Food Chemistry 206 (2016) 3743

Contents lists available at ScienceDirect

Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem

Discrimination of honeys using colorimetric sensor arrays, sensory

analysis and gas chromatography techniques
Haroon Elrasheid Tahir a, Zou Xiaobo a,, Huang Xiaowei a, Shi Jiyong a, Abdalbasit Adam Mariod b,c
a

School of Food and Biological Engineering, Jiangsu university, 301 Xuefu Rd., 212013 Zhenjiang, Jiangsu, China
College of Sciences and Arts-Alkamil, University of Jeddah, P.O. Box 110, Alkamil 21931, Saudi Arabia
c
Department of Food Science & Technology, College of Agricultural Studies, Sudan University of Science & Technology, P.O. Box 71, Khartoum North, Sudan
b

a r t i c l e

i n f o

Article history:
Received 13 September 2015
Received in revised form 5 March 2016
Accepted 10 March 2016
Available online 11 March 2016
Keywords:
Honey
Colorimetric sensor array
SPME-GC/MS
Multivariate analysis
Sensory analysis
PLSR

a b s t r a c t
Aroma profiles of six honey varieties of different botanical origins were investigated using colorimetric
sensor array, gas chromatographymass spectrometry (GCMS) and descriptive sensory analysis.
Fifty-eight aroma compounds were identified, including 2 norisoprenoids, 5 hydrocarbons, 4 terpenes,
6 phenols, 7 ketones, 9 acids, 12 aldehydes and 13 alcohols. Twenty abundant or active compounds were
chosen as key compounds to characterize honey aroma. Discrimination of the honeys was subsequently
implemented using multivariate analysis, including hierarchical clustering analysis (HCA) and principal
component analysis (PCA). Honeys of the same botanical origin were grouped together in the PCA score
plot and HCA dendrogram. SPME-GC/MS and colorimetric sensor array were able to discriminate the
honeys effectively with the advantages of being rapid, simple and low-cost. Moreover, partial least
squares regression (PLSR) was applied to indicate the relationship between sensory descriptors and
aroma compounds.
2016 Elsevier Ltd. All rights reserved.

1. Introduction
More than 600 volatile compounds have been identified in
honey samples. These compounds belong to different chemical
families, including aldehyde, ketone, acid, alcohol, hydrocarbon,
norisoprenoids, terpenes, benzene compounds, esters, furan and
pyran derivatives (Karabagias, Badeka, Kontakos, Karabournioti, &
Kontominas, 2014a; Plutowska, Chmiel, Dymerski, & Wardencki,
2011).
Traditionally, the aroma of honey is determined by sensory
analysis, where 10 or 12 assessors recognise and score welldefined descriptors. However, this method is expensive and
time-consuming (Vaclavik & Christian, 2014). As a substitute,
instrumental techniques, such as solid-phase microextraction couple to gas chromatographymass spectrometry (SPME-GCMS),
can be used to recognise and measure singular aroma components.
Although SPME-GCMS shows good reproducibility, sensitivity
and provides qualitative and quantitative data for these compounds (Cuevas-Glory, Pino, Santiago & Sauri-Duch, 2007), it is relatively high-cost and time-consuming (pnik, Pazitn, ika, &
Szolcsnyi, 2014). Investigation of volatile compounds of honey
up to date has given slight importance to the relationship between
Corresponding author.
E-mail address: zou_xiaobo@ujs.edu.cn (Z. Xiaobo).
http://dx.doi.org/10.1016/j.foodchem.2016.03.032
0308-8146/ 2016 Elsevier Ltd. All rights reserved.

instrumental and sensory analysis (Manyi-Loh, Ndip, & Clarke,

2011). The association of sensory investigation with instrumental
analysis has increased in recent years, as it permits the characterisation of food such as honey according to definite standards.
In the last 20 years, there has been expanding study in achieving a speedy system for surveying food flavours, which has led to
the advancement of gas sensor array systems. These systems,
known as electronic noses, have been applied for the identification
of the botanical origin of Chinese honeys (Huang, Liu, Zhang, & Wu,
2015). Electronic noses based on mass spectrometry, piezoelectric
effects and electrical resistance have been tested on the volatile
compounds of some American (Lammertyn, Veraverbeke, &
Irudayaraj, 2004) and European honeys (Ampuero, Bogdanov, &
Bosset, 2004). These methods need the preceding removal of sugar
and water, which are the major honey constituents (Kakoniene,
Venskutonis, & Ceksteryte, 2008). Hence, it remains a challenge
to identify volatile compounds rapidly and to remove interferences
resulting from changes in humidity (Ouyang, Zhao, Chen, & Lin,
2013). The colorimetric sensor array has stood out as an effective
means to address this problem (Janzen, Ponder, Bailey, Ingison, &
Suslick, 2006). An advantage of this type of sensor is its ability to
sense compounds that cannot be detected by an electrochemical
sensor (Borrs-Linares et al., 2015). Recently, this type of colorimetric sensor array has been used to investigate wine (Ouyang
et al., 2013), monitoring pork sausage spoilage (Salinas et al.,

38

H.E. Tahir et al. / Food Chemistry 206 (2016) 3743

2014) as well as discrimination of Chinese liquors (Li et al., 2014).

Based on the aforementioned studies we can conclude that the colorimetric sensor array has vast potential in the investigation of
honey quality. However, there are no data about the application
of colorimetric sensor arrays for honey discrimination. Additionally, there are few studies on the relationship between sensory
descriptors and volatile compounds identified by SPME-GCMS
in honey.
Multivariate statistical techniques tools, including principal
component analysis (PCA), hierarchical clustering analysis (HCA)
and partial least-squares regression (PLSR) have been specifically
designed for the analysis and visualisation of complex sets of different samples. Several authors have used multivariate analysis
tools to correlate sensory analysis with GCMS data (Aznar,
Lopez, Cacho, & Ferreira, 2003; Castro-Vzquez, Daz-Maroto,
Gonzlez-Vias, & Prez-Coello, 2009; Vilanova, Genisheva, Masa,
& Oliveira, 2010). The aims of this study were: (1) to discriminate
Sudanese honeys using multivariate data analysis coupled with
colorimetric sensor array and SPME-GCMS; (2) to examine the
sensory profile of Sudanese honey in order to simplify its discrimination and; (3) to explain the correlation between aroma compounds and sensory analysis using PLSR.
2. Materials and methods
2.1. Honey samples and chemicals
Six varieties of honey based on floral type, which consist of
Acacia nilotica (n = 10), Acacia seyal (n = 20), Ziziphus spina-christi
(n = 20), Amaranthus graecizan (n = 10), Eucalyptus spp (n = 10)
and multifloral (n = 30) honeys were collected from different
sources across Sudan. Table S1 showed the botanical, geographical
origin and samples codes. The pollen types were placed into four
percentage classes, as determined by Louveaux, Maurizio and
Vorwohl (1978): predominant pollen (>45%); secondary pollen
(645% to >15%); important minor pollen (615% to P3%); and
minor pollen (<3%). The botanical origin of honeys was based on
the pollen spectrum (45% and above), which is the ratio of the frequency of each pollen type in the honey.
5,10,15,20-Tetraphenyl-21H,23H-porphine manganese(III) chloride; 5,10,15,20-tetraphenyl-21H,23H-porphine; 5,10,15,20-tetrakis
(4-sulfonatophenyl)-21H,23H-porphine manganese(III) chloride; 5,
10,15,20-tetraphenyl-21H,23H-porphine; zinc 29H,31H-tetrabenzo
[b,g,l,q]porphine; zinc 5,10,15,20-tetra(4-pyridyl)-21H, 23H-porphine
tetrakis(methochloride) were purchased from SigmaAldrich
(St. Louis, MO). Hydrophobic nanoporous film was obtained from
(Millipore Co, Billerica, MA, USA). Gentian violet (methyl violet
10B); tetrasodium [29H,31H-phthalocyanine-tetrasulfonato(6-)N29,N30,N31,N32] cuprate (4-); bromocresol green; methyl
yellow; nickel(ii) phthalocyanine-tetrasulfonic acid tetrasodium
salt; congo red; methyl orange; screened methyl orange (first transition); sodium chloride and chloroform were purchased from
Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China). A series
n-alkanes (C5C25) was purchased from SigmaAldrich, St. Louis,
MO.
2.2. Colorimetric sensor array preparation
The pattern of the colorimetric sensor array is often grounded
on two essential fundamentals: (1) each chemical responsive dye
contains a centre to interact with analytes, and (2) the interaction
centre must be strongly coupled to a deep chromophore. The prerequisite dyes comprise: (1) Lewis acid/base dyes (i.e., metal ion
containing dyes), (2) Brnsted acidic or basic dyes (i.e., pH indicators), and (3) dyes with large enduring dipoles (i.e., zwitterionic

solvatochromic dyes) (Janzen et al., 2006). Porphyrins and their

metal composites are accepted for identification of analytes with
Lewis acid/base abilities. Metalloporphyrins are ideal for the detection of metal-ligating vapours due to their open coordination sites
for axial ligation, their great spectral changes upon ligand binding,
and their strong coloration. Ordinary pH indicator pigments
change in response to changes in the proton (Brnsted) acidity or
basicity of their environment (Feng, Musto, Kemling, Lim, &
Suslick, 2010; Suslick, Rakow, & Sen, 2004). In this work, we investigated numerous commercially available sensing materials.
Finally, we decided that the abovementioned six metalloporphyrins and eight pH indicators were the best option in this experiment. For developing the colorimetric sensor, every chemically
responsive dye (20 mg) was diluted with 10 mL of chloroform
and sonicated for 2 h at room temperature. A fresh 4
4 colorimetric sensor array was prepared by spotting approximately 0.1 lL of
solution on the surface of hydrophobic nanoporous film membranes. After spotting, the arrays were kept at room temperature
for 15 min prior to use.
2.3. Data acquisition
A flatbed scanner (Epson Perfection 1200S, Seiko Epson
Corporation, Nagano-ken, Japan) was used to obtain the pictures
of the colorimetric sensor array. The initial image was first taken
before exposure to the honey samples, and then the array was
exposed to honey samples. For each type of honey 1.5 g of honey
were placed into a 100 mL beaker and dissolved with 40 mL of
sodium chloride solution (30%). The array was attached to a preservative film, which was used to seal the beaker containing the
honey solutions. Then the sample solutions were partially submerged in an ultrasonic bath (Wuxi Fanbo Biological Engineering,
Wuxi, China) for 35 min at 45 C for extraction (Karabagias et al.,
2014a). After extraction, the sensor arrays were removed from
the preservative film and dried in the fume hood for one hour at
room temperature. Then the sensor was scanned to get the final
image. Based on our previous work (Huang et al., 2015) colour
change plots were obtained from the RGB images by digitally
deducting the initial image from final image using a 314pixel average from the centre of each dye spot as follows:

DR jRa  Rb j

DG jGa  Gb j

DB jBa  Bb j

Here, a, correspond to final; b, correspond to initial.

DR; DG; DB are the colour differences.
The colour change profile is, afterward, simply a 3N-distance
vector (where N = number of dyes) which can be merely studied
by multivariate statistics. It is furthermore appropriate to visually
denote these vectors as colour difference maps which represent
every spot as the absolute value of its colour difference in RGB.
The response of each dye is expressed as the relative difference
of the RGB as follows:

DR DG DB
Rb Gb Bb

2.4. GCMS analysis

The identification and verification of the isolated compounds
was conducted following the method reported by Tahir, Xiaobo,
Zhihua and Yaodi (2015).

H.E. Tahir et al. / Food Chemistry 206 (2016) 3743

2.5. Sensory analysis of honeys

Sensory evaluation was determined following the method
reported by Castro-Vzquez, Daz-Maroto, Gonzlez-Vias and
Prez-Coello (2009). Honeys mentioned in Section 2.1 were introduced at room temperature in 50-mL glass bottles closed with a
twist-off cap to create satisfactory headspace. All coded honeys
were presented to each panellist. Honey samples were evaluated
in duplicate by each evaluator. The panel involved twelve panellists, ranging between 26 and 43 years old with preceding knowledge in sensory analysis. At preliminary meetings, panellists
were trained in descriptive sensory analysis. Then they provided
descriptors independently over the course of numerous sessions.
Seven aroma descriptors including flowery, caramel, fruit,
butter, chemical, grassy and plant-like/green were designated in order to define the differences between the honeys.
Plant-like/green and grassy were grouped together under one
descriptor (green/grassy) during descriptive analysis. Finally
the panellists assessed the intensity of each descriptor according
on scales ranging from 0 to 8 with 02 scarce, 24 light, 46 middle and 68 strong intensity of the attributes (Wei et al., 2015).

39

(Mathworks, Natick, MA) in Windows 7. PLSR mapping was performed using Unscrambler version 10. (Camo Software, Oslo,
Norway).
3. Results and discussions
3.1. Colorimetric sensor array analysis

2.6. Statistical analysis

3.1.1. Sensor responses

The reactions between chemical responsive dyes and volatile
compounds in honey need temperature and a period of time to
achieve equilibrium. The reaction conditions adopted in this work
were 35 min and 45 C. Fig. 1c presents the images of the colorimetric sensor array before initial image and after exposure to
the honey samples final image. As seen in Fig. 1a, the images
and colorific alteration maps of responsive dye dots in the images
of different honey varieties differ from each other. Honey from different botanical origins had distinct images. The colorimetric sensor array was responsive to compounds in honey varieties and not
easily affected by moisture and was able to separate honey from
different botanical origins. However, the colours of some dots were
similar, and variation was not noticeable. Multivariate analysis was
applied to discriminate these samples.

Principal component analysis (PCA) and hieratical cluster analysis (HCA) were applied in the present work. PCA-like methods can
be selected principally for the determination of the correlation
amid data (Gasper, Mijatovic, Bnard, Derenne, Kiss, &
Goormaghtigh, 2010). On the other hand, if we want to study the
clustering of similar data collected from various samples, cluster
analysis must be used (Wang & Mizaikoff, 2008). Related samples
tend to be grouped in the same cluster and the degree of difference
between the groups and the degree of difference between the clusters is stated (Ward Jr, 1963). PLSR was used to find the relationship between sensory analysis results and SPME-GCMS data.
HCA and PCA were performed using Origin Pro version 9.0 (Origin
Lab. Corp., Northampton, MA) and Matlab Version 7.10.0

3.1.2. Multivariate analysis

The colorimetric sensor array was applied to discriminate Sudanese honeys. The colorimetric maps of the six honey varieties are
shown in Fig. 1a. In this work, forty-two values of colour difference
(i.e. 14 red, 14 green, 14 blue colours) for each honey were
extracted from their difference maps, which were used for PCA
analysis. In general, the greater number of PCs is essential for
ensuring degree of discrimination (Zhang, Bailey, & Suslick,
2006). As shown in Fig. S1, the results of PCA demonstrated that
the array needs 10 dimensions to account for 95.72% of the total
variance and 15 dimensions to capture 97.63%. This much high dispersal indicates the wide array of chemical-property space being
examined by our selection of chemo-reactive dyes. The scatter plot

a
b

H1

H2

H6

H7

H3

H8

H4

H5

H9

H10

d
d

Before exposure

After exposure

Color difference

Fig. 1. Colorimetric maps of six honey varieties (a), plot of the first two principal components by PCA with data acquired from the colorimetric sensor (b), image of the
colorimetric sensor array before exposure, after exposure to honey samples, and the difference image (c) and HCA dendrogram of six kinds of honeys (d).

n.d.
n.d.
0.55 0.00
2.82 0.10
n.d.
0.85 0.10
0.39 0.00
2.08 0.20
2.06 0.20
0.46 0.00
n.d.
n.d.
1.24 0.20
n.d.
0.55 0.00
0.62 0.10
n.d.n.d.
n.d.
3.56 0.0
0.40 0.01
n.d.
n.d.
0.54 0.00
3.21 0.00
n.d.
0.20 0.00
n.d.
3.02 0.20
2.35 0.00
n.d.
1.53 0.20
n.d.
n.d.
n.d.
0.32 0.00
n.d.
0.28 0.00
0.37 0.00
5.53 0.2
1.59 0.10
KI, Kovats index; n.d., not detected.

H9
H8

n.d.
n.d.
0.34 0.00
0.47 0.00
n.d.
0.61 0.00
n.d.
3.97 0.30
1.72 0.00
1.24 0.00
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
2.54 0.3
n.d.
n.d.
n.d.
1.33 0.20
4.17 0.00
n.d.
n.d.
n.d.
12.34 1.20
3.47 0.10
1.21 0.00
n.d.
0.40 0.00
n.d.
1.09 0.00
0.43 0.00
n.d.
0.61 0.00
n.d.
1.56 0.1
1.42 0.30

H7
H6

n.d.
0.60 0.00
1.18 0.00
0.97 0.10
n.d.
n.d.
n.d.
16.75 1.30
2.14 0.00
5.22 0.30
1.24 0.10
0.36 0.00
n.d.
0.50 0.10
0.37 0.00
0.48 0.10
2.04 0.10
n.d.
n.d.
1.23 0.00
n.d.
0.98 0.00
1.16 0.10
2.96 0.50
0.88 0.20
0.89 0.10
n.d.
6.54 0.20
5.59 0.20
1.02 0.10
0.63 0.00
1.00 0.10
14.45 1.00
1.86 0.20
0.69 0.10
n.d.
0.52 0.00
1.83 0.30
0.90 0.2
1.67 0.10

H5
H4

n.d.
39.61 1.20
n.d.
0.78 0.00
2.06 0.20
3.17 0.20
6.79 1.10
0.81 0.10
1.92 0.30
n.d.
n.d.
n.d.
n.d.
0.79 0.10
n.d.
n.d.
n.d.
n.d.
4.18 0.4
n.d.
2.33 0.10
n.d.
1.15 0.20
1.91 0.20
3.31 0.00
2.73 0.10
n.d.
14.30 0.50
4.83 0.00
2.91 0.50
n.d.
0.62 0.0
n.d.
0.22 0.00
0.24 0.01
0.10 0.00
1.88 0.10
n.d.
n.d.
1.29 0.04

H3
H2

1.16 0.10
n.d.
0.25 0.00
0.53 0.00
2.24 0.00
3.58 0.00
n.d.
17.73 1.10
2.99 0.00
3.40 0.00
4.00 0.40
n.d.
n.d.
0.61 0.01
0.12 0.00
0.18 0.00
1.74 0.00
n.d.
n.d.
5.54 0.30
1.61 0.10
n.d.
n.d.
n.d.
3.48 0.10
3.55 0.20
n.d.
20.39 1.50
5.82 0.10
4.81 0.00
n.d.
0.57 0.10
n.d.
n.d.
n.d.
0.55 0.01
1.39 0.10
n.d.
n.d.
3.30 0.10
1216
1363
1894
1932
915
918
1086
1472
1536
1587
1664
1573
1677
1857
1189
1272
1516
1668
1618
1717
3-Methyl-1-butanol
1-Hexanol
Benzyl alcohol
Phenyl ethyl alcohol
2-Methylbutanal
3-Methylbutanal
Hexanal
Furfural
Benzaldehyde
5-Methylfurfural
Safranal
2-Methylpropanoic acid
2-Methylbutanoic acid
Hexanoic acid
2-Heptanone
2-Methyldihydro-3(2H)-furanone
2-Acetylfuran
Acetophenone
3,7-Dimethyl-1,5,7-octatrien-3-ol,
4-Oxoisophorone
A1
A2
A3
A4
B1
B2
B3
B4
B5
B6
B7
C1
C2
C3
D1
D2
D3
D4
E1
E2

H1

Relative content (mean standard deviation)

KI

3.2.1. Volatile compounds of honey

The volatile compounds analysed by SPME-GC/MS of six varieties of honey samples are presented in Table S2. There were more
than 55 compounds identified in these honey samples and their
concentrations varied significantly. Regarding the comparison
amongst the analysed honeys, it was clear that only three compounds amid the total compounds were detected in all samples,
whereas a few compounds existed in less than three of the six varieties of samples. The identified compounds consist of of several
families, 2 norisoprenoids, 5 hydrocarbons, 4 terpenes, 6 phenols,
7 ketones, 9 acids, 12 aldehydes and 13 alcohols. These compounds
have been reported as core aroma compounds in honey
(Karabagias, Badeka, Kontakos, Karabournioti, & Kontominas,
2014b). The majority of these volatiles have been reported by several authors (Pasini, Gardini, Marcazzan, & Caboni, 2013;
Plutowska et al., 2011; Tahir et al., 2015). Norisoprenoids such as
2-hydroxy-3,5,5-trimethyl-cyclohex-2-enone and 2,6,6-trimethyl2-cyclohexene-1,4-dione were reported in honey (Bianchi, Careri,
& Musci, 2005). Aroma profile of honey is affected by plant types

Compound

3.2. GC/MS analysis

Code

of data using the first two PCs is shown in Fig. 1b. The contribution
of the first two PCs was 56.21% of the total variance (PC1 = 39.80%,
PC2 = 16.41%). The data are clearly grouped into ten groups based
on the amount of the volatiles in the honey samples. As we can
see from this figure, the data points of H5 and H6 were slightly
more distributed compared to other honey sample groups. This
could be attributed to the potent volatility of the volatile compounds identified in these samples. It can be seen from Fig.1b that
H8 was positioned between acacia honeys H3 and H9, which
belong to different geographical sources. Besides, overlapping
was found between H3 and H8, which may be due to the similarity
of dyevolatile interactions. Additionally, jujube honeys H1 and H6
which also belong to different geographical areas were successfully
discriminated from other samples. However, the clustering of H1
and H6 in the different clusters may be due to the differences of
dyevolatile interactions. Therefore, the composition and sensory
characteristics of honey differ considerably depending on botanical
and geographical origins. Moreover, multifloral honeys (H4, H5 and
H7) from different locations were clearly discriminated. The results
of the colorimetric sensor array indicated that the volatile compositions of honeys analysed varied considerably.
HCA study average between groups linkage method was used
to explore the data (R, G, and B value of each dot and Euclidean distance) obtained from colorimetric reactions. The HCA reveals the
level of similarity of the array reactions between the different compounds by the datas full dimensionality. Additionally, HCA was
able to identify any misclassification between distinct measurements of components. From Fig. 1d, it can be seen that the samples
were obviously separated into two groups: H5 with high carboxylic
acids in group (A) and others honey samples in group (B). According to Janzen et al. (2006), carboxylic acids exhibited strong
response, while alcohol showed weak response just as the mammalian olfactory system. Examination of Fig.1b and d showed that
reasonable clustering was procured in both PCA and HCA, yet, in
both cases, honeys with the same botanical origin did not cluster
closely with each other. Besides, correct clustering of multifloral
honeys (H4, H5 and H7) from different geographical regions by
HCA further proved the sensor to have excellent performance in
discriminating different honeys. This may be due to the fact that
honey properties and composition depend not only on the
nectar-providing plant species, but also on other factors such as
bee species, mode of storage, and even harvest technology and
conditions. These findings are partly in agreement with results of
GCMS data and sensory evaluation analysis.

H10

H.E. Tahir et al. / Food Chemistry 206 (2016) 3743

Table 1
Percentage composition (%) of volatile components used for discrimination of Sudanese honey samples.

40

H.E. Tahir et al. / Food Chemistry 206 (2016) 3743

Table 2
Cumulative loading values of the 20 compounds for the first three principal
components.
Code

PC1

PC2

PC3

A1
A2
A3
A4
B1
B2
B3
B4
B5
B6
B7
C1
C2
C3
D1
D2
D3
D4
E1
E2

0.5253
0.8357
0.1019
0.0796
0.8056
0.7699
0.8333
0.6798
0.1606
0.6375
0.2121
0.0893
0.1561
0.1865
0.0790
0.5987
0.6101
0.1241
0.9110
0.6287

0.5330
0.3849
0.6644
0.8756
0.3930
0.5196
0.3947
0.5895
0.7642
0.0266
0.6658
0.4993
0.5266
0.3325
0.5934
0.4883
0.0895
0.6861
0.0259
0.3574

0.1802
0.0136
0.1022
0.2431
0.2768
0.0747
0.0286
0.1927
0.5696
0.5277
0.3098
0.7923
0.6459
0.7311
0.6579
0.0599
0.3317
0.4362
0.1130
0.0614

and environmental conditions (Anklam, 1998). The results demonstrated critical differences in volatile profiles among honey samples of the same floral source, but from different geographical
areas. For instance, the number of compounds of Jujube honeys
H1 and H6 were found to be different.
3.2.2. GC/MS multivariate analysis
In order to better elucidate the data obtained from SPME-GC/MS,
the 20 abundant volatiles or the active aroma volatiles were manually assimilated and their relative percentages were used for further
multivariate analysis (see Table 1). The first three principal components (PCs) covered 71.33% of the total variance (PC1 = 29.78,
PC2 = 25.87, PC3 = 15.68). Table 2 presented the loadings of the
original variables in the first three PCs. The results indicated that
PC1 positively correlated with furfural, 5-methylfurfural, 2methyldihydro-3(2H)-furanone, 2-furyl methyl ketone and 2,6,6-tri
methyl-2-cyclohexene-1,4-dione; volatiles negatively correlated to
PC1 included 1-hexanol, 2-methylbutanal, 3-methylbutanal, hexanal and 3,7-dimethyl-1,5,7-octatrien-3-ol. Alcohols (benzyl alcohol, phenylethyl alcohol), aldehydes (benzaldehyde, safranal) and
ketones (acetophenone) which are key compounds contributing
to the aroma of honeys showed a positive correlation to PC2, while

41

3-methyl-1-butanol was negatively correlated. Organic acids,

including 2-methylpropanoic acid, 2-methylbutanoic acid and hexanoic acid were positively correlated to PC3, while 2-heptanone
was negatively correlated.
As seen in Fig. 2b, honey samples H3 and H6, were positively
correlated to PC1. Furfural, 5-methylfurfural, 2-methyldihydro-3
(2H)-furanone, 2-furyl methyl ketone and 2,6,6-trimethyl-2cyclohexene-1,4-dione were associated with these samples. The
H4 and H8 honeys were negatively correlated to PC1, these
samples were segregated from other types by 1-hexanol,
2-methylbutanal, 3-methylbutanal, hexanal and 3,7-dimethyl1,5,7-octatrien-3-ol. Volatiles that were positively correlated with
PC2, benzyl alcohol, phenylethyl alcohol and benzaldehyde were
associated with H1 and H2 honeys. H10 and H7 showed statistical
variances from the other studied honeys concerning the key
content of volatiles that positively contribute to PC2: safranal, acetophenone, benzyl alcohol, phenylethyl alcohol and benzaldehyde.
2-Methylpropanoic acid, 2-methyl-butanoic acid and hexanoic
acid were associated with H5, while H9 was negatively correlated
to PC3 and associated with 2-heptanone.
HCA provide a better alternative for graphic illustration of
high-dimensional data. HCA includes a measurement of either
the distance or the similarity among the items to be grouped. In
this experiment, honey similarities were computed according to
the Euclidean distance and average between groups linkage
method based on the selected volatiles to identify honey clusters.
The dendrogram was made in this mode that presented quantitatively the similarities between the several components. As shown
in Fig. 2a the honeys were separated into four main clusters. A
set of samples (H1, H2 and H3) was obviously grouped in one cluster. These honeys are associated with high aldehydes content
(Table 1). The second cluster involves H5 only because of its high
content of acids. This outcome is in agreement with the PCA discrimination. The third cluster comprises H6, H7, H8, H9 and H10,
while the last cluster consists of H4 honeys because of their high
alcohol content as shown in Table 1. HCA confirmed the variability
of aroma compounds in the different honey varieties.

3.3. Sensory analysis

The results of the sensory analysis of honey samples are shown
in Fig. 3. Based on the sensory results a radar plot was generated to
provide a graphic illustration of aroma profiles and variances

Fig. 2. (a) HCA dendrogram of different honeys and (b) plot of the first two principal components showing data obtained from GCMS.

42

H.E. Tahir et al. / Food Chemistry 206 (2016) 3743

3.4. The relationships between sensory analysis and SPME-GCMS

Fig. 3. Radar graphic presenting sensory profile of six honey verities analyzed.

Fig. 4. PLS regression map showing the relationship between GC/MS data correlated with sensory data in the honeys studied. Code of the aroma compounds
correspond to those in Table 1.

In the current study, we have applied PLSR to display the correlation between the aroma compounds identified by GCMS and
sensory attributes. To correlate aroma compounds and sensory
attributes, the 20 selected volatile compounds (Table 1) were
assigned as independent variables and the six sensory data were
assigned as dependent variables. The PLSR modelling between
selected volatile and sensory attributes was presented in a twofactor model elucidating 92% and 58% of the difference in X (designated volatiles) and in Y (sensory attributes) respectively. The
inner ellipse revealed 50% of the explained variance and the outer
ellipse is the unit-circle representing 100% of the explained variance. Fig. 4 shows the resultant correlation loading plot of the first
two factors. As can be seen from this figure, the aroma attributes
fruity and flowery were closed located in the positive region
of the two dimensions and were positively correlated to phenylethyl alcohol, benzaldehyde, safranal, 2-heptanone, acetophenone
and hotrienol. These outcomes exhibited that not all the sensory
attributes could be explained by the measured aroma compounds.
Negative relationships were found between flowery and fruity
descriptors and 3-methyl-1-butanol, furfural, 2-methyldihydro-3
(2H)-furanone, 2-furyl methyl ketone and 4-oxoisophorone. The
positive and negative correlations suggest that the perception of
an aromatic note is influenced not only by the existence of a few
compounds whose aromas form the note, but also by the presence
of other odorants that affect negatively the perception of such aromatic note (Aznar et al., 2003). As can be seen from Fig. 4, green/
grassy was positioned on the positive site of the first factor
and negative site of the second factor and positively correlated to
1-hexanol, 2-methylbutanal, 3-methylbutanal and hexanal. High
correlations were observed between 1-butanol, furfural, and
2-methyldihydro-3(2H)-furanone, 2-furyl methyl ketone and
4-oxoisophorone. Buttery descriptor was positively correlated
to 2-methylpropanoic acid, and 2-methylbutanoic acid. As these
volatiles and buttery descriptor were positioned close to the
centre, they are not well described by factor 1 and factor 2. In
general, good relationships between sensory assessment and
GC/MS data were found.
4. Conclusions

among the honeys for easy identification. The sensory analysis of

six honey varieties revealed substantial differences of the descriptors evaluated. As can be seen in the radar graph, honey samples
from the same botanical origin showed different attribute scales,
for example, H3 and H9 acacia honeys. The different flavour profiles of the tested honeys specified indicated that there are some
major components that separated the honey sample. For example,
H4 had the highest score of green/grassy attribute possibly due
to a high concentration of alcohol such as 1-hexanol, while H5
had the highest score of buttery, because of high carboxylic acid
content. Although H8 and H9 provided comparable scale values for
the fruity and caramel attributes, the aroma profiles of them
varied considerably with respect to the green/grassy and chemical descriptors. Caramel attribute was the specific descriptor
with the highest scores in H1, H2 and H3 honeys. The high
caramel scores for these honeys are related to increased quantities of aldehyde compounds such as benzaldehyde, 5methylfurfural and furfural. H6 honey was characterised with
chemical and Caramel descriptors, while H7 honey was characterized as flowery and fruity with low buttery and chemical notes. In general, some overlaps on the aroma descriptors
were found (Fig. 3 and Table 1). These outcomes were partly in
agreement with the multivariate analysis results mentioned above.

To the best of our knowledge, this is the first work revealing the
discrimination of honey based on the aroma compounds assessed
with colorimetric sensor array, sensory analysis and GCMS analysis combined with multivariate analysis. The outcomes demonstrated that both the colorimetric sensor array and GCMS
effectively discriminated the honeys. Furthermore, good relationships between GCMS and sensory evaluation were found. There
was a decent compatibility between colorimetric sensor array, sensory analysis and GCMS data, indicating that the colorimetric sensor array has the effectiveness to discriminate honeys from
different botanical origins.

Acknowledgment
We acknowledge the financial support of National Science and
technology support program (2015BAD17B04), Chinese 863
Program, (Grant No. 2011AA00807), NSFC (Grant No. 6091079),
the Jiangsu province science fund for distinguished young
scholars (BK20130010), New century excellent talents in university (NCET-11-0986), Specially-appointed professors by Jiangsu
province.

H.E. Tahir et al. / Food Chemistry 206 (2016) 3743

Appendix A. Supplementary data

Supplementary data associated with this article can be found, in
the online version, at http://dx.doi.org/10.1016/j.foodchem.2016.
03.032.
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