ELSEVIER B

CHEMICAL
Sensors and Actuators B 37 (1996) 43-48

Rapid discrimination of meat products and bacterial strains using

semiconductor gas sensors
V&onique Vernat-Rossi a, Carmen Garcia b, R6gine Talon ", Christian Denoyer a, Jean-
Louis Berdagu6 "~

a INRA, Station de Recherches sur la Viande, 63122 St Genbs Chanzpanelle. France

" University of Extremadura, Faculty of Veterinary Science, 10071 Caceres, Spain
Received 25 July 1995; revised 31 May 1996; accepted 5 June 1996

Abstract

The aim of this study is to test, in ambient conditions, the capacities of semiconductor gas sensors to discriminate cured products (dry
sausages of various origins or cured hams of different qualities) as well as aromatic or pathogenic bacterial strains commonly used for curing
or sometimes encountered in meat products. An overall analysis of the dynamic headspace desorbed by the different substrates analysed has
been carried out by six gas sensors with different characteristics. Using factorial discriminant analysis, we classify 94% of dry sausages, 87%
of cured hams and 98% of bacterial strains into their respective groups. Cross-validation of the discriminant functions confirms the stability
of the results. The results of this study show that with a limited number of semiconductor gas sensors, it is possible to classify rapidly and
reliably various products, on the basis of the volatile compounds which they desorb.

Keywords: Bacterial strains; Meat products; Semiconductor gas sensors

1. Introduction 2. Materials and methods

2.1. Semiconductor gas-sensor measurements

Aroma analysis is an established methodology in foodstuff
characterization. The methods of analysis commonly used
involve various extraction techniques such as high-vacuum
distillation [ 11, static headspace [2], dynamic headspace [ 3 ]
and separation techniques such as gas-phase chromatogra-
phy-mass spectrometry [ 4 ]. Recently, th,.se tools were used
to characterize cured products [5] and microorganisms [6],
respectively. This method is time consuming because of the
chromatographic separation and identification stages and can-
not be used routinely in the food industry.
Unlike separative techniques (such as gas chromatogra-
phy), gas sensors provide a non-separative analysis (that is,
a direct analysis of complex gaseous mixtures), which makes
it possible to carry out measurements and obtain very rapid
diagnoses [ 7-9 ].
In this study we shall test the ability of a commercial
semiconductor gas-sensor array to classify products such as
dry sausages, cured hams or bacterial strains. The desorbed
volatile compounds are collected using a particular glass
extractor and discrimination of products will be done after
selecting the relevant variables of the gas-sensor signal.
0925-4005/96/$15.00 © 1996 Elsevier Science S.A. All rights reserved
Pll S 0 9 2 5 - 4 0 0 5 ( 96 ) 0196 8-6
Analyses were performed with an array of six gas sensors
(Fox 2000 Alpha M.O.S.). In the measurement chamber, gas
sensors were n-type tin oxide semiconductors. Their selectiv-
ity depends on the quantity of catalytic metals (platinum,
palladium) incorporated in the tin oxide [ 10]. The principle
of gas detection using sensor units is based on the variation
of electrical resistance when gas molecules are adsorbed on
the sensor surface [ 10]. Resistance variations were converted
into voltage variations (V) and in the calculations 1/Vvalues
were used. Before each aquisition, an autonuli is done. In
order to include the baseline information in the calculation,
the initial voltage values measured one second before the
acquisition have been collected (Vhs~-V,s~).
In order to speed up the surface reactions and reduce water
adsorption on the sensor surface, the operating temperature
is 350°C [ 8 ] Desorption of volatile compounds adsorbed on
the semiconductor depends essentially on the analysed gas,
the humidity, the type and the operating temperature of the
sensor [ 10,11 ]. Semiconductor gas sensors were: TGS822,
TGS812, TGS824, TGS825, TGS880 and TGS800. Accord-
44 V. Vernat-Rossiet al. / Sensorsand Actuators B 37 (1996) 43--48
ambiant
air of the
laboratory
air filter
.f
(active c h a r e o a l ) ~
2 way valve
flexible teflon seam
Fig. I, Analysisof the volatile compoundsof meat products. The analysis consists of two steps dependingon the position of the valves: ( I ) generation of
heads~ce for 10 rain, during which time the gas-sensorsignal returns to baseline; (2) introductionof volatilecompoundsthroughthe measurementchamber,
which is a 200 cm 3 box includingsix gas sensors.
ing to Ref. [ 12], TGS822 and TGS812 are particularly sen-
sitive to alcohols and organic solvents, TGS824 to ammonia,
TGS825 to hydrogen sulphide, TGS880 to vapours from food
and TGS800 to cigarette smoke and gasoline vapours.
2.2. Nature of the samples
Six different types of dry sausages, representative of
French production and purchased from a local shop, were
analysed: Dr, 'saucisson put pore'; D2, 'rosette'; D3, 'saucis-
son pur pore'; D4, 'noisette d'Auvergne'; Ds, 'saucisson de
m6nage'; D~, 'saucisse s~che pur pore'. The sequence for
analysing the six products was D~, D: ..... D~, repeated four
times a day for three consecutive days in order to test the
performance of the sensors over a short period. In total, '72
samples were analysed.
16 Iberian hams were analysed, Two sensory quality
groups were defined by Spanish experts. 10 hams were nor-
mal and six had a defective aroma called 'cala' revealed only
after cutting. Each product was analysed twice on the same
day and the 32 measurements were randomly performed to
approach on-line analytical conditions ~imed at providing a
qualitative diagnosis.
The seven species of Micrococcaceac studied can be iden-
tified in dry meat products. Some are used for aromatization
( + ), others are pathogenic strains ( • ). After inoculating
APT agar medium, tile Petri dishes were inoculated at 30°C
for 48 h and analysed. The sequence for analysing the seven
strains was: Staphylococcas carnosus ( + ), Staphylococcus
xylosus ( + ) , Micrococcus varians ( + ) , Staphylococcus
warned ( + ), Staphylococcus aureus ( * ), Staphylococcus
saprophyticus ( • ), Staphlococcus epidermidis ( • ). This
sequence was repeated six times and 42 inoculations were
analysed. The analyses of Micrococcaceae, like those of
cured hams, were all made on the same day.
i®®®L@
measucernent chamber
/ I®®®1
IL2J
,,~vlio:ad~i~:i
1- " " tlmaS~
S ctor ~tPol~engti°n
signal processing...
~----- solid sample
2.3. Volatile-compounds analysis
2.3.1. Meat products
10 g of dry sausage or 20 g of cured ham were cut into
cubes and put into a glass extractor (Fig. 1). After generating
headspace for 10 min, volatile compounds were pumped at a
flow rate of i 00 ml min- t and injected for 40 s through the
measurement chamber. During the headspace-generation
step, filtered air was blown into the measurement chamber to
make a return to baseline of the gas-sensor signal possible.
Data were acquired at a rate of one measurement per second
per sensor.
2.3.2. Bacterial strains
The headspace generated by bacteria after 48 h of growth
was analysed by a gas-sensor array. Fig. 2(A) describes the
preliminary stage, which trains the sensors. The analysis of
the volatile compounds was divided into two stages
(Fig. 2(B) ). The first consists in extracting the volatile com-
pounds of an empty Petri dish for 10 min to make a return to
baseline of the gas-sensor signal possible. The second con-
sists in pumping the headspace generated by the bacteria for
40 s so as to make it cross the measurement chamber in order
to analyse it. The data were acquired at a rate of one meas-
urement per second and per sensor.
In these examples, in order to avoid unwanted sensor drift
[ 101, measurements were performed on the same day or over
a short period (three consecutive days). We are presently
trying to find a recalibration procedure for long-term analyses
of foodstuffs.
2.4. Statistical analysis
The gas-sensor data were grouped into three matrices.
The first matrix X~ contained 246 variables: 6 sensors X 40
measurements ( s i = l . . . 6 X t j = l . . . 4 0 ) , 6 initial values
 V. Vernat-Rossi et al. I Sensors and Actuators B 37 (1996) 43-48 45

measurement
chamber

A
Io0o I
IOOO I

empty Petri dish non inoculatedPetri dish

measurement
chamber

(]) Iooo I
volatile compound /fl --
ooo,

Inoculated Patti dish measurement

chamber

Iooo i
@ " " ~ 00- , ~

'1
Fig. 2. Analysis of the volatile compounds of bacterial strains. (A) Preliminary analysis to train the gas sensor. Volatile compounds of a non-inoculated Petri
dish are introduced for 40 s through the measurement chamber. (B) The two stages of the volatile-colapound analysis: ( 1 ) return to baseline for the gas-sensor
signal; extraction of the headspace of an empty Petri dish for 10 min; (2) extraction of volatile compounds from an inoculated Petri dish and analysis for 40 s.

(Vbs~ -- 1...6) and 72 observations (6 dry sausages x 12 rep-
etitions, that is 6 groups each :ontaining 12 individuals).
The second matrix X2 contained 246 variables: 6 sen-
sors X 40 measurements, 6 initial values and 32 observations
( 10 hams without any defective aroma and 6 with the 'cala'
defective aroma, each analysed twice, that is 2 groups con-
taining 20 and 12 individuals, respectively).
The third matrix X3 contained 246 variables: 6 sensors x 40
measurements, 6 initial values and 42 observations (7 spe-
c i e s × 6 repetitions, that is 7 groups each containing 6
individuals).
Because the number of descriptive variables was too high
regarding the number of observations, we have to select a
subset of relevant variables. Furthermore, all variables do not
have the same discriminatory power and there is redundancy
and noise. To extract pertinent information from these matri-
ces, a systematic procedure has been used. Stepwise discri-
minant analysis enters (p<0.05) variables into the
discriminatory model according to their Fisher value F from
the analysis of covariance [ 13 ]. At each step, redundancy in
the model is tested and variables can be removed if they do
not satisfy the criterion to stay. The procedure stops when no
more variables can be entered or removed from the rr,odel
[14,15].
Using the subset of selected variables, a linear discriminant
model is elaborated. Linear discriminant analysis is one of
the multidimension reduction techniques [16,17]. It com-
putes i n f ( k - 1, p) linear combinations of the quantitative
selected variables: Y = bO + b l x I + . . . b p x p for k popula-
tions and p variables, in order to maximize between-group
variance and minimize within-group variance. An estimation
of the error rate is calculated in order to evaluate the classifier
performance. To compute a nearly unbiased estimator, the
resampling technique of cross-validation has been used [ 18 ].
This is an attractive technique when the number of observa-
tions is low [19,20]. This procedure classifies each obser-
vation of the data set using the discrimiaant functions
computed from the other n - 1 observations and gives an
estimation of the classification error rate of the discrimination
model.
3. Results and discussion
In the conditions of dynamic analysis of the headspace, the
sensors provided sigmoidal responses (Fig. 3) resulting from
complex phenomena of mass transfer and volatile com-
pound-sensor interactions. As far as the mass transfer was
concerned, the signals could be divided into two parts. The
first, made up of the ascending zone of the curve, was due to
the fact that volatile compounds concentrated in the head-
space came to the surface of the sensors. The second part of
46 V. Vernat-Rossi et al. / Sensors and Actuators B 37 (1996) 43--48

the curve was a steady-state desorption of the volatile com- axis 3

pounds of the analysed sample. The interactions between 0 3.5%
volatile compounds and sensors are the result of complex e~
o
mechanisms of adsorption, desorption, competition and even ee •
%®
of catalysis of the volatile compounds at the surface of the
sensors [ I01. ~P 4. o

Computation on the dry-sausage data set led to the selec- ,g 4.4, el, q ?
I B 4. ~ ..... _~o ° axis !
tion of i5 variables from the possible 246 contained in the a
rli 4. ~4.Oo 86%
initialmatrix X~. The 15 variables selected were: TGS812 O
O
Vb, TGSgl2 ts, TGS812 t6, TGS812 tlo, TGS822 Vb, TGS822 |

t3, TGS822 t4, TGS822 h, TGS825 Vb, TGS825 ts, TGS825 D ! B

re, TGSS00 Vb, TGSS00 t2, TGS824 h, TGS824 t6. The meas-
urements were possible in 10 s. The discriminant functions
calculated made it possible to classify 94% ofthe dry sausages
into their six groups (cross validation: 83% correctly classi-
fied). The results of the classification are given in Fig. 4. The
separation of products according to axis 1 shows that there
are three groups corresponding to the specific aromas of the
dry sausages. D~ and D.s, which make up the first group,
include red wine in their composition. The second group is
made up of D2, D3 and D6, which include garlic and the third
group corresponds to D4, which had only sodium nitrite
added. It was therefore interesting to note that, regardless of
the day of the analysis, the ! 2 repetitions of each product
were able to be grouped together easily. This meant, for this
example, that the use of the ambient air of the laboratory as
a vector gas for the volatile compounds did not disturb the
response of the sensors in such a way that they were no longer
I11
II
J
Fig. 4. Results of fractorial discriminant ananlysis. The plane i-3 explains
89.5% of the total variance of the data. The six groups of dry sausages are :
Dr, C); D 2. C3% D 3, @t; D4, ~ ; D s, + '. D~, Q.
able to be exploited for classification purposes. Different
studies in progress, however, show that it is preferable to
work with reconstituted air to improve in particular the sta-
bility of the baseline and consequently the discriminant
capacity of the sensors.
Computation on matrix X2 (data for cured hams) led to
the selection of two variables (TGS812 t4 and TGS800 t3).
The measurement was possible in 4 s. The discriminant axis
classified 87% of the hams into their sensory groups (cross
validation: 87% correctly classified). The results of classifi-
cation (Fig. 5) show that the 'cala' group had a wide distri-

Voltage (IF'C)
e

Ti
0 _
(seconds)
, l

0 5 to IS 2o as 3o 3S 4o
Fig 3 Time-intensity responses of an army of six gas sensors, represented by: TGS822. l ; TGSgi2, O; "!'GS824. ~ ; TGS825, @; 1"GS880, × ; TGSg00, I-'l.
 V. Vernat-Rossi et al. / Sensors and Actuators B 37 (1996) 43--48 47

number of observations

1 , axis 1
Fig. 5, Classification of hams according to the discriminant function: axis I = - !.7676 (TGS8 2 t4) + 0.9063 (TGS800 h). Quality groups are identified by:
normal hams El, 'cain' hams [[.

bution, perhaps because it is difficult to characterize the
defective aroma 'cain'. It is interesting to note that only two
sensors were sufficient for the classification. The TGS812
sensor selected at the first step of the stepwise discriminant
analysis was more sensitive to alcohols, which is consistent
with the high contents of non-ramified alcohol (from ethanol
to hexanol) measured in the 'eala' hams [ 21 ].
In microorganism classification, 11 variables were selected
from the possible 246 contained in matrix X3. The 11 varia-
bles selected (TGS880 Vb, TGS880 t~ t, TGS880 hT, TGS880
t4o, TGS822 Vt,, TGS822 to, TGS825 t35, TGS825 t4o,
TGS800 tg, TGS~, tls, TGS824 tt2) showed that measure-
ments were possible in 40 s. The discriminant functions clas-
sified 98% of bacteria into their respective groups (cross
validation: 86% correctly classified). Fig. 6 .qhows the plane
!-3 which separates the species S. warneri, S. aureus, S.
saprophyticus and S. epidermidis. It is interestih;3 to note that
in this plane, the pathogenic species S. aureus and S. epider-
midis are perfectly discriminated. The species S. xylosus, S.
carnosus and M. varians, which are confused in plane 1-3,
axis 3
4%
0 for on-line analysis in quality control.
are well discriminated by other axes of the factorial discri-
minant analysis (not presented).
4. Conclusions
The examples of application described here demonstrated
that with a limited number of sensors (six), it was possible
to classify rapidly food products or microorganisms in culture
by a global analysis (non separative) of their volatile
compounds.
Active research is presently being conducted on the anal-
ysis of volatile compounds by multisensor systems. This
research is aimed in particular at improving the met.,',~ of
sampling the gaseous mixture, increasing the specificity and
diversity of the sensors (semiconductors, conducting poly-
mers), improving the stability of the responses over time,
designing specific standards for recalibration procedure and
enabling the signal to be treated in real time. The final aim of
this research is to provide the food industry and analytical
laboratories with robust, easy-try-use and inexpensive systems

axis 1 References
I I ! I I I I
:" ~82%
o-'D
OgB
O i
OD
[ I ] C. Garcia, J.L. Berdagu~, T. Antequera, C. L6pez-Bote, J.J. C6rdoba
• o and J. Ventanas, Volatile cr- ,onents of dry cured iberian ham. Food
Chem., 41 ( 1991) 23-32.
0 [21 H. Hachenbergerand A.P. Schmidt, Gas Chromatography Headspace
$ Analysis, Heyden.Germany, 1977, p. 125.
0
[ 3 ] J.L. Berdagu6, N. Bonnaud, S. Rousset and C. Touraiile. Influenceof
Fig. 6. Results of factorial discriminant analysis. The plane !-3 explains pig crossbreed on the composition, volatile compound content and
86% of the total variance of the data. The seven species are represented by: flavour of dry cured ham, Meat Sci., 34 (1993) ! 19-129.
S. carnosus, ¢t; S. xy!osus, O; S. saprophyticus, ~; S. warneri, i ; S. epi- [4] J. Tranchant, Manuel Pratique de Chromatographie en Phase
dermidis, + ; S. aureus, 0; M. varians, [:3. Gazeuse, Masson, Paris, 1982, p. 504.
48 V. Vernat-Rossiet al. / Sensors and Actuatorv B 37 (1996) 43--48
151 J.L. Berdagu~,P. Monteil,M.C. Monteland R. Talon,Effectsof starter
cultures on the formation of flavour compounds in dry sausage, Meat
Sci., 35 (1993) 275-287.
[61 T.O. Larsenand J.C. Frisvad,A simple methodforcollectionof volatile
metabolites from fungi based on diffusivesamplingfrom Petri dishes,
J. Microbioi. Meth., 19 (1994) 297-305.
[71 T. Aishinm, Aroma discrimination by pattern recognition analysis of
responses fromsemiconductor gas sensor array,3'.Agric. Food Chem.,
39 (1991) 752-756.
[g] P.N. Bartlett and J.W. Gardner, in P.N. Bartlett and J.W. Gardner
(eds.), Sensors and Sensory Systemsfor an Electronic Nose, Kluwer,
Dordrecht, Netherlands, 1992, Ch. 4, pp. 31-51.
I91 V. Rossi, C. Garcia, C. Denoyerand J.L. Berdagu~, Discriminationof
food products using semiconductor gas sensors. Application to meat
product, Syrup.Olfaction and Electronic Noses, Toulouse, France. 26-
27Sept., 1994.
[101 S.R. Morrison, Semiconductor gas sensors. Sensors and Actuators, 2
(1982) 329-341.
[1tl K. Ihokum and J. Watson, The Performance of the Stannic Oxide
Ceramic Gas Sensors, eRe, Boca Raton, FL, USA. 1994, Ch, 2, pp,
49-89.
[121 Figaro Engineering Inc., Figaro Products Catalog, Osaka, Japan,
1990.
1131 SAS Institute lnc. Stepdisc procedure/stepwise method. SAS/STAT
User's Guide, 1988, pp. 909-922.
!!41 M.C.Costanzaand A.A. Afifi,Comparisonof stoppingrules in forward
stepwise discriminant analysis, J. Am. Statist. Assoc., 74 (1979) 777-
785.
II5] B, Burtschy and !=. Nivelle, Un exemple de discrimination: la
reconnaissance d'objets par radar, in G. Celeux (ed.). Analyse
Discriminante sur Variables Continus, lnria, Rocquencourt, France,
1990. Oh. 7. pp. 101-126.
[161 R.O. Duda and P.E. Hart. Pattern Classification and Scene Analysis,
Wiley-lnterscience, New York, 1973.
1171 J.H. Friednmn, Regularized discriminant analysis, J. Am, Statist,
Assoc,. 84 (1989) 165-175.
1181 SAS Institute Inc,, Discrimprocedure, SAS/STATUser's Guide, 1988,
pp. 359-.447.
[191 B, Eft'on, ~timating the error rate of a prediction rule: improvement
on cross validation, ,I, Am, Statist. Assoc,. 78 (1983) 316--331.
[201 B. Efron and G, Gong, A leisurely look at the bootstrap, the jacknife
and cto~ validation, The American Statistician, 37 (1983) 36-48.
[2ll C. Gareia, V. Rossi, C. Vialion. J.L. Berdagu~ and J. Ventanas,
Comparative study of volatile compounds from normal and spoiled
Iberian hams measm'ed by gas chromatography-mass spectrometry
and gas sensors. J, Fm~lSci., (1996) in press,
Biographies
W r o n i q u e Vernat-Rossi is currently in the third year of her
doctoral studies on rapid characterization of food products
by semiconductor gas-sensor analysis of their volatile com-
pounds. She works in the Flavour Laboratory, INRA of Cler-
mont-Ferrand-Theix, France. She obtained her DEA in
alimentation science at the University Clermont Ferrand II
(63000 Clermont Ferrand, France) in 1993.
Carmen Garcia is a professor in the University of Extre-
madura, Caeeres, Spain. She works in the department of
foodstuffs.
R~gine Talon is a researcher in microbiology at the Meat
Research Department of INRA of Clermont-Ferrand-Theix,
France. She was awarded the degree of engineer at CUST in
1978 and a doctorate in food science in 1981 (Clermont-
Ferrand, France). Her main topic is the study of meat starter
cultures. She is involved both in identification of this flora by
different methods (multigas sensors and pyrolysis mass spec-
trometry) and in characterization of the role of this flora in
the production of aromatic compounds in meat fermented
products.
Christian Denoyer is an engineer in the Fiavour Labora-
tory, INRA of Clermont-Ferrand-Theix, France. His work is
focused on volatile-compounds analysis by gas chromatog-
raphy coupled to a flame ionization detector or mass
spectrometer.
Jean-Louis Berdagud is a chemical and food science
researcher. He is the scientific manager of the Flavour Lab-
oratory, INRA of Clermont-Ferrand-Theix, France. He was
awarded the degree of engineer at ENSBANA (Dijon,
France) in 1979 and a doctorate in food science and nutrition
in 1982. His main topics of interest are focused on flavour
analysis of food p:oducts with both classical techniques and
new ones, like multigas sensors, headspace mass spectrom-
etry, pyrolysis mass spectrometry or infrared measurements.