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Meat Science 88 (2011) 638644

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Meat Science
j o u r n a l h o m e p a g e : w w w. e l s ev i e r. c o m / l o c a t e / m e a t s c i

Rapid identication of pork for halal authentication using the electronic nose and gas
chromatography mass spectrometer with headspace analyzer
M. Nurjuliana a, Y.B. Che Man a,, D. Mat Hashim a, A.K.S. Mohamed b
a
b

Halal Products Research Institute, Universiti Putra Malaysia, 43400 UPM, Serdang, Selangor, Malaysia
Department of Food Science, Faculty of Food Science and Technology, Universiti Putra Malaysia, 43400 UPM, Serdang, Selangor, Malaysia

a r t i c l e

i n f o

Article history:
Received 15 July 2010
Received in revised form 31 January 2011
Accepted 22 February 2011
Keywords:
Authentication
Pork
Electronic nose
Surface acoustic wave
Principal component analysis

a b s t r a c t
The volatile compounds of pork, other meats and meat products were studied using an electronic nose and gas
chromatography mass spectrometer with headspace analyzer (GCMS-HS) for halal verication. The zNose
was successfully employed for identication and differentiation of pork and pork sausages from beef, mutton
and chicken meats and sausages which were achieved using a visual odor pattern called VaporPrint, derived
from the frequency of the surface acoustic wave (SAW) detector of the electronic nose. GCMS-HS was
employed to separate and analyze the headspace gasses from samples into peaks corresponding to individual
compounds for the purpose of identication. Principal component analysis (PCA) was applied for data
interpretation. Analysis by PCA was able to cluster and discriminate pork from other types of meats and
sausages. It was shown that PCA could provide a good separation of the samples with 67% of the total variance
accounted by PC1.
2011 Elsevier Ltd. All rights reserved.

1. Introduction
Testing of food products for the purpose of labeling and
authentication is necessary to avoid unfair competition and assure
consumer protection against fraudulent practices in the food industry.
One of the major issues concerning authenticity is where high value
raw materials are substituted with cheaper materials (Al-Jowder,
Kemsley, & Wilson, 1997) and especially in cases involving value
added products, where the potential nancial rewards for substitution
of cheaper ingredients are relatively high (Lai, Kemsley, & Wilson,
1995). Problems related to adulteration of meat species in ground and
comminuted products have been a widespread problem in some retail
markets, while meat species identication is a major global concern
(Murugaiah et al., 2009). Identication of the species of origin in meat
samples is relevant to consumers for several reasons. The fallout from
fraudulent substitution or adulteration will possibly lead to economic
losses, jeopardize the health of consumers who may have specic food
allergies and emotional disturbance due to religious reasons (Asensio,
Gonzlez, Garca, & Martn, 2008; Bonne & Verbeke, 2008; Ghovvati,
Nassiri, Mirhoseini, Moussavi, & Javadmanesh, 2009; Haunshi et al.,
2009).
The advance in food technology has resulted in the issues getting
more complicated where ingredients used in foods are more difcult
to understand by the consumers unless they are directly involved in
the related eld. Additionally, the task of food authentication cannot

Corresponding author. Tel.: +60 3 89430405; fax: +60 3 89439745.


E-mail address: yaakobcm@gmail.com (Y.B. Che Man).
0309-1740/$ see front matter 2011 Elsevier Ltd. All rights reserved.
doi:10.1016/j.meatsci.2011.02.022

totally rely on the expertise of the practitioners of the related eld


alone, but also require the contributions of those who are from other
technical elds for example, chemistry and veterinary science. In halal
authentication, one cannot rely solely on physical inspection and
documentation, but will need complementary evidence of the latest
high technology analytical instrumentation. There are many analytical
techniques that have been successfully applied to detect and identify
adulteration of porcine based ingredients in foods. Che Man and
Mirghani (2001) developed a Fourier-transform infrared (FITR)
spectroscopic method for detecting the presence of lard in certain
animal fat mixtures. Che Man, Syahariza, Mirghani, Jinap and Bakar
(2005) also successfully applied the FTIR spectroscopy technique to
detect adulteration due to lard in cakes. The effectiveness of using
DNA-based technology such as polymerase chain reaction (PCR), for
species identication in meat and fats has also been successfully
carried out by Aida, Che Man, Wong, Raha and Son (2005). The PCR
method for species identication from pork and lard samples has been
demonstrated to be a potentially reliable technique for detection of
pig meat and fat. Chromatographic techniques such as High
Performance Liquid Chromatography (HPLC) have also been
employed in order to distinguish lard from other animal fats
(Marikkar, Ghazali, Che Man, Peiris, & Lai, 2005). However, the
methods mentioned previously are generally sample destructive and
time consuming in order to accomplish a complete analysis. The need
to develop a rapid method for halal screening arose from the fact that
the Electronic nose can offer a non destructive, relatively low cost and
reliable method. The potential use of the electronic nose for detection
of lard adulteration in RBD palm olein has also been successfully
investigated by Che Man, Gan, NorAini, Nazimah and Tan (2005).

M. Nurjuliana et al. / Meat Science 88 (2011) 638644

639

Fig. 1. Typical electronic nose chromatogram of pork. SAW detector response for pork.

The aim of this study was to investigate the use of an electronic


nose based on surface acoustic wave sensor for detection of pork and
its discrimination from other types of meat and meat sausages. For
aroma proling and identication of the components that contribute
to the avor of pork, gas chromatography mass spectrometer with
headspace analyzer (GCMS-HS) was employed.
2. Materials and methods
2.1. Meat samples
Meat samples from sheep, cow, chicken and pork were used in the
experiments. For sausages, two pork, one chicken and one beef sausages
were used. All samples were purchased from the local wet market in
Serdang, Selangor, Malaysia. All samples were stored at 20 C in order
to minimize any deteriorative changes to the samples. The samples were
not subjected to any pre treatment that may have altered their aroma
components.
2.2. The electronic nose apparatus
The electronic nose (7100 vapor analysis system, Electronic Sensor
Technology, New Bury Park, USA) is a portable bench top enclosure for
eld laboratories or xed on-line installations. The zNose is based
upon well known principles of gas chromatography. This electronic nose
uses a single, uncoated, high quartz surface acoustic wave (SAW) sensor
which consists of an uncoated 500 MHz acoustic interferometer or
resonator bonded to a Peltier thermoelectric heat pump with the ability
to heat or cool the quartz crystal. This detector possesses advantages

Table 1
Tentative identication of volatile compounds of raw pork from the electronic nose prole.
Peak

Kovat's indices

Compounds

Odor description

1
2
3
4
5
6
7

612
718
806
903
1000
1104
1207

Diacetyl
3-hydroxy-2-butanone
2-methyl-propanal
Heptanal
Trimethyl pyrazine
Nonanal
Decanal

Buttery
Buttery
Pungent
Fatty
Roasted
Soapy
Soapy

Table 2
Major volatile components of pork by HSGCMS.
Peak

Retention time (min)

Compounds

1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43

0.461
2.295
4.432
6.616
11.821
15.466
16.219
17.083
17.674
18.671
19.264
19.739
20.190
20.837
21.288
21.971
22.909
23.104
23.585
23.704
23.793
23.858
24.013
24.185
25.615
26.410
26.725
26.986
28.043
28.844
29.052
30.986
31.230
31.390
32.755
33.010
33.366
34.500
34.916
35.206
35.984
37.041
38.780

Phenol
Hexanal
2-Butanone
1-methoxyl-2-methyl-2-Pentanone
Heptanal
Benzaldehyde
Heptyl ester 1-heptanol
2-pentyl-furan
Octanal
1-Hexanol
2, 4-Dimethylamphetamine
2-Octenal
1-Octyl-triuroacetate
Butanal
Nonanal
Decyl ester
2-Heptadecenal
Pentasiloxane
Naphtalene
Acetic acid
3-methyl-3,5 tetrahydro-4-thiopyranone
Acetamide
Dodecane
Nitro-L-arginine
2-Decenal
2-Heptadecenal
2H-Pyran
2,4-Decadienal
2-Undecenal
Tetradecane
2, 4-Bis(hydroxylamino)-6-methylpyramidine
Pentadecane
1-Heptadecanamine
Sulfur
Thiophene-3-ol
Dodecane
Hexadecanal
Propenoic acid
Sulfurous acid
Hexadecanal
3, 5-di-tert-Butyl-4-hyroxybenzaldehyde
Octadecanal
2-Octamine

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Fig. 2. Typical electronic nose chromatogram of pork sausage. SAW detector response for pork sausage.

such as high sensitivity, easy handling of signal, low power and long
term stability.

surface of the sensor, the frequency of the SAW will be altered and will in
turn affect the detection signal and allow the identication of the
contaminants. The ow rate of puried helium was xed at 3.0 mL/min.
The total cycle time per sample was 15 s.

2.3. Electronic nose analysis


Five grams of each meat sample was minced and weighed into septasealed vials and prepared in triplicate using the same unit. After precooking in a heated water bath maintained at 60 C for 10 min, the
sample's vapor was pumped for 3 s into the electronic nose with a sideported sampling needle through the septa. The electronic nose analysis
involved a two-step process. For the rst step, the vapors of the sample
were concentrated in a Tenax trap (inlet 200 C) and carefully controlled
to produce a repeatable and accurate collection of ambient vapors for
analysis in the next step. In the second step, the trap was rapidly heated
and the released vapors were re-focused at the head of a relatively low
temperature (40 C) capillary column (DB-5). This system is based on
the principle of gas chromatography. The column temperature was
programmed to heat from 40 to 160 C at a rate of 5 C/s, following a
linear rise to its maximum temperature. This will cause the different
chemical component in the sample to be released, travel through the
column and land on the surface of the SAW. The SAW sensor was
operated at a temperature of 30 C. When volatiles are adsorbed on the

Table 3
Tentative identication of volatile compounds of pork sausage from the electronic
nose prole.
Peak

Kovat's indices

Compounds

Odor description

1
2
3
4
5
6
7
8
9
10
11
12
13

435
646
657
709
754
820
925
1043
1140
1212
1255
1902
2100

Ethanal
2-methybutanal
2-methyl pentan-3-one
Ethyl propionate
2-pentanone
Butanoic acid
Hexanethiol
Phenylmethanol
-a-Prenchyl alcohol
5-methyl-2-furanaldehyde
(t)-(4s)-carvone
Isoeugenol
Octadecanoic acid

Pungent
Pungent
Mint

Ethereal
Rancid
Sulfur

Camphor
Almond
Caraway
Floral

2.4. Gas chromatography mass spectrometer with headspace


analyzer analysis
Five grams of each meat sample was transferred into a 20 ml
headspace vial. The extraction of the volatile compounds of the samples
was performed using a headspace auto sampler (Model G1888, Agilent
Technologies, Palo Alto, CA, USA). The transfer line from the headspace
sampler was directly connected to the injector of the gas chromatograph
(GC). The oven was set at 110 C. The extraction conditions in the
headspace auto sampler were programmed as follows: 20.0 min for vial
equilibration, 0.20 min for vial pressurization, 0.20 min for lling the
injection loop, 0.05 min for loop equilibration and 1.0 min for sample
injection. Helium with a purity of 99.999% was used for vial
pressurization and as carrier gas. The volatile compounds were analyzed
using a GC MS (Model 7890, Agilent Technologies, Palo Alto, CA, USA)
equipped with a non polar column (J&W Scientic DB-5; 30 m, ID
0.25 mm, lm thickness 0.25 m). The column temperature was kept at
40 C for 10 min, increased at 6 C/min to 240 C and isothermally
maintained for 20 min. The mass selective detector (Model MSD 59556,
Agilent Technologies, Palo Alto, CA, USA) was used in electron ionization
mode. A mass range between 30 and 550 m/z was scanned. The mass
spectra obtained were compared to the NIST Mass Spectral Search
Program for compound identication.
2.5. Statistical analysis
Unsupervised multivariate analysis, principal component analysis
(PCA) was used for data processing using the Unsrambler v.9.6
(CAMO AS, Trondheim, Norway) software. They were computed
iteratively in such a way that the rst principle component is the one
that carries the most information (or in statistical terms, the most
variance explained). The second principle component will then carry
the maximum share of the residual information. Therefore, PCA nds
an alternative set of coordinate axis, principal components, of which
the data set may be represented (He, Li, & Shao, 2007; Li & He, 2006;

M. Nurjuliana et al. / Meat Science 88 (2011) 638644

Zhang et al., 2006). The two main aims of PCA are the reduction in the
number of variables and elimination of redundancy.
3. Results and discussion
The electronic nose was used for rapid qualitative detection and
discrimination of pork from other types of meat and meat products
while the gas chromatography mass spectrometer with headspace
analyzer (GCMS-HS) was used for aroma proling of pork and other
meats.
3.1. Volatile compounds of pork
The chromatographic proles of raw meat aroma of pork, beef,
mutton and chicken obtained by electronic nose are shown in Fig. 1.
The chromatogram from the electronic nose is a graphical display of
the derivative of the frequency change versus time. Each peak found
in this derivative plot corresponds to a specic volatile compound and
has a retention time (given in seconds) which is specic to the column
and analysis temperature. The area under the peak was correlated to
the compound concentration and was expressed in counts (cts). There
were 7 (peaks: 1, 2, 3, 4, 5, 6 and 7) common compounds for all meat
samples. However, each sample showed variations in the amount for

641

every compound. Table 1 shows the peaks of the volatile compounds


and their odor description. The identication of the peaks was
tentatively carried out based upon Kovat's indices database stored in
the substance library of the Microsense software using n-alkanes as
standards. In contrast to well known analytical instruments for the
analysis of avor compounds, the electronic nose does not give any
identication of the compounds present whereas it attempts to
integrate measurements of the total headspace volatile compounds
and produce an aroma pattern that will exhibit differences or
similarities among the samples (Arnold & Senter, 1998; Boothe &
Arnold, 2002). Table 2 shows that there were a total of 43 volatile
components of pork identied by the GCMS-HS. The majority of the
compounds are well known lipid oxidation products, including
aldehydes, ketones and alcohols. It was observed from the aroma
analysis of pork using both techniques that the compounds were
positively identied and the most detected were aldehydes and
ketones. Most studies reported that almost all the aldehydes present
in pork such as heptanal and nonanal are oxidation products of oleic
acid and linoleic acids which were the most abundant unsaturated
fatty acids of pork (Meinert, Andersen, Bredie, Bjergegaard, & Aaslyng,
2007; Schiliemann, Wolm, Schrodter, & Ruttloff, 1987; Wettasinghe,
Vasanthan, Temelli, & Swallow, 2001). From these results it can be
shown that the electronic nose is a potentially feasible method for the

Fig. 3. VaporPrint of different meats and sausages. 2 dimensional olfactory images which provide the odor concentration and characteristic shape for each sample.

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M. Nurjuliana et al. / Meat Science 88 (2011) 638644

Fig. 3 (continued).

Fig. 4. Four different meats (score plot) in principal component analysis of the electronic nose data. Abbreviations: p1, p2, p3, Pork1, Pork2, Pork3; c1, c2, c3, Chicken1, Chicken2,
Chicken3; m1, m2, m3, Mutton1, Mutton2, Mutton3; b1, b2, b3, Beef1, Beef2, Beef3.

M. Nurjuliana et al. / Meat Science 88 (2011) 638644

643

Fig. 5. Four different meats and 3 different sausages (score plot) in principal component analysis of the electronic nose data. Abbreviations: p1, p2, p3 Pork1, Pork2, Pork3; c1, c2, c3
Chicken1, Chicken2, Chicken3; m1, m2, m3 Mutton1, Mutton2, Mutton3; b1, b2, b3 Beef1, Beef2, Beef3; ps1, ps2, ps3, ps4, ps5 Pork sausage1, Pork sausage2, Pork sausage3, Pork
sausage4, Pork sausage5; cs1, cs2, cs3, cs4, cs5 Chicken sausage1, Chicken sausage2, Chicken sausage3, Chicken sausage4, Chicken sausage5; bs1, bs2, bs3, bs4, bs5 Beef sausage1,
Beef sausage2, Beef sausage3, Beef sausage4, Beef sausage5.

analysis of aroma of raw materials and can possibly offer a technology


which can rival GCMS-HS.
The gas chromatograms of the aroma of meat products are shown in
Fig. 2. The chromatograms show that pork sausage contained more
aromatic compounds and this observation is based on the number of
peaks. Table 3 shows the set of volatile compounds corresponding to
peaks 113 and their odor descriptions while Fig. 3 displays the aroma
pattern of 4 different types of meat, namely pork (Fig. 3a), chicken meat
(Fig. 3b), mutton (Fig. 3c) and beef (Fig. 3d) and 3 different types of
sausages, namely, pork sausage (Fig. 3e), chicken sausage (Fig.3f) and
beef sausage (Fig. 3g). The VaporPrint is a 2-dimensional olfactory
image which provides the odor concentration and characteristic shapes
(Sim et al., 2003). In this polar format, the display starts at the 0.0
position and follows around the dial with retention times increasing in a
clockwise direction. By measuring the time required for each chemical
to reach the sensor and the amount affects the SAW crystal's vibration

and hence both the identity (retention time) and the quantity (amount)
of the substance can be calculated using the software incorporated in the
device. Different meat and sausage samples showed variations in the
amounts of each compound and generate a unique and easily
recognizable image. The unique nature of this display is subjected to
the relative concentrations of several compounds making up the mix
while also quantifying the strength of each chemical compound within a
sample. However, to get an overall view of the complex data, principal
component analysis was carried out (Figs. 4 and 5).
3.2. Principal component analysis
Principal component analysis (PCA) as an unsupervised classication method to visualize the resemblance and difference among
different measurements in the data sets was used in order to structure
the data matrix. The meat samples were separated along the rst PC

Fig. 6. Seven electronic nose variables (loading plot) in principal component analysis of the electronic data.

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M. Nurjuliana et al. / Meat Science 88 (2011) 638644

which described 67% of the peak variations (Fig. 5) and showed seven
dened groups. Along the PC1 axis, pork, chicken, mutton, beef and
beef sausages were located with high positive scores but on the other
hand along the PC3 axis, chicken sausage had low positive scores
while pork sausage had low negative scores. This percentage appears
to sufciently dene a good model, especially for qualitative purposes.
Fig. 6 shows the loading plot with seven variables. Only four variables
(2, 4, 6 and 7) had a far Euclidean distance from the origin while the
remaining variables were considered as unimportant for discrimination (low loading values along PC1 and close to the origin). The high
positive correlation between peak 4 and PC1 indicated that the
volatile prole of pork contained a higher proportion of heptanal
(peak number 4) (Table 1). This indicates that the heptanal has a
major inuence upon the discrimination of pork from other types of
meats and sausages. This concurred with the observation of Shahidi
(1994) who reported that aldehydes are the major components
identied in the volatiles of cooked pork.
4. Conclusion
The ability of the zNose to qualitatively discriminate and cluster
among 4 common meat samples and 3 types of sausage was
demonstrated in this study. Measurements of the volatile compounds
by GCMS-HS were also employed which indicate that the electronic
nose has adequate selectivity and sensitivity to perform avor
detection in meats. With a total analysis of less than a minute and
requiring less than 5 g of sample, the electronic nose offers a rapid,
accurate, low cost and environmentally friendly tool for detection of
porcine based ingredients in foods and this is especially useful for
halal authentication and verication.
Acknowledgment
This research work was supported by Universiti Putra Malaysia
(Grant No. Research University Grant Scheme: 91033) awarded to
Professor Dr. Yaakob Bin Che Man. The authors are also greatly
indebted to Mr. Tibby Lim for his technical support, Dr. Marina Abdul
Manaf, Miss Syahariza Zainul Abidin and Mdm. Siti Munira Abduk
Razak for their assistance.
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