Journal of Apicultural Research

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Palynological and physicochemical properties of

multifloral honey produced in some regions of
Algeria

Salim Zerrouk & Rayan Bahloul

To cite this article: Salim Zerrouk & Rayan Bahloul (2020): Palynological and physicochemical
properties of multifloral honey produced in some regions of Algeria, Journal of Apicultural
Research, DOI: 10.1080/00218839.2020.1856559

To link to this article: https://doi.org/10.1080/00218839.2020.1856559

Published online: 22 Dec 2020.

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Journal of Apicultural Research, 2020
https://doi.org/10.1080/00218839.2020.1856559

ORIGINAL RESEARCH ARTICLE

Palynological and physicochemical properties of multifloral honey produced in some
regions of Algeria
Salim Zerrouka,b
and Rayan Bahloula,b
a
Faculty of Sciences, Department of Biology, Amar Telidji University, Laghouat, Algeria; bLaboratory of Biological and Agricultural Sciences
(LBAS), Amar Telidji University, Laghouat, Algeria

(Received 25 March 2020; accepted 13 June 2020)

This study aimed to determine the palynological and physicochemical properties of multifloral honey collected from dif-
ferent regions of Algeria. A total of 24 different honey samples have been analysed for various physicochemical parame-
ters (moisture, electrical conductivity, pH, acidity (free, lactone and total), Hydroxymethylfurfural (HMF), proteins,
color, total sugars, fructose, glucose, sucrose, and fructose/glucose ratio). The results obtained in the present study
show the variability of the chemical composition of the honey samples. The results of the study indicated that most of
the honey samples were of good quality, and were found to meet all major international honey specifications. From the
palynological point of view, 62.5% of the samples had a moderate amount of pollen grains per 10 gram of honey (class
II); seven samples (29.16%) were rich in pollen (class III), and only two samples (8.33%) had a low pollen concentration
(class I). A total of 94 pollen types, belonging to 48 botanical families, were identified. The pollen types identified per
sample vary from 15 to 37, with an average of 27 types. The best represented families were Fabaceae, Boraginaceae,
and Asteraceae, which present in all the samples. The best represented secondary pollen types were Echium sp.,
Trifolium sp., Peganum harmala, Eucalyptus sp., Olea europaea, Chenopodium sp., Ononis natrix, and Ziziphus lotus. The quali-
tative pollen analysis demonstrated the important participation of nectarless plants in the makeup of the honeys, espe-
cially Chenopodium sp., and Olea europaea.
Keywords: Algeria; honey; quality; physicochemical; pollen analysis

Introduction source used to collect nectar or honeydew, geograph-

Honey is the natural sweet substance produced by Apis
mellifera bees from the nectar of plants or from secre-
tions of living parts of plants or excretions of plant-
sucking insects on the living parts of plants, which the
bees collect, transform by combining with specific sub-
stances of their own, deposit, dehydrate, store and
leave in honeycombs to ripen and mature (Council
Directive 2001/110/EC, 2001).
Depending on the raw material used by the bees,
honey may be classified as nectar, honeydew, or mixed
nectar–honeydew (Juszczak et al., 2009). The nectar
honey can be classified as uninofloral honey, such as
Eucalyptus, Citrus, and Thymus, or multifloral honey.
Uninofloral honeys are produced by one plant species
containing predominantly its nectar with minor nectar
contributions from other botanical origins. Multifloral
honey is a type of honey made from the nectar of sev-
eral plant species flowering at the same time of year,
none of which is predominant (Vuli
c et al., 2015).
Honey has been used by humans both as food and
medical product from ancient times to the modern civil-
ization, where its use in human nutrition and health is
due to its chemical composition (Alvarez-Suarez et al.,
2018). Honey has a very complex chemical composition
that varies depending on the bee species, the floral

Corresponding author. Email: zerrouksalim@yahoo.fr
ical origin, as well as climatic, processing, and storage
conditions (De-Melo et al., 2018).
From a chemical point of view, honey is composed
mainly of carbohydrates, comprising about 95% of
honey dry weight (predominantly fructose and glucose),
lesser amounts of water, and a great number of minor
components, such as phenolic components, organic
acids, amino acids, volatile compounds, vitamins, pro-
teins, minerals, pigments, waxes, pollen grains, enzymes
and other phytochemicals (Bogdanov, 2011; Geana &
Ciucure, 2020a).
As a natural product with a relatively high price, honey
has been for a long time a target for adulteration by misde-
scription of geographical and botanical origin source, the
addition of commercial sugar syrups, or when honey bees
are fed adulterating substance (Bogdanov & Martin, 2002),
hence adversely affecting the consumers' health along with
the quality of honey (Ghramh et al., 2020).
There are many types of commercially available
honey in Algeria, whether unifloral or multifloral honey,
but consumers prefer some particular honeys more
than others. In addition, they are looking for a trusted
source to buy this valuable product (Zerrouk et al.,
2013). According to the Codex Alimentarius Standard
(Codex Alimentarius Commission, 2001) and the

ß 2020 International Bee Research Association

2 S. Zerrouk and R. Bahloul
Figure 1. Geographical origin of the honey samples.
European Union Council Directive relating to honey
(Council Directive 2001/110/EC, 2001) honey shall not
have added any food ingredient than honey to it nor
shall any particular constituent be removed from it.
Honey shall not have any objectionable matter, flavour,
aroma, or taint from foreign matter during its process-
ing and storage.
Contrary to the unifloral honeys, the multifloral hon-
eys do not exhibit distinct physical or chemical charac-
teristics apart from a huge variability, which makes their
authentication particularly difficult (Ruoff et al., 2006).
Therefore, the aim of the current study was to investi-
gate the palynological and physicochemical properties of
multifloral honey produced in Algeria.
Materials and methods
Sample collection
Twenty-four selected multifloral honey samples from A.
mellifera, were collected during 2017 from bee-keepers
in different regions of Algeria (Laghouat with 5 samples,
Djelfa with 13 samples, and M
ed
ea with 6 samples)
(Figure 1). The honey was obtained from the hives by
centrifugation and stored in hermetically closed glass
jars in dark at 4  C until its analysis.
Melissopalynological analysis
Quantitative analysis
Based on the total number of plant elements (per 10
grams of honey), honeys are placed into one of the fol-
lowing classes (Maurizio, 1939): class I (<20,000), class
II (20,000–100,000), class III (100,000–500,000), class IV
(500,000–1,000,000), and class V (>1,000,000).
Qualitative analysis
For the qualitative analysis, the method without acetoly-
sis described by Louveaux et al. (1978) was followed.
Ten grams of honey were weighed and dissolved in
20 ml of hot distilled water, not above 40  C. The solu-
tion was centrifuged for 10 min (about 3500 rpm, RCF
¼ 2000) and the supernatant was drawn off. Then
another centrifugation was done for 5 min following the
same conditions. The entire sediment was put into a
slide and spread out over an area of about
24 mm 24 mm.
The percentage of representation for each pollen
type was calculated counting at least 800 grains of pol-
len per sample. The corresponding percentages of pol-
len grains were classified in frequency classes as follows:
D: predominant pollen (45%), S: secondary pollen
(15–45%), I: important minor pollen (3–15%), and M:
minor pollen (1–3%).
Physicochemical analyses
Moisture content
Moisture content was determined with a Carl-Zeiss
Jena refractometer, by measuring the refractive indices
at 20  C. The moisture content was calculated using the
Wedmore table and the results were expressed as
percentages.
Electrical conductivity
Electrical conductivity was measured at 20  C in a 20%
(w/v) honey solution (dry matter basis) in CO2-free
deionized distilled water by a EUTECH instrument con-
ductivity meter (Con.520), and the results were
expressed as lS/cm.
pH, free, lactone and total acidity
Free, lactone and total acidity were determined by the
titrimetric method: the addition of 0.05 N NaOH, is
stopped at pH 8.50 (free acidity), immediately a volume
of 10 mL 0.05 N NaOH is added, and without delay,
back-titrated with 0.05 M HCl from 10 mL to pH 8.30
 Palynological and physicochemical properties of Algerian honey
Figure 2. Distribution of the honey samples in Maurizio's classes.
(Lactonic acidity). Total acidity was obtained by adding
free plus lactone acidities. Results were expressed as
meq/kg.
Protein content
Protein content was determined by the method of Azeredo
et al. (2003). A volume of 0.1 mL of protein extract (honey
sample 50% w/v) was added to 5 mL of Coomassie Brillant
Blue. After 2 min of incubation, the quantity of proteins was
estimated at 595 nm in relation to the bovine serum albumin
standard curve (10–100 lg 0.1/mL).
Hydroxymethylfurfural (HMF)
HMF content was determined using the White spectro-
photometric method. The absorbance of the solutions
was measured at 284 and 336 nm with a UV–vis spectro-
photometer (Thermo Scientific Helios Gamma, England).
Color
The color was determined by spectrophotometric
measurement of the absorbance of a 50% (w/v) honey
solution at 635 nm. The honeys were classified accord-
ing to the Pfund scale after the conversion of the
absorbance values (White, 1984).
Sugar
Fructose, glucose, and sucrose were determined by the
HPLC method (Lee & Coates, 2000). Aliquots of honey
were diluted to 100 ml with distilled water, filtered
through a 0.45-lm filter (Albet, Barcelona, Spain), and
immediately injected into a Waters high-performance
liquid chromatography (HPLC) constituted of a 600
Controller pump with a quaternary gradient pump sys-
tem, a 717 plus Autosampler (Waters, Milford, MA,
USA), a 410 Differential Refractometer refractive index
(RI) detector. Separation of sugars was carried out using
a Phenomenex PhenoSphere NH2 80A column (250 mm
 4.6 mm, 5 lm) fitted with a guard cartridge packed
with the same stationary phase. HPLC conditions were
3
isocratic mobile phase, 80% acetonitrile, and 20% water;
flow rate, 1.8 ml/min; injection volume, 20 ll, with the
column temperature maintained at 30  C.
Total sugars were determined using a special refract-
ometer (Carl-Zeiss Jena refractometer) reading at 20  C.
Statistical analysis
All analyses were carried out in triplicate and the data
were presented as means, maximum (Max) and min-
imum (Min) value, and standard deviations.
The data obtained in the study were analysed statis-
tically using principal component analysis (PCA). The
variables included in this procedure were the physico-
chemical parameters. Statistical differences between
honeys samples from different geographic regions were
compared by One-Way ANOVA assuming there were
significant differences among them when the statistical
comparison gave P < 0.05. The statistical analyses were
performed with the STATGRAPHICS Centurion 16.
Results
Melissopalynological analysis
Quantitative analysis
The concentration of pollen grains per 10 gram of
honey varies from 13,500 to 351,250, with a mean value
of 94,709. Quantitative analysis of honeys revealed that
fifteen samples (62.5%) had a moderate amount of pol-
len grains per 10 gram of honey (class II); seven samples
(29.16%) were rich in pollen (class III), and only two
samples (8.33%) had a low pollen concentration (class I)
(Figure 2). Honeydew elements were practically absent.
Qualitative analysis
The results of the honey pollen analysis are shown in
Table 1. A total of 94 pollen types, belonging to 48
botanical families, were identified in the honey samples
analysed. The best represented pollen types were a
total of 62 and correspond to 32 plant families. 22 pol-
len types in the honey samples were classed as present
pollen (1% of pollen spectra) and were not included
in Table 1. Some of the samples contained a great diver-
sity of pollen types varying from 15 to 37, with an aver-
age of 27 pollen types (Figure 3).
The families with a higher incidence of pollen types
were Asteraceae with 11 types followed by Fabaceae
(10 types), Boraginaceae (6 types), and Apiaceae (5
types) botanical families which are frequent in the
studied region. Moreover, in many samples (25%) a
small percentage of unidentified pollen grains were
found, in which their values in the pollen spectra not
more than 1.1% (Table 1).
Pollen from Fabaceae, Boraginaceae, and Asteraceae fam-
ily was present in all the samples. Brassicaceae, Rosaceae,
Myrtaceae, Salicaceae, Cistaceae, Zygophyllaceae, Apiaceae
and Oleaceae were very frequently (75%) in the honey
4 S. Zerrouk and R. Bahloul

Table 1. Principal pollen types and their frequency classes in the honey samples.

Family Pollen types % M (1–3) I (3–15) S (15–45) D  45 Max
Acanthaceae Acanthaceae 8.3 1 0 0 0 1.8
Anacardiaceae Anacardiuma 4.2 0 0 1 0 41.1
Other Anacardiaceae 25 4.2 8.3 0 0 10.2
Apiaceae Eryngiuma 37.5 2 2 0 0 8
Daucus carota 25 0 0 1 0 25
Pimpinella anisum 16.7 0 0 2 0 28
Other Apiaceae 58.3 6 1 0 0 5.3
Araliaceae Hedera helix 20.8 2 0 0 0 1.8
Arecaceae Chamaerops sp. 33.3 2 1 0 0 4.7
Asteraceae Carduus sp. 66.7 7 1 0 0 8
Centaurea sp. 66.7 2 2 0 0 6.9
Taraxacuma 62.5 4 1 0 0 4.8
Matricaria sp. 54.2 2 3 0 0 6.1
Carthamus sp. 33.3 1 0 0 0 2.6
Ambrosia sp. 12.5 1 0 0 0 2
Launea sp. 12.5 1 0 0 0 1.2
Galactite tomentosa 4.2 0 1 0 0 5.1
Other Asteraceae 62.5 4 2 0 0 5
Boraginaceae Echium sp. 100 5 6 4 0 28
Cynoglossum officinale 8.3 1 1 0 0 11.2
Brassicaceae Brassica napusa 33.3 1 3 3 0 32.3
Other Brassicaceae 83.3 5 7 1 0 19.5
Caprifoliaceae Viburnum tinus 25 3 0 0 0 2.7
Casuarinaceae Casuarina sp. 16.7 1 0 0 0 2
Chenopodiaceae Chenopodium sp. 66.7 7 1 1 2 47
Cistaceae Cistus sp. 83.3 7 8 0 0 14.7
Ericaceae Erica arborea 16.7 1 1 0 0 3.7
Euphorbiaceae Euphorbia sp. 33.3 1 0 0 0 2.5
Other Euphorbiaceae 25 1 0 0 0 1.1
Fabaceae Trifolium sp. 91.7 8 5 2 0 20
Lotusa 50 5 4 0 0 12.1
Ononis natrix 50 3 4 3 0 41.8
Vicia sp. 25 1 0 1 0 38.1
Hedysarum coronarium 20.8 0 1 4 0 30.6
Galega officinalis 8.3 0 0 1 0 23.4
Gleditsia sp. 8.3 0 2 0 0 11.7
Ceratonia siliqua 4.2 0 1 0 0 13.3
Other Fabaceae 83.3 11 4 2 0 41.6
Fagaceae Quercus sp. 41.7 3 0 0 0 2.3
Lamiaceae Rosmarinus officinalis 20.8 1 1 0 0 3.1
Lauraceae Lauraceae 4.2 0 1 0 0 9.3
Liliaceae Muscari comosum 16.7 1 1 2 0 22.4
Other Liliaceae 25 0 1 0 0 3.4
Malvaceae Malva sylvestris 33.3 1 0 0 0 1.2
Myrtaceae Eucalyptus sp. 83.3 9 2 3 0 44.4
Oleaceae Olea europaea 75 1 4 2 1 53.7
Plantaginaceae Plantago sp. 33.3 2 0 0 0 1.8
Polygonaceae Rumex sp. 25 1 0 0 0 1.1
Polygonum sp. 12.5 2 0 0 0 2.9
Punicaceae Punica granatum 25 1 1 2 0 42.1
Rafflesiaceae Cytinus hypocistis 8.3 0 1 0 0 4.9
Resedaceae Reseda sp. 41.7 4 4 0 0 6.4
Rhamnaceae Ziziphus lotus 50 1 4 3 0 35.4
Rosaceae Prunusa 54.2 3 4 0 0 6.6
Rubus sp. 16.7 1 0 0 0 1.2
Other Rosaceae 54.2 3 2 0 0 5.8
Rutaceae Citrus sp. 33.3 2 1 0 0 4.3
Salicaceae Salix sp. 87.5 6 4 0 0 4.6
Urticaceae Ulmus sp. 16.7 2 0 0 0 1.4
Urtica sp. 12.5 1 0 0 0 2.2
Zygophyllaceae Peganum harmala 83.3 3 8 7 0 33.6
Other Zygophyllaceae 4.2 0 1 0 0 3.4
Indeterminate 25 1 0 0 0 1.1

Present pollen ( 1% of pollen spectra) was not include in table.
a
The pollen grain had similar morphology for some genus and species.
Notes: %: percentage of presence of each pollen type in the samples. Max: maximum value reached in samples for the pollen type.
 Palynological and physicochemical properties of Algerian honey 5

Table 2. Physicochemical parameters of analyzed honey samples.

Mean Min Max St. Dv.
Fructose % 41.15
34.04 46.90 3.11
Glucose % 32.59
27.74 36.98 2.58
FþG % 73.74 ns 64.33 80.48 3.39
F/G 1.27
0.92 1.60 0.16
Total sugar % 82.55 ns 77.72 84.40 1.53
Sucrose % 4.89 ns 2.74 7.87 1.32
Moisture % 15.66 ns 13.73 20.70 1.60
pH 3.87 ns 3.49 4.44 0.24
Free acidity meq/Kg 24.69 ns 13.72 40.33 7.33
Lactone acidity meq/Kg 5.66
1.66 9.29 2.07
Total acidity meq/Kg 30.34 ns 15.38 49.10 8.74
EC mS/cm 428.79
173 927.33 194.08
Figure 3. Number of pollen types found in honey samples. HMF mg/Kg 8.8 ns 00 39.62 8.32
Proteins mg/g 855.6 ns 546 1806 288.5
Color (mm Pfund) 63 ns 18 119 20.72
(Table 1). Other frequent families were Chenopodiaceae
St. Dv.: Standard deviation; F þ G: Fructose þ glucose; F/G: Fructose/
(67%), Poaceae (63%), Urticaceae (58%), Euphorbiaceae, Glucose ratio; EC: electrical conductivity; HMF: Hydroxymethylfurfural.
and Rhamnaceae (50% each). The best represented second-
Significant differences between regions at p ˂ 0.05; ns: not significant.
ary pollen types were Echium sp., which present in all the
samples; Trifolium sp. (found in 91.7% of the samples), The results obtained for the color were between 18
Peganum harmala, Eucalyptus sp., other Fabaceae and other and 119 mm Pfund, which represents a variation color
Brassicaceae they were found in more than 83% of the between extra white to dark amber, with a mean value
samples, Olea europaea (75%), Chenopodium sp., (66.7%), of 63.28 mm Pfund (light amber).
Ononis natrix and Ziziphus lotus (50% each) (Table 1). The average total sugar content was 82.55% and the
range was 77.72–84.40%. The total content of glucose
Physicochemical parameters and fructose is over 60% of honey (range between 64.33
and 80.48%, mean of 73.74%), in accordance with the
The results of the physicochemical analyses of honey
European Commission for honey, for all samples (Council
samples with the average and standard deviation are
Directive 2001/110/EC, 2001). The value for fructose was
shown in Table 2.
higher (between 34.04 and 46.90%, mean of
Moisture content in analyzed honey samples is
41.15 ± 3.11%) than that observed for glucose (between
between 13.73% and 20.70%, with a mean value of
27.74 and 36.98%, mean of 32.59 ± 2.58%). The level of
15.66 ± 1.60%. These results show that only one sample
sucrose (%) in the analyzed samples ranged from 2.74 to
exceeds the limit (20%) recommended by the inter-
7.87, with a mean value of 4.89%. Except for nine samples
national quality regulations (Council Directive 2001/110/
(37.5% of the honey samples), all other tested Algerian
EC, 2001).
multifloral honeys had sucrose contents below 5%, which
Honey samples showed an appropriate electrical
is the maximum prescribed limit for the sucrose content
conductivity ranging from 173 mS/cm to 927.33 mS/cm,
as per the Codex standard and the European Commission
with an average of 428.79 ± 194.08 mS/cm.
for honey (Codex Alimentarius Commission, 2001;
Honey pH values varied from 3.49 to 4.44, with an
Council Directive 2001/110/EC, 2001).
average of 3.87 ± 0.24. The free acidity of the samples
The fructose/glucose ratio was calculated, and varied
studied was below the allowable maximum (50 meq/kg)
between 0.92 and 1.60, with an average of 1.27 ± 0.16.
and thus conforms to the international regulatory stand-
ards for quality honey (Council Directive 2001/110/EC,
2001). It varied between 13.72 and 40.33 meq/kg, with Principal component analysis (PCA)
a mean value of 24.69 ± 7.33 meq/kg. The lactonic acid- Figure 4 shows the projection of the two first compo-
ity ranges between 1.66 and 9.29 meq/kg; and the total nents among the physicochemical parameters. Statistical
acidity ranges between 15.38 and 49.10 meq/kg. analysis showed that the first five principal components
Usually, honey contains a small amount of proteins. accounted for 86.5% of the variation in the honey sam-
In the investigated samples, the mean value of the pro- ples analysed. The first factor explained the 26.8% of
tein content was 855.6 lg/g (ranging between 546 and the variability, when combined with the second factor,
1806 lg/g). explained 47.4% of the variability of the data.
The Hydroxymethylfurfural (HMF) of the honey sam-
ples lay in the range of 0 and 39.62 mg/Kg (mean value
Discussion
of 8.8 ± 8.32 mg/Kg). It can be observed that it complied
with the limitation of 40 mg/kg established by the Melissopalynological analysis
European Commission (Council Directive 2001/110/ From a quantitative point of view, the pollen content
EC, 2001). varied highly with the type of honey but depend on
6 S. Zerrouk and R. Bahloul
Figure 4. Plot of the two first components of the PCA.
other factors as the procedure used by the beekeeper
to obtain the honey including the honey extraction
from the combs, the filtering system, or the type of
hive used. In the samples studied, additionally to the
botanical origin of the honey, some management practi-
ces as the harvest of honey combs with bee bread
could contribute to increase the pollen content. The
samples which belong to the class III of the Maurizio
classification contain predominant pollen (Chenopodium
sp.,) or secondary pollen, such as Echium sp., Eucalyptus
sp., Peganum harmala, Ononis natrix and Ziziphus lotus.
The variances of the pollen types present in the dif-
ferent honey samples analyzed are the result of its
diverse floral and geographical origin, as well as of cli-
matic conditions (Combarros-Fuertes et al., 2019).
Most of the honey samples (66.66%) had more than 20
pollen types, indicates the abundance of pollen and/or
nectar sources in the study area and the generalist
behaviour of Apis mellifera when searching for resources
(Bosco & Luz, 2018).
Asteraceae along with Fabaceae had the highest pol-
len diversity in the investigated honey samples since
they include various pollen types that reached high fre-
quency in the samples such as Carduus sp., Centaurea
sp., and Taraxacum type or Trifolium sp., Lotus type,
Ononis natrix and Hedysarum coronarium, which confirms
the important of these two families for beekeeping and
for honey production in the studied area.
The qualitative pollen analysis demonstrated the
important participation of non-nectariferous plants in
the makeup of the honeys. Three samples had high pol-
len frequency percentage (>45%) of Chenopodium sp.,
and Olea europaea but were discarded from the unino-
floral honey because these species do not produce any
nectar. These nectarless plants are particularly valuable
in spring, when the colony needs large amounts of the
protein contained in pollen for their brood-rearing
(Zerrouk et al., 2014). O. europaea species is one of the
most important trees for the agricultural economy; it is
a typical tree of the Mediterranean vegetation. Several
authors found high percentages of this pollen type in
their honeys samples analyzed; it was found as predom-
inant pollen in Greek thyme honeys (Karabournioti
et al., 2009), and has also been reported as secondary
pollen in Turkish, Moroccan and Portuguese honeys
(with percentages of 36%, 22%, and 30.6%, respectively)
(Silici & G€ okceoglu, 2007; Silva et al., 2017; Terrab
et al., 2003). This remark leads us to point out that
although olive trees are not considered as melliferous
plant, their flowers are however very visited by bees to
collect abundant pollen (Louveaux & Abed, 1984).
The pollen types from the plants of Echium sp.,
Trifolium sp., Peganum harmala, Eucalyptus sp., Olea euro-
paea, Chenopodium sp., Ononis natrix, and Ziziphus lotus
have been cited as nectar and/or pollen resource of the
bees and it has been found in honey samples from
Algeria (Zerrouk et al., 2014). Other pollen types were
very frequent (>66%) in the samples as important
minor pollen, like Salix sp., Cistus sp., Carduus sp. and
Centaurea sp.
The pollen spectrum found in this study indicates
that the flora is riches, diverse, and most spontaneous,
such as Peganum harmala, Ziziphus lotus, Carduus, Echium,
Euphorbia, and Rosmarinus officinalis. In addition, there is
some pollen from cultivated plants but generally with
low frequency, such as Pimpinella anisum, Ceratonia sili-
qua, Citrus sp., and Cucumus sativus. Many pollen types
found in our study have also been identified in multiflo-
ral honey from Algeria (Makhloufi et al., 2010) and
Spain (Silva et al., 2017).
Physicochemical parameters
Moisture content, a parameter related to the maturity
degree. The main factor causing fermentation in honey
is the high moisture content. For this reason, the mois-
ture content is a highly important quality parameter for
the shelf-life of the honey during storage (El Sohaimy
et al., 2015). In the present study, the moisture content
of 95.83% of the samples indicated a proper degree of
 Palynological and physicochemical properties of Algerian honey
maturity (below than 20%). The variation of moisture in
analyzed honey samples can be explained by production
season, the climate, and the degree of maturity reached
in the hive (Al-Khalifa & Al-Arify, 1999). These results
were similar to those obtained from Algerian multifloral
honey (Makhloufi et al., 2010).
Regarding the electrical conductivity, this parameter
was considered as a quality parameter indicating the
possible botanical origin of honey (Saxena et al., 2010).
These values are lower than those shown by Makhloufi
et al. (2010) for Algerian honey, and in agreement with
those reported by Kasperov
a et al. (2012) and Balos
et al. (2018) for multifloral honey produced in Slovak
and Serbia, respectively. The electrical conductivity of
honey is closely related to the contents of their mineral
salts and organic acids, and is a parameter that shows
the greatest variability according to the physicochemical
parameters of honeys depending on their botanical ori-
gin (Majewska et al., 2019).
All honey samples showed acidic pH values, a typical
characteristic of floral honey (Abdulkhaliq & Swaileh,
2017). These low pH values contribute to honey stabil-
ity during extraction and storage and inhibit the pres-
ence and growth of microorganisms. The free acidity of
the samples studied was lower than those shown by
Terrab et al. (2002). The Free acidity of honey is due to
the variations of some organic acids, particularly the glu-
conic acid, which remain in equilibrium with internal
esters, lactones, and some inorganic ions such as phos-
phate, chloride, and sulfates (Majewska et al., 2019).
The lactonic acidity is considered the reserve of acidity
when the honey becomes alkaline (Terrab et al., 2002).
The variation of the total acidity can be attributed to
geographical origin and the nectar source of the honey
samples. The high value of total acidity may imply that
at some point the honey began to ferment (Majewska
et al., 2019).
The results of protein content in this study are
lower than those obtained for other honeys types from
Algeria (Ouchemoukh et al., 2007). However, honey
proteins may act as markers for authentication (Lee
et al., 2015). The protein content of honey is dependent
on the nectar, honeydew, and mainly pollen of different
regional plants visited by bees, and can be attributed to
the presence of enzymes introduced by salivary glands
of bees themselves (De-Melo et al., 2018; Won et al.,
2009). Therefore, the protein profile could be more
associated with the entomological origin than the botan-
ical origin (Ram
on-Sierra et al., 2020).
Considering the HMF content, generally, this com-
pound not present in fresh honey, and the high levels of
it provide an indication of overheating, storage in poor
conditions or age of the honey (Zappala et al., 2005).
According to Mahrous Rady et al. (2018), honey is con-
sidered adulterated when the value of HMF more than
150 mg/kg of honey. For this reason, HMF is a recog-
nized parameter related to freshness and quality. The
7
data of HMF obtained from investigated honey samples
were higher than those reported by other researchers
(Yilmaz & Yavuz, 1999; Ozcan et al., 2006) and lower
than those shown by Terrab et al. (2002) and Makhloufi
et al. (2010).
Honeys show very different colors varying from
white or pale yellow to dark red or even black (Terrab
et al., 2004). Color is considered, as a quality parameter
for honey, very important for commercial activities, and
has a great influence on customer preferences (Ku
s
et al., 2014). The floral source, mineral content, age of
honey, and storage conditions can influence the honey
color (Zerrouk et al., 2013). In addition, it is related to
the content of the pollen, total phenolic, and antioxi-
dant activity (Bertoncelj et al., 2007). Moreover, recent
research shows that dark honeys (buckwheat, heather,
honeydew) are more healthful than light honeys, and
multifloral honeys are more beneficial to health than
unifloral ones (Go
sli
nski et al., 2020).
Regarding sugars, the honey samples studied had a
total sugar within the values reported for Italian multi-
floral honeys (Conti et al., 2007). Honey is composed
primarily of sugars and the most dominant sugars in
honeys are fructose and glucose, with higher amounts
of fructose compared with glucose (Geana et al.,
2020b), which confirm that all honey samples are genu-
ine honeys (Bentabol et al., 2014). The results of fruc-
tose and glucose in the current study are higher than
those obtained by Ouchemoukh et al. (2010) for
Algerian multifloral honeys. According to Anklam
(1998), the fructose and glucose content of any honey
type depends largely on the nectar source. Moreover,
the fructose/glucose ratio gives information about the
crystallization state of honey, when fructose is higher
than glucose the honey is fluid (Al et al., 2009).
The sucrose content in honey is considered an
important sugar from the legislative point of view. The
high content of sucrose in some samples may be due to
an early harvest of the honey and the origin of the nec-
tar compound of the honey (Kahraman et al., 2010).
Statistical analysis indicated that there were no sig-
nificant differences between honey samples collected
from different geographic regions (Laghouat, Djelfa and
M
ed
ea) for moisture, pH, free acidity, total acidity, total
sugar, the sum of fructose and glucose (F þ G), sucrose,
HMF, color, and proteins content. Significant differences
were only observed between geographic regions only
for fructose, glucose, fructose, and glucose ratio (F/G),
lactone acidity, and electrical conductivity (Table 2).
A statistical analysis by PCA was used to reduce the
dimensionality and to explain the variance-covariance
structure of the data and to find any relationship
between the variables and samples (Zerrouk et al.,
2018). According to Chen et al. (2018), if the first four
PCs explain more than 75% of the total variation, a reli-
able model may be obtained. In the investigated samples
the first four PCs explain more than 79%. The variable
8 S. Zerrouk and R. Bahloul
with the highest weight in the first component was only
total sugar with a positive value, and free acidity, total
acidity, and moisture with a negative value. Concerning
the second factor, fructose, fructose-glucose ratio, and
proteins showed the highest weight (in the positive
part), glucose, and lactone acidity in the negative part.
Some variables are placed close in the plot. Then pro-
teins, color, HMF, and moisture are in the same quad-
rant. The correlation between some parameters may be
due to the nectar source, climatic condition, and geo-
graphical origin of the honey samples.
Conclusions
The present work enriches our knowledge about the
honeys produced in the central region of Algeria. Based
on the results of physicochemical parameters, the multi-
floral honeys analyzed in this study were found to
meet all major international honey specifications, except
for the recommended criteria of sucrose and moisture
content, where 41.66% of the honey samples were of
unacceptable quality. With respect to the color param-
eter, it varies from extra white to dark amber color.
Among the 15 parameters studied, the honey samples
from tree geographic regions did not show any signifi-
cant difference in quality parameters except for fruc-
tose, glucose, fructose/glucose ration, lactone acidity,
and electrical conductivity. Quantitative pollen analysis
shows that most of the samples have a moderate
amount of pollen grains per 10 gram of sample (class II).
For the qualitative analysis, A. mellifera in this area uses
a wide spectrum of nectariferous plants; such as Echium
sp., Eucalyptus sp., Ononis natrix, and Ziziphus lotus; as
well as nectarless plants, that had high pollen frequency
percentage of Chenopodium sp., and Olea europaea.
Acknowledgements
We would like to express our sincere thanks to Hadjoudja
Mustapha, manager of Biology Laboratories from Laghouat
University for providing all the necessary facilities to conduct
this research.
Disclosure statement
No potential conflict of interest was reported by the authors.
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