Alcohol: Breath Analysis

AW Jones, Linköping University, Linköping, Sweden

r 2016 Elsevier Ltd. All rights reserved.

Abstract

The analysis of ethanol in exhaled breath is widely used in forensic science and legal medicine as a rapid
test for overconsumption of alcohol. The police authorities use breath-alcohol instruments to control
sobriety in people suspected of driving under the influence of alcohol. The noninvasive nature of breath
sampling offers a definite advantage over drawing blood for analysis of alcohol at a forensic laboratory.
Two categories of breath-alcohol instrument are currently in use: hand-held electronic instruments that are
used to screen motorists at the roadside and more sophisticated evidential breath analyzers. Results with
the latter are accepted by the court when traffic offenders are prosecuted. This article reviews historical
development, physiological principles, and practical application of breath-alcohol analysis in forensic
science and legal medicine.

Glossary determining the concentration of the same substance

Accuracy A measure of the closeness of
agreement between an analytical result and the
known or assigned value of the quantity measured
(e.g., concentration of analyte). The discrepancy
or difference between the true value and the
analytical result obtained is often referred to as
bias.
Alveolar air Alveolar air is defined as that fraction of
the exhaled breath remaining after air from the dead-
space region of the lungs (mainly nose and upper
airway) has been ventilated out. For determination of
the concentration of alcohol in breath at least 1.5 l of
a prolonged exhalation should be discarded prior to
sampling.
Analytical sensitivity The ability of a method or
instrument to discriminate between samples
containing different concentrations of the target
analyte. The slope of the analytical calibration
function (plot) is one index of the method’s
sensitivity.
Analytical specificity The ability of a method of
analysis to determine the desired compound without
responding or cross-reacting with interfering
substances that might be present in the sample
analyzed.
Calibration A process or sequence of steps in which
an analytical instrument or method is standardized
to allow quantitative or semiquantitative
measurements. This is usually done by analysis of
known strength standards prepared from a pure
substance and used to establish response of the
instrument at increasing concentrations of the target
analyte. The resulting calibration curve and
appropriate response factors are then used for
in the unknown specimens.
Forced vital capacity (FVC) This is the maximum
volume of air exhaled from full inspiration to forced
maximum expiration. The FVC during the first
second of exhalation is another parameter of lung
function.
Gas–liquid chromatography (GLC) An analytical
technique widely used for separating volatile
substances in a sample on the basis of their
solubility and partition between a gas and a
liquid phase.
Henry’s law Deals with the solubility of gases in
liquids and states that for dilute solutions at
equilibrium and at constant temperature, the
solubility of a gas in a liquid is directly proportional to
the partial pressure of the gas above the liquid.
Infrared (IR) A region of the electromagnetic
spectrum extending from approximately 0.78 to
300 mm wavelength. Many evidential breath analyzers
incorporate IR detectors either at wavelengths of
B3.4 mm corresponding to the CH stretching in
ethanol molecules or B9.5 mm corresponding to the
C–O stretching.
Lambert–Beer law The absorbance of a
homogeneous sample containing an absorbing
substance is directly proportional to the concentration
of the absorbing substance.
Precision The closeness of agreement between
independent analytical measurements of the same
substance under prescribed conditions. A measure of
the dispersion or scatter of the repeated measurements
about the mean defined by the standard deviation of
the measured values. A method with high precision
has good repeatability.

Encyclopedia of Forensic and Legal Medicine, Volume 1 doi:10.1016/B978-0-12-800034-2.00011-2 119

120 Alcohol: Breath Analysis
Introduction
The smell of alcohol on breath along with the person’s
general appearance and behavior constituted the first,
albeit nonspecific, clinical test of drunkenness (Borken-
stein, 1985). More specific tests for alcohol influence
required identification of ethanol as the intoxicating
substance and quantitative analysis of the concentration
present in blood, breath, or urine. Many human drink-
ing studies showed that the signs and symptoms of
alcohol intoxication, such as excessive talkativeness,
staggering gait, slurred speech etc., were not always
visible (Brick and Erickson, 2009). A much more sci-
entific test for overconsumption of alcohol is furnished
by measuring the concentration of ethanol in the per-
son’s blood or breath. The impairment of body func-
tions caused by drinking alcohol increases in direct
proportion to the amounts consumed and the concen-
trations in blood or breath, albeit with large intersubject
variations (Mellanby, 1920; Penttila and Tenhu, 1976).
The first scientific studies aimed at investigating the
disposition and fate of ethanol in the body took place in
the mid-nineteenth century, at a time when rapid ad-
vances occurred in analytical chemistry. In Victorian
Britain overconsumption of alcohol and drunkenness was
a major public health problem as it still is today (Porter,
1985; Spring and Buss, 1977). The medical community
was divided as to what actually happened to alcohol after
drinking; was it a food, a poison, or a medicine? Some
physicians considered that all the alcohol consumed was
eliminated unchanged in breath, urine, and sweat,
whereas others were convinced it was broken down and
utilized in the body just like ordinary foodstuffs.
This question was resolved by a British physician
Francis Edmund Anstie (1833–1874) in experiments on
humans and in animals (Anstie, 1874). To investigate
what happened to alcohol in the body, an analytical
method was needed so that the concentration in bio-
logical specimens, including exhaled breath, perspir-
ation, and urine could be determined. For this purpose
Anstie developed an oxidizing agent consisting of a
mixture of potassium dichromate and concentrated
sulfuric acid, which became known as Anstie’s reagent.
This same chromic acid solution, with minor modifi-
cations, was used for many decades afterwards for the
analysis of ethanol in blood and other biological fluids.
The reagent developed by Anstie changed color from
red-brown to emerald green as ethanol in the sample
was oxidized to acetic acid and the intensity of the color
change was proportional to the amount of ethanol in the
sample analyzed. After known amounts of alcohol were
given to healthy volunteers, hospital patients, and ex-
perimental animals, Anstie found that only a small
fraction of the amount administered was recoverable
unchanged in breath, sweat, and urine. He therefore
concluded that the bulk of the ingested dose of ethanol
was oxidized in the body and used as other nutrients.
During his experiments on measuring alcohol in breath,
Anstie warned about the risk of sample contamination by
alcohol in the mouth and oral mucosa from a recent drink
(Anstie, 1867). He wrote “Much caution is necessary
however in applying this test. It must not be tried within
the first quarter hour after a dose has been taken for the
mouth retains the characteristic smell even if the most
moderate dose of any of the stronger smelling drinks for
fully this time.” Now referred to as the ‘mouth alcohol
effect’ this old observation is the reason that at least
15–20 min must elapse after the last drink before breath-
alcohol instruments are used for evidential purposes.
The first practically useful instrument designed for
breath-alcohol analysis was developed in the mid-1950s
by Professor Robert Frank Borkenstein at Indiana
University, in the USA (Lucas, 2000). This instrument
was named the Breathalyzers and by the 1960s its use
was accepted for testing drivers and as evidence for the
prosecution of traffic offenders in the USA, Canada, and
Australia (Borkenstein and Smith, 1961). The person to
be tested had to make a forced exhalation through a tube
and mouthpiece so that a known volume of the breath
was captured inside the Breathalyzer. This breath was
passed through a mixture of potassium dichromate and
sulfuric acid contained in a glass ampoule. The ethanol
content of the breath sample, if any, was determined by
chemical oxidation and the end-point of the reaction was
based on photometry (Dubowski, 1991). After reaction
with alcohol, the change in the intensity of the yellow
color of the oxidizing agent was measured by application
of the Lambert–Beer law. The Breathalyzer instrument
was designed and calibrated to report results of the test as
the estimated blood-alcohol concentration (BAC), which
required assuming a population average blood/breath
ratio (BBR) of alcohol of 2100:1 (Begg et al., 1964).
In the 1970s methods for measuring ethanol in both
blood and breath were improved considerably by use of
more specific analysis, including use of gas–liquid
chromatography (GLC), infrared (IR) spectrometry, and
electrochemical (EC) oxidation principles (Jones, 1996).
Moreover, a lot of research was done concerning the
physiological principles of breath-alcohol analysis and
the factors that influence the blood-breath alcohol rela-
tionship or ratio (Hlastala, 1998; Wright et al., 1975).
Among other things, the BBR was not a constant 2010:1
and instead tended to vary both among subjects and also
within subjects as a function of sampling time after
drinking ended (Alobaidi et al., 1976). The dependence
of BBR on sampling time was mainly a consequence of
arterial-venous differences in BAC and the fact that
breath-alcohol concentration (BrAC) runs closer to the
arterial BAC (Martin et al., 1984).
The noninvasive nature of breath sampling, coupled
with advances in microelectronics and computers led to
the development of more compact and easier to operate
breath-alcohol analyzers. In the 1980s many countries in
Europe began to update their existing drink-driving laws

to include provisions for using evidential breath-alcohol
instruments in addition to blood-alcohol testing. Two
kinds of breath-alcohol testing equipment are available.
First, hand-held instruments became widely used as
preliminary roadside screening tests for people suspected
of driving under the influence of alcohol. The second
type of breath analyzer used in law enforcement is more
sophisticated and the results are used as evidence for
prosecuting traffic offenders.
Since the 1980s, many European countries (UK, Hol-
land, Austria, Norway, and Sweden) have approved the
use of breath-alcohol analysis for evidential purposes and
the results were accepted by the courts for prosecuting
offenders (Emerson et al., 1980). The legal significance of
evidential breath-alcohol test results and consequences
for those found to exceed the prescribed alcohol limit
required a rigorous program of quality assurance. This
included making duplicate determinations, ensuring that
the instrument is properly calibrated when every suspect
is tested, the analysis of ambient air as a blank test as well
as providing a printed record to document the final result
of the testing (Dubowski, 1994).
The results of breath-alcohol analysis are either re-
ported as the concentration of ethanol per unit volume
of breath (mg/l, mg/100 ml, or even g/210 l) or alter-
natively a conversion is made to give the equivalent
concentration in blood. Making such a conversion
requires use of a calibration factor, the so-called blood-
breath ratio, which is subject to considerable inter-
and intrasubject variation. Because of biological and
physiological variations in the BBR of alcohol, for legal
purposes it was considered more rational to define
statutory BrAC limits separate from pre-existing blood-
alcohol limits (Mason and Dubowski, 1976). This
meant that drunk drivers could be prosecuted based on
the concentration of alcohol in breath without the need
for additional evidence obtained from the analysis of a
blood sample. The statutory BrAC limits were derived
from the pre-existing statutory BAC by assuming a
certain population average BBR of alcohol. Unfortu-
nately, different countries opted to use different BBRs
when statutory BrAC limits were calculated, and these
BBRs ranged from 2000:1 to 2400:1 (Jones, 2011).
This chapter deals with the physiological principles
and the practical application of breath-alcohol analysis
when used in forensic science and legal medicine to
control sobriety of drivers and evidence for prosecution
of traffic offenders. The words alcohol and ethanol are
used interchangeably in this account and refer to ethyl
alcohol (CH3CH2OH), the main pharmacologically
active constituent of alcoholic beverages.
Alcohol Traffic Legislation
Different countries have their own structure and tradi-
tions when it comes to dealing with the problem of
Alcohol: Breath Analysis 121
drunk driving. Factors that influence government policy
and legislation include statistics about drunk driving
and road-traffic fatalities, attitudes of the media and
the general public towards drinking and driving, and the
activity of temperance movements and the way courts
interpret the law. Other things that impact on the
prevalence of drunk driving include the relative cost and
availability of alcoholic beverages, the opening hours of
bars and restaurants, and the resources available to the
police authorities for apprehending and prosecuting of-
fenders. In most countries statutory limits of BAC and/or
BrAC exist above which it is an offence to drive a motor
vehicle on the public roads.
The threshold blood-alcohol limits vary from 20 to
80 mg/100 ml in EU countries and this fourfold differ-
ence seems to depend more on political attitudes rather
than traffic safety research. The statutory alcohol limits
for driving in a particular country should balance the
dangers of drunk driving with police powers to enforce
the law and public attitudes and acceptability of routine
alcohol testing of motorists.
Many epidemiological surveys verify that the risk of
involvement in a traffic crash increases in direct pro-
portion to the driver’s BAC or BrAC (Blomberg et al.,
2009). At a BAC between 20 and 50 mg/100 ml the risk
of a crash is only marginally higher than at zero BAC,
but as BAC passes 80 mg/100 ml a significant and ex-
ponential increase in risk of a crash occurs. Further
evidence for the dangers of drunk driving comes from
evaluation of autopsy reports of drivers fatally injured in
crashes. Statistics show that in 20–50% of victims the
BAC at autopsy was above the statutory alcohol limit
(Romano et al., 2014). The percentage of drivers killed
with BAC above the legal limit was appreciably higher
for single vehicle crashes compared with multiple vehicle
crashes (Jones et al., 2009).
Statutory Limits of Blood-Alcohol Concentration
Norway was the first country to introduce a statutory
BAC limit for driving. It was originally set at 0.50 mg/g
blood (B50 mg/100 ml), but has been revised since then
to 0.20 mg/g (B20 mg/100 ml). Sweden introduced a
punishable BAC limit of 0.80 mg/g (B80 mg/100 ml) in
1941, and this was subsequently lowered to 0.50 mg/g
in 1956 and stands at 0.20 mg/g (B20 mg/100 ml)
today. Enhanced penalties are imposed if drivers have
a BAC 4100 mg/100 ml and this includes having to
retake a driving test and being subjected to a medical
examination for substance abuse disorder. How to deal
with hard-core drinking drivers is a dilemma because
many still continue to drive even without a valid driving
permit. This has led to the mandatory use of ignition
interlock devices, so that a driver has to undergo and
pass a breath-alcohol test before the vehicle will start
(Marques et al., 2003).
122 Alcohol: Breath Analysis

A compilation of statutory BAC and BrAC limits
currently enforced in various countries is shown in
Table 1. Note the different concentration units used to
report BAC and BrAC and the various BBRs used to
derive the threshold BrAC from the pre-existing BAC
limits. Also shown are the evidential breath-alcohol
analyzers approved and the analytical principle they
incorporate. Manufacturers of evidential breath-alcohol
analyzers are continuously updating and improving their
products in various ways, so the actual model currently
available might not be exactly the same as those in
Table 1.

Table 1 Statutory concentration limits of alcohol in blood (BAC) and breath (BrAC) in different countries and the blood-breath ratios (BBR) of
alcohol used to derive statutory BrAC limits

Country Statutory BAC Statutory BrAC BBR Evidential breath- Analytical principle for
alcohol analyzer detection of ethanol

Austria 0.50 g/l 0.25 mg/l 2000 Alcotest 7110 IR at 9.5 mm and EC
Belgium 0.50 g/l 0.22 mg/l 2300 Ethylometer IR 9.5 mm
Alcotest 7110 IR 9.5 mm and EC
Alcotest 8510 IR 9.5 mm
France 0.50 g/l 0.25 mg/l 2000 Ethylometer IR 9.5 mm
Greece 0.50 g/l 0.25 mg/l 2000 Alcolmeter SD-400 EC oxidation
Italy 0.50 g/l 0.25 mg/l 2000 Alcotest 7110 IR (9.5 mm) and EC
Intoxilyzer 8000C IR 9.5 mm
Ethylometer IR 9.5 mm
Poland 0.20 g/l 0.10 mg/l 2000 AlcoSensor IV EC oxidation EC
Alcotest 7410 oxidation
Portugal 0.50 g/l 0.25 mg/l 2300 Alcotest 7110 IR (9.5 mm) and EC
Ireland 50 mg/100 ml 22 mg/100 ml 2300 Evidenzer IR multiple wavelengths
3.3–3.5 mm
Spain 0.50 g/l 0.25 mg/l 2000 Alcotest 7110 IR (9.5 mm) and EC
Netherlands 0.50 mg/ml 220 mg/l 2300 DataMaster IR 3.4 mm
UK 80 mg/100 ml 35 mg/100 ml 2300 Intoximeter EC-IR EC oxidation
Intoxilyzer 6000 IR multiple wavlengths
in 3.3–3.4 mm range
Australia 0.05 g/100 ml 0.25 mg/l 2100 Alcotest 7110 IR (9.5 mm) and EC
DataMaster IR 3.4 mm
Canada 80 mg/100 ml 0.08 g/210 l 2100 Intoxilyzer 5000C IR 3.3–3.5 mm
Intoxilyzer 8000C IR 9.5 mm
Intox EC/IR II EC detector (primary)
New Zealand 80 mg/100 ml 400 mg/l 2300 Seres Ethylometer IR 9.5 mm
Intoxilyzer 5000
Dräger Alcotest 9510 IR 3.4 mm
IR 9.5 mm and EC
USA 0.08 g/100 ml 0.08 g/210 l 2100 Intoxilyzer 8000 Intox IR 3.4 mm and 9.4 mm.
EC/IR DataMaster EC detector (primary)
DMC
Alcotest 7110 IR (3.4 and 9.5 mm) IR
(9.5 mm) and EC
Alcotest 9510 IR (9.5 mm) and EC
In the countries below statutory BAC limits are reported in mass/mass units (mg/g), which must be converted to units of mass/volume (mg/ml)a when
blood-breath ratios (BBR) of alcohol are calculated in the 4th column
Denmark 0.50 mg/g (0.53 mg/ml)a 0.25 mg/l 2100 Evidenzer IR absorption at
multiple wavelengths
3.3–3.4 mm
Finland 0.50 mg/g (0.53 mg/ml)a 0.22 mg/l 2400 Alcotest 7110 IR (9.5 mm) and EC
Evidenzer IR multiple wavelengths
3.3–34 mm
Germany 0.50 mg/g (0.53 mg/ml)a 0.25 mg/l 2100 Alcotest 7110 IR (9.5 mm) and EC
Norway 0.20 mg/g (0.21 mg/ml)a 0.10 mg/l 2100 Intoxilyzer 5000 N IR multiple wavelengths
Evidenzer 3.3–3.4 mm
Sweden 0.20 mg/g (0.21 mg/ml)a 0.10 mg/l 2100 Evidenzer IR multiple wavelengths
3.3–3.4 mm
a
The blood-alcohol concentrations expressed in mg/ml equals the concentration in mg/g multiplied by the density of whole blood (1.055 g/ml).
Note: The approved instruments for evidential breath testing and the analytical principle for determination of ethanol are also shown where EC ¼ electrochemical oxidation and
IR ¼ infrared spectrometry.

In the USA and UK there is a lot of discussion and
debate about lowering the statutory blood-alcohol limit
from 80 mg/100 ml to 50 mg/100 ml (Fell and Voas,
2014). Such a change in the law will probably not deter
hard-core offenders and traffic delinquents from driving
after drinking, because many of these individuals are
problem drinkers or alcoholics and would benefit
more from treatment rather than the usual punishments
for drunk driving. However, lowering alcohol limits
draws public and media attention to the problem of
drunk driving, which will hopefully have a pedagogic
influence on young people learning to drive. Besides
lowering BAC limits, the police authorities should be
given powers to conduct random (mandatory) breath
testing of drivers for alcohol influence, because signs and
symptoms of impairment are not always evident at a
BAC of 50 mg/100 ml.
Statutory Limits of Breath-Alcohol Concentration
In most countries punishable BAC limits for driving
existed long before the use of evidential breath-alcohol
instruments was considered feasible. An exception
was the USA because of constitutional issues about un-
reasonable search and seizure and self-incrimination it
was more problematic to obtain blood samples for an-
alysis in criminal cases. When the first drink-driving
laws were enforced the suspects were expected to pro-
vide samples of breath or urine for analysis of alcohol
concentration.
When in the 1980s European nations began to use
evidential breath-alcohol instruments, it was considered
more scientifically sound to establish a threshold BrAC
instead of converting the result into a BAC. Unfortu-
nately, different countries opted to use different BBRs
to arrive at statutory BrAC limits ranging from 2000:1
to 2400:1 as indicated by the information in Table 1.
The UK Road Transport Act of 1981 stipulated
35 mg/100 ml as the punishable BrAC limit and
this was derived from the pre-existing BAC limit
of 80 mg/100 ml. This assumed a BBR of 2300:1
(80/35  1000¼2286:1, which rounds up to 2300:1).
The Republic of Ireland, the Netherlands, and Belgium
also adopted a 2300:1 BBR when setting their respective
BrAC limits. Other European nations such as France and
Spain approved a BBR of 2000:1.
The situation in Germany and the Nordic countries
is more complicated because the statutory BAC limit
is reported in mass/mass concentration units (e.g.,
0.50 mg/g or 0.50 g/kg), which is the same as 0.527 mg/
ml (density of blood¼ 1.055 g/ma). Dividing 0.527 mg/
ma by 2100 (BAC/BrAC ratio) gives a threshold
BrAC of 0.25 mg/l ((0.50/2100)  1000¼ 0.25 mg/l).
The statutory BAC and BrAC limits that operate in
Germany and the Nordic countries are also shown in
Table 1.
Alcohol: Breath Analysis 123
Alcohol in the Body
The fate of alcohol in the body is usually considered as
three separate processes: absorption, distribution, and
elimination depending on time elapsed after drinking
starts. However, in reality these three processes occur
simultaneously but at different rates depending on the
pattern of drinking and the time elapsed after end of
drinking until blood samples were taken (Norberg et al.,
2003). Immediately after drinking, the ethanol in alco-
hol beverages starts to become absorbed from the
stomach and intestines, whereas the rate of absorption
depends primarily on dose administered, speed of
drinking, and the efficacy of gastric emptying.
The absorption phase refers to the increase in BAC
that takes place during and shortly after the end of
drinking. The distribution phase refers to the change in
BAC that occurs as more and more alcohol is absorbed
and passes from the bloodstream into the various body
organs and tissues. The speed of distribution is different
for different tissues depending on the ratios of blood
flow to tissue mass. Accordingly, organs with a rich
blood supply and a high ratio of flow to mass (e.g., brain
and kidney) reach equilibrium with alcohol in the ar-
terial blood more rapidly, whereas bulky skeletal mus-
cles, which have a much smaller ratio of blood flow to
tissue mass, require up to 60 min. The elimination phase
refers to the linearly decreasing BAC mainly reflecting
metabolism of ethanol in the liver and excretion in
breath and urine.
Absorption Phase
After drinking, the alcohol contained in alcoholic bev-
erages starts to become absorbed through the gastric
mucosa although the bulk of the dose ingested is ab-
sorbed in the duodenal and jejunal mucosa, owing to the
much larger absorption surface area available. Factors
that delay or accelerate gastric emptying play a major
role for the absorption of ethanol. This impacts on both
the time of reaching a peak concentration in blood
and the maximum concentration reached (Jones and
Jonsson, 1994). Many factors influence gastric emptying
including splanchnic blood flow and importantly pres-
ence and amount of food in the stomach, the alcoholic
beverage consumed, smoking cigarettes, and various
medications.
The time elapsed after end of drinking before reach-
ing a peak concentration in blood usually ranges from 5
to 120 min (Mitchell et al., 2014). On reaching the
portal venous blood the alcohol gets transported to the
liver, then on to the heart through the hepatic vein, via
the heart to the lungs, and then throughout the entire
body and back to the heart with the deoxygenated
venous blood. There is some evidence that a small
amount of ingested alcohol gets metabolized in the gut
124 Alcohol: Breath Analysis
mucosa or the liver before reaching the systemic circu-
lation (Baraona et al., 1994). This phenomenon is
referred to as first-pass metabolism (FPM) and can be
avoided by giving alcohol intravenously. The extent of
FPM is also negligible when alcohol is consumed as a
bolus dose on an empty stomach, because ethanol can be
used as a marker to determine total body water in the
same way as radio- or stable isotopes of water are used
(Endres and Gruner, 1994).
Distribution Phase
From the hepatic vein alcohol reaches the inferior vena
cava and mixes with systemic venous blood from the
superior vena cava and the coronary sinus in the right
atrium. The blood is then pumped from the right ven-
tricle through the pulmonary circulation and back to the
left ventricle. The left ventricle of the heart supplies
all organs and tissues with oxygenated arterial blood.
Alcohol distributes into the water compartment of the
body, which corresponds to 50–60% of body weight,
and there is no evidence that ethanol binds to plasma
proteins or concentrates in any body organs or tissues.
The speed of distribution of alcohol into body fluids
and tissues depends on the ratio of blood flow to tissue
mass. Organs and tissues with a rich supply of blood
equilibrate rapidly with arterial blood concentrations of
alcohol, whereas equilibration over the capillary beds in
the bulky muscle tissue takes a longer time. The con-
centration of alcohol in arterial blood is therefore higher
than in venous blood returning to the heart during the
absorption phase of the blood-alcohol curve (Jones
et al., 2004).
The arterial (A) blood contains a higher concen-
tration than the venous (V) blood for about the first
60–90 min after drinking, as can be seen from the con-
centration–time plot in Figure 1. When the distribution
phase ends, the concentrations of ethanol are the same in
A and V blood but only for an instant. At all later times
the concentration of alcohol in the venous return from
skeletal muscles is slightly higher than the arterial blood
concentration as ethanol continues to be cleared from
the central compartment in the liver. This temporal
variation in arterial and venous BAC has relevance for
the relationship between BrAC and venous BAC.
Elimination phase
Most of the alcohol consumed (95–98%) is eliminated
from the body by metabolism which occurs primarily in
the liver by the action of enzymes. The remaining frac-
tion of the dose (2–5%) is excreted unchanged in the
kidneys, the lungs, and a small amount through the skin
in sweat (Drummer, 2014). The principal oxidative en-
zyme is alcohol dehydrogenase (ADH), which is located
Blood alcohol concentration (mg/100 ml)
140
Arterial blood
120 Venous blood
100
80
60
40
20
0
0 100 200 300 400 500
Time (min)
Figure 1 Concentration–time profiles of ethanol in venous blood
(cubital vein) and arterial blood (radial artery) in one male subject
who consumed 0.80 g ethanol per kg body weight on an empty
stomach.
in the cytosol fraction of the hepatocytes (liver cells)
and which converts ethanol into acetaldehyde. This
toxic metabolite is then oxidized further into acetic
acid by the enzyme aldehyde dehydrogenase (ALDH),
which is located in mitochondria. The acetate produced
from oxidation of acetaldehyde enters the citric acid
cycle as acetyl-CoA and is converted in peripheral
tissues into the end products carbon dioxide and
water. After drinking ethanol labeled with 14C, such as
14
CH2CH2OH, then 14CO2 is almost immediately
afterwards measurable in breath.
The rate of ethanol metabolism depends on many
factors, including gender, age, ethnicity, circadian
rhythm, liver blood flow, food intake, and habituation
so that metabolic tolerance develops. After consumption
of a moderate dose of alcohol and when BAC exceeds
20 mg/100 ml the pharmacokinetics of ethanol is a zero-
order process, meaning that a constant amount is elim-
inated from the body per unit of time, independent of the
concentration present in blood. A good average rate of
ethanol elimination from blood is 15 mg/100 ml per h,
ranging among individuals from 10 to 25 mg/100 ml per h
(Jones, 2010).
Classification of Breath-Alcohol Instruments
The many instruments available for breath-alcohol an-
alysis can be classified in various ways, such as by their
size, purchase cost, method of ethanol analysis, or the
intended area of application. A useful method of clas-
sification is according to the area of application, whether
for self-testing or as a preliminary roadside screening test
for alcohol or as a quantitative evidential analyzer when
results are used as evidence for prosecution of traffic
offenders. Hand-held screening instruments are used
by police officers to test drivers on the street or at the
 Alcohol: Breath Analysis 125

roadside or even as they are sitting behind the wheel of
their vehicles. By contrast, tests with evidential breath
analyzers are done at police stations and the procedure
involves many more quality control requirements com-
pared with roadside testing.
Table 2 presents a historical time line showing the
development of handheld breath analyzers used for
screening motorists at the roadside. The latest gener-
ation of breath-alcohol screening instruments incorpor-
ates an electrochemical sensor for analysis of ethanol
in breath. The instruments available have options for
digital display of results, which provides a pass, warn,
and fail indication. The sampling of breath is done
automatically as the test subject exhales into a dis-
posable mouth piece fitted to the instrument and makes
a prolonged exhalation. Provided blow time and pres-
sure restraints are met, a sample is captured auto-
matically. However, the hand-held screening units
cannot detect presence of mouth alcohol and there is
currently no requirement to make a calibration control
check with every subject tested. The results from 50 up
to 100 consecutive tests can be stored in an internal
memory of the analyzer and downloaded later along
with date and time of the test. Alternatively, results can
be printed out directly on-site.
Although manufacturers of breath-alcohol analyzers
claim certain levels of accuracy and precision and
specificity, it is always advisable to verify these claims by
independent testing. Such testing should involve both
in vitro conditions using known concentrations of
ethanol in air and also in vivo testing with drinking
subjects. The in vitro testing is usually done using a
breath-alcohol simulator by passing a stream of air
through a known strength ethanol solution kept at a
constant temperature of 34 1C. More commonly today
dry-gas ethanol standards in pressurized tanks are used
for the in vitro performance tests. The final stage of
testing is done with human subjects, both male and fe-
male of various ages (20–60 years), who consumed
known amounts of ethanol under laboratory conditions
to reach a BAC above the legal alcohol limits for driving.
When evaluating results of the controlled drinking
studies, it is important to distinguish the precision and
accuracy of measuring alcohol in breath from the pre-
cision and accuracy of estimating BAC. The latter re-
quires use of a calibration factor so that breath test
results are reported as the presumed BAC. The closeness
of agreement (accuracy) of the breath analyzer results
and the actual BAC depends on the value of the BBR
used for calibration of the instrument.
Besides the roadside screening tests discussed above,
another category of breath-alcohol instruments is used
that are accepted by the courts as evidence when traffic
offenders are prosecuted. These instruments are highly

Table 2 Historical developments in breath-alcohol instruments used as preliminary roadside tests and the analytical principle incorporated for
determination of ethanol

First appearancea Instrument/device (country of origin) Brief details of the analytical principle used for
measuring alcohol

1953 Alcotest tube and bag (Germany) Chemical oxidation with K2CrO4 þ H2SO4 adsorbed on
silica gel
1956 Kitagawa chemical detector tube (Japan) Oxidation with chromic acid
1969 Sober-meter chemical detector tube (USA) Oxidation with K2CrO4 þ H2SO4 adsorption on silica
gel for later verification analysis
1969 Alcolyser tubes (Wales, UK) Oxidation with K2CrO4 þ H2SO4 adsorbed on silica gel
carrier
1974 Alcolmeter (Wales, UK)b Electrochemical oxidation
1975 AlcoSensor (USA)c Electrochemical oxidation
1975 ALERT (Canada) Taguchi cell, solid state stannic oxide semiconductor
detector
1978 Alcytron (Germany) Infrared absorption at 3.3–3.4 mm
1980 Alcotest (Germany) Infrared absorption at 3.3–3.4 mm
1988 Alcotest 7410 (Germany) Electrochemical oxidation
Breathalyzer 7410 (USA)
1995 Alcomat S (Germany) Electrochemical oxidation
1995 Alcoodose 2 (France) Infrared absorption at 9.5 μm
2000 Lifeloc (USA) Electrochemical oxidation
2000 Alcotest 6510 and 6810 (Germany) Electrochemical oxidation
2000 Intoxilyzer S-D5 (USA) Electrochemical oxidation
2000  present A large number of hand-held instruments intended Solid state stannic oxide semiconductor (Taguchi cell)
primarily for self-testing by the general public are
available
a
The dates shown are approximate and represent the time when the first published report appeared.
b
The electrochemical sensor incorporated in the Alcolmeter device was also used in the first model of the AlcoSensor marketed in USA.
c
Over the years different versions of the AlcoSensor instrument have appeared (model IV, FST, VXL etc.).
126 Alcohol: Breath Analysis

sophisticated and incorporate slope detectors to ensure a Infrared Breath-Alcohol Analyzers

proper sample for analysis, more specific identification
of ethanol, calibration control with every subject test,
and a printed record of the results are obtained. Table 3
gives details of the historical breath-alcohol instruments
for evidential testing.
When the first breath-alcohol instruments appeared
in the USA in the 1950s (e.g., the Breathalyzer) statutory
limits of alcohol in blood for driving in most states was
150 mg/100 ml. The results of the breath-alcohol test
were mainly used as supportive evidence that a driver
was under the influence of alcohol. The major thrust of
any prosecution case was the circumstances surrounding
the driving incident and the results of clinical tests of
drunkenness. Suspects were examined by a police sur-
geon or a forensic physician and questions asked about
drinking and drug habits and simple cognitive and
psychomotor tests were performed. The demands for
high accuracy and precision of the breath-test results
were less stringent and were used as supporting evi-
dence. The introduction of a concentration per se law
meant that a person could be prosecuted based on the
result of alcohol analysis in blood or breath and without
the need for any clinical evidence of drunkenness. The
enactment of the so-called per se laws shifted the focus
toward quality assurance and reliability of the chemical
analysis of ethanol in samples of blood or breath and
away from clinical evidence of impairment of the driver.
Most of the breath-alcohol analyzers currently used for
evidential purposes use infrared (IR) spectrometry as the
analytical principle for quantitative analysis and identi-
fication of ethanol (Harding and Zettl, 2008). Infrared
analysis is a nondestructive technology and calibration
of the instruments shows good long-term stability. The
theory and practice of infrared spectrometry is well es-
tablished and is widely used in analytical chemistry for
identifying functional groups in organic compounds.
Infrared spectrometry applied to breath-alcohol analysis
monitors the absorption of selected wavelengths of
infrared radiation after passage through a known vol-
ume of the breath sample. The higher the concentration
of ethanol in the sample the more infrared energy is
absorbed and the lower is the percent transmittance.
When organic compounds absorb IR energy at cer-
tain wavelengths, the vibration and rotational motion of
chemical bonds in the molecule become amplified and
more intense. Alcohols have characteristic IR absorp-
tions associated with O–H, C–O, and C–H bond
stretching vibrations. The C–H bond vibrations are not
unique for alcohols, because many other simple organic
molecules, such as hydrocarbons and acetone, also have
C–H bonds that can absorb IR radiation at the 3.3–
3.5 mm wavelength. Nevertheless, using several narrow
band width optical filters the specificity for detecting just

Table 3 Historical developments since the 1950s in breath-alcohol instruments used for evidential purposes and the analytical principle
incorporated for determination of ethanol

Time period Breath-alcohol instrument Brief details of the analytical principle for measuring ethanol

1953  1970 Breathalyzer A solution of K2CrO4 þ sulfuric acid is contained in a glass ampoule through which a
Photo-Electric Intoximeter known volume of breath passes. Any ethanol present is oxidized and the end-point
Ethanographe of the reaction is determined by photometry
1969 GC Intoximeter Gas chromatography with a flame ionization detector (FID) for the quantitative
analysis of ethanol and retention time for qualitative analysis
1971 Alco-Analyzer Gas chromatography with thermal conductivity detector for quantitative analysis and
retention time for qualitative analysis
1971 Omicron Intoxilyzer The first breath analyzer based on infrared absorption at a single wavelength of
3.39 mm
1973 ALERT (screening test) Metal oxide semiconductor Taguchi (T-cell)
1975 Intoximeter 3000 Single wavelength IR (3.4 mm) for ethanol and a Taguchi T-cell to detect interfering
substances
1979 Alcomat Absorption of IR energy at wavelengths of 3.4 mm (C–H stretching) or 9.5 mm (C–O
Alcotest stretching)
1976 Intoxilyzer 5000 Dual wavelength IR analyzer (3.39 mm and 3.48 mm)
1979 BAC DataMaster Dual wavelength IR analyzer (3.37 mm and 3.44 mm)
1986 Alcotest 7110 IR analyzer operating at a single wavelength of 9.5 mm
Alcomat
1992 Intoxilyzer 6000 Multi-wavelength IR analyzer in the 3.3–3.5 mm region of the infrared spectrum
1994 Intoximeter EC/IR Electrochemical fuel cell detector for analysis of alcohol and IR detector for
monitoring the CO2 profile during exhalation
1995 Alcotest 7110 Mark III Both IR spectrometry and electrochemical (fuel cell) detector
2000–present DataMaster DMT Multiple wavelength IR absorption at 3.34, 3.37, and 3.50 mm
2000–present Alcotest 9510 Combined IR (9.5 mm) and electrochemical (fuel cell) detector for alcohol detection
2000–present Evidenzer Multiple (5) wavelengths in the 3.5–3.9 mm region of the IR spectrum
2000–present Intoxilyzer 9000 IR detection at wavelengths of 3.4 mm and 9.5 mm

2.5 3.5 4.5 5.5
100
Transmittance (%)
H C
C–H stretch
3.4 µm
0
Figure 2 Infrared absorption spectrum of ethanol molecules in the gas phase showing the parts of the spectrum that correspond to vibrations of
the C–H bond (3.4 mm) and C–O bond (9.5 mm).
ethanol is improved considerably. Several infrared
breath analyzers incorporate between three and five fil-
ters that absorb IR light at fixed wavelengths of 3.32,
3.40, and 3.49 mm and a reference wavelength common
to several potential interfering substances (Jones and
Andersson, 2008). A typical vapor phase infrared ab-
sorption spectrum for ethanol is shown in Figure 2 with
frequency shown both as wave number (cm1) and in
micrometers (mm).
Ethanol molecules absorb IR light at wave number
3391 cm1 (2.95 mm) and this corresponds to the O–H
stretching vibrations, whereas the C–O stretching
is prominent at 1102 and 1055 cm1 (9.1–9.5 mm). To
enhance the analytical selectivity some infrared breath
analyzers are designed with optical filters that absorb
infrared energy at 3.4 and 9.5 mm wavelengths, corres-
ponding to C–H stretch and C–O stretch.
Electrochemical (Fuel Cell) Instruments
The other major analytical technology used to measure
ethanol in breath for forensic purposes is electro-
chemical oxidation. These devices incorporate a fuel cell
consisting of two platinum conductor electrodes separ-
ated by an electrolyte, such as phosphoric acid, that
conducts ions between anode and cathode parts of the
cell. The ethanol contained in a known volume of breath
(e.g., 0.5–1 ml) enters the chamber of the cell and acts as
the combustible fuel, whereas atmospheric oxygen
functions as the oxidant. In a two-stage oxidation re-
action ethanol is converted to acetaldehyde and the
latter is further oxidized to acetic acid, thus four elec-
trons are released and the flow of current is directly
proportional to the concentration of ethanol in the
breath sample. After amplification the response from the
fuel cell can be followed on a chart recorder (Figure 3)
or displayed as a digital or analog signal. The peak
Alcohol: Breath Analysis 127
Wavelength (µm)
6.5 7.5 8.5 9.5 10.5 11.5 12.5
H H
C OH
H H C–O stretching
9.5 µm
response is directly proportional to the concentration of
ethanol in the sample of breath or vapor analyzed.
The electrode potential of the fuel cell is selected so
that neither water vapor nor oxygen in the breath reacts
to any measurable amount. The breath from healthy
individuals does not contain any volatiles besides etha-
nol after a person drinks alcoholic beverages that are
oxidized by the electrochemical cell. Acetone might be at
elevated concentrations in the breath of diabetics or after
a prolonged fast, although this ketone is not oxidized at
the electrode potential of the fuel cell. If other volatile
alcohols existed in blood, such as methanol or iso-
propanol, from drinking denatured alcohol then they
theoretically increase the response attributed to ethanol.
Even acetaldehyde, the main metabolite of ethanol, is a
potential interfering substance. However, the concen-
trations of other volatile substances in blood and breath
after drinking alcoholic beverages, including acetalde-
hyde, are far too low to constitute an interference
problem.
Lung physiology and Gas Exchange
The lung is an amazingly efficient body organ for the
purpose of gas exchange. The trachea connects the
throat (glottis) to the lungs and subdivides into the left
and right bronchi forming the left and right lobes of the
lung. Each of the two bronchi undergoes further sub-
division about 23 times until the alveoli air sacks
are reached. The inspired air comes into contact with the
pulmonary capillary blood in the small air sacs on the
terminal ends of the respiratory bronchioles. Each lung
contains about 300 million alveoli and the thickness
of the alveolar-capillary membrane is only 0.001–
0.02 mm. Because of the large number of alveoli, the
surface area for gas exchange is about 70 m2, approxi-
mately the size of half a tennis court. Accordingly,
128 Alcohol: Breath Analysis

1.2

Peak response
1.0

Fuel cell response

0.8 Electrochemical oxidation
CH3CH2OH —> CH3CHO —> CH3COOH
0.6

0.4

0.2

0.0
0 20 40 60 80 100
Time after breath sampling (s)
Figure 3 Response of an electrochemical fuel cell detector to ethanol vapor (breath) showing peak response at 10–15 s after sampling and
recovery to baseline in 90–100 s.

alcohol or any other volatiles in the pulmonary blood Table 4 Anatomical and physiological characteristics of the
make intimate contact with the air in the lungs and human lung important for respiratory gas exchange
diffusion occurs across the alveolar-capillary membrane
Pulmonary gas exchange Average values in healthy
interface. The basic anatomical and physiological fea- adults
tures of the human lung can be gleaned from infor-
mation given in Table 4. Pulmonary blood flow 5–6 l per min
During breathing, air enters through the nose and Alveolar ventilation 6–7 l per min
Breathing rate 12–14/min
mouth and passes down the trachea into the two main
Tidal volume at rest B500 ml
bronchi and after multiple branches to the respiratory Number of alveoli B300–500 million
bronchioles and alveoli (air sacs) where most of the gas Forced vital capacitya Men 4.5 l average
exchange occurs as depicted in Figure 4. Women 3.2 l average
During inspiration, the ambient air is warned to body Total lung capacity Men 6.0 l
temperature (37 1C) and humidified, actually saturated Women 4.2 l
with water vapor, as it passes over the watery mucosa Anatomical dead-space volume 150 ml
Residual volume Men 1.2 l
lining the upper airways. During exhalation, these pro-
Women 1.1 l
cesses occur in reverse and now the alveolar air, which is Ventilation-perfusion ratiob 0.8 on average
saturated with water vapor at body temperature, is Volume of blood in alveolar 60–140 ml
cooled as it passes over the airway mucosa and water capillaries at one time
vapor condenses at the lower temperature in the upper Thickness of alveolar-capillary 0.001–0.002 mm
airways. The high solubility of ethanol in water means membrane
that this volatile substance also undergoes exchanges Surface area for gas exchange B50–100 m2
End-expiratory breath temperature 34.5 1C on average
between inspired and expired air and the mucosa. After
Blood/air partition ratio for ethanol at 1800:1c
drinking alcohol, the watery mucosa will contain some 37 1C
ethanol provided by the blood circulation to the
a
airways. Depends on the person’s age, gender, body size, posture, and physical condition
(smoking).
The concentration of ethanol in exhaled breath is b
Differs for different parts of the lung.
lower than the concentration in the alveolar air, because c
Determined by experiments in vitro.
during a prolonged exhalation the air cools. The core
body temperature in healthy individuals is at 37 1C
whereas breath leaves the mouth at 34.5 1C. Further- equilibration depends on several factors including
more, during exhalation the content of the alveolar air breathing pattern of the subject prior to expiration.
equilibrates with the watery mucosa membranes that The concentration of ethanol in the exhaled breath is
cover the upper airways. This interaction between dependent on a number of physiological variables that
ethanol and the mucosa and the completeness of the differ among individuals, such as their age, height,
 Alcohol: Breath Analysis 129

Trachea

Bronchi

Bronchiole

Alveoli

Figure 4 Schematic illustration of the human lung, showing trachea, left and right bronchi, and subdivision into the bronchioles before reaching
the alveoli sacs where gas exchange occurs. Also shown (inset) is the rich network of blood capillaries covering the alveoli air sacs.

gender, and lung function. Nevertheless, many direct
comparisons show a high correlation between the con-
centration of ethanol in venous blood and in the end-
expired breath.
The scientific basis supporting the use of breath-al-
cohol analysis as a way to estimate the concentration in
blood depends on Henry’s law, which was named after a
British chemist William Henry (1774–1836) who in-
vestigated the solubility of gases. Henry’s law can be
stated in several ways, such as “When a solution of gas
or somewhat volatile solvent in water is brought into
equilibrium with air in a closed container, there is a fixed
relationship or ratio between the concentration of the
substance in the air phase and in the water phase at
constant temperature and pressure.”
The above relationship holds for dilute solutions of
ethanol in water and other media and the ratio between
the concentrations in the liquid and air phases at equi-
librium is often referred to as the Ostwald solubility
coefficient. However, the sampling and analysis of
breath is a dynamic process and extensive exchanges of
ethanol and water vapor occur with the mucous mem-
branes during inhalation and exhalation, which means
that the principle of Henry’s law is less applicable to the
analysis of alcohol in breath.
Blood–Breath Ratios of Alcohol
The Breathalyzer instrument, which was developed in the
mid-1950s, was calibrated in such a way that the results
were reported as the concentration of alcohol present in
blood. This conversion required use of a calibration fac-
tor, the so-called blood/breath ratio, which was assumed
to be 2100:1 (Begg et al., 1964). During the 1950s–
1070s, the analysis of alcohol in the breath was used as a
surrogate for sampling blood for the analysis of ethanol.
The assumption of a constant BBR was difficult to
defend because many drinking studies showed that the
ratio varied both among and within individuals (Jones,
1978). The BBR tended to be less than 2100:1 during the
absorption phase of the blood-alcohol curve and greater
than 2100 in the post-absorptive elimination phase (Jaffe
et al., 2013). In tests made 15–30 min after end of drinking
the BBR was closer to 1800:1 and then rose to 2100:1
by about 90 min post-drinking. As time after drinking
increased, the BBR increased further reaching 2300:1 or
2400:1 by 120 min and at later times to ratios greater than
3000:1 where possible as BAC decreased toward zero.
Investigations of the BBR of alcohol in drinking
subjects and in apprehended drivers using modern in-
strumental methods of analysis suggest that the mean
ratio is 2400:1 instead of 2100:1 (Stowell et al., 2008). If
a person is tested on a breath-alcohol instrument cali-
brated on the basis of 2100:1, whereas in reality the
actual BBR is 2400:1, this introduces a 14% bias (Jones
and Andersson, 1996). The venous BAC is accordingly
underestimated by this amount if an instrument is cali-
brated on the basis of a 2100:1 ratio.
To avoid legal arguments about physiological vari-
ations in the BBR, when evidential breath analyzers were
approved for use in European countries in the 1980s, the
results were reported directly as BrAC without any as-
sumption about a BBR (Jones, 2011). The notion of re-
porting BrACs separately from BAC was originally
introduced in the USA (Mason and Dubowski, 1976).
Their statutory blood-alcohol limit for driving was
0.08 g/100 ml and the corresponding breath-alcohol limit
was 0.08 g/210 l breath, the ratio of the two (BAC/BrAC)
being 2100:1, although in any individual case there was
no conversion made from breath to blood alcohol.
The relationship between BrAC and venous BAC
is shown in Figure 5 for one subject who drank a
moderate dose of alcohol (0.68 g/kg) on an empty
130 Alcohol: Breath Analysis
3000
Ratio BAC/BrAC
Ratio BAC/BrAC
2000
1000
0 60 120 180 240
150
BAC or BrAC (mg/100 ml)
100
50
0
0 60 120 180 240
Time after start of drinking (min)
Figure 5 The concentration  time profiles of alcohol in venous
blood (BAC) and end-expired breath (BrAC  2100) in one male
subject who drank 0.68 g ethanol per kg body weight on an empty
stomach. The upper plot shows how the BAC/BrAC ratios of ethanol
change as a function of sampling time after drinking.
stomach. The upper plot shows changes in the venous
BBR of alcohol in the relation to time after drinking.
Role of Arterial-Venous differences
During absorption into the blood, the concentration of
ethanol is highest in the arterial blood (A-BAC) com-
pared with venous blood (V-BAC) returning to the heart
after circulation throughout the peripheral tissues. The
concentration of alcohol in pulmonary capillary blood
equilibrates instantaneously with the alveolar air so
BrAC runs closer to the A-BAC rather than the V-BAC.
However, it is the V-BAC that is always used in forensic
practice when drunk drivers are prosecuted.
The concentration-time profiles of ethanol in venous
blood and end-expired breath are compared in Figure 6
for one subject who drank a moderate dose of ethanol.
The higher BrAC compared with venous BAC early after
end of drinking and lower BrAC in the post-absorptive
phase are remarkably similar to the arterial BAC and
venous BAC relationships shown in Figure 1.
The existence of arterial-venous differences in ethanol
concentration and how these change in relation to time
after drinking represents a physiological explanation for
variations in the venous blood to breath-alcohol ratio
(Lindberg et al., 2007). The difference between A-BAC
and V-BAC was greatest early after end of drinking then
120
Alcohol concentration (mg/100 ml)
100 Venous blood
Breath × 2100
80
60
40
300 360 420 480
20
0
BAC
0 60 120 180 240 300 360 420 480
BrAC × 2100
Time from start of drinking (min)
Figure 6 The concentration  time profile of ethanol in venous blood
and breath in one male subject who drank 0.68 g ethanol per kg body
weight on an empty stomach. The concentrations of ethanol in breath
were multiplied by a factor of 2100:1 (blood-breath ratio) to allow
direct comparison with blood-alcohol concentrations.
300 360 420 480 gradually decreased to decreasing towards zero by
90 min after end of drinking. At all later times the
concentration of alcohol in venous blood was slightly
higher than in the arterial blood, because ethanol is
cleared by metabolism in the central (liver) compartment
and not in peripheral tissues (Jones et al., 2004).
Mouth Alcohol Effect
The concentration of alcohol in beer (3–5 g/100 ml),
wine (8–12 g/100 ml), or spirits (35–45 g/100 ml) is
considerably higher than the concentration of ethanol in
blood or exhaled breath. If a breath sample is taken for
analysis shortly after drinking an alcoholic beverage the
concentration of alcohol is higher than in the alveolar air
because of contamination with the much higher alcohol
content in the mouth and oral mucosa. The problem
posed by this mouth alcohol (MA) effect was recognized
very early in the history of breath-alcohol testing
(Bogen, 1927). Studies have shown that it takes about
10–15 min before the higher concentration of ethanol
in the oral mucosa dissipates and this has led to a
mandatory waiting period before making an evidential
breath-alcohol test.
A mouth alcohol effect might also occur if a person
burps or regurgitates stomach contents when alcohol is
still being absorbed from the stomach and intestines, a
process that might last up to 90 min after drinking ends.
Furthermore, use of oral hygiene products, such as
mouth washes, medicines, and certain foodstuffs that
contain alcohol can artificially increase BrAC if testing is
done within 15–20 min afterwards (Sterling, 2012).
Examples from the latest generation of infrared breath-
alcohol instruments are fitted with slope detectors,
such that the exhaled ethanol profile is continuously
 Alcohol: Breath Analysis 131

monitored. If the shape of this exhalation profile in the exhalation profile following by a decrease in
deviates significantly from that expected without the concentration on reaching an end exhalation. Another
confounding influence of mouth alcohol, the test can algorithm looks at the waviness of the BrAC profile
be aborted and the instrument should indicate in the or the change in BrAC after 1 s and 5 s of exhalation.
display and on the printed test record “mouth alcohol Another test for the presence of MA is to make two
detected.” measurements not less than 6 min apart and evaluate the
Figure 7 shows exhalation profiles of ethanol deter- difference in concentration in relation to known elim-
mined with an Evidenzer instrument in one subject at ination kinetics of MA and rate of ethanol metabolism.
various times after washing the mouth with whisky but The simplest alternative is to observe the suspect for a
without swallowing any alcohol. time of 15–20 min and to ensure that nothing was
The Evidenzer and other infrared-based breath- placed in the mouth (Gullberg, 1992). The concentration
alcohol analyzers incorporate algorithms to monitor the of alcohol in the mouth after drinking dissipates rapidly
integrity of the exhaled ethanol breath profile. One such and by 10–15 min the sample is no longer contaminated
algorithm relies on detecting an early peak concentration with alcohol from a recent drink or use of mouthwash or
mouth spray. Some of the more modern evidential
breath analyzers monitor not only ethanol in the exhaled
0.6 air but also CO2 and water vapor and the known phy-
Breath alcohol concentration (mg/l)

siology of these exhaled gases and relationships to

0.5 ethanol exhalation can help to detect MA.
10 min
In some jurisdictions there is interest in conducting
0.4 the evidential breath-alcohol tests at the roadside and
therefore closer to the time of driving. This change in
0.3
procedure requires use of an instrument that incorpor-
0.2 ates a reliable slope detector so as to distinguish a proper
8.5 min
exhaled ethanol profile from a sample contaminated
0.1 17.5 min with alcohol from a recent drink or other sources of
ethanol (Wigmore and Leslie, 2001).
0 2.0 4.0 6.0 8.0 10.0 12.0
Figure 8 shows an exponential decrease in breath-
Exhalation time (s)
alcohol concentration after subjects ate alcohol-
containing chocolates or used an alcohol-containing
Figure 7 Single breath-alcohol exhalation profiles monitored using
mouth spray; it took 5–8 min to return to zero BrAC.
an evidential breath-alcohol analyzer (the Evidenzer) at 8.5 min,
10 min, and 17.5 min after drinking whisky. The wavy curves after 8.5
This experiment supports 15 min as an appropriate
and 10 min illustrate the effect of mouth alcohol on the exhalation waiting period after having alcohol in the mouth
profile, whereas the 17.5 min profile is uncontaminated end-exhaled before the result of an evidential breath-alcohol test is
breath. considered valid.

3
BrAC (mg/l)

Cherry chocolate

2 Mouth spray

Brandy chocolate

0
0 1 2 3 4 5 6 7 8
Time (min)
Figure 8 Elimination rate of mouth alcohol after subjects had eaten alcohol-containing chocolates or used a mouth spray that contained alcohol
just before tests were made. Courtesy of Draeger Safety AG, Lübeck, Germany.
132 Alcohol: Breath Analysis
Physiological Factors and Medical Conditions
The use of breath-alcohol analysis is subject to more
preanalytical factors and physiological variations com-
pared with blood sampling and analysis of the latter at a
forensic laboratory. Therefore, it is important to stand-
ardize the breath-sampling procedure so that a minimum
exhaled volume, time of exhalation, and pressure (flow
rate) to reduce as far as possible variations caused by
different breathing patterns. Studies have shown that the
mean temperature of exhaled breath is close to 34.5 1C
and the temperature coefficient of alcohol solubility is
6.5% per degree. The temperature of the exhaled breath
and the exhaled volume, as well as temperature and
humidity of ambient air, are other variables that impact
on the concentrations of ethanol in the breath.
The most common medical conditions that impact on
a person’s ability to provide a proper breath-alcohol test
are asthma and chronic obstructive pulmonary disease
(COPD). In a controlled drinking study, people with
COPD had higher BBR of alcohol compared with people
with healthy lungs (Hahn et al., 1991). When asthmatics
were tested on two different evidential instruments, they
were unable to provide a proper breath sample, that is,
to make a continuous exhalation at a fixed pressure for
sufficiently long time to satisfy the requirements of
modern breath-alcohol analyzers.
Nevertheless, as shown in Figure 9, even after a few
seconds of exhalation the concentration of alcohol
reached 66% of the final end-exhaled concentration.
Another factor to consider in connection with breath
analysis is the high solubility of alcohol in the mucous
membranes covering the upper airways of the lungs.
Some scientists believe that the breath-alcohol concen-
tration arises exclusively from equilibration of ethanol
in the upper airway and not as originally postulated at
the alveolar-capillary membrane in the alveoli (Hlastala,
2010). Wherever the final alcohol equilibrium occurs,
one cannot deny the fact that breath and blood-alcohol
180
Breath alcohol concentration (µg/l)
160
140
120
100
80
60
40
20
0
0 1
Figure 9 Single breath-alcohol exhalation profile produced with an evidential infrared analyzer (the Evidenzer) showing ethanol content of partial
exhalations expressed in percent of the highest concentration after an end exhalation lasting 10 s.
concentrations are highly correlated over a wide range
of drinking conditions (Jones, 2000).
The medical condition known as gastroesophageal
reflux disease (GERD) was thought to falsify the results
of breath-alcohol analysis, owing to alcohol erupting
from the stomach into the mouth just before making the
test (Duffus and Dunbar, 1984). However, a controlled
human alcohol dosing study with patients suffering from
GERD failed to verify that this medical complaint lead to
falsely high BrAC results compared with BAC (Kechagias
et al., 1999). However, more studies are needed to in-
vestigate whether GERD is indeed a bogus defense ar-
gument. Such experiments should include more subjects
diagnosed with this condition and different doses of al-
cohol consumed on an empty stomach and after a meal.
Response to Interfering Substances
Human breath consists of a mixture of oxygen, nitrogen,
carbon dioxide, and water vapor and traces amounts of
other volatile substance, either produced endogenously
or inhaled with the ambient air (Spanel and Smith,
2011). Acetone is the main endogenous volatile in ex-
haled in the breath and this substance absorbs infrared
radiation in the 3.3–3.5 mm wavelength range currently
used to monitor ethanol in breath (Spanel et al., 2011).
However, the use of breath-test instruments that moni-
tor absorption of infrared radiation at two or more
wavelengths can discriminate acetone from ethanol.
Some of the latest generation of infrared analyzers are
fitted with five filters in the range of 3.3–3.5 mm (e.g.,
Evidenzer) and can discriminate between several poten-
tially interfering substances. In apprehended drivers
tested with Evidenzer, some of the breath tests were
aborted because the instrument detected an interfering
substance – a control analysis of blood in these indi-
viduals showed elevated concentrations of acetone and/
or isopropanol (Jones and Andersson, 2008).
98%
94% 97% 164
87% 91%
85%
81%
75%
66%
End exhalation
2 3 4 5 6 7 8 9 10
Exhalation time (s)

Breath analyzers fitted with electrochemical detectors
respond to other alcohols such as methanol and iso-
propanol, which are also oxidized but at different rates
compared with ethanol. Monitoring the rate of the oxi-
dation reaction using algorithms is a way to enhance
selectivity of electrochemical analysis of ethanol. Acetone
is not oxidized at the same electrode potential as ethanol
and does not cause interference problems when fuel cell
instruments are used. Examples of other gases and
volatiles produced endogenously include acetone, me-
thane, isoprene as well as acetaldehyde, the first product
of ethanol metabolism. Substances inhaled with the am-
bient air (gasoline fumes, toluene, butane) are also po-
tential interfering substances. People might be exposed to
these substances in the workplace or in connection with
volatile organic compound (VOC).
On the whole the problem posed by interfering sub-
stances is much exaggerated, because apart from acetone
human breath does not contain other substances in
sufficiently high concentrations to pose a problem for
the analysis of ethanol. The hand held fuel-cell breath
analyzers do not oxidize acetone at the electrode po-
tential used for ethanol analysis and so do not respond
to elevated breath-acetone. The infrared analyzers cur-
rently used for legal purposes incorporate multiple in-
frared wavelength filters to detect the presence of an
interfering substance, such as acetone, in the breath
sample. Either a correction is made to compensate for
the concentration of acetone or the evidential breath test
is aborted and a sample of blood or urine taken instead.
Interfering substances might represent a problem in
people who abuse organic solvents by sniffing from a rag
or inhalation by other means. More studies are needed
to test the effect of VOCs on the response of breath-
alcohol analyzers.
Breath Analyzers for Self-Testing
It is widely accepted that motor vehicles require a
speedometer to inform the driver whether the speed limit
is being exceeded. Many believe drivers should have
access to a personal breath-alcohol tester to indicate
whether the statutory breath-alcohol limit for driving is
exceeded after an evening’s social drinking. Some traffic
safety authorities would like to see all newly produced
cars fitted with a device to analyze the driver’s breath for
alcohol (Radun et al., 2014). So far this radical idea has
not been put into practice although a number of low-
cost breath-test instrument are available for purchase by
the public. However, some such instruments have ac-
quired a dubious reputation with regard to analytical
specificity and overall reliability of the results.
The first breath-alcohol analyzers manufactured and
sold for self-testing appeared in the 1970s. The simplest
of these was the chemical tube and bag (e.g., Alcotest or
Alcolyser), a once-only test involving the oxidation of
Alcohol: Breath Analysis 133
ethanol by potassium dichromate. Also available were
small electronic instruments that incorporated a metal
oxide semiconductor or Taguchi Gas Sensor (TGS) as
the analytical principle. The TGS comprises a tiny bead
of tin oxide which, when heated in ‘clean air’ absorbs
oxygen onto its surface until a characteristic conduct-
ance is obtained. When a combustible gas (such as
ethanol, propane, or methane) is also present in breath,
it gets absorbed onto the bead surface, reacts with
oxygen, and electrons are released changing the con-
ductance of the bead in direct proportion to the
concentrations of the combustible gas. The TGS is
commonly used in smoke alarm detectors in homes, but
is also incorporated into some devices for determination
of BrAC. Semiconductors are not only used for identi-
fying ethanol, but also respond to other types of alco-
hols, hydrocarbons, and combustible gases such as
carbon monoxide contained in cigarette smoke. The
response to ethanol concentration is nonlinear and the
calibration drifts over time. The effective working life of
the TGS is limited, depending to a large extent on how
often they are used. It appears that the surface effect by
which they operate is dependent on the atmospheric
partial pressure of oxygen, and semiconductors have
been found to vary in sensitivity to alcohol depending on
the weather and the altitude at which they are operated.
Caution is necessary when breath-alcohol analyzers
are used for self-testing, because it is not a standard
practice to check the calibration with each test and there
is no way to monitor the presence of mouth alcohol. A
test should not be made until at least 15–20 min have
elapsed since the last drink to ensure extraneous sources
of alcohol have disappeared from the mouth and oral
mucosa. The most appropriate use for self-testers would
be in the morning after an evening of heavy drinking to
ensure that alcohol has been eliminated from the body.
Ignition Interlock Devices
Most drunk drivers have a substance abuse problem and
could probably be diagnosed clinically as being alcohol
dependent. Studies have shown that repeat offending is a
common occurrence among people arrested for drunk
driving in most nations. Dealing with this problem is a
major concern for traffic safety organizations (Marques
et al., 2001). Mandatory fitting of an interlock device is
one approach to reduce recidivism rates among con-
victed drinking drivers. Most interlocks incorporate a
fuel cell type of breath analyzer connected to the vehicle
ignition system and a driver must pass the breath test
before the vehicle can be started in the conventional way
(Fulkerson, 2003). Most interlock devices are set to read
as ‘fail’ at low threshold BrAC corresponding to 10 or
20 mg/100 ml blood-alcohol concentration.
A number of safeguards are incorporated into inter-
locks to prevent a driver from circumventing the breath
134 Alcohol: Breath Analysis

test, such as sensing temperature and pressure of Concluding Remarks

the exhaled breath entering the instrument. Some people
have resorted to using a hand-pump to provide alcohol- During the 1970s the technology available for breath-
free ambient air into the breath analyzer and in this alcohol analysis underwent radical changes and a new
way pass the test and start the vehicle. Many ignition generation of analytical instruments appeared. Unlike
interlocks are programmed to request a second test the Breathalyzers which measures the ethanol in breath
after the vehicle is in motion, which is a safeguard by chemical oxidation, this new generation of instru-
to ensure that somebody other than the driver has ments utilized physical-chemical methods of analysis
provided the negative breath test needed to start the such as electrochemical oxidation and infrared spec-
vehicle. trometry. These new instruments were easier to operate
The stability of the calibration of interlock breath- and the entire sequence of tests was controlled by
alcohol analyzers requires close attention and monthly microprocessors. The breath sample was captured
or bimonthly maintenance checks should always be in- automatically as the test subject reached end exhalation,
cluded as a safeguard. Another problem sometimes en- which usually required a discarded volume of 1.5 l and
countered is from interfering substances in the ambient continuous exhalation for at least 6 s. Use of infrared
air, such as solvents used as windscreen cleaners, many instruments allowed the increase in ethanol concen-
of which contain isopropanol. Furthermore, people with tration to be monitored as a function of exhalation time
an interlock device fitted to their vehicle should be and curve analysis which made it possible to detect an
warned about extraneous sources of ethanol, such as abnormal ethanol profile that might indicate contamin-
that contained in cough medicines, vitamin tonics, and ation with mouth alcohol.
mouthwash preparations. The concentrations of ethanol in end-exhaled breath
Nevertheless, ignition interlock technology has ad- and in venous blood are highly correlated over a wide
vanced considerably and one finds that these devices range of concentrations of ethanol and drinking pat-
are fitted to public transport vehicles, including buses, terns. This is illustrated by the scatter plot in Figure 10
taxies, etc. in some nations (Willis et al., 2004). obtained from tests with people apprehended by the
If a convicted drunk driver agrees to having an inter- police for drunk driving in Sweden. Note that the BAC
lock device installed in his or her vehicle, this is one ranges from zero to 400 mg/100 ml and the correlation
way to speed-up the re-granting of a driving permit. coefficient between blood- and breath-alcohol was
On the whole the use of breath-activated interlock statistically highly significant (r ¼ 0.98, po.001). The
devices represents an effective way to reduce rates of dotted diagonal line in Figure 10 shows the line of
drunk driving recidivism by hard-core offenders (Elder identity. Most of the points (blood-breath pairs) are
et al., 2011). below this line, which means that results from the breath

400
N = 793
Estimated breath alcohol concentration (mg/100 ml)

Pearson’s r = 0.98
Residual SD = 15 mg/100 ml
300

200

100

0
0 100 200 300 400
Venous blood alcohol concentration (mg/100 ml)
Figure 10 High correlation between venous blood and exhaled breath-alcohol concentrations in tests carried out on drink drivers. Venous blood
was analyzed by headspace gas chromatography and breath-alcohol was determined with an infrared analyzer (the Intoxilyzer 5000).

test are lower than the actual venous BAC in the vast
majority of cases.
The main advantage of breath-alcohol analysis com-
pared with blood-alcohol analysis includes noninvasive
nature of the sampling and the fact that results of the test
are obtained immediately afterwards. A positive roadside
breath-alcohol test of a driver needs to be verified
by making a more reliable evidential test at a police
station. The seriousness of an impaired driving charge
and the consequences for those found guilty of this crime
demands rigorous quality assurance procedures. Some of
the most important requirements are listed below:
• The evidential breath-alcohol analyzer used must be
certified for its intended purpose by an appropriate
government agency or other legal authority.
• The instrument should be operated by a trained and
certified operator and not by the arresting police
officer.
• A minimum time of 15–20 min must have elapsed
since the last consumption of alcohol. The suspect is
not allowed any food, water, or medication during
this pre-test observation and deprivation period.
• All evidential breath-alcohol tests should be done in
duplicate with a minimum of 5 min and maximum of
15 min between the two tests. The two test results
should agree within certain predefined limits (e.g.,
710%) depending on concentration of ethanol in the
sample analyzed.
• In conjunction with every subject test, a known
concentration of ethanol in gas or vapor should
be analyzed as a check on accuracy. The target
concentration of alcohol in the standard is usually
at or near the statutory alcohol limit for driving.
The accuracy test result should be within specified
limits, such as 75–10% of the known target
concentration.
• Samples of the ambient room air should be analyzed
before and after every subject is tested and these
should register zero alcohol concentration.
• A printed record of the test results and time of testing
should be generated including any error messages.
This printout should be signed by the responsible
police officer and made available to the defense at-
torney if and when required.
Compared with the results of blood-alcohol analysis,
the sampling and analysis of breath are more prone
to biological variations, such as lung physiology. The
ethanol concentration in exhaled breath depends to
some extent on the person’s pattern of breathing before
exhalation is made into the instrument. The volume of
breath exhaled prior to sampling and the temperature of
the breath sample also influence the ethanol content.
Other variables are the temperature and humidity of
ambient air breathed and evidential testing should be
done in an air-conditioned room or other controlled
Alcohol: Breath Analysis 135
environment. Obtaining a proper breath sample might
not be feasible depending on the person’s age, gender,
and body stature. There is considerable evidence that
people with pulmonary dysfunction, such as asthma and
COPD, are unable to provide a proper sample with most
breath-alcohol analyzers.
The analytical specificity of instruments for identifi-
cation of ethanol in breath is lower than for blood
analysis by gas chromatography (GC), because GC
methods separate interfering substances prior to quan-
titative analysis. Several GC instruments were developed
as breath-alcohol analyzers (see Table 3), but were less
practical for use by police officers outside of a labora-
tory environment. The problem of interfering substances
when IR and EC methods are used for ethanol analysis
even without GC separation is much exaggerated. Apart
from acetone, the human breath does not contain vola-
tiles in sufficient amounts to interfere with ethanol ex-
cept in rare instances when people abuse gasoline-type
solvents by sniffing. The absorption of infrared radiation
at a number of different wavelengths – 3.4 mm (C–H
stretch) and 9.5 mm (C–O stretch) – enhances selectivity
further. In the C–H stretching region of the IR spectrum
use of multiple IR wavelength filters (e.g., 3.37, 3.41,
3.47, and 3.53 mm) allow the detection of interfering
substances.
Besides forensics, analysis of volatile organic com-
pounds (VOCs) in breath has found applications in
clinical and diagnostic medicine as biomarkers of dis-
eased states, such as acetone (diabetics), carbon mon-
oxide (smokers), methane (malabsorption), and nitric
oxide (asthma and lung disease). Methods involving
liquid nitrogen freeze-traps have been developed to
analyze an exhaled breath condensate for various en-
dogenous and exogenous substances. The condensate
is later analyzed by gas chromatography–mass spec-
trometry (GC–MS) or liquid chromatography–mass
spectrometry (LC–MS) and provide useful and inter-
esting metabolic profiles with diagnostic potential.
The sampling of breath has shown promise as a way
to detect the presence of drugs other than alcohol and as
a practical alternative to sampling and analysis of saliva
or urine. Trace amounts of substances such as amphet-
amine, cannabis, and cocaine are seemingly expelled in
the breath and can be trapped on a specially prepared
filter for later desorption and analysis at a forensic or
clinical laboratory. The drugs attached to the filter are
identified by highly sensitive analytical equipment, such
as LC–MS. GC–MS or LC–MS instruments that permit
the sampling and analysis of drugs in breath at the
roadside are not feasible at the present time.
Without doubt, the main forensic application of
breath analysis is to test the sobriety of drivers and
generate evidence for prosecution of traffic offenders.
However, because some traffic offenders refuse to co-
operate with the police in providing an approved sample
of breath, there will always be a need for alternative
136 Alcohol: Breath Analysis
specimens, such as blood, saliva, or urine to verify
overconsumption of alcohol.
The concentration of alcohol in the breath is a better
indication of brain exposure to ethanol than is the
venous blood. Studies have shown higher concentrations
of ethanol in arterial blood compared with venous blood
during the absorption phase of the BAC curve. Because
arterial blood transports alcohol to the brain, this sug-
gests that breath-alcohol is a better indicator of the
pharmacological and behavioral effects of ethanol than
venous blood, especially during the absorption phase of
ethanol metabolism.
The enforcement of drunk driving laws by means of
breath-alcohol instruments has several practical advan-
tages. The immediate result of analysis means that sus-
pects can be confronted with information on whether
they are in breach of the law at a time when they are
most aware of their excessive drinking. Moreover, the
results from the breath test, if above the statutory alco-
hol limit means that immediate sanctions can be im-
posed, such as confiscation of the driving permit.
The technology currently available for breath-alcohol
analysis is impressive and will be hard to improve upon
in terms of ease of use and accuracy and precision of the
test results. The latest generation of breath-testing in-
struments are more robust and portable and this has led
to the notion of conducting the evidential test at the
roadside in closer proximity to the actual driving. Time
will tell whether this change in operating procedure in
use of breath-alcohol analyzers is practical and feasible
and whether the results are deemed acceptable for legal
purposes.
See also: Alcohol: Acute and Chronic Use and Postmortem
Findings. Alcohol: Blood and Body Fluid Analysis. Backtracking
Calculations for Alcohol and Drugs. Road Traffic: Global Overview
of Drug and Alcohol Statistics
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