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Physicochemical profile, antioxidant activity and mineral contents of honey

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Physicochemical profile, antioxidant activity and

mineral contents of honey from stingless bee and
honey bee species

Nur Irma Izzati Muhammad & Norizah M. Sarbon

To cite this article: Nur Irma Izzati Muhammad & Norizah M. Sarbon (2021): Physicochemical
profile, antioxidant activity and mineral contents of honey from stingless bee and honey bee
species, Journal of Apicultural Research, DOI: 10.1080/00218839.2021.1896214

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Journal of Apicultural Research, 2021
https://doi.org/10.1080/00218839.2021.1896214

ORIGINAL RESEARCH ARTICLE

Physicochemical profile, antioxidant activity and mineral contents of honey from
stingless bee and honey bee species
Nur Irma Izzati Muhammad and Norizah M. Sarbon

Faculty of Fisheries and Food Science, Universiti Malaysia Terengganu, Terengganu, Malaysia

(Received 4 February 2019; accepted 17 August 2020)

This study aims to investigate the physicochemical profile, antioxidant activity, and mineral contents of honey from
selected honey bee species namely Kelulut, Tualang, and Acacia honey. The findings show that Kelulut honey is superior
due to its high antioxidant and total acidity, low pH, and high colour intensity. The total phenolic content of Kelulut,
Tualang and Acacia honey varied between 12.15 and 26.45 mg GAE/kg, and FRAP value for Kelulut, Tualang, and Acacia
honey ranged between 49.54 and 139.47 mg Fe (II) per kilogram. The total mineral contents founded in all honey sam-
ples ranging from 545.76 to 570.66 mg/kg, with potassium, sodium, and calcium being the most abundant elements.
Keywords: honey bees; stingless bees; Kelulut honey; Tualang honey; Acacia honey; Malaysian bees

Introduction The qualities of honey are mostly centred on physi-

Malaysia is a Southeast Asian country rich in natural for-
est resources such as honey, which is known as a nat-
ural food among consumers and naturally consists of
sugar and other constituents with a broad range of bio-
logical effects and natural antioxidants (Alqarni et al.,
2012). These properties are identified by the mineral
components within the honey, which are usually
dependent on its type and origin (Moniruzzaman et al.,
2014). Even though the compositions of each type of
honey differ according to the plant species on which
the bees forage, the primary constituents within the
honey remain constant for all types (Shahnawaz
et al., 2013).
Honey composition, however, can also be influenced
by the type of bee. Historically, the honey bee is any
bee member of the genus Apis from the tribe of Apini.
The most well-known subspecies is Apis mellifera (Kek
et al., 2014). Other subspecies such as Apis dorsata, Apis
cerana, and Apis florea are mostly found in South and
South-East Asia (Guerrini et al., 2009). One type of
honey produced by Apis dorsata in Malaysia is Tualang
honey, while another type produced by Apis Mellifera is
Acacia honey. Stingless bees are from the tribe
Meliponini and have been reported to consist of more
than one genus, including Melipona, Scaptotrigona, and
Trigona (Kek et al., 2014). The stingless bee species
commonly found in Malaysia is Trigona, known as
Kelulut among Malaysians. The quality and composition
of stingless bee honey have been reported to differ
markedly in terms of colour, taste, and viscosity from
Apis honey bee (Almeida-Muradian et al., 2014).

Corresponding author. Email: norizah@umt.edu.my; norizahsarbon@yahoo.com
cochemical properties including pH, total acidity, colour
intensity, ash, moisture content, and
Hydroxymethylfurfural. Honey content can differ greatly
even within a single area, and this can make honey mar-
keting very difficult for exporters. Furthermore, inter-
national standards for honey quality are still lacking and
sometimes are not suitable for a certain country
(Bradbear, 2009). In addition, honey is well recognized
for its own antioxidant properties due to the presence
of phenolic acids, flavonoids, catalase, peroxides, carote-
noids derivatives, and ascorbic acid as well as peroxides
(da Silva et al., 2013).
Numerous elements have been reported to appear
in different types of honey. These elements are catego-
rized as major elements, trace elements, and ultra-trace
elements. Each element is required in certain amounts
by humans (Alqarni et al., 2012). Major elements should
be present at >50 mg/d; meanwhile, trace elements are
required in concentrations of <50 mg/d (Solayman
et al., 2016). The mineral content in honey is typically
around 0.04–0.2%. Considering the lack of information
on the composition of Malaysian honey, this study aims
to investigate the physicochemical profile and bioactivity
of three types of Malaysian honey (Kelulut, Tualang, and
Acacia honey) originating from different bee species
(stingless bee and honey bee).
Materials and methods
Honey samples
Three types of Malaysian honey samples representing
the Apis spp. or Trigona spp. honey were selected.

ß 2021 International Bee Research Association

2 N.I.I. Muhammad and N.M. Sarbon
Tualang (Koompassia excelsa) and Acacia represented
the Apis spp. Meanwhile, Kelulut honey represented
Trigona spp. All Kelulut, Tualang, and Acacia honey were
purchased fresh from a local supplier in Marang,
Terengganu, Malaysia. All honeys were stored under
refrigeration at 11  C in airtight containers until fur-
ther analysis. All chemicals and reagents used were of
analytical grade.
Physicochemical properties of honey bee and
stingless bee honey
pH determination
The pH of each honey was measured using a pH meter
(Cyberscan series 600, New York). Each honey (5 g)
was dissolved in 20 ml of distilled water. These samples
were measured in triplicate and the results obtained
were expressed as mean ± SD (Chua et al., 2012).
Total acidity determination
The total acidity of the honey by titration method was
measured based on AOAC 962.19 (2006).
Approximately 10 g of honey were dissolved in 75 ml
distilled water and this solution was titrated with 0.05 M
NaOH solution until the pH reached 8.5. About 10 ml
of 0.05 M NaOH was added immediately and back-
titrated with 0.05 M HCl solution until the pH reached
8.3 (lactone acidity) in order to determine the acidity.
Total acidity was determined as follows:
Total acidity ¼ Free acidity þ lactone
Free acidity ¼ ðtitre of 0:05M NaOH  blankÞ  0:05
 ð1000=10Þ
Lactone ¼ ð10:00 – titre of 0:05M HCLÞ  0:05
 ð1000=10Þ
Colour determination
The colour intensity of honey was measured per Kek
et al. (2017). The honey solutions (50%, w/v) were pre-
pared with warm milli-Q water at 45  C–50  C and fil-
tered in order to remove any course particles. Net
absorbance was determined as the difference between
the absorbance at 450 and 720 nm using a UV-VIS spec-
trophotometer (Thermo Fisher Scientific, Massachusetts,
US). Measurements were performed in triplicate and the
results expressed in terms of mAU.
Moisture content determination
The moisture content of honey samples was measured
referring to the AOAC (1990). Approximately 5 g of
each sample was weighed and placed into a pre-weighed
aluminium drying dish (UNB500, Memmert, Germany).
Then, the sample was dried to constant weight in an
oven at 105  C overnight under a vacuum. The
following equation was used to calculate the moisture
content of honey:
Moisture content ð%Þ ¼ ðM1 – M2 Þ=ðM1 – M0 Þ x 100
Where,
M0 ¼ weight of the aluminium dish
M1 ¼ weight of the fresh sample þ dish
M2 ¼ weight of the dried sample þ dish
Antioxidant activity of honey bee and stingless
bee honey
Total phenolic content (TPC) determination
The concentration of phenolics in the honey samples
was estimated using the spectrophotometric Folin-
Ciocalteu method. Briefly, 1 ml of honey solution (0.1 g/
ml) was prepared. The Folin-Ciocalteu phenol reagent
(2 ml) and 2.0 ml of 7.5% sodium carbonate solution
were added. The mixture then was agitated and left to
stand at room temperature for 2 h, after which the
absorbance was read at 760 nm using UV/VIS spectro-
photometer (Thermo Fisher Scientific, Massachusetts,
US). Gallic acid was used to calculate a standard curve
with concentrations of 10, 20, 30, 40, and 50 lg/ml; r2
¼ 0.9986. The equations obtained from the standard
curve were used to calculate the concentration of total
phenolic content of honey sample. The results were
reported as the mean ± standard deviation and
expressed as mg of Gallic acid equivalents (GAEs) per
kilogram of honey (Khomsug et al., 2010).
Ferric reducing-antioxidant power (FRAP) assay determination
FRAP assays were conducted following Moniruzzaman,
Sulaiman, et al. (2013). Approximately 200 mL (0.1 g/ml)
was mixed with 1.5 ml FRAP reagent. Then, the reaction
mixture was incubated for 4 min at 37  C and its
absorbance was read at 593 nm against a blank prepared
with distilled water. Fresh sample reagent was prepared
by mixing 10 volumes of 300 mM/L acetate buffer (pH
3.6) with 1 volume of 10 mM TPTZ solution in 40 mM/L
HCL containing 1 volume of 20 mM ferric chloride
(FeCL3.6H2O). A calibration curve was prepared using
an aqueous solution of ferrous sulphate (FeSO4.7H20)
at 0.1  1.0 mmol/L. The equation obtained from the
standard curve was used to calculate the FRAP values
of honey and expressed in terms of micrograms of fer-
rous equivalent per kilogram of honey.
Mineral properties of honey bee and stingless
bee honey
Sample preparation
Sample preparation for mineral analysis was conducted
following Shadan et al. (2018). Honey (1 g) was diluted
with 10 ml of HNO3 (65%, v/v). Then, each solution
was sonicated at 5000 rpm for 10 min followed by
 Physicochemical profile, antioxidant activity and mineral contents of honey
Table 1. Physicochemical properties of selected Malaysian honey (Kelulut, Tualang, and Acacia honey) from different bee species.
Honey pH Total acidity (meq/kg)
Kelulut 3.81 ± 0.02c 89.33 ± 1.04a
Tualang 4.13 ± 0.02b 41.50 ± 1.00b
Acacia 4.20 ± 0.02a 31.17 ± 1.04c
Note. Means were compared using one-way ANOVA with post hoc multiple comparisons. In each column, values with different letter (a-c) indicate
significant differences (p < 0.05); means followed by the same letter in the column do not differ statistically (p > 0.05); all values are expressed as
means of triplicate determinations ± standard deviation.
centrifugation for 15 min. In order to remove the sus-
pension, the pellet was filtered using a syringe filter.
Then, the cleaned pallet was transferred into a clean fal-
con tube, sealed with parafilm, and kept at room tem-
perature prior to analysis. Using the same sample
preparation method, a standard solution containing
10 ml of HNO3 (65%, v/v) was prepared.
Mineral content determination
For quantitation, standard solutions of multi-element stand-
ards (1000 mg/l, Perkin Elmer, USA) were prepared by
diluting with suitable amounts of HNO3 (65%) in serial
dilution. The concentrations of the standard solutions
were 0.5, 1.0, 5, and 10 mg/l. The analytical parameters
standardized for ICP-OES (Perkin Elmer, USA) used were
radio frequency power (0.7–1.5 kW, 1.2–1.3 kW for axial),
plasma gas flow rate (Ar) (radial: 10.5–15 l/min, axial: 15 l/
min), auxiliary gas flow rate (Ar: 1.5 l/min) and viewing
height (5–12 mm) (Shadan et al., 2018).
Data analysis
All analyses were conducted in triplicate and the data
are presented as mean ± standard deviations. Analysis of
variance (ANOVA) was used in order to compare the
quantified variables in the samples of honey. All statis-
tical analyses were performed using SPSS Statistics and
a p-value  0.05 was considered to be significant.
Results
Physicochemical properties of honey bee and
stingless bee honey
pH properties of honey
The pH values of the honey samples are presented in
Table 1. Kelulut honey was found to have the most
acidic taste with the lowest pH value. There were sig-
nificant differences in pH values among all samples
(p < 0.05). The pH value variation between the Kelulut,
Tualang, and Acacia honey can be explained by the
presence of numerous organic acids. These organic
acids play an important role in the characteristic fla-
vours and aromas of honey.
Total acidity of honey
The total acidity levels of the honey samples are pre-
sented in Table 1. The total acidity of Kelulut honey
3
Colour intensity (mAU) Moisture contents (%)
0.44 ± 0.00a 29.20 ± 0.10a
0.11 ± 0.00b 23.60 ± 0.78b
0.10 ± 0.00b 21.73 ± 1.48b
was significantly higher than those of Tualang and
Acacia honey (p < 0.05). The variation of total acidity of
honey is strongly associated with the balance of organic
acids present in honey, which vary in terms of floral
composition and the bee species (de Sousa et al., 2016).
The present results showed only Tualang and Acacia
honey were in an acceptable range while Kelulut honey
exceed the standards set by the Codex Alimentarius
(8.7–59.5 meq/kg) (codex, 2001).
Colour intensity of honey
The colour intensities of the honey samples are pre-
sented in Table 1. There were significant differences in
colour intensity between Apis spp. and Trigona spp.
(p < 0.05). However, there were no significant differen-
ces in colour intensity between honeys from the same
genus (genus Apis). Based on the results obtained, the
colour intensities of Kelulut, Tualang, and Acacia honey
vary, even though all samples were obtained from the
same region. Kelulut honey had the highest value of col-
our intensity as compared to Tualang and Acacia honey,
indicating its high antioxidant potential.
Moisture content of honey
The moisture contents of different honey samples are
presented in Table 1. The highest moisture content was
shown by Kelulut honey, with significant differences
between Apis spp. and Trigona spp. (p < 0.05). However,
there were no significant differences in moisture con-
tent between honeys from the same genus (genus Apis).
Based on the results obtained, all three types of
Malaysian honey exceeded the maximum permitted
value of the moisture content for honey.
Antioxidant activity of honey bee and stingless
bee honey
Total phenolic content (TPC) of honey
The total phenolic content obtained for three types of
Malaysian honey (Kelulut, Tualang, Acacia honey) is
shown in Table 2. There was a significant difference in
total phenolic content between honeys (p < 0.05).
Based on the present results, the honey produced by
Trigona spp. (Kelulut honey) showed the higher values
of total phenolic content than honey produced by Apis
spp. (Tualang and Acacia honey).
4 N.I.I. Muhammad and N.M. Sarbon

Note. Means were compared using one-way ANOVA with post hoc multiple comparisons. In each column, values with different letter (a-b,) indicate significant differences (p < 0.05); means followed by the
0.46 ± 0.11b
0.41 ± 0.21b

same letter in the column do not differ statistically (p > 0.05); all values are expressed as means of triplicate determinations ± standard deviation; Na (sodium); Mg (magnesium); Ca (calcium); K (potassium);
Table 2. Antioxidant activity of selected Malaysian honey

0.95 ± 0.23a
Table 3. Total mineral contents, major elements, trace elements, and heavy metals of selected Malaysian honey samples (Kelulut, Tualang, and Acacia honey) from different bee species.

(mg/kg)
(Kelulut, Tualang, and Acacia honey) from different

Cu
bee species.
TPC FRAP
(mg GAE/ (mg of Fe (II)

Trace element and heavy metal of honey

Honey kg honey) equivalent/kg honey)
26.45 ± 1.65a 139.47 ± 11.53a

0.03 ± 0.00a
0.03 ± 0.01a
0.02 ± 0.00a
Kelulut

(mg/kg)
Tualang 16.32 ± 0.44b 89.29 ± 5.40b

Cd
Acacia 12.15 ± 0.74c 49.54 ± 3.98c
Note. Means were compared using one-way ANOVA with post hoc
multiple comparisons. In each column, values with different letter (a-c)
indicate significant differences (p < 0.05); all values are expressed as
means of triplicate determinations ± standard deviation; TPC (total

1.43 ± 0.23a
1.55 ± 0.89a
1.17 ± 0.30a
phenolic content); FRAP (ferric reducing antioxidant potential).

(mg/kg)
Zn
Ferric reducing-antioxidant power assay (FRAP assay)
The Ferric Reducing-Antioxidant Power (FRAP) Assay
that obtained Kelulut, Tualang, Acacia honey is shown

10.90 ± 3.92a
10.62 ± 1.54a
12.83 ± 4.69a
(mg/kg)
in Table 2. There was a significant difference in FRAP

Fe
assay between Kelulut, Tualang, and Acacia honey
(p < 0.05). Meanwhile, the lowest value of FRAP assay
was exhibited by Acacia honey, showing its low antioxi-
dant potential. FRAP values indirectly indicate a larger

370.65 ± 25.19b
429.21 ± 22.90a
reduction of ferric ions to ferrous ions. This phenom-

466.56 ± 5.17a
(mg/kg)
enon was further described by El-Haskoury et al.

K
(2018), as the presence of reducing agents in the honey
reduces the ferric ions to ferrous ions.

Mineral properties of Kelulut, Tualang, and Acacia honey

The total elements obtained for three types of 46.23 ± 10.34b
40.61 ± 1.64b
59.513 ± 4.01a
(mg/kg)

Malaysian honey (Kelulut, Tualang, Acacia honey) are

Major elements of honey
Ca

shown in Table 3. There was a significant difference in

mineral content between honey (p > 0.05). Overall, the
total mineral content, including major elements, trace
elements, and heavy metals found in examined honey
samples decreased in the following order:
10.09 ± 0.574a

6.34 ± 0.09b
7.60 ± 0.72b

Acacia > Kelulut > Tualang.

(mg/kg)
Mg

Discussion
A major factor that might contribute to the sour taste
with the high level of acidity in Kelulut honey is its
50.08 ± 0.79b
42.72 ± 1.88b
108.78 ± 6.40a

moisture content. It has been proven that Kelulut

(mg/kg)

honey from stingless bees has higher moisture content

Na

Fe (iron); Zn (zinc); Cd (cadmium); Cu (copper).

than honey from honey bees, such as Tualang and

Acacia honey. This high moisture content strongly cor-
relates with the fermentation process by the osmophilic
yeasts. This process leads to the breakdown of alcohol
to acetic acid and water, which eventually gives a sour
contents (mg/kg)
562.35 ± 35.53a
545.76 ± 12.18a
Total mineral

570.66 ± 8.66a

taste to the honey (Prica et al., 2015). The present

results are in agreement with those of Chuttong et al.
(2016), who reported that the honey produced from
stingless bee had a pH value, with a range of 3.1–3.9.
Similarly, this finding was supported by another study by
Biluca et al. (2016) who found that the pH value of
stingless bee honey collected from Brazil ranged from
Tualang
Kelulut
Honey

Acacia

3.33 to 6.56. The pH values for all the honey samples

were in accordance with the acceptable range suggested
 Physicochemical profile, antioxidant activity and mineral contents of honey
by the Codex Alimentarius (3.4  6.1) for honey (codex,
2001). This range can be used to determine the fresh-
ness of honey. Thus, the acidic honey with the lowest
pH value was that produced by the stingless bee spe-
cies, namely Kelulut honey.
Honey within this acidity range is capable of enhanc-
ing the shelve stability of the product and eventually
lead to the prevention of spoilage by microorganisms
(Lawal et al., 2009). Sari and Ayyildiz (2012) explained
that high total acidity may indicate that the honey has
fermented at some time and this fermentation was
done by the osmophilic yeasts on the glucose and fruc-
tose, forming alcohol and carbon dioxide. This process
eventually leads to the breaking down of alcohol into
organic acid with the presence of oxygen. To date,
studies conducted on stingless bee honey remain scarce,
as only two studies have reported on the total acidity
in stingless bee honey which was in Brazil and Thailand
(Biluca et al., 2016). According to Biluca et al. (2016),
the total acidity of stingless bee honey obtained from
the Brazil region lay in the range from 16.2 to 139 meq/
kg. These results were in agreement with the current
study, which showed that the total acidity for Kelulut
honey produced by Trigona spp. was 89.33 ± 1.04 meq/
kg. Stingless bee species (Kelulut honey) obtained the
highest total acidity.
Theoretically, the color intensity of honey was con-
sidered as a reliable parameter that represents the
existence of pigments which represent antioxidant activ-
ity, such as carotenoids, phenolic, and flavonoids. All
these compounds can be easily found in pollen, nectars,
resin, and oil collected by bees as food (Abeshu &
Geleta, 2016). One of the factors that may contribute
to the higher colour intensity of Kelulut honey is the
size of Trigona spp. itself. Trigona spp. are naturally
smaller in size compared to other honey bees like Apis
dorsata and Apis mellifera. Due to that, they are capable
of taking flower nectar from the deepest region of blos-
soms. As a result, Kelulut honey possessed higher nutri-
tional values and antioxidant activities as compared to
honey produced by the Apis spp. The present results
are in agreement with the study done by Kek et al.
(2014), in which Trigona spp. had a higher value of col-
our intensity as compared to the Apis spp. This finding
proved that Kelulut honey from the Trigona spp. has
higher levels of colour intensity than honey from the
Apis spp. This colour measurement can be a good indi-
cator for estimating the total phenolic content
of honeys.
Pursuant to valid Regulation by Codex Alimentarius,
the maximum moisture content in honey allowed in the
market is fixed to 20%. Nevertheless, 20% only repre-
sents the limit and does not indicate the preferred or
appropriate moisture content for honey. This is because
even though the honey contains moisture content at
around 17% if it contains a sufficient amount of yeast
spores, it still will ferment (White & Doner, 1980).
5
Technically, the moisture content of honey is consid-
ered as the most crucial parameter in determining the
quality of honey, since it determines the stability and
resistance of honey towards microbial spoilage during
storage. The high moisture content in honey represents
a higher probability for the fermentation of yeasts to
occur and this will lead to the changes of honey flavour
which was become sourer than usual (Prica et al.,
2015). Generally, honey samples from tropical countries
like Malaysia possessed high moisture content. Because
of that, the chances for fermentation to occur were
quite high and it was hard to be avoided. In order to
improve the quality of honey, the fermented honey can
sometimes be reclaimed by heating it to 65  C for a
short time. This method will stop the fermentation pro-
cess and at the same time expel some of the off-flavour.
This finding is in agreement with a previous study done
by Chuttong et al. (2016) in which the range of mois-
ture content found in stingless bee honey in Thailand
was 25–47%. Similarly, stingless bee honey from Brazil
had moisture content values in the range between
25.99 and 36.89% (do Nascimento et al., 2015). Thus,
the findings are in agreement with all previous studies,
as all the stingless bee honeys showed values higher
than those recommended by the Codex Alimentarius,
indicating that the essential regulations are geared
towards stingless bee honeys, with their own physico-
chemical characteristics (Biluca et al., 2016).
The variation in the total phenolic content of exam-
ined honey samples was due to its dependence upon
floral sources of honey and its collection region (Khalil
et al., 2011). In addition, the colour intensity of honey
was described as a valid parameter to determine the
presence of pigments which has antioxidant activities,
for instance, carotenoids and flavonoids (Moniruzzaman,
Khalil, et al., 2013). According to Blasa et al. (2006),
generally, darker honey tended to possess higher phen-
olic contents as compared to lighter coloured honey.
The present results are in agreement with the study
reported by Blasa et al. (2006) in which Kelulut honey
with higher colour intensity also possessed a higher
value of total phenolic contents. The total phenolic con-
tents of the Kelulut, Tualang, and Acacia honey meas-
ured in this study were similar to those reported by
Kek et al. (2014) where the highest total phenolic con-
tent in their study was shown by Kelulut honey fol-
lowed by other Malaysian Apis honey. However, the
total phenolic content recorded in this study was too
low when compared to the study by Kek et al. (2014),
which found a range of 206.3–791.5 mg GAE/kg honey.
The Kelulut honey from the Trigona spp. contains higher
phenolic content as well as higher colour intensity than
honeys from the Apis spp. Thus, the colour measure-
ment may be used as a reliable indicator of total phen-
olic contents in honey.
Based on the present result, the higher FRAP values
of Kelulut honey may be due to its stronger antioxidant
6 N.I.I. Muhammad and N.M. Sarbon
properties as compared to other honey, indicating a
larger reduction of Fe3þ to Fe2þ ions corresponding to
samples with a higher reducing power that increase in
absorbance. The FRAP value recorded in this study was
too low as compared to a previous study by
Moniruzzaman, Khalil, et al. (2013) with the range of
87.47–576.91 mM Fe (II)/100 g. This was expected, as
the properties and compositions of honey are greatly
influenced by various factors such as nectar sources,
botanical origins, and type of bee. Additionally, in com-
parison to Tualang honey in other places in Malaysia,
the FRAP value recorded in this study was substantially
lower than those reported by Mohamed et al. (2010),
at 322.1 mM Fe(II) per 100 g. This indicates that honey
collected from Marang, Terengganu has lower antioxi-
dant potential when compared to other states
in Malaysia.
The present results are not in agreement with the
study conducted by Moniruzzaman et al. (2014), which
stated that both antioxidant activity and colour intensity
strongly correlated with the mineral content of honeys.
This level of minerals within honey was not in accord-
ance with what it should be because, in this study, only
8 minerals were tested. Meanwhile, in general, around
54 minerals have been reported in honey around the
world (Ruschioni et al., 2013).
Based on Table 3, all honey samples showed a broad
range of potassium (370.65–466.56 mg/kg). Acacia honey
showed the highest amount of potassium, followed by
Tualang and Acacia honeys. The high amount of potas-
sium within the examined honey samples is in agree-
ment with a study by Moniruzzaman et al. (2014), who
reported that potassium was the most abundant mineral
found in fourteen samples of different honeys collected
from different regions in Malaysia. In contrast, the con-
centrations of potassium recorded in this study were
quite low as compared to a study conducted by
Moniruzzaman et al. (2014). The huge difference in
terms of concentration between the present study and
previous study was due to geographical origin and flo-
ral sources.
Similarly, the present study showed all the honey
samples possessed high concentrations of sodium, rang-
ing from 42.72 to 108.78 mg/kg. Kelulut honey is rich in
sodium. This may be explained by the presence of
greater amounts of this mineral in the plants from
which the bees collected the pollen. In comparison, the
concentration of sodium in the present study was
higher than honey from Poland (21.6–28.4 mg/kg)
(Chudzinska & Baralkiewicz, 2010). This discrepancy
might be from the geographical variation in the sources
of honey. Additionally, the mean concentrations of Na
in the Malaysian honey samples including Kelulut,
Tualang, and Acacia in the present investigation were
higher as compared to Irish (31 mg/kg), and Turkish
(33 mg/kg) honey (Moniruzzaman et al., 2014).
High concentrations of calcium (40.610–59.513 mg/
kg) were also observed among all the examined honey
samples. In comparison to other countries, the concen-
tration of calcium in Malaysian honey (Kelulut, Tualang,
and Acacia honey) was higher than that reported by
Yilmaz and Yavuz (1999) for honeys from Italy at
around 47.7 mg/kg.
Besides major elements, a few trace elements (cad-
mium, copper, and zinc) which are also heavy metals
were detected in all honey samples (Table 3). From a
nutritional standpoint, it is essential to investigate the
contents of trace elements within honey samples due to
the potential for toxicity. Based on the present result,
all the heavy metals were present less than 1 ppm
except for zinc that presents 1 ppm. The mean concen-
trations of heavy metals found in examined honey sam-
ples like Tualang, Kelulut, and Acacia honey decreased
in the following order: Zinc > Copper > Cadmium.
In the present study, the concentrations of copper
(Cu) ranged between 0.40 and 0.95 mg/kg. In compari-
son to other countries, concentrations of copper in the
present study were higher than honeys from Greece
(01.4–0.48 mg/kg) (Ioannidou et al., 2005), and the Lazio
region (0.3 mg/kg) (Conti, 2000), respectively.
Moreover, levels of cadmium (Cd) in the Kelulut,
Tualang, and Acacia honey ranged from 0.02 to 0.03 mg/
kg. In comparison to honeys from other countries, the
concentrations of cadmium were much lower as com-
pared to Manuka honey with a value of 1.01 mg/kg.
However, this is in accordance with a previous study
done by Moniruzzaman et al. (2014), as the levels of
cadmium in those honey ranged from 0.00 to 1.03 mg/
kg. These values are quite similar with the present
results. In addition, among all honeys investigated,
Tualang honey was the richest in zinc. Even though the
concentrations were highest among the trace elements
detected in the honey samples, the concentration of
zinc can still be considered as low in comparison with
honey from other countries such as Spain (3.9 mg/kg),
Turkey (2.7 mg/kg), and India (12.69 mg/kg)
(Moniruzzaman et al., 2014).
Even though several trace elements were detected in
the Kelulut, Tualang, and Acacia honey, the concentra-
tions of these trace elements were still in the provi-
sional tolerable daily intake (3 mg). This value is the
limit for metal intake based on the body weight of an
average 60 kg adult (Biland
zic et al., 2011). In general,
this study has shown that all three types of honey were
rich in minerals content.
Conclusions
In conclusion, Kelulut honey (Trigona spp.) was generally
superior to Tualang (Koompassia excelsa) and Acacia
(Apis spp.) honey due to its high antioxidant and total
acidity, colour intensity value, standard of the minerals,
and heavy metal low pH levels compared to Tualang
and Acacia honey. Acacia honeys possessed a moderate
 Physicochemical profile, antioxidant activity and mineral contents of honey
amount of total phenolic content as compared to other
studies. Results indicate that the honeys are all safe
to consume.
Acknowledgement
This research did not receive any specific grant from funding
agencies in the public, commercial or not for profit sectors.
There was no conflict of interest between authors.
Disclosure statement
No potential conflict of interest was reported by the authors.
ORCID
Norizah M. Sarbon http://orcid.org/0000-0003-0904-1039
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