WISE PROJECT: DEVELOPMENT OF A NEW TECHNOLOGY FOR INSTRUMENTAL

MEASUREMENT OF ODOROUS COMPOUNDS

Jean-Michel GUILLOT1, Thierry LIVACHE2, Tristan ROUSELLE2

1 Ecole des Mines d’Alès, Environmental Engineering Laboratory, 6 avenue de Clavières, 30319 Alès cedex, FRANCE, jean-
michel.guillot@mines-ales.fr;

2 Aryballe Technologies SA, CEA/MINATEC BHT – Bat 52, 7 Parvis Louis Néel, CS 20050, 38040 Grenoble Cedex 09,
FRANCE, t.livache@aryballe.com, tristan@aryballe.com;

Key words- E-nose, Odour, SPR, Wellmess, Detection, Diagnosis

Abstract

The project is linked to the development of a new type of instrument to measure odorous volatile compounds. This development
carried out by Aryballe Company, in based on a combination of technologies such as Nanotechnology, Biotechnology, Data
processing… The heart of the device is based on Surface Plasmon Resonance (SPR) imaging, technology generally used in liquid
samples.

Such analytical device must be very helpful for different potential applications as for example:
 For people suffering from anosmia (loss of smelling), the device is able to recognize 30 to 50 different day to day odours;
 For odour panels that can refer to value of the device as a reference to protect original sensorial recipes;
 For pollution monitoring and also disagreement assessment caused by malodorous compounds;
 For gustatory and safety quality of high added value food with intelligent kitchen equipment including cooking assistance;
 For some disease detection in the case this disease can be characterized by a low specific odour and when trained dogs are
generally used for this kind of diagnosis;
 …

The WISE Project (Wellness & medical dIagnostics olfactory SEnsors) is typically to improve this new measurement device and the
application fields. It concerns technical aspects such as the size reduction, the cost decrease and the simplicity to use. It’s also linked
to real applications to prove the efficiency of the instrument as an alternative to other measurement methods.

1. Introduction

Developments of measurement devices for odorous volatile compounds such as electronic noses are generally based on assembly of
non-specific sensors. Such an approach differs from lock/key-recognition-based approach in the bio-analytical field. Surface
Plasmon Resonance Imaging of biological interactions is commonly used in liquid phase and the development of such a technique in
gas phase opens applications on volatile organic compounds (VOCs) and odorous compounds.
On an array of bioinspired receptors, the binding of VOCs produces signals that can be seen as 3D continuous dynamic images or
movies. Finally, complex data obtained from interactions between compounds and receptors are analyzed as simple images via a
specific database. By developments on miniaturization and down-scaling of all the components, a hand-portable analyzer is now
proposed and called Neose. The interactions of odorous compounds with receptors are converted into an image that gives a visual
representation of an odorous atmosphere.

2. Description

A lot of sensors and sensor arrays as e-noses have been developed during las decades. The objective is always to try to measure
odours (and mainly odorous atmospheres) as better as possible. Even if some instruments are efficient, it’s extremely difficult to
obtain a direct sensitivity like human nose. For qualitative aspect, strong algorithms for data treatment can also perform
recognition/distinction of odours after system training, but the human brain is also a wonderful treatment process to analyze signals
sent by nasal receptors. Developments are carried out for various applications such as odorous chemicals 1, food industries2, medicinal
plants3, environment4…
Progress in research fields such as microtechnology, biotechnology, fine chemistry are fast and especially Important steps have been
taken during last decades. With miniaturization developments, the possibility to graft biomolecules on metals has been carried out.
This kind of engraving opened the development of SPRimaging (Surface Plasmon Resonance) allowing the real time and label free
measurement of biological interactions on a microarray format 5.
With this technique, lots of biological applications exist in different fields from fundamental biological researches 6 to more applied
fields. As an example, the detection of a very low number of bacteria in complex samples 7 is now allowed with real time monitoring
of bacteria growth.
As shown on Figure 1, the key element of the gas-imaging device is a SPR prism. This prism is chemically functionalized with a
specific set of several tenths of biosensors which can react with a wide range of molecules and then can allow the identification of
odorous mixtures. The measurement device is completed by microfluidic system, image acquisition and processing software.

Figure 1: From left to right - Binding of VOCs on the chemical ligands arrayed on prism covered by a gold layer; interrogating the
surface by SPRi; image analysis and data transfer.

Figure 2: Global scheme of measurement device: 1- Pumping of air and odorous molecules; 2- Transport of compounds to the
chamber; 3- Interaction of compounds with chemical sensors arrayed on a prism; 4- Lighting the prism with LED lamp; 5-
Recording optical signal transduction with a camera; 6- Storage (database) and analysis of results.

Generally, e-noses (and e-tongues) are mainly based on non-specific interactions between compounds and sensors 4. Of course sensors
can be selected because of stronger interactions with some components than others but the individual selectivity remains limited.
Ideally, the selectivity can be obtained by copying specific bio-recognition via a lock-key approach using receptors, antibodies or
aptamers... So, a new approach of SPRi-based on optical e-tongue and using an assembly of non-specific combinatorial sensors was
developed8. For all sensor arrays, the combination of information, produced by sensors, gives a fingerprint that corresponds to the
analyzed mixture. The idea to improve such a measurement is based on a combinatorial approach to simplify the preparation of
sensing materials as e-noses or e-tongues. The syntheses of different receptors represent time and cost consuming actions. In order to
avoid this problem, the preparation of combinatorial assemblies behaving as cross-reactive receptors can be retained. This
combination as array is based on self-assembly of a handful of building blocks mixed in varying and controlled proportions.
Measurement is then carried out by real-time optical detection system (SPR imaging); In that case, 3D continuous evolution patterns,
produced by the e-tongue, are like real-time landscape images 8.
The technological gap for e-nose application was to adapt SPRi for gas phase. One problem was the limitation of SPR to detect small
compounds (and lots of odorous volatile compounds are concerned). SPR imaging (after optical measurement) consists on a 2D map
where signals are roughly proportional to the mass of the compounds adsorbed on the SPR gold layer. Based on a saturated 2D
assembly model, the typical mass density of a small volatile compound to detect would be in the range of 1 to 500pg/mm2. But with
a specific setup, an SPR signal just needs an optical contrast between an adsorbed molecular layer and air.

The binding of odorous and volatile compounds on sensors must be reversible. So sensors are selected among small organic
molecules (or biological molecules) like small oligosaccharides. These organic molecules are arrayed on the gold prism using a two
dimensions (x-y) spotter as reported by Hou et al 8. Of course, developments on miniaturization and optimization are important to
propose a portable measurement device. The Figure 3 illustrates the two last generation of measurement devices with a down-scaling
of the device to improve portability. In this last development, the camera that collects SPR images is connected by Wifi or Bluetooth
to a smartphone integrated in the casing of the measurement device.

When volatile organic compounds bind on the sensors, dynamic images are measured. This record corresponds to
association/dissociation phases of the interaction/recognition process between compounds and sensors. With training of know odours
(real mixtures of compounds), new measurements and new images are compared with stored images in the database. If the mixture is
known, identification and quantitation can then be carried out. Results are given as a normalized radar chart allowing for a
straightforward recognition process or can be used in another format. Of course, the sample is not modified by a measurement cycle
because the interaction is not a chemical reaction and the measurement is carried out at ambient temperature.

Figure 3: Two generations of Neose instruments - On the top: the device (until end 2016); top right, typical image generated by the
sensors and down, the more recent device (2017) that includes a smartphone as interface

This device based on SPR imaging detects a wide panel of compounds with threshold detection around ppmv level. If currently,
around 50 sensors are present on the prism, the number can be extended to 100 allowing a precision improvement of the
measurement. It’s typically a great increase of sensor numbers comparatively to e-noses used during last decades. It’s by such an
increase that precision can be improved because human receptors are hundreds.

3. Conclusion

This new measurement device open a new way for odorous atmosphere analysis. It is of course a complement of identification of
odorous compounds (with GC/MS or CG/MS-O) and odour concentration measurement (by olfactometry). As all technologies based
on sensors, the objective is multiple: to propose possibility for continuous measurement; to identify known odours (comparison of
mixture fingerprints); to detect abnormal situations… But, this approach opens a new way to analyze an Odour by the translation of a
complex chemical mixture (characteristic of an odour), into an image (by SPRi). Then such an image can be analyzed by a classical
image recognition process. The complex multidimensional correlated images, generated by interactions of compounds with sensors,
are also representations of odour complexity. It can be considered that this device, an opto-nose is a tool to visualize odours and then
to represent one sensation (olfaction) by another one (view).
4. Acknowledgement

The authors thank all national and regional supports that finance, in the frame of FUI program, developments of this new Technology
and the WISE Project. This project is based on a wide consortium with 8 partners: Aryballe Technologies (leader), CEA-INAC,
GIPSA-Lab, GIP Mind, ARMINES / Centre LGEI Mines-Alès, Prestodiag, Cemag Consulting, SEB.

5. References

1. Gebicki J., Application of electrochemical sensors and sensor matrixes for measurement of odorous chemical compounds,
Trends Anal. Chem. 2016, 77, pp 1-13
2. Kiani S., Minaei S. and Ghasemi-Varnamkhasti M., Application of electronic nose systems for assessing quality of medicinal
and aromatic plant products: A review, JARMAP, 2016, 3, pp 1-9
3. Peris M. and Escuder-Gilabert L., Electronic noses and tongues to assess food authenticity and adulteration, Trends Food Sci.
Technol., 2016, 58, pp 40-54
4. Wilson A.D.. Review of electronic-nose technologies and algorithms to detect hazardous chemicals in the environment.
Procedia Technology, 2012, 1, pp 453-463.
5. Laplatine L.; Leroy L.; Calemczuk R.; Baganizi D.; Marche P. N.; Roupioz Y. and Livache T. Spatial resolution in prism-
based surface plasmon resonance microscopy. Optics Express, 2014, 22, pp 22771-22785.
6. Sage J, Mallèvre F, Barbarin-Costes F, Samsonov SA, Gehrcke JP, Pisabarro MT, Perrier E, Schnebert S, Roget A, Livache T,
Nizard C, Lalmanach G, Lecaille F. Binding of Chondroitin 4-Sulfate to Cathepsin S Regulates Its Enzymatic Activity.
Biochemistry, 2013, 52, pp 6487-6498.
7. Bulard E., Bouchet-Spinelli A., Chaud P., Roget A., Calemczuk R., Fort S., and Livache T. Carbohydrates as New Probes for
the Identification of Closely Related Escherichia coli Strains Using Surface Plasmon Resonance Imaging. Anal. Chem., 2015,
87, pp 1804-1811.
8. Hou Y., Genua M., Tada Batista D., Calemczuk R., Buhot A., Fornarelli P., Koubachi J., Bonnaffé D., Saesen E., Laguri C.,
Lortat-Jacob H. and Livache T. Continuous evolution profiles for electronic tongue based analysis, Angew. Chem. Int. Ed.,
2012, 51, 10394-10398