas Coca‐Cola plant provides about 5–7% of total bottled water beverages
for the country (Pani, 2017). As bottled water is not strictly regulated in Mexico,
both small and large companies use water from multiple sources including aqui-
fers, groundwater, rainwater and river water. Since 2000, Coca‐Cola alone has
used water from 19 different aquifers and 15 different rivers (Franco et al., 2015).
Sources of agricultural water, which inform the isotopic signature of locally
produced foods, differ by HAR. In Mexico, the majority of agricultural water
comes largely from surface water sources (64%), which are subject to

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 348    Forensic science and humanitarian action

evaporative effects (CONAGUA, 2015; McCulligh and Tetreault, 2017). However,

Veracruz, Oaxaca and Mexico State have significant contributions from ground-
water. Given the prevalence of bottled water consumption in Mexico, and the
fact that bottled water is not always drawn from local sources, we cannot assume
that the isotopic composition of agricultural water and drinking water are related.

23.3  Tuning parameters in mathematical models used

for provenance analysis

Practically, the mathematical models proposed for provenance analysis based on

hair and drinking water presume a near‐linear relationship between hair isotopes
and drinking water. A simple regression on isotopic data indicated this assumption
to be valid for paired hair–water data from modern US populations. This led
Ehleringer et al. (2008) to posit that drinking water accounts for a substantial part
of the geographical variation in hair isotopes, and to derive a semi‐mechanistic
framework to codify the hair–water isotope relationship. In doing so, they identi-
fied a small number of parameters that were unconstrained by prior empirical
observation. The authors treated the fraction of non‐exchangeable H in keratin
that was fixed in vivo as a fitting parameter. Likewise, the parameters αow, αhw and
g were tuned to data consisting of paired hair–water samples. These parameters
represent, respectively: (i) the isotope fractionation of carbonyl oxygen in hair
relative to gut water; (ii) isotope fractionation of H in de novo synthesized amino
acids relative to hair follicle water; and (iii) the fraction of O in the gut that derives
from gastric juices rather than the water content of food.

23.4  Extension to analysis of hair isotopes in the absence

of paired water samples

A subsequent application of the model to mid‐twentieth century indigenous pop-

ulations motivated two key modifications (Bowen et al., 2009). Firstly, the authors
posited a fraction of the diet to be derived from local sources, as the populations
in question lived a pre‐globalization lifestyle and were geographically dispersed
across several continents. Thus, a dietary isotopic input based on relatively homo-
geneous twenty‐first century supermarket foods would likely be inappropriate in
representing the consumption patterns of these indigenous groups. Instead, a
locally produced dietary component was assumed, and the relevant food isotopes
were modeled as an offset from the average heavy isotope content of regional pre-
cipitation. Use of these empirically determined offset values implicitly presumes a
stable relationship between precipitation and the water used in agricultural
endeavors (e.g. via groundwater). Secondly, as no tap water data were available
for these anthropological samples, precipitation values were used as a proxy

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 Stable isotope ratios of tap water and human hair in Mexico    349

for  drinking water isotopes. With the model thus modified, parameters were
adjusted to provide a good fit to data from several geographically disparate
­indigenous groups.
The success of the Bowen model in describing hair isotope variations as a
function of drinking water (in this case, regional precipitation) is interesting in
that it suggests that l, the fraction of diet derived from local sources, can be treated
as a global constant. Hair samples from nearly all the indigenous groups fell along
a common line indicating the same fraction of food being locally derived.
Furthermore, the non‐local dietary contribution was still described using data
representing the modern continental United States. Yet the hair samples origi-
nated from populations in regions ranging from Canada to Pakistan to Botswana,
and a timespan from the 1930s to 1950s. Similarly, fs, a physiological parameter
likely to be subject to inter‐individual variability, was treated as constant over all
samples and individuals.
We aim herein to investigate the impact on model quality of using different
approaches to represent the contributions of non‐local and local diet to hair iso-
topes. Essentially, l is likely to differ based on socioeconomic, cultural and biological
differences between individuals, and that difference is critical to making
quantitative links between hair and region of origin.

23.5  Materials and methods

For this study, we used a subset of paired (N = 62) human hair, (N = 76) tap water
and (N = 73) bottled water samples from a larger dataset of unpaired samples
(N = 290 for tap water, N = 156 bottled water, and N = 118 hair). For samples to
be considered paired, they needed to be from the same location (e.g. city or town).
Sample locations were only included in this subset if at least two hair and water
samples were available for analysis. All hair samples were obtained from anony-
mous confirmed residents from ten cities in six Mexican states (Table  23.1).
Samples consisting of beard or scalp hair, representing several milligrams of
material, were collected on site, in clean coin envelopes. Data on city and state of
residence was collected for all samples.
Tap water samples were collected from the homes of private donors and local
restaurants. All water samples were collected in acid‐washed 300 ml HDPE bottles
following the recommendation of Spangenberg (2012) with no head space. Water
and hair samples were prepared and measured for δ2H and δ18O isotopes at the
SIRFER lab at the University of Utah following the methods outlined in Thompson
et al. (2010) and Bowen et al. (2005). Briefly, hair samples were equilibrated for
a 7‐day period with hair reference materials. Following equilibration, weighed
hair samples and water samples were pyrolysed in a temperature conversion
­elemental analyser (TC/EA) and the H and O isotope ratios of hair and water sam-
ples were measured using an isotope ratio mass spectrometer (IRMS) operated in

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 Table 23.1  Summary of hair, and tap water and bottled water isotope ratio data, by state and city.

State City Mean Mean Mean Mean Mean Mean

δ18Oh‰ δ2Hh‰ δ18Otw‰ δ2Htw‰ δ18Owb‰ δ2Hwb‰

Mexico 12.1‰ −78.7‰ −9.5‰ −68‰ −10.7‰ −75.4‰

SD ±1.1‰ ±5.9‰ ±0.6‰ ±3.9‰ ±0.4‰ ±2.6‰
N 15 15 20 20 10 10
Mexico Mexico City 12.3‰ −76.9‰ −9.5‰ −68‰ −10.7‰ −75.4‰
SD ±1.1‰ ±5.9‰ ±0.6‰ ±3.9‰ ±0.4‰ ±2.6‰
N 10 10 15 15 5 5
Mexico Tultepec 11.7‰ −83.3‰ −9.3‰ −66.6‰ −10.7‰ −75.4‰
SD ±1.0‰ ±1.8‰ ±0.4‰ ±3.0‰ ±0.4‰ ±2.6‰
N 5 5 5 5 5 5
Oaxaca 14.5‰ −78.4‰ −8.2‰ −60.5‰ −10.1‰ −71.7‰
SD ±1.0 ±7.8 ±2.9 ±16.0 ±0.9 ±6.6
N 20 20 50 50 24 24
Oaxaca Oaxaca City 12.5‰ −75.2‰ −9.4‰ −68.3‰ −10.1‰ −71.7‰
SD ±1.0‰ ±10.7‰ ±0.3‰ ±2.7‰ ±0.9‰ ±6.6‰
N 15 15 22 22 12 12
Michoacán 14.5‰ −74.5‰ −9.1‰ −66.1‰ −9.7‰ −69.2‰
SD ±0.2‰ ±4.7‰ ±1.0‰ ±6.8‰ ±1.3‰ ±7.8‰
N 15 15 20 20 10 10
Michoacán Tzintzuntan 14.5‰ −74.5‰ −10.‰ −73.4‰ −9.7‰ −69.9‰
SD ±0.2‰ ±4.7‰ ±1.0‰ ±6.8‰ ±1.3‰ ±7.8‰
N 5 5 5 5 5 5
Michoacán Pátzcuaro 14.8‰ −69.2‰ −9.3‰ −67.3‰ −9.7‰ −69.9‰
SD ±1.2‰ ±2.2‰ ±1.0‰ ±4.8‰ ±1.3‰ ±7.8‰
N 5 5 4 4 5 5

c23.indd 350 11/26/2019 8:28:04 PM

 State City Mean Mean Mean Mean Mean Mean
δ18Oh‰ δ2Hh‰ δ18Otw‰ δ2Htw‰ δ18Owb‰ δ2Hwb‰

Michoacán Quiroga 14.5‰ −78.4‰ −9.3‰ −67.5‰ −9.7‰ −69.9‰

SD ±1.0‰ ±2.5‰ ±1.8‰ ±4.4‰ ±1.3‰ ±7.8‰
N 5 5 4 4 5 5
Morelos 12.5‰ −81.1‰ −11.1‰ −77.4‰ −10.4‰ −73.7‰
SD ±0.6‰ ±7.2‰ ±0.3 ±2.4 ±0.6 ±4.4
N 8 8 12 12 18 18
Morelos Cuernavaca 12.5‰ −78.4‰ −11.1‰ −77.4‰ −10.4‰ −73.7‰
SD ±0.6‰ ±7.2‰ ±0.3‰ ±2.4‰ ±0.6‰ ±4.4‰
N 5 5 8 8 9 9
Chiapas 14.3‰ −66.1‰ −7.2‰ −41.0‰ −4.2‰ −20.9‰
SD ±0.2 ±0.8 ±1.8 ±15.2 ±0.0 ±0.3
N 4 4 14 14 6 6
Chiapas Palenque 14.5‰ −66.1‰ −4.6‰ −24.5‰ −4.2‰ −20.9‰
SD ±0.2‰ ±0.8‰ ±1.8‰ ±15.2‰ ±0.05‰ ±0.3‰
N 4 4 4 4 3 3
Veracruz 15.2‰ −63.4‰ −5.6‰ −32.7‰ −9.1‰ −63.9‰
SD ±0.4‰ ±12.9‰ ±1.3‰ ±10.9‰ ±1.8‰ ±14.9‰
N 8 8 25 25 24 24
Veracruz Papantla 15.1‰ −56‰ −5.0‰ −27.7‰ −9.1‰ −63.9‰
SD ±0.5‰ ±2.3‰ ±0.7‰ ±5.4‰ ±1.8‰ ±14.8‰
N 4 4 4 4 12 12
Veracruz Gutierrez Zamora 15.4‰ −78.3‰ −5‰ −26.5‰ −9.1‰ −63.9‰
SD ±0.3‰ ±2.0‰ ±0.1‰ ±1.7‰ ±1.8‰ ±14.8‰
N 4 4 5 5 12 12

, hair; tw, tap water; wb, bottled water.

h

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 352    Forensic science and humanitarian action

continuous‐flow (CF) mode. Analytical precision for water samples was 0.3‰ and
0.1‰ for hydrogen and oxygen isotopes respectively. Analytical precision for hair
samples was 2.2‰ and 0.3‰ for δ2H and δ18O respectively. All data are reported
using the standard delta notation δX = [Rstandard/Rsample − 1] × 1000, and all data are
reported against the internationally accepted standard VSMOW. General statistical
analysis was conducted in SPSS version 25. Parameter estimation for model fitting
was performed in Matlab using the approach of approximate Bayesian
computation.

23.5.1  Isotope mapping procedure

Analysis of spatial patterns in the hair isotope data were conducted using ESRI’s
ArcGIS 10.3.1 and following the procedures specified in Bowen et  al. (2007).
Spatial patterning of isotope data was identified using the Spatial Autocorrelation
tool (Spatial Statistics Toolbox, ArcGIS 10.3.1). Moran’s I statistics were calculated
for δ2H and δ18O values using unstandardized weights derived from inverse‐
squared Euclidean distances between tap water sample data points. Isotope value
prediction maps for δ2H and δ18O were produced through spatial interpolation of
the raw hair and water isotope survey data. Interpolation was accomplished
through ordinary kriging (Spatial Analyst Toolbox, ArcGIS 10.3.1) utilizing
ArcGIS’s Geostatistical Analyst extension. All kriging in this analysis was accom-
plished using a spherical semivariogram model.

23.5.1.1 Results
Figures  23.1 to 23.3 and Table  23.1 show the spatial variation in the complete
sample of δ18O and δ2H isotopes in Mexican tap water, bottled water and hair sam-
ples. Tap water values spanned a range from −12.7‰ to +0.4‰ and −91.7‰ to
−4.2‰ for δ18O and δ2H, respectively. Tap water samples differed in their standard
deviation by collection location. Standard deviations ranged from 0.1–2.9‰ for
δ18O and 1.7–15.2‰ for δ2H. For 93% of samples, standard deviations for repli-
cated water samples from the same locations were small, at <1‰ and <6‰ for
oxygen and hydrogen values respectively. Samples from Papantla Veracruz
showed an unusually high level of deviation, indicating inconsistency in water
sources. The calculated local meteoric water line (LMW) for tap waters was
δ2H = 7.54 δ18O + 5.0‰, r2 = 0.91, P < 0.001. Bottled water values spanned a range
of −11.5‰ to 0.5‰ and −80.2‰ to 1.3‰ for δ18O and δ2H, respectively. Bottled
water samples differed minimally in their standard deviation by collection loca-
tion. Standard deviations ranged from 0.03–1.3‰ for δ18O and 0.3–14.9‰ for δ2H.
For 62% of bottled water samples, standard deviations for replicated water
­samples from the same locations were small, at <1‰ and <6‰ for oxygen and
hydrogen values respectively. Samples from Veracruz showed an unusually high
level of deviation (±1.8‰ and ±14.9‰) for O and H respectively, indicating
­multiple sources for bottled waters. The calculated water line for bottled waters
was δ2H = 7.34 δ18O + 2.8‰, r2 = 0.97, P < 0.001. Wassenaar and colleagues (2009)

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 Stable isotope ratios of tap water and human hair in Mexico    353

Interpolated δ18O values in Hair Samples

11.8 – 12.1
12.1 – 12.4
12.4 – 12.7
12.7 – 12.9
12.9 – 13.2
13.2 – 13.5
13.5 – 13.8
13.8 – 14.1 0 125 250 500 750 1,000
Miles
14.1 – 14.4

Figure 23.1  Geographical information system (GIS)‐generated maps of the predicted

average O isotope ratios δ18O In Mexican hair samples.

Interpolated δ2H values in Hair Samples

–80.6 – –78.9
–78.9 – –77.2
–77.2 – –75.5
–75.5 – –73.8
–73.8 – –72.0
–72.0 – –70.3
–70.3 – –68.6
–68.6 – –66.9 0 125 250 500 750 1,000
Miles
–66.9 – –65.2

Figure 23.2  Geographical information system (GIS)‐generated maps of the predicted

average H isotope ratios δ2H In Mexican hair samples.

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 354    Forensic science and humanitarian action

0.0
Tap water Bottled water
–10.0 δ2 H = 7.54 × δ18 O + 5.0‰ δ2 H = 7.34 × δ18 O + 2.8‰
R2 = 0.91 R2 = 0.97
–20.0
A
B
D
–30.0 C
δ2 H vs. VSMOW (‰)

–40.0
Hair
–50.0 δ2 H = 2.98 × δ18 O – 133.3‰
R2 = 0.21

–60.0

–70.0

–80.0

–90.0

–100.0
–14.0 –12.0 –10.0 –8.0 –6.0 –4.0 –2.0 0.0 2.0 4.0 6.0 8.0 10.0 12.0 14.0 16.0
δ18O vs. VSMOW (‰)

Figure 23.3  A plot of the δ2H and δ18O isotope ratios for tap water samples marked by
dots and bottled water samples marked by dashes, and hair (solid line). Points marked
by letters A–B are isotopically enriched points discussed in the text. Points C–D are
isotopically depleted points discussed in the text.

previously reported two water lines for Mexico, the Mexican meteoric water line
δ2H = 7.5 δ18O + 6.1‰ defined by precipitation IAEA stations from three locations
(Mexico City, Veracruz and Chihuahua), and a groundwater meteoric water line
δ2H = 7.9 δ18O + 6.4, r2 = 0.95 based on their collected dataset. The LMW for tap
waters is virtually indistinguishable from that of groundwater. However, the water
line calculated for bottled waters has a lower slope, possibly indicating evapora-
tive effects in some of the bottled water samples.
Hair values ranged from 9.5‰ to 15.5‰ and −90.8‰ to −54.4‰ for δ18O and
δ H, respectively. This range is slightly smaller than those previously reported by
2

Bowen et al. (2009), Ehleringer et al. (2008) and Thompson et al. (2010). The
most depleted δ18O and δ2H hair values came from individuals in the state of
Morelos. Like the water samples, hair samples differed in their standard devia-
tions by collection location. Standard deviations ranged from 0.2‰ to 1.1‰ and
0.8‰ to 12.9‰ for δ18O and δ2H, respectively. Excluding hair samples from
Veracruz, all remaining locations demonstrated a standard deviation that was less
than that for the entire sample. A linear regression through the hair dataset pro-
duced a weak correlation, δ2H = 2.9 δ18O − 113.3, r2 = 0.21, P < 0.002. A subset of
paired N = 62 hair and N = 76 water samples was culled from the larger dataset to
investigate the impact of locality on samples (Table 23.1). Stable isotope composi-
tion for the subset of paired water samples ranged from −11.1‰ to −4.6‰ for δ18O

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 Stable isotope ratios of tap water and human hair in Mexico    355

and −77.4‰ to −24‰ for δ2H, and ranged from 12.1‰ to 15.4‰ for δ18O
and −81.1‰ to −56‰ for δ2H for paired hair samples. Cook’s distance identified
both the Veracruz groups in the subset as outliers. The Gutierrez Zamora population
had a particularly irregular relationship between δ2H isotopes in hair and water.
The water values for both Veracruz locations were enriched relative to the rest of
the sample subset, which make sense for its coastal location, but the δ2H isotopes
in hair were among the most depleted. A linear regression through the subset of
hair data produced a weak correlation δ2H = 3.26 δ18O − 118.6, r2 = 0.31, P < 0.07.
The weak correlations between O and H of both the individual hair samples and
the subset of group means is in stark contrast to the strong correlations reported
by Bowen et al. (2009), r2 = 0.75 and r2 = 0.88 for samples and means, Ehleringer
et al. (2008) for samples r2 = 0.87, or Thompson et al. (2010) for group means
r2 = 0.80. The following relationships between hair (h), tap water (tw) and bottled
water (wb) from the data subset were produced using least squares regression:
δ18Oh = 0.35 δ18Otw + 16.6‰, r2 = 0.38, P = 0.05; δ2Hh = 0.229 δ2Htw − 60.8‰, r2 = 0.36,
P = 0.06; and δ18Oh = 0.33 δ18Owb + 16.8‰, δ2Hh = 0.229 δ2Hwb − 58.6‰, r2 = 0.22,
P = 0.18 (Figures 23.4 and 23.5). These paired group means for tap water and hair
were entered into the process‐based model published by Bowen and colleagues
(2009), in order to compare model outcomes, and determine whether the implicit
assumptions of the model were appropriate for this population.

16.5
δ18Oh = 0.357 × δ18Ow + 16.7‰
R2 = 0.380 Tultepec
15.5
Oaxaca City

Tzintzuntan
14.5
Pátzcuaro
δ18O Hair (‰)

Mexico City
13.5
Quiroga

Cuernavaca
12.5
Palengue

Papantla
11.5
Guiterrez
Zamora
10.5
–12 –11 –10 –9 –8 –7 –6 –5 –4 –3 –2
δ18O Water vs. VSMOW (‰)

Figure 23.4  Mean δ O values measured in paired hair and tap water samples, data points
18

are mean per site.

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 356    Forensic science and humanitarian action

–50

–55
δ2Hh = 0.221 × δ2Hw – 61.3‰ Mexico City
R2 = 0.34
–60 Tultepec

Oaxaca City
–65
δ2H Hair (‰)

Tzintzuntan

–70 Pátzcuaro

Quiroga
–75
Cuernavaca

–80 Palengue

Papantla
–85
Guiterrez
Zamora
–90
–90 –80 –70 –60 –50 –40 –30 –20 –10
δ2H Water vs. VSMOW (‰)

Figure 23.5  Mean δ2H values measured in paired hair and tap water samples, data points
are mean per site.

23.5.2  Analysis and discussion

23.5.2.1  Water samples
The most 2H‐ and 18O‐depleted tap water samples were distributed over the inland
high‐altitude regions of central Mexico, in the states of Puebla, Zacatecas, Jalisco,
Oaxaca and Morelos (Figure 23.3). The slope and d‐excess parameter of the cal-
culated LMW constructed of tap water samples was slightly lower than the global
meteoric water line calculated by Craig (1961). This was due to isotopically
enriched samples from Oaxaca and Jalisco (marked A–B) (Figure  23.3). Two
­additional points were outliers with low δ18O and high δ2H: C is tap water from
highland Chiapas, demonstrating depletion from altitude effects, while point D is
tap water from inland Campeche. The range for Mexican groundwater values
reported by Wassenaar et al. (2009), δ18O = −10.8‰ to −2.3‰ and δ2H = −77.5‰
to −13‰, was smaller than the range for tap water reported here. Significant dif-
ferences in mean δ2H values between tap and groundwater were identified in
Guanajuato where median values for tap water (−73.5‰) were lower than
groundwater (−69.4‰) U = 0, Z = −2.952, P = 0.002. This suggests that for most
of the states tested by both studies, groundwater rather than surface water is the
major source of municipal supply. Wilcoxon signed‐rank tests demonstrated that
median bottled water values for O and H were indistinguishable from median tap
water values from the same locations for O and H respectively: z = −1.8, P = 0.65;
z = −1.24, P = 0.20.

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 Stable isotope ratios of tap water and human hair in Mexico    357

23.5.2.2  Hair samples

H and O measured in hair were weakly correlated (r2 = 0.21), and had a slope
of 2.9, far lower than the slope of 8.6 previously reported for mid‐twentieth
century populations or the slope associated with modern Americans (5.7) or
Asian samples (5.6) (Thompson et  al., 2010; Bowen et  al., 2009). This sug-
gests that controls on these isotope systems were not coupled across this
sample. Likely it is the case that drinking water is not tied to either regional
food or location. The most depleted water and hair values were found in the
state of Morelos, while the most enriched water and hair samples were found
in Veracruz. The relationships b­ etween paired hair and water from the same
locations were analysed. The ­relationship between O and H in hair between
tap and bottled water was ­indistinguishable by region: δ18Oh = 0.35 δ18Otw
+ 16.7‰, r2 = 0.38, P = 0.05; δ2Hh = 0.22 δ2Htw − 61.8‰, r2 = 0.36, P = 0.06,
δ18Oh = 0.33 δ18Owb + 16.8‰, δ2Hh = 0.229 δ2Hwb − 58.6‰, r2 = 0.22, p = 0.18.
According to these relationships, 35% and 22.9% of the variation in hair was
explained by oxygen and hydrogen isotopes in drinking water respectively
(Figures 23.4 and 23.5). These slopes for both O and H were lower than those
reported by either Bowen et al. (2009) or Thompson et al. (2012). However,
the slopes and intercepts for O and the slope for H overlap directly with values
for modern American populations from Ehleringer et  al. (2008), and those
previously reported by Sharp et  al. (2003). Rather than interpret this as a
­similarity in drinking water behavior between this study population and US
residents, we suggest that this is a reflection of the geospatial distribution of
regional food and food water, which in the Mexican case is uncoupled from
drinking water.

23.5.2.3  Analysis via the semi‐mechanistic model

Figure 23.6 is a pathway diagram showing how hair isotopes are conceptually
related to consumed food and drinking water in the Ehleringer and Bowen
models. Primary inputs are denoted by double‐lined boundaries. All other com-
partments are intermediate calculations used to fractionate isotopes into the pri-
mary output, which is the hair. Drinking water is typically an empirical input
determined from samples taken from each location of interest. Given the
significant water stress in southern Mexico and the possibility that supply of and
access to safe drinking water has consequently been decoupled from local aqui-
fers, rivers and lakes, we chose to estimate the likely isotopic signature of Mexican
drinking water. Thus, δ2Hwe and δ18Owe, the H and O content of drinking water,
were treated as model tuning parameters. By estimating a single, shared value
for all locations, we have assumed that the drinking water signature is relatively
homogeneous across these southern Mexican States and can be treated as
independent of location.

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 358    Forensic science and humanitarian action

drinking food food

water (locally derived) (non-locally
derived)

absorbed through
drinking water dietary breathing
contribution contribution (O only)

loss through
body water breathing

hair

Figure 23.6  Pathway diagram showing how hair isotopes are conceptually related to the
key inputs of food and water in the Ehleringer et al. (2008) and Bowen et al. (2009)
models.

23.5.2.4  Representing local and non‐local water and dietary contributions

Following the approach of Bowen and colleagues, the isotopic signature of locally
derived food was expressed as an offset from the drinking water signature.
However, the additive offsets given in their publication were empirically deter-
mined based on US supermarket meats and water supplies local to US feedlot
operations. We decoupled our model implementation from that prior approach by
estimating plausible values for the Mexican food supply, adding Δ2Hd,l and Δ18Od,l
to our parameter set. Similarly, we replaced the non‐locally derived US super-
market diet with estimable parameters δ2Hd and δ18Od, representing that portion
of the Mexican diet that is composed of international sources rather than foods of
local origin. As the drinking water in our model is being estimated as a global
parameter, the “local” food isotopic signature, expressed as an offset from drinking
water, represents a Southern Mexican regional diet.
Given the economic stratification of this region and the poor correlation bet-
ween hair and water samples, we additionally chose to consider the possibility
that l and fs are local parameters for this population. Essentially, we assumed that
both the fraction of diet that is regionally derived and the associated fraction of
hydrogen that is fixed in vivo will differ across the subregion. Both parameters will
depend on the relative affluence of the area, the sources of dietary protein, and
the rural/urban divide. Taking that to be the case, we estimated different values of
l and fs for each location. This represents another key departure from the approach
of Bowen et al., where all parameters were treated as global (i.e. common across
all populations), despite broad geographical dispersion. That assumption is likely
a valid approximation for the mid‐twentieth century context, where it was
unlikely that indigenous populations would have received their diet primarily

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 Stable isotope ratios of tap water and human hair in Mexico    359

from large‐scale agricultural operations and processed foods. More heterogeneity

is expected in this modern Mexican population, where access to both food and
water is subject to acute variations in geographical and socioeconomic factors.

23.6  Estimation of credible parameter values

by approximate Bayesian computation

For the ten hair isotope datasets reported in Table 23.1, we must then estimate a
total of 26 parameter values – 6 characterizing inputs assumed to be shared by all
ten locations, and 20 describing the location‐specific properties l and fs. With only
two data points for each location (i.e. one for each isotope), this model represents
an underdetermined system. Approximate Bayesian computation (ABC) is a
statistical approach that can be used to infer credible parameter intervals for com-
plex models when faced with the challenge of very small datasets (Csilléry et al.,
2010). The approach requires a generative model to approximate the likelihood
function – essentially a probability distribution indicating how likely the experi-
mental dataset would be, given a set of beliefs about the distribution of model
parameter values. The ABC method starts with sampling parameter values, θ,
from distributions P(θ) that represent any existing knowledge about the possible
parameter values. In our case, we know little more than the upper and lower
bounds of the parameter values (e.g. 0 < l, fs < 1), so we used bounded uniform
distributions as uninformative a priori assumptions for all parameters. These were
implemented as scaled beta distributions (α = 1, β = 1) to take advantage of that
distribution’s ability to evolve from uniform to a variety of unimodal intermediate
shapes as the hyperparameters α and β are increased.
Having sampled values for each of the 26 parameter values, the isotope semi‐
mechanistic model was used to generate predictions for 2H and 18O in hair. We then
compared the predictions with the hair isotope data and preserved the parameter
values if the prediction adequately reflected the data (i.e. storing to the posterior
distribution ∝ P(θ|x)). After repeating this sample‐predict‐compare process for
1000 different samples, we used the cumulative degree of success in predicting
the data as a means of updating our prior assumptions about the parameter distri-
bution. With this improved knowledge about the system, we repeat the sampling
process. Several similar iterations yield increasingly credible parameter distributions
by narrowing our sampling domain of the 26‐dimensional parameter space to
those regions that yield the best fit to the data, that is, those values that have the
greatest probability of giving rise to the observed data. Expressing the mean and
appropriate quantiles of these distributions provides both an estimate of each
parameter’s value and a 95% credibility interval. Thus, instead of a single value
with confidence bounds, one obtains a distribution of values for each parameter
that can be interpreted as representing uncertainty in the estimated parameter
value (Tavaré et al., 1997).

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 360    Forensic science and humanitarian action

23.7  Results obtained using the established

US supermarket diet

Prior applications of the Bowen and Ehleringer models assumed tap water to be a
valid proxy for drinking water. Using our local tap water measurements and
an international food contribution identical to the modern US supermarket diet
(δ2Hd = −115, δ18Od = 36), we fail to find values for l and fs that map to the Mexican
hair samples. Figure 23.7 demonstrates the poor quality of fit achieved via this
approach, wherein l and fs are assumed to vary with location, and the local
food  contribution is expressed as a fixed offset from tap water (Δ2Hd,l = −50‰,
Δ18Od,l = 35.4‰).
Given the significant water stress in the region and the prevalence of bottled
water use in both drinking and cooking, we considered whether estimating the
drinking water isotopic signature might improve the quality of the model fit.

(a) (b)
18 –50

16 –60
δ18Ohair

δ 2Hhair

14 –70

12 –80
ε = 0.6 ε = 0.4
10 –90
–15 –10 –5 0 –80 –60 –40 –20
δ18O tw δ 2Htw
(c) (d)
18 –50

16 –60
δ18O hair

δ2H hair

14 –70

12 –80
ε = 0.4 ε = 0.3
10 –90
–15 –10 –5 0 –80 –60 –40 –20
δ18O water δ 2H water

Figure 23.7  Model fits using American dietary parameters. Models assume established
USA values for non‐local dietary contribution and local diet offset from drinking water.
(A–B) Local tap water is used for the isotopic composition of drinking water. (C–D)
Drinking water is assumed to be homogeneous for this region and its isotopic composi-
tion is estimated as part of the Bayesian fitting algorithm. Data and model predictions are
shown as square markers and circle markers respectively. The model reported by Bowen
is also shown as a dotted line.

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 Stable isotope ratios of tap water and human hair in Mexico    361

Figure 23.7 parts C and D report the modestly improved fit. However, the δ18Oh
values predicted now appear to be independent of location. Indeed, the estimated
drinking water signature (δ2Hwe = −61‰, δ2Hwe = −5.9‰) has been treated as a
global parameter. As drinking water is the basis for expressing the local food iso-
topes, the local food signature is now also independent of location. The only var-
iable that can account for the variation in hair 18O is the local value of l. This
proves to be an inadequate tuning parameter. This is a logical outcome, as the
local contribution to diet, when calculated from the estimated drinking water, is
δ2Hd,l = −111‰ and δ18Od,l = 29.5‰. That is nearly identical to the US supermarket
diet and consequently does not allow the model to have the degrees of freedom
necessary to fit the hair isotopes.

23.8  Results achieved by estimating international diet,

drinking water, and regional diet isotopes

Figure 23.8 shows the quality of fit achieved using our Bayesian approach and esti-
mating not just drinking water, but also new values for the local and non‐local die-
tary contributions. The 18O and 2H data are concurrently modeled with similar
accuracy, as indicated by the mean summary statistic values (ε = 0.009 and 0.01 for
18
O and 2H respectively). The estimated values for the Mexican non‐local dietary sig-
nature and local diet offset were (mean [95% intervals]): δ2Hd = −136‰ [−135,−137],
δ18Od = 17.8‰ [17.6,18.2], δ2Hd,l = −84‰ [−83,−86] and δ18Od,l = 48‰ [47,48]. Those
for drinking water were δ2Hwe = −65‰ [−63,−68] and δ18Owe = −10‰ [−10,−11].
Interestingly, this estimated isotopic signature for regional drinking water is very sim-
ilar to the mean value for our bottle water samples, δ2Hwb = −65.5‰, δ18Owe = −9.4‰.

model hair samples Bowen model

(a) (b)
18 –50

16 –60
δ18Ohair

δ2Hhair

14 –70

12 –80
ε = 0.009 ε = 0.01
10 –90
–15 –10 –5 0 –80 –60 –40 –20
δ18Otw δ2Htw

Figure 23.8  Model fits using estimated Mexican dietary parameters. Drinking water is
assumed to be homogeneous for this region and its isotopic composition is estimated as
part of the Bayesian fitting algorithm. (A) δ2H; (B) δ18O. Data and model predictions are
shown as square markers and circle markers respectively. The model reported by Bowen
is shown as a dotted line.

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 362    Forensic science and humanitarian action

0.3
Cuernavaca Patzcuaro Gutierrez
Palenque Zamora
0.2 Mexico Tzintzuntzan
Tultepec City
P(I)

Oaxaca Quiroga Papantla

0.1 City

0
0.3 0.35 0.4 0.45 0.5 0.55 0.6
I

0.3
Quiroga Tzintzuntzan Mexico City Papantla
Patzcuaro
0.2 Cuernavaca Palenque
Oaxaca
P(fs)

Gutierrez Tultepec City

0.1 Zamora

0
0.1 0.15 0.2 0.25 0.3 0.35 0.4
fs

Figure 23.9  Intervals estimated for l and fs in the ten locations for which hair samples
were analysed.

Figure 23.9 shows the location‐specific estimates for l and fs as histograms for

the credible distributions of each parameter. The ten locations from which hair
was sampled readily segregate into two groups with respect to l, the fraction of
diet that is derived from the Southern Mexican region. This largely reflects the
rural/urban divide, with large urban centers like Mexico City, Oaxaca City and
Cuernavaca appearing at l < 0.4, while more rural centers like Tzintzuntzan and
Patzcuaro are centered at l > 0.5. Although SES plays a role in determining access
to local versus non‐local foods, it is not a sole determinant. In rural areas, fresh
local foods are likely to be readily available and inexpensive, while in urban zones
local foods may actually be more expensive than non‐local, processed snacks
(Ponce et  al., 2006; Fernald, 2007; Batis et  al. 2011; Sparks and Sparks, 2012;
López‐Olmedo et al., 2018). It follows then that parameter l tells us little about the
socioeconomic stratification of the region. The complete overlap of distributions
for Oaxaca City and Cuernavaca is interesting, because data on socioeconomic
status in Mexico identifies these two regions as economic polar opposites. In the
absence of specific data on food and hair from these regions it is not possible to
assess the validity of the estimated l values for each location. However, a possible
interpretation of this apparent divide is that urban residents have increased access
to non‐local foods compared with their rural counterparts.
One interpretation of fs is as an indicator of protein consumption, with lower
values of fs associated with increased protein consumption. Studies on obesity and
health in Mexico indicate that the majority of calories in the daily diet of Mexicans
come from carbohydrates (>60%), while proteins make up only ~11% of caloric
intake (Heien et al., 1989; Fernald et al. 2004; Ruiz‐Arregui et al., 2007; Ortiz‐
Hernández and Gómez‐Tello, 2008). Access to high‐quality protein sources (fs)

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 Stable isotope ratios of tap water and human hair in Mexico    363

may be more closely tied to socioeconomic status, coastal access, and regional
animal husbandry practices than the relative contributions of international and
regionally derived foods (l). Indeed, the three highest values for fs correspond to
Palenque, Oaxaca City and Papantla. Two of these locations, (Oaxaca City and
Palenque) are classified as having high to very high marginality indices. The
Consejo Nacional de Población (CONAPO, the Mexican national council on
population) generates marginality indices every ten years based on census data
(Vertiz, 2011). The marginality index (MI) is a summary measure that differenti-
ates states and municipalities according to socioeconomic deficiencies such as lack
of access to education, inadequate housing, insufficient monetary income, and
rural living. Although MI tells us nothing about the circumstances of any individual
person, it serves as a coarse characterization of SES differences between locations.
Thus, individuals in areas of high MI might be expected to have limited access to
animal protein, and their hair would reflect H fixed from body water to a greater
extent (i.e. high fs).
The lowest fs value is obtained for the coastal location of Gutierrez Zamora,
where marine protein consumption is likely high. Readily available local seafood
will tend to produce markedly different isotopic signatures and distinctly different
relationships between l and fs than might be observed for inland locations.
Michoacán is one of the major pork‐producing regions in Mexico and a location
where consumption of pork is affordable for local populations (Hernandez et al.,
2014). Non‐urban locations like Quiroga, Tzintzuntzan and Patzcuaro having fs
values interspersed with more affluent urban centers is likely consistent with the
state of Michoacán hosting an industry that offers relatively low‐cost and
­abundant access to high‐quality protein.

23.9 Conclusions

We quantified the relationship between modern Mexican hair and water samples
using a modified approach to the semi‐mechanistic framework established by
Bowen et al. (2009). This framework was developed based on the assumption that
individuals consume water that comes from sources near their place of residence,
and that the isotopic signature of that water is quickly reflected in hair isotopes.
That assumption is not likely to hold true for our Southern Mexican population.
The paired hair–water data presented herein does not represent a consistent trend
between hair and tap water. Instead, we found a rather more nuanced connection
between what Mexicans are drinking and what shows up in their hair. After con-
sidering multiple factors that might alter the quality of the model fit, we found
that our data are consistent with a population whose drinking water has no con-
nection to place of residence. Instead, the drinking water isotopes match almost
exactly with the mean of bottled water samples. The bottled water and beverage
industry likely supplies most of what is consumed by Mexicans, as tap water is

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 364    Forensic science and humanitarian action

either not available or unsafe to drink. Beverage suppliers homogenize the

drinking water signature by pulling water from a variety of widely distributed
aquifers, lakes and rivers. Many of the large producers of bottled water also pro-
duce and distribute sodas, utilizing the same water sources for both products. As a
result, the high rates of soda consumption in Mexico likely mimic the homoge-
nized isotopic composition estimated for drinking water. The lack of a clear corre-
lation between locally derived water and hair is consistent with the complex
relationships that modern Mexicans have with drinking water, where upwards of
80% of the population reports bottled water use.
While the drinking water source appears homogeneous across this region, we
were still able to use the semi‐mechanistic model to map regional variations in
hair isotopes to different food consumption patterns. By estimating Mexican
values for international and regional dietary contributions, we obtained values of
l and fs that cleanly segregated urban areas like Mexico City from rural towns like
Quiroga. Likewise, the coastal city of Gutierrez Zamora is clearly distinguished.
The very low value of fs estimated for this location can readily be interpreted as
evidence of an easily accessible, high‐protein, seafood‐based diet. We should note,
however, that the low fs value may also reflect an artefact in the model‐based
analysis. Marine protein will have a distinctly different isotopic signature from
terrestrial animal food sources. If the seafood consumed by inhabitants of Gutierrez
Zamora is characterized by a significantly higher value for δ2Hd,l than what we
have estimated for the regional diet, the corresponding estimate for fs would
appear lower than its true value.
With SES impacting both food and water access, it is likely that increasingly
complex models and analytical approaches that quantify variability and uncer-
tainty will be needed to support provenance estimation. While models capturing
the mechanistic ties between hair and drinking water isotopes hold much promise
for tracing place of origin and migratory behavior, caution must be exercised
in  cases where water stress and its interplay with socioeconomic stratification
obfuscate the relationship between what we consume and where we live.

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