Sensors and Actuators B 171–172 (2012) 1–17

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Sensors and Actuators B: Chemical

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Review

Biology and applications of olfactory sensing system: A review

Sindhuja Sankaran a,b,∗ , Lav R. Khot a,b , Suranjan Panigrahi b,c,∗
a
Citrus Research and Education Center, IFAS, University of Florida, 700 Experiment Station Road, Lake Alfred, FL 33850, USA
b
Bioimaging and Sensing Center, Department of Agricultural and Biosystems Engineering, North Dakota State University, Fargo, ND 58102, USA
c
Department of Electrical and Computer Engineering Technology, Purdue University, West Lafayette, IN 47907, USA

a r t i c l e i n f o a b s t r a c t

Article history: Various metal oxide and conducting polymer-based sensors have been used in electronic nose systems
Received 27 September 2011 for the detection of volatile organic compounds (VOCs). Some of the major constrains of these sensor
Received in revised form 8 March 2012 materials are their sensitivity and selectivity. Recently, much research has been towards the develop-
Accepted 10 March 2012
ment of biosensors that mimic biological olfactory mechanism. The sensing materials based on biological
Available online 19 March 2012
olfactory system can be well adapted to detect low concentrations of VOCs as it exhibits high selectiv-
ity/specificity, fast response time, high sensitivity, simplicity in fabrication, and are not toxic resulting
Keywords:
from their biocompatibility and rapid biodegradability. A better understanding of human olfactory system
Mammalian olfactory system
Olfactory receptor
can aid in the development of an artificial olfactory sensing system. The application domain of such sen-
Odorant binding protein sors/systems could be potentially expanded for rapid and accurate detection of VOCs in environment and
Volatile organic compounds other biological media. Therefore, this paper provides a brief overview of the biology, associated mecha-
Gas sensors nisms, and the functional principles of the olfactory sensing system in humans. Moreover, it summarizes
E-nose the recent research on the development of an olfactory sensor utilizing olfactory receptors, odorant bind-
ing proteins and olfactory epithelium for the detection of different VOCs, and research needs in this area.

© 2012 Elsevier B.V. All rights reserved.

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
2. Biology and mechanism of olfaction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
2.1. Structure of olfactory system . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
2.1.1. Olfactory region. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
2.1.2. Olfactory receptor cells and olfactory receptors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
2.1.3. Other receptors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
2.1.4. Regions of olfactory signaling . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
2.2. Mechanism of olfaction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
2.2.1. Odorants and odorant binding proteins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
2.2.2. Signal initiation and termination . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
2.2.3. Signal processing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
3. Development of olfactory sensing system . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
3.1. Electronic noses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
3.2. New generation biomimetic olfactory sensors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
3.2.1. Olfactory receptors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
3.2.2. Odorant binding proteins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
3.2.3. Olfactory neurons . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
3.2.4. Synthetic polypeptides . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
3.2.5. Process overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
3.3. Pattern recognition in olfactory sensing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12

∗ Corresponding authors. Tel.: +1 863 956 8771/765 494 6908; fax: +1 863 956 4631.
E-mail addresses: sindhu@ufl.edu (S. Sankaran), spanigr@purdue.edu (S. Panigrahi).

0925-4005/$ – see front matter © 2012 Elsevier B.V. All rights reserved.
doi:10.1016/j.snb.2012.03.029
2 S. Sankaran et al. / Sensors and Actuators B 171–172 (2012) 1–17

4. Comparison of natural and artificial (e-noses) olfaction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13

5. Summary and conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
Acknowledgements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
Biographies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17

1. Introduction 2. Biology and mechanism of olfaction

Vertebrate olfactory systems can identify and distinguish 2.1. Structure of olfactory system
volatile compounds (odorants) of diverse molecular structures with
high accuracy. The mammalian nose can detect certain compounds 2.1.1. Olfactory region
in concentrations as low as few parts per trillion [1]. Such perfor- The human olfactory system comprises of an external nose and
mances are due to numerous olfactory receptors (ORs) expressed inner nasal cavity. The nasal cavity consists of two chambers divided
by olfactory sensory neurons and their subsequent neuronal pro- by a septum [34]. Inside the nostril is a slight dilation called the
cessing. Each of the ORs can bind to numerous odorants with vestibule, lined with skin containing hairs and sebaceous glands.
specific affinities, although some receptors are relatively restricted Entire nasal cavity, above and behind the vestibule is covered by
to a set of few chemically related compounds [2]. In the pro- mucous membrane. Human nose consists of three turbinates/nasal
cess of sensing the smell, the binding of specific odorants to the concha (superior, middle and inferior). Turbinates are long, narrow
OR proteins is the initiation step in odor recognition and trigger- and curled bone shelf protruding into the nasal passage that direct
ing of signal transduction in a cell. Gomila et al. [2] stated that the air towards the olfactory epithelium. The mucous membrane
“Given the fantastic odor space detected by the olfactory recep- can be divided into two parts: an olfactory region, above the supe-
tors, it is tempting to harness them to some generic electronic rior turbinate and corresponding to nasal septum, and a respiratory
devices that could be endowed with some of the most prominent region, which comprises of rest of the nasal cavity. Ciliary action
properties of animal olfaction: discrimination, specificity and sen- in the respiratory region aids in active drainage back and down
sitivity.” into the nasopharynx. The highly vascular surface of nasal mucosa
Recent studies have led to a more refined understanding of also helps in warming and humidifying the inspired air [34,35].
olfactory neurons and the mechanisms involving odorant detec- Olfactory regions are the sensory regions that sense the smell.
tion. Joint efforts of biologist and biochemists have revealed that The mucous membrane lining the olfactory region is termed as
olfactory receptor achieve odorant identification and signal trans- the olfactory epithelium (Fig. 1). It is a specialized epithelium tis-
duction by employing molecular elements [3]. An olfactory system sue inside the nasal cavity that plays an important role in olfaction.
plays an important role in identifying food and recognizing envi- Olfactory epithelium consists of three major types of cells: (i) the
ronmental conditions. Olfactory sensing can be used for detecting basal cells, (ii) the olfactory supporting cells, and (iii) the olfactory
human diseases [4–7], food contamination [8–11], or hazardous receptor cells/olfactory neuron cells. The basal cells are the stem cells
agents [12,13]. Currently, olfactory research is focused towards the that give rise to the olfactory receptor cells. Supporting cells have
discovery of potential commercial applications. numerous microvilli and secretory granules, which provide the
Biomimetic design of an electronic nose on the principle of epithelium with mucous. The olfactory mucosa bathes the olfac-
the mammalian olfactory system can aid in increased sensitivity tory epithelium protecting the sensory cells incorporated in them.
and selectivity [14] for various trace level odorant detection appli- The mucus layer contains molecules such as mucopolysaccharides,
cations. Different components of the biological olfactory system immunoglobulins, proteins (such as lysozymes, odorant binding
are being used for fabricating sensors. Researchers are investi- proteins), pigmented cells and various enzymes (such as pepti-
gating the development of olfactory sensors based on olfactory dases). Epithelium color depth is a factor that could be correlated
receptors [15], odorant binding proteins (OBP) [10,12,16], olfac- with the sensitivity of the olfactory system, although their exact
tory neurons [14], and insect antennae [17–19] among others. For function in enhanced sensitivity is unknown. This color is light
example, Lin et al. [4] simulated the three-dimensional structure yellow in human, whereas dark yellow/brown in dogs.
of the OR protein and synthesized a peptide sequence for the
breath analysis for uremia diagnosis. However, research on arti- 2.1.2. Olfactory receptor cells and olfactory receptors
ficial olfactory sensor development has not reached to the point Olfactory receptor cells are principally the most important sen-
where one can mimic the sensitivity and selectivity of biologi- sory cells with receptors that sense odor. The olfactory epithelium
cal olfactory receptors [14]. Thus, it is imperative to understand has more than 6–10 million olfactory receptor cells [20,21]. These
the biological olfactory system for developing artificial olfactory cells are bipolar neurons, consisting of dendrites at one end (pro-
sensors. jecting into the mucus above the surface of the epithelium) and
Some reviews on olfactory systems [20–23], biosensors axons at the other end (projecting into the olfactory bulb). Recep-
[7,24–29] and electronic nose [30–33] are available in literature. tor cells are distinct from other sensory cells, due to their ability
There is a need to integrate the engineering and science for a better to regenerate and replace themselves upon injury [20]. Another
understanding for the development of olfactory sensors. There- unique aspect of these cells is that as the age of the organism
fore, this study was aimed to develop an in-depth understanding increases, cells tend to live longer [20]. The axons of olfactory recep-
of the biological olfactory sensing system, in order to develop suit- tor cells form bundles that are collectively known as olfactory nerve
able novel techniques of engineered olfactory sensors for odorant or the first cranial nerve that carries the sensory information from
detection applications. Specifically, this review paper focuses on the olfactory epithelium to the olfactory bulb.
(i) conducting a state-of-the-art review of the biology and mecha- The dendritic end of the olfactory receptor cells ends in the form
nisms of biological olfactory sensing systems and (ii) establishing of an apical dendritic olfactory knob into the mucous of the olfactory
a framework for developing intelligent sensors based on the bio- epithelium. Each knob contains five to fifty long specialized cilia
logical olfactory system and their applications in volatile organic that extend into the olfactory epithelium [20,21,36,37]. The olfac-
compound (VOC) sensing. tory cilia are cylindrical with diameter ranging from 0.25 to 0.5 ␮m
 S. Sankaran et al. / Sensors and Actuators B 171–172 (2012) 1–17
Fig. 1. Structure and mechanism of human olfactory system.
and length of 30–200 ␮m and consisting of basal body and tubulin
with 9 + 2 microtubular arrangement covered within a lipid bilayer
sheath [20,36]. These non-motile cilia of the olfactory receptor cells
are much longer than the motile cilia of the cells in the respiratory
epithelium. The surface area of the cilia projecting into the mucus
layer in the nasal epithelium is important for olfactory sensitivity
as these cilia possess the ORs that are involved in transduction of
odorous stimuli [38]. Assuming six million olfactory receptor cells
have 25 cilia each and each cilium having diameter of 0.5 ␮m and
height of 30 ␮m, the total surface area is about 71 cm2 .
The cilia contains seven domain transmembrane receptors (G-
protein coupled receptors), termed the olfactory receptors (Fig. 1)
that interact with the odorants. Olfactory receptor cells contain
about a million receptors on its cilia. The chemosensory mem-
brane lining the cilia of cells consists of a phospholipid bilayer,
with hydrophilic phosphate groups and hydrophobic hydrocarbon
groups. In this membrane, the hydrocarbon tails face one another
and the hydrophilic ends are towards extra- and intra-cellular
surfaces. Major function of this membrane is to differentiate the
olfactory receptor cells, control the pathway of elements and ions
from and into the cell and finally, to enable detection of odorant
molecules and allow cell-to-cell communication.
ORs in the chemosensory membranes are the chemoreceptors
for odor detection in nasal cavity. These receptors are expressed
by about 1% of the vertebrate gene family [39]. About 900 olfac-
tory receptor genes are present in human genome, out of which
two-thirds are non-functional (pseudogenes). In mouse, out of
1300 olfactory receptor genes, few hundreds are pseudogenes
[1]. In vertebrates, olfactory receptor genes have been discov-
ered in rats, mice, fish, frogs and humans. For invertebrates, these
receptors have been discovered only in round worm (nematode)
Caenorhabditis elegans and fruit-fly Drosophila melanogaster. Olfac-
tory receptors in Drosophila belong to a diverse family, having
3
marginal similarity in the sequence within their members and with
no similarity in sequence of olfactory receptors of vertebrates and
nematodes [22].
ORs in vertebrates are one of the largest groups of the G-protein
coupled receptor family. The G-protein linked ORs have seven
hydrophobic helical transmembrane domains connected by dif-
ferent lengths of intra- and extracellular loops and several short
protein sequences [1]. In the three dimensional structure of G-
protein coupled ORs, the major regions of divergence occur in the
sequence of third, fourth and fifth transmembrane domain [1]. The
cavity formed at one-third the distance from membrane by these
three membrane domains possesses possible binding sites for the
odorant molecules. The variation in the binding site of olfactory
receptors allows the binding of wide range of odorant, and their
identification and classification. Olfactory G-protein linked recep-
tors trigger the biochemical synthesis of neurotransmitters upon
activation (binding of the odorant molecules). Each olfactory recep-
tor neuron contains one type of receptor, and each receptor detects
limited number of odorant substances.
The surface area of the epithelium is a factor related to the acu-
ity of an organism’s olfactory system. The area of the epithelium
and surface area of the cilia in dogs is much larger than in humans
[20,40], making them more sensitive to odors than humans due to
the presence of larger number of olfactory receptors per unit area
[41].
2.1.3. Other receptors
In addition to the OR protein, another group of receptors was
discovered in 2006, termed as Trace Amine-Associated Receptors
(TAAR) by Liberles and Buck. Initially, these receptors were found
in mouse olfactory epithelium and genes coding TAAR are found
in humans, mouse and fish. TAAR are receptor protein specifically
sensitive to volatile trace amines in urine. Observations such as
4 S. Sankaran et al. / Sensors and Actuators B 171–172 (2012) 1–17
absence of G-protein in small percent of olfactory receptor cells,
stimulation of olfactory receptor cells by peptide-bound histocom-
patibility complex protein, and indication of presence of olfactory
structures with receptors different from odorant receptor; lead to
the discovery of TAAR. In mouse, TAAR detects compounds found
in urine, which are a part of its social cues. They help in pheromone
detection related to reproductive development in mice. Evidence in
expression of TAAR in olfactory epithelium of fishes indicates that
they might serve as olfactory receptors in humans [42].
2.1.4. Regions of olfactory signaling
Signaling begins in the ORs present in the olfactory recep-
tor cells. The OR converts the olfactory signals into electri-
cal/neurological signals and sends it to brain. The primary
information about the odorant is processed in the olfactory receptor
cells and sent to the olfactory bulb, which distributes the informa-
tion to different parts of brain. The cells involved in the transfer of
secondary information are termed as secondary sensory cells. Ten
to hundreds of axons (of the olfactory receptor cells) bundle up and
penetrate the ethmoid cribriform plate and terminate in the olfac-
tory bulb, converging on synaptic glomerulus [43]. The glomeruli are
spherical conglomerates of about 50–200 ␮m diameter. The mam-
malian glomeruli in the olfactory system receives the input from
the ‘axons of the olfactory receptor cells’ and output connections
are to ‘the dendrites of the principle neurons in the olfactory bulb,
the mitral cells and tufted cells’ [21,43]. Rats contain about 2000
glomeruli in their olfactory bulb, each of which receives informa-
tion from about 1000 to 3000 olfactory receptor cells and these
olfactory signals are transmitted over 25 mitral cells and 50 tufted
cells in each glomerulus [21,43].
The mitral cells have an apical dendrite that extends from the
glomerulus and their axons merge to form the lateral olfactory tract,
which passes the olfactory signal to the olfactory cortex. The axons of
the mitral and tufted cells (major second-order neurons) sharpen
or alter the neural information. Periglomerular cells are involved
in lateral inhibition at the level of the glomeruli. Granule cells are
also inhibitory interneurones. The lateral olfactory tract terminates
in the pyriform and prepyriform (primary cortex), from where pri-
mary projections go into the thalamus (symmetric part of brain).
Axons project from thalamus into the neocortex (part of mam-
malian brain).
The olfactory cortex has five regions, with specific functions,
involved in olfaction. The regions are: anterior olfactory nucleus
(transfers odor information between regions such as piriform cor-
tex, olfactory bulb, etc.), prepiriform cortex (associated with odor
perception), lateral entorhinal cortex (linked with odor memory,
emotional and autonomic responses), periamygdaloid cortex (pro-
cesses olfactory information), and cortical nucleus of amygdala
(provides sense of smell) [20]. Information from olfactory bulb gets
transmitted to frontal cortex (for conscious perception of smell),
hypothalamus amygdale (for motivational and emotional aspects
of smell) and hippocampus (for odor memory). Savic [44] studied
the brain image upon activation by odorants. It was suggested that
neuroimaging of brain to odorant signals can be beneficial as a the-
oretic as well as diagnostic tool to study various olfactory disorders.
2.2. Mechanism of olfaction
2.2.1. Odorants and odorant binding proteins
Process of olfaction in air-breathing vertebrates begins when
volatile odorous molecules or odorants are inhaled. The odor-
ants are low molecular weight VOCs [38] that have low water
solubility and polarity, and relatively high vapor pressure. Odor-
ant compounds have relative molecular mass not more than
300 Daltons. Normal average breathing rate of about 250 mL/s [45]
provides a fast and moderately turbulent air flow that carries the
odorants/VOCs into the olfactory epithelium. The odorants of low
concentration traverse through an aqueous medium, mucus layer
covering the nasal epithelium, prior to its contact with the olfac-
tory epithelium. Recently, abundant small, water soluble proteins,
homologous to carrier proteins (of hydrophobic ligand) in other
body fluids, were discovered in the olfactory epithelium. These
proteins are thought to shuttle hydrophobic odorants towards
olfactory cilia and possibly act as chemostimulants and/or initiators
of signal transmission process. These proteins, termed as odorant
binding proteins bind to odorants reversibly. The OBPs are secreted
by non-neural support cells [46], and are present in both verte-
brates and insects, although morphologically different [3]. The OBPs
are present in the sensory lymph of insects and mucous of olfac-
tory epithelium in humans [22]. The presence of OBPs in terrestrial
vertebrates as well as insects suggests molecular adaptation of the
organisms to terrestrial environment [1].
The vertebrate OBPs belong to lipocalin family, while inverte-
brate OBPs constitute a unique protein family. Lipocalins are carrier
proteins for hydrophobic molecules in many biological fluids
[12,47]. Various OBP structures have been well studied in inver-
tebrates as well as vertebrates [47–49]. The three-dimensional
structure of the bovine OBP is a ␤-barrel, while insect OBPs mainly
contains ␣-helix motifs [47]. D’Auria et al. [12] stated that ver-
tebrate OBP had conserved folding pattern with an 8-stranded
␤-barrel flanked by an ␣-helix at C-terminal of the polypeptide
chain. The ␤-barrel has a central non-polar cavity (calyx) that bind
to hydrophobic odorant molecules. OBP of various species such as
pig, rat, mouse, cow and rabbit have been identified. Few OBPs
tuned to specific odorant groups have also been investigated. For
example, rat OBP-1 binds with heterocyclic compounds such as
pyrazine derivates, OBP-2 is specific for aliphatic aldehydes and car-
boxylic acids, and OBP-3 interacts with saturated and unsaturated
ring structures [12].
OBPs are important components of the olfactory signaling pro-
cess, however, their exact functional role in uncertain [3,38,46].
Potential functions are thought to be: help in the transfer of
lipophilic odorants across the olfactory mucus layer to the olfactory
receptors, concentration of odorants in the mucus layer relative
to air, removal of the utilized odorants for degradation, thereby
allowing another odorant molecule to interact with the receptor,
and protecting receptors by preventing excessive amounts of the
odorant from reaching the receptors. The differences between the
vertebrate and invertebrate OBPs are that vertebrate OBPs belong to
lipocalin family, consisting of hydrophobic molecular transporters;
while invertebrate OBPs (such as in insects) are an independent
gene family [46], and vertebrate OBPs attach to the ligand at the
interface of dimers, while invertebrate OBPs attach to the odorants
as monomers. Similar to vertebrate OBPs, the function of inver-
tebrate OBPs is unknown [46]. Possible functions in invertebrates
may be that they transport hydrophobic odorants, prevent degrada-
tion before activating the olfactory receptors, act as co-factors for
activation of transduction, and degrade odorants after activation
and removing them from lymph [49].
The mucous covering the olfactory epithelium provides the opti-
mum ionic environment for signal generation in olfactory receptor
cells. The OBP is believed to play a role in transferring the odorant
molecule through the mucous layer to the ORs. Deactivation and
elimination of the odorants is required for continuous procuring
odorant signals in the olfactory system. Enzymes in mucus inacti-
vate odorants making them lipid insoluble and incapable for signal
activation [1,3].
There are several theories describing the mechanism and speci-
ficity by which odorants bind to the olfactory receptors. Turin and
Yoshii [50] proposed theories based on shape, vibration and tun-
neling on odorant binding to receptors. Some of these theories are
summarized in Table 1.
 S. Sankaran et al. / Sensors and Actuators B 171–172 (2012) 1–17 5

Table 1
Theories of structure-odorant relationship.

Theories Description

Shape based theories (Odotopes)
Vibration theories
Biological spectroscope
The detection of odor associated with a molecule is due to the pattern such as relative number of receptors to which odorant binds;
part of molecular shape of odorant (odotopes) along with the measure of intensity of excitation allows vast detection of odorants.
Functional groups can be differentiated based on specific vibration frequencies; except enantiomers that have same vibrational
spectra but different odors. Thus, the theory of electron tunneling was proposed as possible mechanism of facilitating proteins to
behave as vibrational spectroscope.
Electron tunneling occurs when molecular vibrational energy equivalent to energy difference between energy level of both donor
and acceptor. Under these circumstances, the olfactory receptors operate as spectroscope that permits the detection of a single
well-defined energy.

2.2.2. Signal initiation and termination
The mechanism of chemical olfactory transduction
[3,20,21,38,40] has been well understood in humans. The OR
proteins are linked to guanine nucleotide-binding protein, G-
protein. When the odorants bind to the olfactory receptors, their
attachment induces a change in the shape of the olfactory receptor.
Thus, the modification in the structure of the receptor induces
their binding to the G-protein. The G-proteins are comprised of
three subunits: alpha (␣) subunit being active subunit, and beta
(␤) and gamma (␥) (Fig. 1). In absence of a signal, ␣ subunit binds
to guanine diphosphate (GDP). When activated by an odorant, the
olfactory receptor couples with G-protein and GDP is replaced by
guanine triphosphate (GTP). Once GTP is bound to the olfactory
receptor, ␣ subunit protein dissociates from the ␤ and ␥ sub-
units, and attaches to the enzyme adenylyl cyclase (AC). The AC
converts adenosine triphosphate (ATP) into cyclic-3 ,5 -Adenosyl
monophosphate (cAMP). cAMP is a neurotransmitter (Fig. 1). As
the intracellular concentration of cAMP increases, it acts as a
hormone and moves through the cell cytoplasm activating the
gated ion-protein channel (cyclic nucleotide-gated, CNG channel)
which permits the entry of extracellular inorganic ions (Na+ and
Ca2+ ). The resting potential of the olfactory plasma membrane
is −65 mV. Movement of ions across the cell membrane into the
olfactory neuronal cell generates a membrane potential leading to
electrical signal that represents the transfer of the chemosensory
information to the olfactory bulb via axons. The influx of Na+ and
Ca2+ causes the inside of cell to become less negative. The cAMP
concentration diminishes as it is hydrolyzed to AMP (Adenosine
monophsophate). In some mammals, Inositol tri phosphate (IP3) is
also known to participate in olfactory transduction.
Olfactory signal transduction in invertebrates is less under-
stood than vertebrates [21]. Among the invertebrates phyla studied
(crustacean-lobster, nematode-C. elegans and insects-Drosophila),
the cation channel is either activated by cyclic nucleotide (CN, Trans-
duction pathway I) or by IP3 (Transduction pathway II) or both. In
transduction pathway I, the G-protein activates the enzyme AC,
which results in formation of cAMP. In transduction pathway II, the
G-protein activates the enzyme phospholipase-C, which hydrolyzes
membrane phosphatidylinositol to form IP3 and diacylglycerol (DAG).
In lobster, odorant activates the receptors leading to formation of
both cAMP and IP3 in the outer dendrites of olfactory receptor cells.
The cAMP leads to activation of K+ channels and hyperpolarization
of cell, while IP3 leads to depolarization of the olfactory neuron.
For insects, IP3 pathway is more prevalent. Although there is some
indication that cAMP pathway may also play a role in insect olfac-
tion, the exact evidence remains to be unexplored. In nematode,
both pathways of signal transduction occur [22].
In general, most odors are composed of multiple odorant
molecules, and each odorant activates several odorant receptors.
This leads to a combinatorial code forming an “odorant pattern”.
This is the basis for ability of human to recognize and form memo-
ries of approximately 10,000 different odors. As quoted by Buck [40]
“Just like letters of the alphabet are used in different combinations
to form different words, the odorant receptors are used in different
combinations to detect different odorants and encode their unique
identities.”
The olfactory receptor cell contains single type of OR, however,
each OR is sensitive to few odorant molecules. The nerve end-
ing of olfactory receptor cells terminates in the microdomain of
glomeruli and olfactory domains, from where the signal is transmit-
ted towards olfactory bulb and parts of brain. A single glomerulus
receives signals from the cells containing same type of OR [40]. The
combinatory approach enables the detection of numerous odors
inspite of having about few thousand ORs.
Many odorant substances are recognized by more than one
receptor and most receptors recognize several odorant molecules,
probably related by chemical property. Receptors identify spe-
cific characteristics (epitopes/determinants) of odorant molecules.
Odorant molecules may contain a number of ‘epitopes’. Thus, the
recognition of an odorant molecule depends on which receptors
are activated and extent of activation. In the olfactory bulb, broad
regions of sensitivity to different features (for example, functional
group or molecular length) of odorant molecules are observed
[21,51]. Firestein [21] has explained the combination pattern of
odor recognition in pictorial presentation.
Another important aspect of the olfactory system is the rapid
recovery of the odor sensing ability. OBP act as scavengers remov-
ing odorants from the olfactory mucous. In addition, within the
transduction pathway, there are mechanisms that aid in signal ter-
mination. Protein kinase (such as protein kinase A or protein kinase
C) or specialized G-protein receptor kinase (such as rhodopsin
kinase) phosphorylate the ORs and inactivate them [38]. Odor-
ants can also induce phosphorylation of immunoprecipitable ORs
[38,52]. Another signal inactivation method in olfactory system is
through CNG pathway. The influx of Ca2+ into the cell membrane
increases the intracellular Ca2+ concentration, which inhibits the
ion channels. The Ca2+ concentration reduce the cAMP concentra-
tion that eventually inhibits the signal generation [38].
2.2.3. Signal processing
In general, the binding of odorant molecules to the olfactory
receptors in cilia results inwardly depolarizing current that acti-
vates the action potentials of the receptor cell, and this process
provides the neural signals to be decoded in the higher parts of brain
[20]. The olfactory bulb processes the molecular signal obtained
from olfactory receptor cells in the olfactory system. The olfactory
bulb is segregated into zones in rabbits and mice; while in human
it is exhibited as glomeruli convergence [43]. A glomerulus may
respond to few odorant molecules as it receives information from
a particular olfactory receptor that reacts with different odorant
molecules. As the neurons of similar olfactory receptor converge
in the same glomerulus, each glomerulus is devoted to single type
of olfactory receptors. Sensory information received from olfactory
receptors is processed in olfactory bulb before being transmitted
to the olfactory cortex. The periglomerular cells in the glomeru-
lus form synaptic association with mitral and tufted cells upon
excitation [38].
6 S. Sankaran et al. / Sensors and Actuators B 171–172 (2012) 1–17
Fig. 2. Artificial olfactory sensing systems and their applications.
Activation of tufted and mitral cells results in feedback inhibi-
tion of neighboring mitral and tufted cells. The dendrite of these two
cells forms dendrodendritic reciprocal synapses with periglomeru-
lar cells. Periglomerular cells send inhibitory projections to the
dendrites of neighboring mitral and tufted cells, aiding in lateral
inhibition. Thus, these interneurons play a key role in processing
olfactory information. The lateral inhibition is tuned through den-
drodendritic reciprocal synapses with granule cells that enhance
the contrast between mild and strong signals. This increases the
selectivity of each mitral and tufted cells to odorant molecules
[43].
The signals separated in nose and olfactory bulb appears to
be combined in individual neurons in the olfactory cortex [53].
The cortical neurons integrate the signals from different olfactory
receptors that detect the same odorant and thus, perform initial
step of odor image construction. Zou and Buck [53] experimentally
found out that the dual odorant mixes activates number of cortical
neurons than each odorant compound. The olfactory cortex con-
trasts the olfactory bulb response, where the mitral cells respond
to each odorant compound. The finding indicated that the olfactory
cortex exhibit an integrative capability that is absent in olfactory
bulb. Therefore, humans display a distinct sense of smell for odorant
mixtures, while individual odorants are sensed as different odors.
3. Development of olfactory sensing system
3.1. Electronic noses
Olfactory sensor systems are beneficial in many fields includ-
ing food industry for regularly monitoring food quality and safety
(Fig. 2). The physiological and psychological status affects the
human sensory evaluations and decision-making ability. In con-
trast, an olfactory sensing system can provide a qualitative as
well as quantitative estimate of odor perception without any
bias [54,55]. Advanced research has been conducted to develop
‘electronic nose (e-nose)’ that can mimic mammalian nose. The
electronic noses are being fabricated using different operating prin-
ciples with sensitivity towards defined target VOCs. The elementary
steps of signal transduction, signal processing, and identification of
chemical patterns differ drastically from those that exist in natural
nose. An e-nose system makes use of an array of detectors, each
designed to match the odorant selectivity of single type of olfac-
tory receptor. These detectors provide multi-dimensional response
when in contact with certain VOCs. The multi-detector responses
can further be analyzed by suitable pattern recognition techniques
to categorize specific perceptions related to VOCs.
The sensing materials of the e-nose employ metal oxide detec-
tors, quartz resonators, and materials such as conducting polymers
(CPs) for detecting different VOCs. An ideal sensing material to be
integrated in an e-nose should fulfill the following criteria: (i) high
sensitivity to chemical compounds; (ii) low sensitivity to humidity
and temperature; (iii) high selectivity; (iv) high stability; (v) high
reproducibility; (vi) high reliability; (vii) short reaction and recov-
ery period; (viii) robust and durable; (ix) easy calibration, and (x)
small dimensions [32].
There are number of books [55–59] and journal articles
[24,28,32,33] available on e-nose systems. Table 2 summarizes few
applications of e-nose systems and some feature extraction meth-
ods. Conducting polymers and metal oxides are commonly used
sensing materials in such systems. Arshak et al. [65] reviewed
some of the sensing materials used in e-nose systems. Bai and
Shi [66] reviewed CP-based sensing materials such as polyani-
line, polypyrrole and polythiophene, and their properties that are
currently being investigated for gas sensing applications. These
sensing materials have also been used in commercial e-nose sys-
tems. System with 32 polymer composites sensor array has been
commercialized by Cyrano Sciences (Smiths Detection). Young et al.
[63] reports use of commercial e-nose systems, from Karlsruhe
Research Center, Air Sense, and Daimler-Benz Aerospac, that con-
sists of about 38, 7 and 5 metal oxide sensors, respectively for air
contamination detection in space shuttles. Röck et al. [31] sum-
marized approaches used in e-nose systems and commercially
available products. Considering the wide application domain of
e-noses, Table 3 summarizes few recent applications of e-nose sys-
tems in different fields.
E-noses have also been used for monitoring plant diseases and
toxins. Falasconi et al. [82] used an olfactory sensor for detect-
ing mycotoxin (Fusarium verticillioides) contamination in maize.
 S. Sankaran et al. / Sensors and Actuators B 171–172 (2012) 1–17 7

Table 2
Applications of electronic nose systems and associated feature extraction methods.

Application VOCs detected Sensor array details Classification/prediction model References

Detection of beef spoilage
Detection of microbial
contamination in
processed tomatoes
Quality assessment of
modified atmosphere
packaged poultry meat
Evaluation of cheddar
cheese quality
Air contaminants in space
shuttle
Evaluation of sardine fish
freshness
Methane Custom built e-nose Features: AUC [8]
Hydrogen sulfide Sensor type: MO Classification: PCA, LDA, QDA
Alcohols No. of sensors: 8
Organic solvents
Carbon dioxide
Dimethyl sulfide Commercial e-nose Features: Rminimun /Rbaseline [60]
3-Methyl-furan EOS835 (Sacmi Imola scarl, Italy) Classification: PCA, kNN
Acetone Sensor type: MO
Ethanol No. of sensors: 6
3-Methyl-1-butanol
Hydrogen sulfide Commercial e-nose Features: not detailed [61]
Dimethyl sulfide NST 3320 instrument (Applied Sensor, Used NST Senstool software (Ver.
Sweden) 2.7.4.27, Applied Sensor, Sweden)
Sensor type: MO, MOSFET Classification: PCA, PLS, ANN
No. of sensors: 24
Acetone Commercial e-nose Features: mass units [62]
Butanone Chem Sensor 4400 (Agilent Technology Classification: CA, PCA
Acetic acid Kenner, LA)
Butanol Sensor type: mass selective detector
Dimethyl sulfide and gas chromatograph
4-Mercapto-4-methyl-pentan-2-on No. of sensors: -
Acetone Commercial e-nose Features: maximum initial slope, AUC, [63]
Ammonia KAMINA (Karlsruhe research center, FFT coefficient
Nitrogen dioxide Germany) Classification: Linear classifier with RU
Toluene Sensor type: MO estimator
Methylethyl ketone No. of sensors: 38 RU is average of classification achieved
Isopropyl alcohol i-Pen2 (AirSense analytics, Germany) using leave-one-out & using
Xylene Sensor type: MO resubstitution approach
Trichloroethylene No. of sensors: 7
Hydrazine Cyranose (Cyrano sciences, Inc. CA): 32
Mono methyl- hydrazine polymer sensors
Sensor type: polymer
No. of sensors: 32
Custom built e-nose
NASA Jet propulsion laboratory
Sensor type: CP
No. of sensors: 32
Alcohol Custom built e-nose Features: initial & steady-state [64]
Organic solvents Sensor type: MO conductance, dynamic slope of
Hydrogen sulfide No. of sensors: 6 response curve, AUC
Ammonia Classification: PCA, SVM
Chlorofluorocarbons

ANN: Artificial neural network; AUC: Area under curve; CA: Cluster analysis; CP: Conducting polymer; FFT: Fast Fourier transform; kNN: k-nearest neighbors; LDA: Linear
discriminant analysis; MO: Metal oxide; MOSFET: Metal oxide semiconductor field-effect transistor; PCA: Principal component analysis; PLS: Partial least squares regression;
QDA: Quadratic discriminant analysis; R: Resistance; SVM: Support vector machine.

Commercial (Model EOS 835, Scami, Italy) system with six thin film
metal oxide semiconductor sensors was used for the analysis. The
e-nose was found to be effective in screening the maize bulk so as
to prevent the entry of mycotoxins into the food chain.
E-nose systems offer several benefits, especially in the indus-
tries where aroma is an important component of the product
quality, such as perfume industry and food industry [86]. Such
systems are also applied for environmental air quality monitoring
and toxic vapor detection. A simple example of e-nose system is
the fire alarms. Although e-nose systems are widely explored for
many different applications, some limitations of such systems are:
high sensitivity to temperature and humidity changes with many
sensors requiring high operating temperature, complex interface
circuitry, baseline signal drift over time, low signal to noise ratio,
non-specific or partially specific response, moderate reproducibil-
ity, and high costs.
Few constraints of specific type of sensors (such as electro-
chemical and piezoelectric sensors) can be addressed using hybrid
sensors. Swann et al. [87] developed polyethylene oxide-based
hybrid sensor (in combination with carbon black) to detect vapor
densities of various VOCs such as cyclohaxane, hexanol, toluene,
chloroform, dichloromethane, etc. They determined sensitivity of
sensing materials using two different approaches, conductimet-
ric (change in resistance of an interdigitated electrode sensor) and
piezoelectric (change in frequency of a quartz crystal microbalance,
QCM sensor), and combined these sensitivities together to relate
the sensor response to the molecular density. Similar approach was
utilized by Ulmer et al. [88] where different sensing mechanisms
such as conductimetric, piezoelectric, and calorimetric techniques
were combined to determine flavors and odors. Cui et al. [89] com-
bined conductimetric and piezoelectric methods to develop VOC
odor map. Several researchers have also investigated the integra-
tion of multiple sensing materials [90–94].
With recent technological developments, limitations of e-nose
systems are being addressed using different approaches. Recent
findings have led to the development and application of new
conducting polymers, molecular imprinted polymer, stationary
phase material of gas chromatography, metalloporphyrins, nano-
dimensional sensors, and plasma polymerized films [95]. For
example, in molecular imprinted sensor, formation of specific
active sites in the macromolecules, allows the co-polymerizaion
of functional monomers in the presence of the VOC of interest.
These imprinted polymers can contain fluorescein that allows flu-
orescence sensing [96]. Bunte et al. [97] developed such molecular
imprinted polymer for detecting 2,4,6-trinitrotoluene vapors. Sev-
eral applications of molecular imprinted sensors are being explored
[98–101]. Shimizu and Stephenson [102] have summarized the
properties and applications of molecular imprinted polymer sensor
8 S. Sankaran et al. / Sensors and Actuators B 171–172 (2012) 1–17

Table 3
Applications of metal oxide and polymer based olfactory sensors in various research fields.

Sensor materials Applications References

Food Quality and Safety

Polymer Identification of spoiled beef [8]
Metal oxide Saccharomyces cerevisiae and Escherichia coli detection in tomato [60]
Metal oxide Aroma of virgin olive oil [67]
Metal oxide, polymer Antioxidants and aroma of infant cereal [68]
Metal oxide Assessment of meat freshness [69]
Metal oxide Monitoring of milk [70]
Metal oxide Coffee ripening analysis [71]
Synthetic oligopeptides Dioxin detection in food [72]

Environmental monitoring
Polymer Amine vapor detection [73]
Metal oxide Carbon monoxide and nitrogen oxide detection [74]
Metal oxide Odors from wastewater treatment plant [75]
Metal oxide Assessing odors from landfill [76]
Polymer Emissions from polluted river [77]
Metal oxide Monitoring benzene in urban environment [78]
Metal oxide Hydrogen sulfide and nitrous oxide detection [79]

Plant diseases
Polymer Basal stem rot disease in oil palm [80]
Polymer Blueberry fruit disease [81]
Metal oxide Fusarium verticillioides in corn [82]

Human diseases
Quartz microbalance sensors- porphyrins films Discrimination between lung cancer and other lung diseases [83]
Polymer Discrimination between lung cancer and chronic obstructive pulmonary disease [84,85]

arrays. Such polymers offer better stability at variable pH, pres-
sure, temperature and radiation conditions, and are compatible
with micromachining techniques than natural biomolecules [96].
Challenges remain in the area of reliable design procedures, devel-
opment of effective immobilization procedures, improvement in
polymer binding affinity, and preventing the cross-reactivity with
improved specificity [96,102]. Metalloporphyrins are also recently
being explored for VOC sensing applications. Paske et al. [103] and
Chevallier et al. [104] developed interfacially polymerized metallo-
porphyrin thin film for calorimetric detection of alcohols, ketones
and chlorocarbon vapors, and metalloporphyrin-functionalized
diamond nanoparticles for nitroaromatic vapor detection, respec-
tively.
E-nose systems perform satisfactorily with moderate selectiv-
ity and sensitivity. However, sensors with higher sensitivity and
selectivity are desired to further widen and improve their appli-
cation domain. The new generation olfactory sensors thus need to
be developed simulating the biological olfactory system (Fig. 2).
Progress in the current development of e-noses and in our under-
standing of biochemical, molecular, biological, physiological, and
behavioral details of odor sensation makes it possible to fabricate
new generation olfactory sensors, termed as “bioelectronic noses”
[28]. The following section elaborates on the possible new genera-
tion intelligent biomimetic olfactory sensors.
3.2. New generation biomimetic olfactory sensors
Human olfactory system can distinguish various odorants with
high accuracy and at low detection limits. If components of such
advanced olfactory system can be understood and utilized, it
is possible to develop a robust system with high accuracy and
precision to detect very low concentrations of VOCs. Several bio-
logical olfactory system components have been used for fabricating
sensors that detect different odorants. The commonly used com-
ponents/biomolecules in developing sensors are: ORs, OBPs, and
olfactory neurons [16,105,106]. Artificial sensing materials have
also been synthesized that mimic olfactory-based components
[107,108].
Despite several advantages, there exist some concerns related
to the application of new generation olfactory sensors such as: (i)
deposition of the biomolecules to a sensor substrate can be chal-
lenging, (ii) short life of the biosensor, and (iii) challenges related to
the sensor reproducibility. The biomolecules may be difficult to iso-
late from the olfactory system, cultivate and treat; moreover, the
heterologous expression of the olfactory sensors is also not easy
[106]. Some aspects related to different biological sensor mate-
rials with biomimetic capabilities are discussed in the following
sections.
3.2.1. Olfactory receptors
Olfactory receptor cells in the olfactory epithelium contain OR
protein in their cilia that interact with odor molecules and transmit
the signal to the higher level of sensory organelles. An important
function of the G-protein linked olfactory receptors is to convert
the chemical signal (by linking to the odorant molecules) to elec-
trical signal and transmit it to the brain. Thus, these receptors can
be the ideal and most effective tool as a sensor (Fig. 3). Other charac-
teristics that make olfactory receptors appropriate for sensing are:
(i) they generate change in electric properties upon odorant bind-
ing; (ii) their molecular events can be detected by opto-electronic
devices; and (iii) they are endowed by genetic engineering, i.e., their
detection, grafting, and orientation in sensor devices are possible
using specific tags [2]. They can also respond to range of VOCs that
share specific molecular feature and discriminate small discrep-
ancies in odorant structure, size and concentration. In addition,
olfactory receptors can detect concentrations as low as 10 pM to
10 nM [109]. Table 4 summarizes some of the results obtained from
studies on sensors based on olfactory receptors.
Two initial steps involved in obtaining ORs are extraction of
biomaterials from animals and expressing them in a microor-
ganism or cells to confirm the existence of the receptor protein.
Table 5 presents the organisms from which the olfactory recep-
tors were extracted/isolated and the biomolecules used to express
the isolated olfactory receptor in some studies. Wetzel et al. [113]
used a heterogeneous system to characterize the selectivity and
sensitivity of human receptor protein (OR 17-40). The receptor
proteins were functionally expressed in both human embryonic
 S. Sankaran et al. / Sensors and Actuators B 171–172 (2012) 1–17 9

Fig. 3. Development of olfactory receptors-based biomimetic sensors.

kidney (HEK)-293 cells and Xenopus Laevis (frog) oocytes. The OR
17-40 expressed in both these cells could be activated by micromo-
lar concentration of helional and heliotroplyacetone. Recombinant
expressions with HEK-293 cells have issues namely the responses
are not fully reversible which often saturates after few applica-
tions, and there are also reports of difficulty in establishing the
dose–response relationships. In these aspects, proteins expressed
in oocytes provide a response for many hours, allowing different
mixtures to be applied during the time for and quantitative quali-
tative evaluation.
Vidic et al. [109] obtained the genes encoding the OR (University
of Nottingham, UK) and co-expressed mammalian olfactory recep-
tors and an appropriate G-protein in Saccharmyces cerevisiae, from
which membrane nanosomes were produced. Carboxy-methyl
modified dextran polymers were used for linking the nanosomes
to the gold sensor chip. This adhering technique was followed

Table 4
Studies on olfactory sensors of biological origin.

Detectors Odorants Odorant concentration References

Rat olfactory receptor expressed on
human embryonic kidney cells
integrated with piezoelectric
material
Isolated olfactory receptor proteins
from bullfrogs coated onto the
surface of a piezoelectric
electrode
Olfactory receptor protein of C.
elegans, expressed in Escherichia
coli membrane fraction. Quartz
crystal microbalance coated with
membrane extract integrated
with piezoelectric material
Mouse trace amine-associated
receptor 5 expressed in Xenopus
laevis melanophore
Olfactory receptor OR17 expressed
in yeast
Hexanal, Heptanal, Octanal, Nonanal, Decanal Responded to 1 mM hexanal, heptanal, and [16]
octanal. Response was stronger to octanal.
10−8 –10 mM octanal
LOD: 10−8 mM octanal
n-Decyl alcohol, n-Caproic acid, Isoamyl acetate, 10−6 g n-Caproic acid, 10−7 g Isoamyl acetate, [105]
n-Octyl alcohol, ␤-Ionone n-Octyl alcohol and ␤-Ionone
Hexanal, Heptanal, Octanal, Decanal, Diacetyl Responded to 10−5 M diacetyl, hexanal, heptanal, [106]
octanal and decanal. Response was stronger to
diacetyl.
10−12 –10−5 M diacetyl
LOD: 10−12 M diacetyl
Trimethyl amine and other amines 1–100 ppm [110]
LOD: 1 ppm
Heptanal, Octanal, Nanonal, Helional Responded to 10−5 to 10−13 M helional, heptanal, [111]
octanal and nanonal.
Response was stronger to heptanal and octanal.
LOD: 3 × 10−9 M heptanal, 7 × 10−9 M octanal,
5 × 10−8 M nanonal

LOD-Lower limit of detection.

10 S. Sankaran et al. / Sensors and Actuators B 171–172 (2012) 1–17
Table 5
Animals used for extracting olfactory system based biomaterials.
Olfactory component
Olfactory receptor prepared using I7 receptor gene
Olfactory receptor protein (5 fractions obtained)
Olfactory receptor protein ODR-10
Olfactory receptor and appropriate G protein- plasmids encoding OR17
from rat, OR 1740 from humans and GPA1-G protein
Trace amine-associated receptor 5
Olfactory receptor I7
because of the: (i) lesser risk of receptor alteration and activ-
ity loss, (ii) requirement of low olfactory receptors concentration,
(iii) receptor binding sites remain accessible and functional, and
(iv) measurements of receptor activity were easier. The VOCs
that were sensed using surface plasma resonance (SPR) technique
were: octanal, nonanal, heptanal, diacetyl, cyclohexyl-acetate,
octanol, octanone, octanoic acid, isoamylacetate, pyridine, helional,
cassione, piperonyl acetate, 3,4-methylenedioxyphenyl acetone
and 3,4-methylenedioxypropiophenone. Sensor responses were
directly proportional to the molecular weight, thus the low molecu-
lar weight compounds could not be detected. VOCs such as octanal,
helional, cassione, piperonyl acetate, 3,4-methylenedioxyphenyl
acetone, and 3,4-methylenedioxypropiophenone showed good
sensor response at 10−6 M concentrations. Seven cycles of exper-
iment, for reproducibility test, with an hour interval produced
repeatable signals. However, after about eight cycles, the system
did not show any detectable response. This signifies the importance
of repeatability and recovery of such biological olfactory sensors.
Sung et al. [106] coated the quartz crystal (electrode) with
E. coli membrane fraction containing olfactory receptor by simply
introducing 1 ␮L aliquot on the substrate and drying it for 4 h at
room temperature. Contrary to Vidic et al. [109], Sung et al. [106]
found that the response of olfactory receptor based QCM sensor
was inversely proportional to the number of carbons in the odor-
ant molecules (hexanal, heptanal, octanal and decanal), although
the variation in sensor response was high. However, diacetyl (2,3-
butanedione) gave a good response, possibly due to the presence of
two C=O group when compared to other VOCs (aldehydes). Prob-
ably, sensors response depends on the selected olfactory receptor
and the detection method. It should be noted that Vidic et al. [109]
used a higher molecular weight compounds compared to Sung et al.
[106]. Minic et al. [111] developed olfactory sensor based on yeast
expressed olfactory receptor I7 similar to research performed by
Vidic et al. [109], and found that the sensor was more sensitive
in terms of differential bioluminescence to octanal compared to
nonanal and heptanal. Relationship between molecular weight and
sensitivity was not found as reported by Vidic et al. [109].
Wu [105] also used a simple deposition technique, by evenly
spreading the solution containing olfactory receptor protein, using
a glass rod and allowing it to dry in desiccator for 4 h. The cilia
from the trout (bull frog) olfactory rosettes and mollusk gills were
isolated to extract the olfactory mucous. Gel electrophoresis was
performed to separate the olfactory receptors from the extracted
cells and five different fractions were, each possibly represent-
ing a type of olfactory receptors. Each fraction was concentrated
and deposited on an electrode surface. For comparison, coating
bovine serum albumin (protein) and two phospholipid materials
(phosphotidylcholine and phosphotidylinosital) were coated. In
this study, although the olfactory receptor was isolated, presence of
olfactory receptors was not confirmed. Considering these aspects,
expression of an olfactory receptor on biological cells may be useful
for reliable results. Many studies have used HEK cells in compar-
ison to other cells [16,112,113]. Sung et al. [106] indicated the E.
coli cells do not require an import sequence for the expression of
the olfactory receptor protein on the cell membrane as required
Extracted from animals Expressed in organisms References
Rat Human embryo kidney-293 cell [16]
Bull frog – [105]
Caenorhabditis elegans (nematode) Bacteria Escherichia coli [106]
Rat, Human Yeast Saccharomyces cerevisiae [109]
Mouse Xenopus laevis melanophores [110]
Mice, Rats Human embryo kidney-293 cell [112]
for HEK-293 cells. This makes the expression of olfactory receptor
protein in E. coli much easier than the HEK-293 cells. Also, HEK-
293 cells require a stable cell line to be used as a biosensor, which
is tedious, time-consuming and expensive process. Wetzel et al.
[113] also preferred frog oocyte compared to HEK-293 cells.
Wu [105] investigated the reproducibility of olfactory receptor
protein based sensor and found that the sensor sensitivity towards
n-Caproic acid did not reduce after eleven test runs. These runs
were performed over a period of two weeks in dry chamber so
that the humidity will not affect the piezoelectric crystal. Sensor
response did not change even after 5 months, when stored in dessi-
cator. Although Wu [105] used a crude method for extracting the
olfactory cells, a good reproducibility was observed.
3.2.2. Odorant binding proteins
OBPs, similar to ORs, can also be used as an efficient tool as
sensing material. OBPs are low molecular weight soluble proteins,
serving as carriers for conveying odorants through the aqueous
nasal mucus in the olfactory system. OBPs reversibly bind to dis-
tinct chemical classes of odorants with dissociation constants in the
micromolar range, making them a potential sensing material in the
design of artificial olfactory systems [15]. Similar to other proteins,
OBPs have a well-defined structure.
Molecular interactions defining the specific alcohol-binding site
in the structure of OBP LUSH (Drosophila) have also been inves-
tigated and identified [49]. Thus, OBP such as LUSH could be
potential sensor materials for detecting alcohols and other odor-
ant molecules. Sankaran et al. [10] developed a QCM sensor based
on LUSH OBP to detect alcohols such as 3-methyl-1-butanol and 1-
hexanol utilizing the knowledge of LUSH binding site. Similarly,
D’Auria et al. [12] also worked on OBP-based olfactory sensor.
OBP was acquired from the dog’s nasal epithelium for developing
olfactory sensors. Details of OBP extraction were not specified in
the paper. Developed sensor was sensitive to 10–100% of 0.2 mM
pyrazine solution in presence of 0.75 mg/mL of dog OBP.
Hou et al. [15] analyzed the deposition techniques for deposit-
ing recombinant rat OBP-1F. A mixed Langmuir (Langmuir and
Langmuir-Blodgett, LB) technique was used to develop OBP based
biosensor. The OBP was expressed in yeast, Pichia pastoris. The films
were characterized using scanning electron microscopy (SEM),
atomic force microscopy (AFM) and electrochemical impedance
spectroscopy (EIS). As reported, the films deposited by mixed Lang-
muir method were reproducible, stable and well-ordered. Analysis
using EIS before and after the contact of an odorant (isoamyl alco-
hol) to the mixed Langmuir films suggested a decrease in resistance
from about 1.18 M (before contact) to 25 k (after contact).
3.2.3. Olfactory neurons
Olfactory neurons possess thousands of olfactory receptor cells
responding to different types of odorants. Thus, olfactory neu-
rons based olfactory sensors can exhibit exquisite specificity, high
sensitivity and fast response, making them an ideal biomaterial
for olfactory cell based sensors [114]. Liu et al. [14] used olfac-
tory receptor neurons and cells of olfactory bulb from rat pups to
develop olfactory sensor. They used poly-l-orithine and laminin
 S. Sankaran et al. / Sensors and Actuators B 171–172 (2012) 1–17
mixture prior to seeding the cells into the surface of the silicon chip
to improve the adhesion of the cells. Once prepared, the cells were
maintained in the chip for one week at 37 ◦ C under 5% humidity,
with continuous feeding of cells every two days. An electrochemi-
cal technique, light addressable potentiometric sensor (LAPS) was
used to measure the extracellular potential of olfactory neuron cells
deposited on the semiconductor chip. The extracellular potential
changed when the cells were exposed to neurotransmitters. Sen-
sor responded to 25 ␮M glutamic acid, while had a low response
to 1 ␮M and 50 ␮M glutamic acid. Probable reason for the absence
of the response at 50 ␮M could be the toxicity of glutamic acid
to the neuron cells. When 1 ␮M, 25 ␮M and 50 ␮M acetic acid
were measured, the amplitude of the frequency increased in a
concentration-dependent manner. However, the amplitude disap-
peared 10 min after discontinuing the odorant stimulation. This
signifies toxic effect of high odorant concentrations on the biolog-
ical cells.
In a recent study [115,116], researchers extracted the olfac-
tory mucosa from rats and deposited on the LAPS surface. SEM
image demonstrated presence of olfactory cilia, where the olfac-
tory receptors were located. Developed sensor was used to detect
25 ␮M/mL concentrations of VOCs, butanedione and acetic acid
at room temperature. Sensitivity of blowfly’s olfactory receptor
neurons to VOCs has also been investigated [117,17] towards 1-
hexanol, butyric acid and 1,4-diaminobutane.
3.2.4. Synthetic polypeptides
Another unique way of developing olfactory sensors is by syn-
thesizing artificial peptide sequences mimicking the binding site
of OR or OBP (Fig. 3). Although the biological olfactory system-
based biomolecules have been used as sensing material, the process
poses several challenges during extraction. Moreover, identifying
olfactory receptors specific to an odorant may be time-consuming,
labor-intensive and expensive [106]. Therefore, developing olfac-
tory receptor-based polypeptide sequences as sensing materials
offers a simple alternative with desirable sensitivity and selectivity
for odorant detection. In addition, it offers several benefits such
as enhanced stability, better reproducibility, predictable output,
cost-effectiveness and better shelf-life [72,118]. This reduces the
chances of degradation in comparison to the biological cells. The
amino acid residues may also have a higher affinity to the VOC of
interest.
Blanco et al. [119] studied the binding energies and conduc-
tance responses of putative sensing peptides to ammonia and acetic
acid and found that these peptide sequences offer high sensitivity.
Such peptide sequences have also been used for detecting chemical
compounds such as herbicides and dioxins [120]. Wu and Lo [118]
established the structure of dog’s olfactory receptor and its binding
site between receptor and odorant molecule (trimethylamine-
TMA) using computer simulations. Two oligopeptide sequences,
orp61 and orp188 were identified for odorant sensing, synthe-
sized, purified, and deposited on piezoelectric gold electrodes.
Sensitivity of the probes was improved with site-specific modifi-
cation using amino acid sequence. Response of synthetic olfactory
receptor-based peptide sequences to TMA (45%), ammonia (33%),
acetic acid (100%), ethyl acetate (99.5%), methanol (99.9%), and
benzene (99.7%) were measured. The orp188 responded to all the
VOCs, while orp61 responded to all but benzene. The relative sen-
sitivity of olfactory receptor probes with site-specific modification
increased TMA sensitivity of orp61. Sensor response to other odor-
ant molecules displayed a marginal variation before and after
site-specific modification.
Lin et al. [4] synthesized peptides by simulating olfactory
receptor protein related to a target odorant molecule. Recep-
tor structure was simulated to obtain secondary and 3-D
structure. Once the structure was modeled, the reaction sites
11
between odorant molecule and olfactory receptor was simulated
using software programs. Finally, the peptides and reaction sites
were synthesized (although the method of synthesis was not
described). QCM technique was employed to measure the response
for the non-invasive diagnosis of uremia. Breath analysis of patients
with uremia, chronic renal failure (CRF), chronic renal insufficien-
cies (CRI) and healthy individuals indicated that the sensor was able
to differentiate healthy individuals, uremia patients and patients
with CRI/CRF with a classification of up to 87%. The healthy indi-
viduals were classified with 100% accuracy, while CRI/CRF patients
were classified with 90% accuracy. Thus, the sensor was appli-
cable for diagnosing diseases at an early stage. Similar to Lin
et al. [4], Wu et al., [108] synthesized peptides similar to human
olfactory receptor proteins. The peptides were modeled using
molecular simulation method, and possible binding sites between
model structure and target VOCs (trimethylamine, ammonia, acetic
acid and o-xylene) were identified. The series of polypeptide
sequence (probes) for TMA, ammonia, acetic acid and o-xylene
were horp61, horp109, horp193 and horp103, respectively. Polypep-
tide probes were synthesized by fluorenylmethoxycarbonyl (Fmoc)
method. Self assembled monolayer (SAM) technique was used to
deposit/immobilize synthesized peptides into the electrode. The
probes displayed maximum sensitivity to their respective odorant
compound. Authors suggested that the probes sensitive to ammo-
nia and TMA can be applied to identify kidney failure patients (as
they exhale significant amounts of ammonia and organic amines),
and probes sensitive to acetic acid and o-xylene can be applied
for specific environmental monitoring [108]. Panigrahi et al. [121]
further assessed the horp193 sequence for detection of lower
concentrations of acetic acid using a modified sensor sensitivity
characterization system. An estimated detection limit for acetic
acid detection was about 1–3 ppm. Study on stability of the QCM
sensor with time indicated that the sensor was sensitive to acetic
acid even after a period of one month. Lu et al. [122] developed
similar polypeptide probes for the detection of acetic acid, butyric
acid, dimethyl amine and chlorobenzene using piezoelectric sensor
chips. A multi-array platform was developed to detect a range of
VOCs. Although, the sensor sensitivities were investigated, details
of probe selection was not reported. The polypeptide probes were
sensitive to range of VOCs such as acetic acid, butyric acid, ammo-
nia, dimethyl amine, benzene, chlorobenzene and their mixtures.
Principal component analysis (PCA) was performed to classify the
different compounds. Study reported the potential of using such
polypeptide probes for VOC detection.
Sankaran et al. [10] utilized a synthetic polypeptide sequence
based on LUSH OBP binding site as a sensing material in a QCM
sensor to detect alcohols. The sensor was sensitive to low con-
centrations (with estimated detection limit of less than 5 ppm)
of alcohols such as 3-methyl-1-butanol and 1-hexanol. Sankaran
et al. [11] also developed QCM-based polypeptide sensors based on
simulation studies. Tripos/Sybyl® , a molecular simulation program
was used to predict the binding site of mouse olfactory receptor
774 using a modeled 3-D structure. A polypeptide sequence incor-
porating the binding site amino acid residues was then selected
and synthesized as the possible sensing material. This material was
deposited on a QCM crystal to assess the sensor sensitivity to alco-
hols. The sensor was developed for the detection of alcohols that
could be related to Salmonella contamination in packaged beef. It
was reported that the developed sensor was sensitive to 1-pentanol
and 1-hexanol with an estimated lower limit of detection of about
3–5 ppm and 2–3 ppm, respectively.
Jaworski et al. [123] found that such approach of fabricating
peptide-based receptor was sensitive and selective to VOCs such
trinitrotoluene and 2,4-dinitrotoluene. Similar to these studies,
Izumi et al. [54] fabricated a water membrane based olfactory sens-
ing system that mimics the biological olfactory system. The partial
12 S. Sankaran et al. / Sensors and Actuators B 171–172 (2012) 1–17
structures of odorant molecules (based on odotope hypothesis) are
thought to induce the receptor molecules. The partial structures
of odorants were indirectly measured using olfactory sensing sys-
tem. The adsorption of odorant molecules depends on its (odorant)
polarity and the roughness of the surface. By controlling the sur-
face polarity of the electrode, adsorption interaction of the partial
structures was controlled in the study. Platinum electrode were
used as transducers to test different types aromatic alcohols and
other compounds (acetone and acetic acid) using the sensor. The
alcohols exhibited better response compared to other compounds,
as they are strongly hydrophilic in nature. The results were fur-
ther analyzed by PCA and regression models. Both techniques were
successful in discriminating the odorants.
Although the polypeptide-based sensing materials have been
used for sensing applications, there has been no study where the
length of the peptide sequence has been optimized. Studies dis-
cussed in here use a peptide chain with five to sixteen amino
acid residues. A secondary structure mimicking the receptor can
be established with a longer chain length. However, the effect of
chain length needs to be studied further to determine the optimum
length of peptide sequence required for acquiring high sensitivity
and selectivity.
3.2.5. Process overview
In summary, odorant specific/non-specific biological olfactory
system based biomolecules can either be extracted from the
olfactory system of an organism or receptor-based polypeptide
sequences can be synthesized. These biomolecules are expressed
prior to its application in olfactory sensors. The olfactory cells are
commonly expressed in cells such as Escherichia coli [106], HEK-
293 [16] or Saccharmyces cerevisiae [109]. The sensing materials
are deposited on the inert sensor chip using different techniques.
The deposition technique can range from simple dip-coating/drop-
coating technique to advanced techniques such as SAM or LB
method. In LB technique, one or more monolayers of organic mate-
rial can be deposited on a solid surface (substrate is hydrophilic)
sequentially. The monolayers are usually composed of polar
molecules with both hydrophobic and hydrophilic end [124]. In
SAM technique, monolayers of organic molecules can be bound to
the substrate by spontaneous chemisorption of organic molecules
into the substrate [125]. The LB technique offer advantages such
as good reproducibility and speed, controlled amount of biomate-
rials, homogenous deposition, can produce multilayer structures
with varying layer composition, and process also displays a good
preservation of activities and recognition properties of the bioma-
terials. Meanwhile, the advantages of SAM are that they are simple,
stable, adaptable systems offering a good control of surface ori-
entation of biomolecules [15]. It is important to characterize the
deposition process by techniques such as AFM or SEM.
Another important aspect during the biomaterial deposition
process is the selection of suitable substrates. Generally, ceramic
and glass based substrates are used; however, other organic as
well as inorganic substrates may also be appropriate. Lakard
et al. [126] investigated the suitable biocompatible substrates such
as: polyethyleneimine, polypropyleneimine, and polypyrrole. The
adhesion and proliferation rates of rat neuronal cell cultures to
these substrates were studied. Immunohistochemical analysis and
confocal microscopy was used to characterize the morphology of
the adhesion. The authors concluded that polyethyleneimine and
polypropyleneimine were best suitable for culturing the olfactory
cells.
Optical methods are commonly being used for sensing odorants.
The QCM [4,105,106,108] and SPR [12,16,109] are often used for
quantitative analysis of odorant molecules. In QCM based sensing,
the odorant molecules are adsorbed onto the surface that results
in the alteration of resonance frequency of the quartz crystal. QCM
offers advantages such as simplicity, low cost, high sensitivity and
convenience [16]. In SPR technique, the contrast between states
before and after the surface reaction is determined. Usually, the
changes in refractive index caused by alternation are measured
[27].
3.3. Pattern recognition in olfactory sensing
Data obtained from olfactory sensor systems needs appro-
priate pattern recognition techniques for classification/prediction
of odorants. Pattern recognition component of e-nose systems
involves (i) sensor signal preprocessing, (ii) feature extraction,
and (iii) development of a suitable classification/prediction mod-
els. These pattern recognition techniques can also be applied for
new generation biomimetic olfactory sensors systems. Some of the
methods are described below.
A variety of features representing sensor signals could be
extracted from a typical olfactory sensor signal such as (i) ini-
tial and steady-state response, (ii) dynamic slope of the response
curve, (iii) area under the response curve (AUC), (iv) fast Fourier
transform (FFT) coefficients, and (v) wavelet transform coefficients
[9,60,63,64].
Researchers have also used the PCA technique to classify the
olfactory sensor sensitivity data. PCA reduces the correlation
(redundancy) within the sensor sensitivities for each class and
orthogonally projects the sensitivity data on a few uncorrelated
dimensions [107] to obtain the maximum variation components
that can be used to classify the groups or odorants. For example,
Si et al. [127] performed PCA on sensitivity data of the QCM sen-
sors to classify eight different VOCs. It was found that two principal
components (PCs) were able to explain about 97% of the variation
within the sensitivity data. The eight VOCs sensitivity data pro-
jected on principal planes confirmed distinguishable groups of the
eight VOCs. Lu et al. [122] performed PCA on QCM sensors sensi-
tivities for 15 VOCs to observe the discrimination of VOCs. Two PCs
accounted for 95% variation in sensitivity data and the projection
of the two PCs demonstrated the clear discrimination of VOCs.
The pattern recognition algorithms need to be developed for
discriminating the extracted features from the e-nose systems for
VOCs and odor recognition. Linear discriminant analysis (LDA) and
quadratic discriminant analysis (QDA) based statistical, and arti-
ficial neural network (ANN) based nonlinear pattern recognition
models have been applied previously for food contamination clas-
sification [8,128–133]. ANN is a biologically inspired classification
technique used to classify and predict complex non-linear systems.
Back propogation neural network (BPNN), radial basis function
network (RBFN), probabilistic neural network (PNN), and support
vector machine (SVM) are commonly used for VOCs classification
[134–136]. The simplest ANN topology involves input feature set
(input layer), decision making (hidden) layer, and decision (out-
put) layer. The input layer connects the input data patterns to the
node of the hidden layer. The hidden layer nodes use supervised,
unsupervised (or both) learning on input data. Generally, the num-
bers of hidden layer nodes depends on the complexity of the input
data [137]. The supervised learning of ANNs involves estimation of
the optimal weights connecting hidden layer neurons and the out-
put layer [128]. In ANN training, the networks can be trained using
rules such as step, momentum, quick propagation, delta bar delta,
conjugate gradient, and Levenberg-Marquardt (LM). The network
training based on back propagation rules involves gradient infor-
mation based weight change [138]. The LM method uses the second
order of the gradient allowing networks to converge faster and does
not depend on the initial weights for network convergence [138].
Therefore, LM method is more stable, efficient and fast in learning
compared to back propagation based learning [139].
 S. Sankaran et al. / Sensors and Actuators B 171–172 (2012) 1–17
DAQ Board
Fig. 4. Comparison of biological olfactory sensing to the artificial olfactory sensing mechanism.
The major concern with ANN is that it needs relatively large
training sets to develop reliable classification and prediction mod-
els [140–142]. For small datasets, BPNN and RBFN are preferred
over other networks. Specifically, RBFNs are preferred over BPNNs
and other network topologies as the RBFN networks train faster,
have better approximation capability and simple network struc-
ture [136]. Moreover, RBFNs do not get trapped into local minima
like BPNN [143]. RBFNs use radial basis functions in the hidden layer
as neurons. Radial basis functions are local in the network, mean-
ing the change in the parameters of one of the neurons does not
affect the inputs that are not associated with that neuron [134].
RBFN also provides better evolution of decision boundaries; the
much needed functionality for the classification problem. In con-
trast to multilayer perceptron neural network, RBFNs tend to give
superior performance on uneven training samples from each class
[144]. Additional information on various pattern recognition tech-
niques used in e-noses can be found in Pearce [145], Distante et al.
[146], and Gutierrez-Osuna [147].
4. Comparison of natural and artificial (e-noses) olfaction
The role of natural and artificial olfaction in real-world is unique
and different. While the development of natural olfaction is for pro-
viding animals cues about their environment (such as food safety,
defense, and mating); the artificial olfaction was developed to pro-
vide information about flavor (e.g. food industry) and aroma (e.g.
perfumes). The application domain of artificial olfaction is being
further extended to detection of pollutants, hazardous chemicals,
etc. Natural and artificial olfaction is analogous to each other with
different unique components within each module performing spe-
cific functionalities (Fig. 4). The major components of olfactory
system are odor delivery system, array of sensors sensitive to range
of VOCs, signal processing, pattern recognition and odor represen-
tation schemes [145].
In natural olfaction, the nasal cavity acts as an odorant delivery
systems; while in e-nose, this function is performed by compo-
nents such as vapor inlet tubing, pumps, and related units. The
major difference between the two systems is that natural olfaction
occurs under high humidity conditions and near room temperature
of nasal cavity; whereas the sensitivity of the artificial sensors to
the humidity and requirement of the high operating temperature
are some of the major challenges. Moreover, physical separation of
odorant molecules in natural olfaction is facilitated by molecules
in the nasal mucosa such as OBPs. If such segregation of odor-
ous compounds could be integrated with e-nose systems (e.g. gas
13
Sensors
Olfactory receptor/
Sensing element
Signal Transduction
Olfactory bulb/
Transducer
Signal Processing
Cerebral hemisphere/
Pattern recognition
Odor Detection
chromatography, other preconcentration units), we can further
improve the capabilities of the artificial olfactory systems [145].
Olfactory receptors serve as the sensing materials in the natu-
ral olfactory system. An array of materials, mostly semiconductors
such as metal oxides and conducting polymers are used in e-nose
systems. The olfactory receptors are known for their very high sen-
sitivity as well as selectivity, capable of detecting up to parts per
trillion, and even some stereoisomeric compounds [3]. In natural
system, the signal to noise ratio can be up to 30 times and more
at the first level of signal processing. Pearce [145] terms the high
sensitivity of the receptors as ‘hyperacuity’. Sensor array in the e-
noses displays a partially specific response (broad range selectivity)
to a range of compounds. Due to this, it is very difficult to pin-
point the specific VOC generating the sensor response. There is also
possibility of acquiring a sensor response to non-odorous chem-
ical compounds, which may be either considered as strength or
limitation of the system depending on its application (e.g. carbon-
di-oxide detection). As a result, selection of the sensors and use of
suitable pattern recognition technique becomes critical in artificial
olfaction. In natural olfaction, potentiometric transduction occurs
when the olfactory signal is activated and transferred through the
change in membrane potential. In e-nose systems, although con-
ductimetric approach is common, it is possible to select other
mechanisms such as piezoelectric and optical transduction. The
advantage of optical transduction is its high sensitivity resulting
from zero background rather than requiring a stable baseline signal.
However, the alignment of optical components can be challenging.
The signal processing in biological olfactory systems occurs
in the glomeruli and brain, and through combinatorial approach,
many different odors can be identified. The biological olfactory sys-
tem is capable to relating emotions to the odors; that cannot be
harnessed in an e-nose system. The signal processing in e-nose
system is performed through application of pattern recognition
techniques and machine learning algorithms. Final odorant signal
representation in natural olfaction occurs in brain memory.
The greatest benefit of artificial olfaction is its ability to quan-
tify and provide non-biased response against human perception
and experience in odor detection. If we can mimic and integrate
the sensitivity and selectivity of natural olfaction components such
as olfactory receptors and odorant binding proteins, with that
of mechanics and control of the e-nose systems, we can further
develop a reliable, robust, accurate e-nose system. The olfactory
receptors, odorant binding proteins could replace the semiconduc-
tor sensing materials in the e-nose systems, and the sensitivity of
these olfactory materials can be utilized to improve the efficiency
of the e-nose systems.
14 S. Sankaran et al. / Sensors and Actuators B 171–172 (2012) 1–17
Table 6
Comparison of sensing materials in natural and artificial olfactory systems.
Properties Sensors from biological olfactory system (e.g. olfactory
receptors, odorant binding proteins)
Detection limit Low ppm-ppt
Sensitivity Very high sensitivity
Selectivity Very high selectivity
Operating temperature Room temperature
Sensitivity to humidity Low
Response time Rapid (in milliseconds)
Safety Biodegradable and safe to use
Shelf life Low (for use and throw)
Table 6 provides a brief comparison of biomimetic sensing mate-
rials with those of inorganic materials used in olfactory systems.
The natural sensing materials such as olfactory receptors and odor-
ant binding proteins offer several benefits such as hyperacuity and
low limit of detection similar to the natural (biological) olfactory
system. Moreover, the biosafety of biological sensing materials is
high with easy biodegradability, in comparison to potentially harm-
ful and unknown effects of inorganic sensing materials such as
nanostructured metal oxides and other materials based on heavy
metals. One of the major limitations in the application of natural
sensing materials is their shelf-life. However, single-use strip or
sensor modules could be fabricated to address this issue.
5. Summary and conclusions
This paper reports the biology and mechanism of the olfactory
system and various new techniques to engineer olfactory sensors.
The olfactory mechanism is the least understood sensing mecha-
nism in mammalian systems. Recent findings and discoveries in
comprehending the mechanism of the human olfactory system,
along with the advancements in technology have increased the
potential for developing new generation olfactory sensors that
mimic the biology of mammalian olfactory system. The intelligent
olfactory sensors can be used to detect low concentrations of VOCs
for assessing food quality and safety, environmental quality, and
plant and animal diseases.
Although the biomimetic olfactory sensors have shown promis-
ing results, number of challenges need to addressed for the
development of intelligent sensor systems. Studies on the possible
substrates and other surface modification need to be conducted to
determine the ideal substrate for developing biomimetic olfactory
sensors. In general, PCA and ANNs has been used for determining
the discriminability of sensor sensitivity to different VOCs. One of
the major challenges in the development of any biosensors is the
shelf-life. As the biological materials are biodegradable, the sensor
life may be restricted upon fabrication. This also limits the repro-
ducibility of the sensor. One of the ways to address this issue could
be to eliminate the presence of moisture from the sensor system
and use a low cost, portable and one-use sensing module. Another
approach to address this issue could be combining the olfactory
sensing materials with other non-biological sensing materials such
as nanowires [148], carbon nanotube and metal oxides.
Acknowledgements
We extend our special thanks to U.S. Department of Agricul-
ture (USDA) – Cooperative State Research, Education, and Extension
Service (CSREES) for their funding and support. This work was a
component of review and Ph.D. research conducted at North Dakota
State University, Fargo, ND.
Sensors in artificial olfactory system (e.g. metal oxides, conducting
polymers)
In ppm
Moderate sensitivity
Partial selectivity
Moderate to high operating temperatures
High
Moderate (in seconds to minutes)
May be hazardous and inert
Moderate
References
[1] H. Breer, Olfactory receptors: molecular basis for recognition and discrimina-
tion of odors, Anal. Bioanal. Chem. 377 (2003) 427–433.
[2] G. Gomila, I. Casuso, A. Errachid, O. Ruiz, E. Pajot, J. Minic, T. Gorojankina,
M.A. Persuy, J. Aioun, R. Salesse, J. Bausells, G. Villanueva, G. Rius, Y. Hou, N.
Jaffrezic, C. Pennetta, E. Alfinito, V. Akimov, L. Reggiani, G. Ferrari, L. Fumagalli,
M. Sampietro, J. Samitier, Advances in the production, immobilization, and
electrical characterization of olfactory receptors for olfactory nanobiosensor
development, Sens. Actuat. B 116 (2006) 66–71.
[3] H. Breer, Sense of smell: Signal recognition and transduction in olfactory
receptor neurons, in: E. Kress-Rogers (Ed.), Handbook of Biosensors and Elec-
tronic Noses: Medicine, Food and the Environment, CRC Press, Massachusetts,
1997.
[4] Y.J. Lin, H.R. Guo, Y.H. Chang, M.T. Kao, H.H. Wang, R.I. Hong, Applica-
tion of the electronic nose for uremia diagnosis, Sens. Actuat. B 76 (2001)
177–180.
[5] D. Pickel, G.P. Manucy, D.B. Walker, S.B. Hall, J.C. Walker, Evidence for canine
olfactory detection of melanoma, Appl. Anim. Behav. Sci. 89 (2004) 107–116.
[6] C.M. Willis, S.M. Church, C.M. Guest, W.A. Cook, N. McCarthy, A.J. Bransbury,
A.R.T. Church, J.C.T. Church, Olfactory detection of human bladder cancer by
dogs: proof of principle study, Br. Med. J. 329 (2004) 712–715.
[7] E.H. Oh, H.S. Song, T.H. Park, Recent advances in electronic and bioelectronic
noses and their biomedical applications, Enzyme Microb. Tech. 48 (2011)
427–437.
[8] S. Panigrahi, S. Balasubramanian, H. Gu, C. Logue, M. Marchello. Neural-
network-integrated electronic nose system for identification of spoiled beef
LWT 39 (2006) 135–145.
[9] S. Panigrahi, S. Balasubramanian, H. Gu, C. Logue, M. Marchello, Design and
development of a metal oxide based electronic nose for spoilage classification
of beef, Sens. Actuat. B 119 (2006) 2–14.
[10] S. Sankaran, S. Panigrahi, S. Mallik, Odorant binding protein based biomimetic
sensors for detection of alcohols associated with Salmonella contamination
in packaged beef, Biosens. Bioelectron. 26 (2011) 3103–3109.
[11] S. Sankaran, S. Panigrahi, S. Mallik, Olfactory receptor based piezoelectric
biosensors for detection of alcohols related to food safety applications, Sens.
Actuat. B 155 (2011) 8–18.
[12] S. D’Auria, V. Scognamiglio, M. Rossi, M. Staiano, S. Campopoani, N. Cennamo,
L. Zeni. Odor binding as probe for a refractive index-based biosensor: New per-
spectives in biohazard assessment, In: A. Mahadevan-Jansen (Ed.). Conference
on Biohazard Detection Techno (Eds.), Biomedical Vibrational Spectroscopy
and Biohazard Detection Technologies. Society of Photo-Optical Instrumen-
tation Engineering, California (2004).
[13] K.S. Mead, Using lobster noses to inspire robot sensor design, Trends Biotech-
nol. 20 (2002) 276–277.
[14] Q. Liu, H. Cai, Y. Xu, Y. Li, R. Li, P. Wang, Olfactory cell-based biosensor: A first
step towards neurochip of bioelectronic nose, Biosens. Bioelectron. 22 (2006)
318–322.
[15] Y. Hou, N. Jaffrezic-Renault, C. Martelet, C. Tlili, A. Zhang, J.C. Pernollet, L.
Briand, G. Gomila, A. Errachid, J. Samitier, L. Salvagnac, B. Torbiéro, P. Temple-
Boyer, Study of Langmuir and Langmuir-Blodgett films of odorant-binding
protein/amphiphile for odorant biosensors, Langmuir 21 (2005) 4058–4065.
[16] H.J. Ko, T.H. Park, Piezoelectric olfactory biosensor: Ligand specificity and
dose-dependence of an olfactory receptor expressed in a heterologous cell
system, Biosens. Bioelectron. 20 (2005) 1327–1332.
[17] M. Huotari, V. Lantto, Measurements of odours based on response analysis of
insect olfactory receptor neurons, Sens. Actuat. B 127 (2007) 284–287.
[18] M. Huotari, M. Mela, Blowfly olfactory biosensor’s sensitivity and specificity,
Sens. Actuat. B 34 (1996) 240–244.
[19] S. Schv̈tz, M.J. Schŏning, P. Schroth, Ü. Malkoc, B. Weißbecker, P. Kordos,
H. Lv̈th, H.E. Hummel, An insect-based BioFET as a bioelectronic nose, Sens.
Actuat. B 65 (2000) 291–295.
[20] R.L. Doty, Olfaction, Annu. Rev. Psychol. 52 (2001) 423–452.
[21] S. Firestein, How the olfactory system makes sense of scents, Nature 413
(2001) 211–218.
[22] J. Krieger, H. Breer, Olfactory reception in invertebrates, Science 286 (1999)
720–723.
[23] U. Stockhorst, R. Pietrowsky, Olfactory perception, communication, and the
nose-to-brain pathway, Physiol. Behav. 83 (2004) 3–11.
 S. Sankaran et al. / Sensors and Actuators B 171–172 (2012) 1–17
[24] Ch. Ziegler, W. Göpel, H. Hämmerle, H. Hatt, G. Jung, L. Laxhuber, H.L. Schmidt,
S. Schütz, F. Vögtle, A. Zell, Bioelectronic noses: a status report Part I, Biosens.
Bioelectron. 13 (1998) 539–571.
[25] L. Su, W. Jia, C. Hou, Y. Lei, Microbial biosensors: A review, Biosens. Bioelectron.
26 (2011) 1788–1799.
[26] K. Länge, B.E. Rapp, M. Rapp, Surface acoustic wave biosensors: a review, Anal.
Bioanal. Chem. 391 (2008) 1509–1519.
[27] O. Lazcka, F.J.D. Campo, F.X. Muňoz, Pathogen detection: A perspective of tra-
ditional methods and biosensors, Biosens. Bioelectron. 22 (2007) 1205–1217.
[28] W. Göpel, C. Ziegler, H. Breer, D. Schild, R. Apfelbach, J. Joerges, R. Malaka,
Bioelectronic noses: A status report Part I, Biosens. Bioelectron. 13 (1998)
479–493.
[29] R. Glatz, K. Bailey-Hill, Mimicking nature’s noses: from receptor deorphaning
to olfactory biosensing, Prog. Neurobiol. 93 (2011) 270–296.
[30] R.P. Arasaradnam, N. Quraishi, I. Kyrou, C.U. Nwokolo, M. Joseph, S. Kumar,
K.D. Bardhan, J.A. Covington, Insights into ‘fermentonomics’: evaluation of
volatile organic compounds (VOCs) in human disease using an electronic ‘e-
nose’, J. Med. Eng. Technol. 35 (2011) 87–91.
[31] F. Röck, N. Barsan, U. Weimar, Electronic nose: current status and future
trends, Chem. Rev. 108 (2008) 705–725.
[32] E. Schaller, J.O. Bosset, F. Escher, Electronic noses and their application to food,
Lebensm.-Wiss. U. Technol. 31 (1998) 305–316.
[33] D.T. Win, The electronic nose: A big part of our future, AU J. Technol. 9 (2005)
1–6.
[34] H. Gray, Anatomy of the human body. 20th ed., in: Warren H. Lewis (Ed.). Lea
& Febiger, Philadelphia (1918).
[35] R. O’Rahilly, F. Müller, S., Carpenter, R., Swenson, A. Basic Human Anatomy,
Darmouth Medical School, Available at: http://www.dartmouth.edu/
∼humananatomy/index.html (2004).
[36] D. Lancet, Vertebrate olfactory reception, Ann. Rev. Neurosci. 9 (1986)
329–355.
[37] H.K. Walker. Cranial Nerve I: The olfactory nerve, In Clinical Methods: The
History, Physical, and Laboratory Examinations, 3rd ed. In: H.K. Walker, W.D.
Hall, J.W. Hurst (Eds.), Butterworths, Boston (1990).
[38] B. Buck, Information coding in the vertebrate olfactory system, Annual Rev.
Neurosci. 19 (1996) 517–544.
[39] L. Buck, R. Axel, A novel multigene family may encode odorant receptors: A
molecular basis for odor recognition, Cell 65 (1991) 175–187.
[40] R. Axel, L.B. Buck, Available at http://nobelprize.org/nobel prizes/medicine/
laureates/2004/press.html. Accessed on 17 July 2006 (2004).
[41] M.F. Bear, B.W. Connors, M.A. Paradiso, Neuroscience Exploring the Brain,
Willimas and Wilkins, Baltimore, 1996.
[42] S.D. Liberles, L.B. Buck, A second class of chemosensory receptors in the olfac-
tory epithelium, Nature 442 (2006) 645–650.
[43] K. Mori, H. Nagao, Y. Yoshihara, The Olfactory bulb: Coding and processing of
odor molecule information, Science 286 (1999) 711–715.
[44] I. Savic, Imaging of brain activation by odorants in humans, Curr. Opin. Neu-
robiol. 12 (2002) 455–461.
[45] P. Whitfield, D.M. Stoddard, Hearing, Taste, and Smell: Pathways of Perception
(Human body), Scribner Books, Inc, New York, 1984.
[46] M.S. Kim, D.P. Smith, The invertebrate odorant-binding protein LUSH is
required for normal olfactory behavior in Drosophila, Chem. Sens. 26 (2001)
195–199.
[47] P. Pelosi, Odorant-binding proteins: structural aspects, Ann. N.Y. Acad. Sci.
855 (1998) 281–293.
[48] M.A. Bianchet, G. Bains, P. Pelosi, J. Pevsner, S.H. Snyder, H.L. Monaco, L.M.
Amzel, The three-dimensional structure of bovine odorant binding pro-
tein and its mechanism of odor recognition, Nat. Struct. Biol. 3 (1996)
934–939.
[49] S.W. Kruse, R. Zhao, D.P. Smith, D.N.M. Jones, Structure of a specific alcohol-
binding site defined by the odorant binding protein LUSH from Drosophila
melanogaster, Nature Struct. Biol. 10 (2003) 694–700.
[50] L. Turin, F. Yoshii, Structure-odor relations: A modern perspective, in: R. Doty
(Ed.), Handbook of Olfaction and Gustation, Marcel Dekker, Inc, New York,
2002.
[51] B. Malnic, J. Hirono, T. Sato, L.B. Buck, Combinatorial receptor codes for odors,
Cell 96 (1999) 713–723.
[52] J. Krieger, S. Schleicher, J. Strotmann, I. Wanner, I. Boekhoff, K. Raming, P. De
Geus, H. Breer, Probing olfactory receptors with sequence-specific antibodies,
Eur. J. Biochem. 219 (1994) 829–835.
[53] Z. Zou, L.B. Buck, Combinatorial effects of odorant mixes in olfactory cortex,
Science 311 (2006) 1477–1481.
[54] R. Izumi, K. Hayashi, K. Toko, Odor sensor with water membrane using surface
polarity controlling method and analysis of responses to partial structures of
odor molecules, Sens. Actuat. B 99 (2004) 315–322.
[55] K. Toko, Biomimetic Sensor Technology, Cambridge University Press, Madrid,
Spain, 2000.
[56] E. Kress-Rogers, Handbook of Biosensors and Electronic Noses: Medicine,
Food, and the Environment, 1st ed., CRC, Press, Inc, Massachusetts,
1997.
[57] W.J. Hurst, Electronic Noses and Sensor Array Based Systems: Design and
Applications, Technomic Publishing Co., Inc, Lancaster, 1999.
[58] J.W. Gardner, J. Yinon, Electronic Noses and Sensors for the Detection of Explo-
sives, Kluwer Academic Publishers, Norwell, 2004.
[59] E.B. Goldstein, Sensation and Perception, Thomson Wadsworth, Belmont,
2002.
15
[60] I. Concina, M. Falasconi, E. Gobbi, F. Bianchi, M. Musci, M. Mattarozzi, M.
Pardo, A. Mangia, M. Careri, G. Sberveglieri, Early detection of microbial con-
tamination in processed tomatoes by electronic nose, Food Contr. 20 (2009)
873–880.
[61] T. Rajamäki, H.L. Alakomi, T. Ritvanen, E. Skyttä, M. Smolander, R. Ahve-
nainen, Application of an electronic nose for quality assessment of modified
atmosphere packaged poultry meat, Food Contr. 17 (2006) 5–13.
[62] M.A. Drake, P.D. Gerard, J.P. Kleinhenz, W.J. Harper, Application of an elec-
tronic nose to correlate with descriptive sensory analysis of Cheddar cheese,
LWT Food Sci. Technol. 36 (2003) 13–20.
[63] R.C. Young, W.J. Buttner, B.R. Linnell, R. Ramesham, Electronic nose for space
program applications, Sens. Actuat. B 93 (2003) 7–16.
[64] N.E. Barbri, E. Llobet, N. El Bari, X. Correig, B. Bouchikhi, Application of a
portable electronic nose system to assess the freshness of Moroccan sardines,
Mater. Sci. Eng. C 28 (2008) 666–670.
[65] K. Arshak, E. Moore, G.M. Lyons, J. Harris, S. Clifford, A review of gas sensors
employed in electronic nose applications, Sensor Rev. 24 (2004) 181–198.
[66] H. Bai, G. Shi, Gas sensors based on conducting polymers, Sensors 7 (2007)
267–307.
[67] D.L. García-González, R. Aparicio, Coupling MOS sensors and gas chromatog-
raphy to interpret the sensor responses to complex food aroma: Application
to virgin olive oil, Food Chem. 120 (2010) 572–579.
[68] W. Li, J. Friel, T. Beta, An evaluation of the antioxidant properties and aroma
quality of infant cereals, Food Chem. 121 (2010) 1095–1102.
[69] V.Yu. Musatov, V.V. Sysoev, M. Sommer, I. Kiselev, Assessment of meat
freshness with metal oxide sensor microarray electronic nose: A practical
approach, Sens. Actuat. B 144 (2010), 99-10.
[70] J.E. Haugen, K. Rudi, S. Langsrud, S. Bredholt, Application of gas-sensor array
technology for detection and monitoring of growth of spoilage bacteria in
milk: A model study, Anal. Chim. Acta 565 (2006) 10–16.
[71] M. Falasconi, M. Pardo, G. Sberveglieri, I. Riccò, A. Bresciani, The novel EOS835
electronic nose and data analysis for evaluating coffee ripening, Sens. Actuat.
B 110 (2005) 73–80.
[72] M. Mascini, A. Macagnano, G. Scortichini, M. Del Carlo, G. Diletti, A. D’Amico,
C. Di Natale, D. Compagnone, Biomimetic sensors for dioxins detection in food
samples, Sens. Actuat. B 111-112 (2005) 376–384.
[73] F. Liao, S. Yin, M.F. Toney, V. Subramanian, Physical discrimination of amine
vapor mixtures using polythiophene gas sensor arrays, Sens. Actuat. B 150
(2010) 254–263.
[74] S.D. Vito, M. Piga, L. Martinotto, G.D. Francia, CO, NO2 and;1; NOx urban
pollution monitoring with on-field calibrated electronic nose by automatic
bayesian regularization, Sens. Actuat. B 143 (2009) 182–191.
[75] L. Capelli, S. Sironi, P. Céntola, R.D. Rosso, M. Grande Il, Electronic noses for
the continuous monitoring of odours from a wastewater treatment plant at
specific receptors: Focus on training methods, Sens. Actuat. B 131 (2008)
53–62.
[76] L. Capelli, S. Sironi, R.D. Rosso, P. Céntola, M. Grande, A comparative and criti-
cal evaluation of odour assessment methods on a landfill site, Atmos. Environ.
42 (2008) 7050–7058.
[77] A. Lamagna, S. Reich, D. Rodríguez, A. Boselli, D. Cicerone, The use of an
electronic nose to characterize emissions from a highly polluted river, Sens.
Actuat. B 131 (2008) 121–124.
[78] S. De Vito, E. Massera, M. Piga, L. Martinotto, G. Di Francia, On field calibration
of an electronic nose for benzene estimation in an urban pollution monitoring
scenario, Sens. Actuat. B 129 (2008) 750–757.
[79] O. Helli, M. Siadat, M. Lumbreras, Qualitative and quantitative identification
of H2 S/NO2 gaseous components in different reference atmospheres using a
metal oxide sensor array, Sens. Actuat. B 103 (2004) 403–408.
[80] M.A. Markom, A.Y. Md Shakaff, A.H. Adom, M.N. Ahmad, Wahyu Hidayat, A.H.
Abdullah, N. Ahmad Fikri, Intelligent electronic nose system for basal stem
rot disease detection, Comput. Electron. Agr. 66 (2009) 140–146.
[81] C. Li, G.W. Krewer, P. Ji, H. Scherm, S.J. Kays, Gas sensor array for blueberry
fruit disease detection and classification, Postharvest Biol. Technol. 55 (2010)
144–149.
[82] M. Falasconi, E. Gobbi, M. Pardo, M.D. Torre, A. Bresciani, G. Sberveglieri,
Detection of toxigenic strains of Fusarium verticillioides in corn by electronic
olfactory system, Sens. Actuat. B 108 (2005) 250–257.
[83] A. D’Amico, G. Pennazza, M. Santonico, E. Martinelli, C. Roscioni, G. Galluccio,
R. Paolesse, C.D. Natale, An investigation on electronic nose diagnosis of lung
cancer, Lung Cancer 68 (2010) 170–176.
[84] Silvano Dragonieri, Jouke T. Annema, Robert Schot, Marc P.C. van der Schee,
Antonio Spanevello, Pierluigi Carratú, Onofrio Resta, Klaus F. Rabe, Peter J.
Sterk, An electronic nose in the discrimination of patients with non-small
cell lung cancer and COPD, Lung Cancer 64 (2009) 166–170.
[85] S. Dragonieri, R. Schot, B.J.A. Mertens, S.L. Cessie, S.A. Gauw, A. Spanevello, O.
Resta, N.P. Willard, T.J. Vink, K.F. Rabe, E.H. Bel, P.J. Sterk, An electronic nose
in the discrimination of patients with asthma and controls, J. Allergy Clin.
Immunol. 120 (2007) 856–862.
[86] A.D. Wilson, M. Baietto, Applications and advances in electronic-nose tech-
nology, Sensory 9 (2009) 5099–5148.
[87] M.J. Swann, A. Glidle, L. Cui, J.R. Barker, J.M. Cooper, The determination of
gaseous molecular density using a hybrid vapour sensor, Chem. Commun. 24
(1998) 2753–2754.
[88] H. Ulmer, J. Mitrovics, G. Noetzel, U. Weimar, W. Gopel, Odours and flavours
identified with hybrid modular sensor systems, Sens. Actuat. B 43 (1997)
24–33.
16 S. Sankaran et al. / Sensors and Actuators B 171–172 (2012) 1–17
[89] L. Cui, M.J. Swann, A. Glidle, J.R. Barker, J.M. Cooper, Odour mapping using
microresistor and piezo-electric sensor pairs, Sens. Actuat. B 66 (2000) 94–97.
[90] K. Parikh, K. Cattanach, R. Rao, D.S. Suh, A. Wu, S.K. Manohar, Flexible vapour
sensors using single walled carbon nanotubes, Sens. Actuat. B 113 (2006)
55–63.
[91] N. Stevens, D.L. Akins, Dye-doped inorganic/organic composite films as fluo-
rescence sensors for methanol vapor, Sens. Actuat. B 123 (2007) 59–64.
[92] M. Castro, J.B. Lu, S. Bruzaud, B. Kumar, J.F. Feller, Carbon
nanotubes/poly(epsilon-caprolactone) composite vapour sensors, Carbon 47
(2009) 1930–1942.
[93] S.S. Barkade, J.B. Naik, S.H. Sonawane, Ultrasound assisted miniemulsion syn-
thesis of polyaniline/Ag nanocomposite and its application for ethanol vapor
sensing, Colloids Surf. A 378 (2011) 94–98.
[94] J. Im, S.K. Sengupta, M.F. Baruch, C.D. Granz, S. Ammu, S.K. Manohar, J.E. Whit-
ten, A hybrid chemiresistive sensor system for the detection of organic vapors,
Sens. Actuat. B 156 (2011) 715–722.
[95] D. James, S.M. Scott, Z. Ali, W.T. O’Hare, Review chemical sensors for electronic
nose systems, Microchim. Acta 149 (2005) 1–17.
[96] S.A. Piletsky, A.P.F. Turner, Optical Biosensors: Present and Future. In: F.S.
Ligler, C.A. Rowe Taitt (Eds.), 2002, pp. 397–425.
[97] G. Bunte, J. Hürttlen, H. Pontius, K. Hartlieb, H. Krause, Gas phase detection of
explosives such as 2,4,6-trinitrotoluene by molecularly imprinted polymers,
Anal. Chim. Acta 591 (2007) 49–56.
[98] F.L. Dickert, O. Hayden, K.P. Halikias, Synthetic receptors as sensor coatings
for molecules and living cells, Analyst 126 (2001) 766–771.
[99] F.L. Dickert, O. Hayden, R. Bindeus, K.J. Mann, D. Blass, E. Waigmann, Bioim-
printed QCM sensors for virus detection–screening of plant sap, Anal. Bioanal.
Chem. 378 (2004) 1929–1934.
[100] M. Matsuguchi, T. Uno, Molecular imprinting strategy for solvent molecules
and its application for QCM-based VOC vapor sensing, Sens. Actuat. B 113
(2006) 94–99.
[101] N. Iqbal, G. Mustafa, A. Rehman, A. Biedermann, B. Najafi, P.A. Lieberzeit, F.L.
Dickert, QCM-arrays for sensing terpenes in fresh and dried herbs via bio-
mimetic MIP layers, Sensors 10 (2010) 6361–6376.
[102] K.D. Shimizu, C.J. Stephenson, Molecularly imprinted polymer sensor arrays,
Curr. Opin. Chem. Biol. 14 (2010) 743–750.
[103] A.C. Paske, L.D. Earl, J.L. O’Donnell, Interfacially polymerized metallopor-
phyrin thin films for colorimetric sensing of organic vapors, Sens. Actuat. B
155 (2011) 687–691.
[104] E. Chevallier, E. Scorsone, H.A. Girard, V. Pichot, D. Spitzer, P. Bergonzo,
Metalloporphyrin-functionalised diamond nano-particles as sensitive layer
for nitroaromatic vapours detection at room-temperature, Sens. Actuat. B 151
(2010) 191–197.
[105] T.Z. Wu, A piezoelectric biosensor as an olfactory receptor for odor detection:
electronic nose, Biosens. Bioelectron. 14 (1999) 9–18.
[106] J.H. Sung, H.J. Ko, T.H. Park, Piezoelectric biosensor using olfactory recep-
tor protein expressed in Escherichia coli, Biosens. Bioelectron. 21 (2006)
1981–1986.
[107] B. Li, S. Santhanam, L. Schultz, M. Jeffries-E.L., M.C. Iovu, G. Sauvé, J. Cooper,
R. Zhang, J.C. Revelli, A.G. Kusne, J.L. Snyder, T. Kowalewski, L.E. Weiss, R.D.
McCullough, G.K. Fedder, D.N. Lambeth, Inkjet printed chemical sensor array
based on polythiophene conductive polymers, Sens. Actuat. B 123 (2007)
651–660.
[108] T.Z. Wu, Y.R. Lo, E.C. Chan, Exploring the recognized bio-mimicry materials
for gas sensing, Biosens. Bioelectron. 16 (2001) 945–953.
[109] J.M. Vidic, J. Grosclaude, M.A. Persuy, J. Aioun, R. Salesse, E.P. Augy,
Quantitative assessment of olfactory receptors activity in immobilized
nanosomes: a novel concept for bioelectronic nose, Lab on a Chip 6 (2006)
1026–1032.
[110] A. Suska, A.B. Ibáñez, P. Preechaburana, I. Lundström, A. Berghard, G
protein-coupled receptor mediated sensing of TMA, Procedia Chem. 1 (2009)
321–324.
[111] J. Minic, M.A. Persuy, E. Godel, J. Aioun, I. Connerton, R. Salesse, E. Pajot-Augy,
Functional expression of olfactory receptors in yeast and development of a
bioassay for odorant screening, FEBS J. 272 (2005) 524–537.
[112] D. Krautwurst, K.W. Yau, R.R. Reed, Identification of ligands for olfactory
receptors by functional expression of a receptor library, Cell 95 (1998)
917–926.
[113] C.H. Wetzel, M., Oles, C., Wellerdieck, M., Kuczkowiak, G., Gisselmann, H.,
Hatt, Specificity and sensitivity of a human olfactory receptor functionally
expressed in human embryonic kidney 293 cells and Xenopus Laevis oocytes,
J. Neurosci. 19 (1999) 7426–7433.
[114] P. Wang, G. Xu, L. Qin, Y. Xu, Y. Li, R. Li, Cell-based biosensors and its application
in biomedicine, Sens. Actuat. B 108 (2007) 576–584.
[115] Q. Liu, W. Ye, L. Xiao, L. Du, N. Hu, P. Wang, Extracellular potentials recording in
intact olfactory epithelium by microelectrode array for a bioelectronic nose,
Biosens. Bioelectron. 25 (2010) 2212–2217.
[116] Q. Liu, W. Ye, H. Yu, N. Hu, L. Du, P. Wang, M. Yang, Olfactory mucosa tissue-
based biosensor: A bioelectronic nose with receptor cells in intact olfactory
epithelium, Sens. Actuat. B 146 (2010) 527–533.
[117] M. Huotari, Biosensing by insect olfactory receptor neurons, Sens. Actuat. 71
(2000) 212–222.
[118] T.Z. Wu, Y.R. Lo, Synthetic peptide mimicking of binding sites on olfactory
receptor protein for use in electronic nose, J. Biotech. 80 (2000) 63–73.
[119] M. Blanco, M.C. McAlpine, J.R. Heath, First principles molecular modeling
of sensing material selection for hybrid biomimetic nano-sensors, in: M.A.
Ryan, A.V. Shevade, C.J. Taylor, M.L. Homer, M. Blanco (Eds.), Computa-
tional Methods for Sensor Material Selection, Springer, New York, 2009,
pp. 135–148.
[120] C. Nakamura, J. Miyake, Combinatorially developed peptide receptors for
biosensors, in: R.A. Potyrailo, V.M. Mirsky (Eds.), Combinatorial Methods for
Chemical and Biological Sensors, Integrated Analytical Systems, Springer,
New York, 2009, pp. 201–221.
[121] S. Panigrahi, S. Sankaran, S. Mallik, B. Gaddam, A.A. Hanson, Olfactory
receptor-based polypeptide sensor for acetic acid VOC detection, Mat. Sci.
Engg. C, http://dx.doi.org/10.1016/j.msec.2011.11.003.
[122] H.-H. Lu, Y.K. Rao, T.-Z. Wu, Y.-M. Tzeng, Direct characterization and quantifi-
cation of volatile organic compounds by piezoelectric module chips sensor,
Sens. Actuat. B 137 (2009) 741–746.
[123] J.W. Jaworski, D. Raorane, J.H. Huh, A. Majumdar, S.W. Lee, Evolutionary
screening of biomimetic coatings for selective detection of explosives, Lang-
muir 24 (2008) 4938–4943.
[124] M.C. Petty, Langmuir-Blodgett Films: An Introduction, Cambridge University
Press, New York, 1996.
[125] T. Wink, S.J. Zuilen, A. Bult, W.P. Van Bennekom, Self-assembled monolayers
for biosensors, Analyst 122 (1997) 43R–50R.
[126] S. Lakard, G. Herlem, N. Valles-Villareal, G. Michel, A. Propper, T. Gharbi,
B. Fahys, Culture of neural cells on polymers coated surfaces for biosensor
applications, Biosens. Bioelectron. 20 (2005) 1946–1954.
[127] P.C. Si, J. Mortensen, A. Komolov, J. Denborg, P.J. Møller, Polymer coated quartz
crystal microbalance sensors for detection of volatile organic compounds in
gas mixtures, Anal. Chim. Acta 597 (2007) 223–230.
[128] S. Balasubramanian, S. Panigrahi, C.M. Logue, H. Gu, M. Marchello, Neural
networks-integrated metal oxide-based artificial olfactory system for meat
spoilage identification, J. Food Eng. 91 (2009) 91–98.
[129] S. Balasubramanian, S. Panigrahi, C.M. Logue, H. Gu, M. Marchello, J. Sherwood,
Identification of Salmonella-inoculated beef using a portable electronic nose
system, J. Rapid Methods Autom. Microbiol. 13 (2005) 71–95.
[130] K. Brudzewski, J. Ulaczyk, An effective method for analysis of dynamic elec-
tronic nose responses, Sens. Actuat. B 140 (2009) 43–50.
[131] U. Siripatrawan, J. Linz, B.R. Harte, Solid-phase microextraction, gas
chromatography, and mass spectrometry coupled with discriminant fac-
tor analysis and multilayer perceptron neural network for detection of
Escherichia coli, J. Food Protect. 67 (2004) 1597–1603.
[132] U. Siripatrawan, J.E. Linz, B.R. Harte, Electronic sensor array coupled with arti-
ficial neural network for detection of Salmonella typhimurium, Sens. Actuat. B
119 (2006) 64–69.
[133] S. Woznica, MS Dissertation, North Dakota State University, Department of
Agricultural and Biosystems Engineering. Fargo, ND (2006).
[134] F. Marini, Artificial neural networks in foodstuff analyses: Trends and per-
spectives A review, Anal. Chim. Acta 635 (2009) 121–131.
[135] E.Z. Panagou, N. Sahgal, N. Magan, G.-J.E. Nychas, Table olives volatile finger-
prints: Potential of an electronic nose for quality discrimination, Sens. Actuat.
B 134 (2008) 902–907.
[136] Nan Qu, Hong Mi, Bin Wang, Yulin Ren Application of GA-RBF networks
to the nondestructive determination of active component in pharmaceu-
tical powder by NIR spectroscopy, J. Taiwan Inst. Chem. Eng. 40 (2009)
162–167.
[137] B. Tudu, A. Jana, A. Metla, D. Ghosh, N. Bhattacharyya, R. Bandyopadhyay, Elec-
tronic nose for black tea quality evaluation by an incremental RBF network,
Sens. Actuat. B 138 (2009) 90–95.
[138] J.C. Principe, N.R. Euliano, W.C. Lefebvre, Neural and Adaptive Systems, John
Wiley and Sons., Inc, New York, 2000.
[139] Z. Yuhong, H. Wenxin, Application of artificial neural network to predict the
friction factor of open channel flow, Comm. Nonlinear Sci. Num. Simul. 14
(2009) 2373–2378.
[140] R.M.H. Zhu, L., Zhang, A. Chen, A new method to assist small data set neu-
ral network learning, Proc. 6th Intl. Conf. on Intelligent System Design and
Applications (ISDA ‘06), IEEE Computer Society, Washington, 2006.
[141] C.F. Huang, C. Moraga, A diffusion neural network for learning from small
samples, Int. J. Approx. Reason 35 (2004) 137–161.
[142] D.C. Li, C.S. Wu, T.I. Tsai, F.M. Chang, Using mega-fuzzification and data trend
estimation in small data set learning for early FMS scheduling knowledge,
Comput. Oper. Res. 33 (2006) 1857–1869.
[143] P. Evans, K.C. Persaud, A.S. McNeish, R.W. Sneath, N. Hobson, N. Magan,
Evaluation of a radial basis function neural network for the determina-
tion of wheat quality from electronic nose data, Sens. Actuat. B 69 (2000)
348–358.
[144] R. Dutta, D. Morgan, N. Baker, J.W. Gardner, E.L. Hines, Identification of Staphy-
lococcus aureus infections in hospital environment: electronic nose based
approach, Sens. Actuat. B 109 (2005) 355–362.
[145] T.C. Pearce, Computational parallels between the biological olfactory pathway
and its analogue ‘The Electronic nose’: Part I. Biological olfaction, BioSystems
41 (1997) 69–90.
[146] C. Distante, M. Leo, P. Siciliano, K.C. Persaud, On the study of feature
extraction methods for an electronic nose, Sens. Actuat. B 87 (2002) 274–
288.
[147] R. Gutierrez-Osuna, Pattern analysis for machine olfaction: a review, IEEE
Sens. J. 2 (2002) 189–202.
[148] M.C. McAlpine, H.D. Agnew, R.D. Rohde, M. Blanco, H. Ahmad, A.D. Stuparu,
W.A. Goddard III, J.R. Heath, Peptide-nanowire hybrid materials for selective
sensing of small molecules, J. Am. Chem. Soc. 130 (2008) 9583–9589.
 S. Sankaran et al. / Sensors and Actuators B 171–172 (2012) 1–17
Biographies
Sindhuja Sankaran received her M.S. in Environmental Engineering from Iowa
State University, Ames, IA and Ph.D. in Agricultural and Biosystems Engineering,
from Bio-imaging and Sensing Center, North Dakota State University (NDSU), Fargo,
ND, USA. Her Ph.D. research was focused towards the development and evalua-
tion of novel sensing materials for detecting contamination in food products. She is
currently working as a postdoctoral research associate at Citrus Research and Edu-
cation Center, University of Florida with major research emphasis on optical sensing
techniques for disease detection in citrus trees and other specialty crops.
Lav R. Khot received M.E. in Agricultural Systems and Engineering from Asian Insti-
tute of Technology, Thailand (2004). He also received a M.S. in Agricultural and
Biosystems Engineering from Iowa State University (2006). He joined Bio-imaging
and Sensing Center of North Dakota State University to pursue his Ph.D. focusing on
17
characterization and pattern recognition of selected sensors for food safety applica-
tions. He received Ph.D. from Agricultural and Biosystems Engineering department
of NDSU (2009). At present, he is working as a postdoctoral researcher at University
of Florida, Citrus Research and Education Center with major research emphasis on
spray and sensor technologies for agricultural production.
Suranjan Panigrahi received his Ph.D. from Iowa State University, Ames, IA, USA.
He served as a Professor in the Agricultural and Biosystems Engineering Depart-
ment, NDSU, Fargo, ND (until Fall 2009). He was also the director of Bio-imaging
and Sensing Center, North Dakota State University, Fargo, ND, USA. He is currently
a Professor in the Department of Electrical and Computer Engineering Technology,
Purdue University, West Lafayette, IN, USA and leads the “Integrated Smart Sensing
and Solutions Laboratory”. His research focuses on machine systems engineering,
and development/evaluation of intelligent sensors/sensing systems for different
biological applications.