CSJ 15(1): June, 2024 ISSN: 2276 – 707X, eISSN: 2384 – 6208 Ahmed and Musa

ChemSearch Journal 15(1): 162 – 173, June, 2024

Publication of Chemical Society of Nigeria, Kano Chapter

Received: 02/05/2024 Accepted: 09/06/2024

http://www.ajol.info/index.php/csj

Determination of some Heavy Metals and Selected Phenolic Compounds in

Commercial Honeys obtained from Northwestern States of Nigeria
1Ahmed, U.U. and 2Musa, M.S.
1
Department of Pharmaceutical Technology, School of Technology, Kano State Polytechnic, Kano, Nigeria
2
Department of Pure and Industrial Chemistry, Bayero University, P.M.B 3011 BUK, Kano, Nigeria
*Correspondence Email: [email protected]

ABSTRACT
Natural honey is one of the highly needed products because of its exclusive, high nutritive and medicinal
properties, it is among the most adulterated products globally. Honey adulteration is a global concern, it has
negative effects on the nutrition and health of consumers and has become a common practice because of the
high demand and limited availability of the product. This research aims to evaluate the heavy metals
concentrations and the phenolic compounds in honey samples obtained from local sellers and compare the levels
with honeys from credible beekeepers in Northwestern states of Nigeria (Kano, Kaduna, Jigawa, Katsina,
Sokoto, Zamfara and Kebbi) in order to detect possible adulteration of the products. Results indicated that most
of the heavy metals detected Cd (ND-0.10±0.16 µg/g), Pb (0.20±0.11-1.62±1.60 µg/g), Ni (0.03±0.04-
0.53±0.46 µg/g), Cu (0.47±0.23-2.61±2.11 µg/g), Zn (5.75±4.04-97.63±131.74 µg/g) and Fe (6.74±1.92-
61.43±98.12 µg/g) in honeys from local sellers were above the permissible range set by the International Honey
Commission (IHC) and NAFDAC while the levels in samples obtained directly from beekeepers closely
complied with the standards. The non-compliance of the metal levels with IHC standards in locally sold honeys
could possibly be due to adulteration of the products using metal-contaminated adulterants. However, lower
values were recorded for Phenolic compounds: flavonoids (154.5-521.7 mg/kg) and Phenolic acids (399.6-
1075.0 mg/kg) in most of the honeys from local sellers except in honeys from Zamfara state. Generally, results
from this study indicate that large percentage of the honey products sold locally in the Northwest Nigeria are
suspected to be adulterated mostly with sweeteners while honeys obtained directly from beekeepers were found
to agree with IHC standards. It is therefore recommended that pure honeys should be obtained directly from
credible beekeepers.

Keywords: Adulteration, Flavonoids, Heavy Metals, Honeys, Phenolic Acids

INTRODUCTION modern. It is one of the products most widely

Honey produced by the honey bee is a sorted for due to its unique nutritional and
natural super saturated sugar solution which has medicinal properties.
been seen as a highly nutritive food and is Honey contains a variety of macro and
composed of a complex mixture of carbohydrates, micro minerals that are the minor constituents of
minerals, vitamins, aromatic compounds, honey present in the range of 0.02–2.03%. Honey
flavouring and enzymes with the water content of contains high levels of potassium, sodium, calcium
about 17 – 20% (Haouam et al., 2016). The major and magnesium, the sources of which were shown
composition of honey are carbohydrates and water to be pollen and nectar (Altun et al., 2017; Batista
(Agbajor and Otache, 2020). It is a high-energy et al., 2012). Apart from the mentioned minerals,
carbohydrate food as the honey sugars are easily honey may contain Fe, Cr, Se, Cu, Mn, Zn, Al, Pb,
digestible as those in many fruits (El-Sohaimy et As, Cd and Hg, depending mainly on
al., 2015). environmental factors (Mutlu et al., 2017). Trace
Honey is one of the major bee products, a elements are mainly the ash content of honey
semi liquid, sweet and flavored food stuff produced (Altun et al., 2017).
from nectar of nectarines of flowers, or secretion of The presence of metals in honey has the
plant-sucking insects which the bees collect, potential to threaten the health of humans as
transform by addition of specific substances of consumers. Lead, arsenic, cadmium and mercury
their own, deposit, dehydrate, store and leave in are among the most abundant heavy metals and are
honeycomb to ripen and mature (Majewska et al., particularly toxic. The excessive amount of these
2019). Natural honey is a liquid mentioned in all metals in food is associated with etiology of a
religious books, and accepted by all generations, number of diseases especially with cardiovascular,
traditions and civilizations, both ancient and

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kidney, respiratory, nervous and as well as bone world several million years ago (Eteraf-Oskouei
diseases (Ernest et al., 2018). and Najafi, 2012).
Honey also contains phytochemicals such
as flavonoids and other polyphenols that makes it a MATERIALS AND METHODS
potential functional ingredient and an antibacterial The materials used and methods adopted
agent (Ndife et al., 2014). Of the polyphenols, in this research are as described:
phenolic acids are likely to be the major group in Pure standards of Rutin (RUE), Gallic acid (GAE)
honey (Chua et al., 2013). It is well known as a and Folin-Ciocalteu were purchased from Sigma-
natural dietary antioxidant. The components Aldrich Chemical Company (Steinheim, Germany).
responsible for the redox properties of honeys are Similarly, HNO3 and NaNO3 reagents were sourced
likely to be phenolic acids, flavonoids, vitamins from Sigma-Aldrich. All other materials and
and enzymes, as well as small amount of mineral solvents used were of analytical grade.
content, particularly copper and iron (Gul and
Pehlivan, 2018). Interestingly, they have been Sampling
given considerable attention to be an eligible One hundred and five (105) honey
parameter for honey quality assessment (Lewoyehu samples were randomly purchased from open
et al., 2019). The use of natural honey as food and markets within Northwestern states (Kano, Jigawa,
medicine by mankind has been in existence since Kaduna, Katsina, Sokoto, Zamfara and Kebbi),
ancient times. Nigeria. Each state was divided into three
Raw honey is the most ancient sweetener, senatorial districts: Central, North and South. Five
and it was noted to have been in use. The samples (control sample inclusive) were collected
components responsible for the redox properties of from each district making a total of 15 samples
honeys are likely to be phenolic acids, flavonoids, from each state. The samples from central
vitamins and enzymes, as well as small amount of senatorial districts in Kano state were labeled as
mineral content, particularly copper and iron (Chua KN CI, KN C2, KN C3 and KN C4, while the
et al., 2013). Interestingly, they have been given control sample was designated KN CC. Also, the
considerable attention to be an eligible parameter samples from Kano North were labeled as KN N1,
for honey quality assessment (Lewoyehu et al., KN N2, KN N3 and KN N4 with the control
2019). Raw honey contains an enzyme called sample as KN NC. Likewise, samples from south
glucose oxidase which combines with water to districts were labelled as KN SI, KN S2, KN S3
produce hydrogen peroxide (H2O2), a mild and KN S4, while the control sample as KN SC.
antiseptic substance (Ndife et al., 2014). Honey This identification trend was used in all other
also contains phytochemicals such as flavonoids States. The honey samples were obtained
and other polyphenols that makes it a potential commercially while the pure honey samples used
functional ingredient and as an antibacterial agent as control were obtained directly from bee keepers
(Ramanauskiene et al., 2012). of the polyphenols, in each State. All the samples were collected in
phenolic acids are likely to be the major group in sterile containers, labeled and stored in a
honey, they are well known as a natural dietary refrigerator in airtight plastic containers until
antioxidant (Chua et al., 2013). throughout the analysis.

Fig. 1: Map of the Sampling Sites

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METHODOLOGY The results were expressed as mg Gallic acid
Extract of Hoey Sample equivalent (GAEs) per kg honey (Abu-Bakar et al.,
The honey sample (6 cm3) was dissolved 2017).
in 2 cm methanol and made up to 60 cm3 with
3

deionized water and left overnight. The mixture RESULTS AND DISCUSSION
was filtered using Whatmann filter paper and Statistical Analysis
stored in a refrigerator until analysis All the tests were done in triplicate and the
data were expressed as mean ± standard deviation
Digestion Procedure (SD). Statistical significance of differences was
Honey sample (5 g) was weighed into a determined using a one-way Analysis of Variance
clean pre-weighed porcelain crucible. It was placed (ANOVA) and the Duncan Multiple range test with
on a hot plate and heated at 300ºC until sample was significant level at 95% (P<0.05) were considered
dried in order to prevent loss by foaming. It was significant.
then ashed to constant weight in a muffle furnace at
a temperature of 550ºC. Constant weight was Heavy Metals Concentration in Honey Samples
obtained by continually heating the sample, cooling The mean concentration of heavy metals
in a desiccator and weighing at 2–3 hours intervals varied significantly across most of the studied
for about 8 hours until no change in weight was states with no specific pattern and were in
observed. The ash obtained was then dissolved in decreasing order of Zn > Fe > Cu > Pb > Ni > Cd.
50 cm3 of 0.5M HNO3. The same procedure was The mean cadmium concentrations in all
carried out for all the samples. All the digests and the studied honey samples obtained from all study
sample blank were analysed for Zn, Cd, Fe, Cu, Ni areas ranged from ND µg/g in honey samples from
and Pb using Microwave Plasma Atomic Emission source to the highest 0.10 µg/g in honey samples
Spectrophotometer (The agilent 4210). from Kano and Katsina (Fig. 2). The concentrations
of cadmium (0.10, 0.10, 0.07 µg/g) in honey
Determination of Total Flavonoid Content samples collected from Kano, Katsina and Kaduna
The total flavonoid content in each sample respectively had exceeded the permissible limit of
was measured using the colorimetric assay. Honey 0.05 µg/g set by World Health Organization
extract (1 cm3) was mixed with 4 cm3 of deionized (WHO, 2015) and were significantly higher
water. Then 0.3 cm3 of NaNO2 (5% w/v) was (p<0.05) compared with honey samples obtained
added. After five minute, 0.3 cm3 of AlCl3 (10% from source (control) and other states. The high
w/v) was added followed by 2 cm3 of NaOH (1M) concentration of cadmium might be attributed to
and left for 6 min. The volume was made up to 10 the high traffic emissions, poor sewage system and
cm3 with deionized water. The mixture was use of fertilizers. Cadmium is released into the
vigorously shaken to ensure adequate mixing and environment through its use in various industrial
the absorbance was read at 510 nm using UV/VIS processes, and enters the food chain from uptake by
Spectrophotometer. A calibration curve (Fig. 9) plants from contaminated soil or water. Therefore,
was created using a standard solution of Rutin 20; the cadmium concentration in various places
40; 60; 80; 100;120;140; 160; 180 and 200 µg/ml. depends on many variables, leading to its different
The results were expressed as mg Rutin equivalents concentration in honey samples in the studied
(RUE) per kg of honey (Abu-Bakar et al., 2017). place. The mean concentration of cadmium in the
present study was comparable with < 0.3 µg/g
Determination of Total Phenolic Acid Content reported by Chukwujindu et al. ( 2015) in honey
The concentration of phenolic in honey consumed in Nigeria, 0.07 – 0.1 µg/g in honeys
samples was estimated using modified from Brinin-Gwari, Nigeria (Idoko et al., 2018),
spectrophotometric Folin-Ciocalteu method. Honey 0.028 – 0.07 µg/g in some states in Northern part
extract (1 cm3) was mixed with 1 cm3 of Folin- of Nigeria (Odoh et al., 2015), 0.001 – 0.1 µg/g in
Ciocalteu reagent (1:1). After 3 min, 1 cm 3 of 10% and around the university of Ilorin Environ., Kwara
Na2CO3 solution was added to the mixture and State, Nigeria (Okeola et al., 2020) and 0.02 – 0.05
made to 10 cm3 with deionized water. The reaction µg/g from selected villages in five Local
was kept in the dark for 15 min, after which the Government areas of Adamawa state, Nigeria
absorbance was read at 725 nm using UV/VIS (Toma et al., 2020). Cadmium concentrations in
Spectrophotometer. Gallic acid was used to plot a this study were lower than 0.05 – 0.76 µg/g from
standard calibration curve at 20, 40, 60, 80; 100; different places in Karnataka (Singh et al., 2014).
120; 140; 160; 180; 200 and 220 µg/ml (Fig. 8).

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0.12

Concentration (µg/g)
0.1 0.1
0.1
0.08 0.07

0.06 0.05
0.04
0.04 0.03 0.03
0.02
0.02
0
0
Kano Jigawa Katsina Kaduna Kebbi Sokoto Zamfara Control IHC
Limits
Study Areas
Fig 2: Mean Concentration of Cd (µg/g)

The mean concentration of lead in honey pollutants that can reach human system through air,
samples obtained from the study areas varied from water and food. This metal has no beneficial role in
the lowest of 0.20 µg/g in control samples from human metabolism and produces a progressive
source to the highest of 1.62 µg/g in honey samples toxicity and can cause health disorders (Singh et
from Kano (Fig. 3). The mean concentrations of al., 2014). The mean concentration of lead in the
lead in all the honey samples obtained from all the analyzed samples is in agreement with 0.8 – 1.2
study areas have exceeded the permissible limit of µg/g reported by Mahmoudi et al. (2015) in honey
0.5 µg/g set by WHO (2015) with the exception of from North-Western region of Iran. It is however
samples from source (0.20 µg/g) and Jigawa state higher than 0.041 – 0.087 µg/g previously reported
(0.30 µg/g). The mean concentration of lead in for honey from some of the states in Northern part
honey samples collected from Kano, Sokoto, and of Nigeria (Odoh et al., 2015), 0.175 – 0.35 µg/g in
Zamfara study areas differed significantly (p<0.05) honey produced within Nsukka and Enugu
with the control (0.20 µg/g). More so, the metropolis (Ernest et al., 2018). Similarly, it is
maximum lead concentration in honey samples lower than 1.72 – 2.97 µg/g for honeys from Brinin
obtained from Kano was significantly higher Gwari, Nigeria (Idoko et al., 2018), 0.23 – 2.53
(p<0.05) compared to mean concentrations in µg/g in honey from different regions of Ethiopia
honey samples collected from all the study areas. (Esubalew et al., 2020), 0.28 – 4.93 µg/g in honey
The possible source of high lead concentration in consumed in Nigeria (Chukwujindu et al., 2015)
these honey samples may be associated with and 0.2 – 4.2 µg/g for honeys from different places
vehicular emission and illegal lead mining in those in Karnataka (Singh et al., 2014).
regions. Lead is one of the most widespread metal

1.8 1.62
Concentration (µg/g)

1.6
1.33
1.4 1.18
1.2
1 0.79 0.78 0.84
0.8
0.6
0.33 0.3
0.4 0.2
0.2
0
Kano Jigawa Katsina Kaduna Kebbi Sokoto Zamfara Control IHC
Limits
Study Areas
Fig 3: Mean Concentration of Pb (µg/g)

Nickel (Ni) was detected in all the were higher than the permissible limits set by
samples of honey analyzed. The mean WHO (2015) of 0.25 µg/g with the exception of
concentration of nickel in honey samples obtained samples obtained from Jigawa (0.03 µg/g), Katsina
from the study areas ranged from the minimum of (0.06 µg/g) and source (0.17 µg/g). The mean
0.03 µg/g in honey samples from Jigawa to concentrations of nickel in honey samples across
maximum of 0.53 µg/g in honey samples from most of the study areas varied significantly
Kaduna (Fig. 4). The concentration of nickel in (p<0.05), however nickel concentrations in honey
honey samples obtained from all the study areas samples (0.53, 0.51, and 0.51 µg/g) from Kaduna,

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Kano and Zamfara respectively, were significantly concentration range of nickel in honey samples has
higher compared with concentration in honey been reported in the literature (Chukwujindu et al.,
samples from source (control). Nickel originates 2015). The mean concentrations of nickel in the
most of the times from the combustion of fossil present work are similar to 0.25 – 0.56 µg/g
fuels, source of the emission of ultrafine metal- reported by Tutun et al. (2019) but were however,
containing particles. These airborne particles lower than 0.25 – 6.98 µg/g and 1. 36 – 3.92 µg/g
eventually deposit on vegetation, soil or surface previously reported by Chukwujindu et al. (2015)
water where bees take up heavy metals from the and Salihaj and Bani (2017) respectively
environment (Costa et al., 2019). A wide

0.6 0.53
0.51 0.51 0.5
Concentration (µg/g)

0.5 0.45
0.39
0.4
0.3
0.2 0.17

0.1 0.06
0.03
0
Kano Jigawa Katsina Kaduna Kebbi Sokoto Zamfara Control IHC
Limits
Study Areas
Fig 4: Mean Concentration of Ni (µg/g)

The mean concentration of copper in all adverse health effects in the body (Aghamirlou et
honey samples analyzed ranged from the lowest of al., 2015). The results obtained are found to be
0.47 µg/g in honey samples from Jigawa to the consistent with 0.72–2.57 µg/g reported by
highest of 2.61 µg/g in honey samples from Kano Njokuocha et al. (2019) for honeys from different
(Fig. 5). The mean concentration of copper in all locations in Nigeria, 0.31–2.482 µg/g reported by
honey samples obtained in this study were below Chuleeporn et al. (2018) for honeys obtained from
the permissible limits set by WHO (2015) of 5.0 different regions of Thailand and 1.85–2.35 µg/g)
µg/g. The mean concentration of copper in honey revealed by Beshaw et al. (2022). Esubalew et al.
samples (2.61, 2.26, 2.46 µg/g) from Kano, (2020) and Stecka et al. (2014) reported lower
Katsina, Kebbi were significantly (p<0.05) higher concentrations to those obtained in the present
compared to honey samples collected from source study, 0.02–1.15 µg/g and 0.01–1.42 µg/g for
(control) and all the study areas. The presence of content of copper in honey respectively. Higher Cu
heavy metals in soils is not only due to external concentrations (0.95–9.24 µg/g and 0.25–71.25
contamination, but can also be of geochemical in µg/g) were recorded by Chukwujindi et al. (2015)
origin. Indeed, high copper contents can occur due and Iwegbue et al. (2015) respectively in honey
to mixed causes, such as abnormal native consumed in Nigeria and several times higher than
geochemical contents being complemented by the level noted in honey varieties produced in
mining contaminants. Copper is a vital element to Turkey (0.223–198.361 µg/g) (Altunatmaz et
the health of all living things and in humans. al.,2018).
However, too much ingestion of copper can lead to

6
5
Concentration (µg/g)

5
4
3 2.61 2.46
2.26
2 1.24 1.33
0.9
1 0.47 0.52
0
Kano Jigawa Katsina Kaduna Kebbi Sokoto Zamfara Control IHC
Limits
Study Areas
Fig 5: Mean Concentration of Cu (µg/g)

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The mean concentration of zinc in all is an essential element for the organism. It plays a
honey samples analyzed ranged from the lowest critical role for the structural and functional
5.75 µg/g in honey samples from Kebbi to the integrity of cells. It has functions in gene
maximum of 97.63 µg/g in honey samples from expression and growth. It protects from ultraviolet
Katsina (Fig. 6). The mean concentration of zinc in radiation, facilitates wound healing, contributes to
honey samples from Kano (12.33 µg/g), Jigawa immune and reduces the risk of cancer and
(19.78 µg/g), Katsina (97.63 µg/g) and Sokoto cardiovascular disease. In most cases excess zinc
(11.04 µg/g) obtained in this study had exceeded generates reactive oxygen species and/or displaces
the permissible limits of 10 µg/g set by WHO other metals from active sites in proteins
(2015). The mean concentration of zinc (97.63 (Marschner, 2012). The most important sources of
µg/g) in honey samples from Katsina was anthropogenic zinc in soil come from discharges of
significantly (p<0.05) higher compared with the smelter slags and wastes, mine tailings and the use
concentration of honey samples from the source of galvanized containers that contain zinc
and all the study areas. Usually, the use of (Aghamirlou et al., 2015). The average Zn
galvanized containers is the most prominent source concentration was higher than those found in
of contamination of honey as reported by Esubalew Nigerian honeys, (1.0 – 31.0 µg/g, 2.12 – 3.56
et al. (2020) and this could be the reason why µg/g, 1.77 – 3.99 µg/g and 6.62 µg/g) reported by
Katsina honey that is sometimes sold in such Iwegbu et al. (2015), Toma et al. (2020), Okeola et
containers are highly contaminated with zinc al. (2020) and Idoko et al. (2018) respectively.
possibly due to leaching. Some researchers have Other reported levels of Zn in honeys across the
expressed that diverse metal concentrations in world include those from Ethiopia (9.96 – 16.03
honeys is extremely reliant on the kind of flowers µg/g and 1.98 – 2.04 µg/g) reported by Esubalew et
utilized by bees and it can be the chief source of Zn al. (2020) and Tibebe et al. (2022) respectively,
contamination (Aghamirlou et al., 2015). Although, 0.12 – 6.63 µg/g from Iran (Aghamirlou, 2015) and
zinc is an essential element for human body, high 4.70 – 173.77 µg/g from Malaysia (Moniruzzaman
Zn intake may lead to adverse health effects. Zinc et al., 2014).

120
Concentration (µg/g)

97.63
100

80

60

40
19.78 18.63
20 12.33 11.04 10
5.75 6.53 4.38
0
Kano Jigawa Katsina Kaduna Kebbi Sokoto Zamfara Control IHC
Limits
Study Areas
Fig 6: Mean Concentration of Zn (µg/g)

The mean concentration of iron in all the – 40.77 µg/g) reported by Njokuocha et al. (2019)
honey samples analyzed ranged from 6.74 µg/g in in honey from different locations in Nigeria and
honey samples from Jigawa to 61.43 µg/g in honey higher than Fe concentrations reported in honey
samples from Kano (Fig. 7). Mean concentration of samples from Ethiopia, 0.56 – 18.69 µg/g (Melaku
iron in all the honey samples obtained in this study and Tefera, 2022), Algeria, 4.22 – 8.96 µg/g
had exceeded the permissible limit set by World (Chafik and Adnene, 2022) , Saudi Arabia, 2.08 –
Health Organization, WHO (2015) of 15 µg/g 8.79 µg/g (Aljedani, 2022) and Brazil, 0.12 – 8.76
except in honey samples from Jigawa (6.74 µg/g), µg/g (Maria et al., 2013). The Fe concentrations of
Katsina (13.10 µg/g) and source (12.05 µg/g). The the study were lower than the previously detected
mean concentration of iron (61.43 µg/g) in honey Fe concentrations in honey samples from Nigeria,
samples from Kano was significantly (p<0.05) 5.0 – 163.2 µg/g (Iwegbue et al., 2015) and
higher compared with the concentration of honey Turkey, 3.506 – 1278.779 µg/g (Altunatmaz et al.,
samples from the source and all the study areas. A 2018). Fe is an essential element for the
wide concentration range of Fe in honey samples production of red blood cells. It has an ability to
has been reported in literature and the mediate electron transfer in the catalysis of
concentrations of Fe in our samples (6.74 – 59.76 enzymatic reactions which is also potentially toxic
µg/g) were comparable with Fe concentrations (ND

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because it can catalyse the conversion of hydrogen into free radicals (Altun et al., 2017).

70 61.43

Concentration (µg/g)
60
50
40
30 24.97 23.86 24.43
19.29
20 13.1 12.05 15
6.74
10
0
Kano Jigawa Katsina Kaduna Kebbi Sokoto Zamfara Control IHC
Limits
Study Areas
Fig 7: Mean Concentration of Fe (µg/g)

2.5
y = 0.010x
2 R² = 0.9944
Absorbance

1.5

0.5

0
0 50 100 150 200 250
GAE Conc. (mg/L)

Fig.8: Calibration Curve for Gallic Acid Equivalent (GAE).

0.3
y = 0.0015x
0.25 R² = 0.9998
Absorbance

0.2
0.15
0.1
0.05
0
0 50 100 150 200
RUE Conc. (mg/L)
Fig 9: Calibration Curve for Rutin Equivalent (RUE).

Using the calibration plot of RUE, the 2011). In addition, some previous studies have also
flavonoid contents (mg/kg of honey) were found the following total flavonoid content (426.0
determined for all the samples and found to be in to 9504.1 mg/kg) in various honeys from Algeria
the range of 154.5 in samples from Kaduna to (Bakchiche et al., 2020). Pham-Nhut et al. (2022)
294.4 mg RUE/kg in samples from Zamfara states, reported 49 to 89 mg QE/kg) from different
while the control samples contain the highest mean botanical and geographical sources, 416.7 to
value of 521.7 mg RUE/kg (Fig. 10). The average 12497.4 mg/kg was reported by Kaya and Yildrim
quantity for total flavonoids content in the honey (2021) of honey of five different regions of Bingol
samples were found to be lower than 4052 to 9661 province, 180 to 422 mg/kg by Sime et al. (2015)
mg/kg in honey from Southern Nigeria (Ukom et of natural honeys from different geographical
al., 2019) and relatively higher than 25.2 to 272.1 regions of Ethiopia and 656.5 mg/kg by
mg/kg in honey from the Southern rain forest and Moniruzzaman et al. (2013) of Malaysian honeys.
Northern savannah ecosystem in Nigeria (Ita,
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600

Flavonoid (mg RUE/kg)

521.7
500
400
294.4
300
188.8 198.8 212.3
163.2 183.8
200 154.5

100
0
Kano Jigawa Katsina Kaduna Sokoto Zamfara Kebbi Control

Study Areas
Fig 10: Variation of Mean Concentration of Flavonoid (mg/kg) in Honey Samples Analyzed

The total phenolic compound varied from Bangladesh honeys (470-980 mgGAE/kg) reported
399.6 mgGAE/kg in samples from Kaduna state to by Pham-Nhut et al. (2022) and Alzaharani et al.
1075.0 mgGAE/kg in samples from Zamfara with (2012) respectively, as well as from Romanian
an average value of 1580.0 mg/kg in honey honey (230-1250 mg GAE/kg) reported by Islam et
samples from source (control) (Fig. 11). The mean al. (2012). Wabaidur et al. (2020) reported that the
concentrations of (399.6, 431.5, 433.1, 457.2 and content of phenolic acids in Yemeni honey ranged
461.1 mg GAE/kg) in honey samples obtained at level of 10.74-86.80 mg GAE/100g. Chaturvedi
from Kaduna, Sokoto, Jigawa, Kano and Katsina et al. (2014) reported the phenolic content of
states respectively were significantly (P<0.05) processed honey samples from central India in the
lower compared with honey samples obtained from range of 80.5 to 147.5 mg GAE/kg while the
Zamfara and source. It was observed that the unprocessed samples showed the highest content of
commercial honey samples had a lower phenolic 765.3 and 814 mg GAE/kg. Previous studies have
content when compared with control samples mentioned a high gallic acid contents in many
obtained directly from the known beekeepers. honeys. Gallic acid was also well documented
These differences might be attributed to the effect phenolic acid responsible for the antioxidant
of adulteration which removes most of the phenolic activity of honeys (Cheung et al., 2019). General
content in honey samples. In the present study, the observation can be made that dark honeys were
phenolic acid results were lower than those found characterized by considerably higher phenolic
in Nigerian honeys: 60.47-73.41 mg GAE/g (Ukom content than light coloured honeys.
et al., 2019) but similar to the results obtained from
Vietnamese honey (890-1110 mg GAE/kg), and
Phenolic Acid (mg GAE/kg)

1800
1580
1600
1400
1200 1075
1000
800 617.7
600 457.2 433.1 461.1 399.6 431.5
400
200
0
Kano Jigawa Katsina Kaduna Sokoto Zamfara Kebbi Control

Study Areas

Fig 11: Variation of Mean Concentration of Phenolic Acid (mg/kg) in Honey Samples Analyzed

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Table 1: Correlation Coefficient between Flavonoid, Phenolic Acid and Heavy Metals Concentration observed
in the Honey Samples.
Heavy Metals Flavonoid Phenolic Acid
Zn -0.09 -0.110
Cd -0.193* -0.096
Fe -0.132* -0.093
Cu -0.039 -0.073
Ni -0.078 -0.022
Pb -0.058 0.068
NB: Correlation coefficient values with asterisk are significant at P<0.05

Pearson product moment correlation Aghamirlou, H. M., Khadem, M., Rahmani, A.,
analysis between flavonoid, phenolic acid and Sadeghian, M., Mahvi, A.H., Akbarzadeh,
heavy metals (Table:1) in the honey samples A. and Nazmara, S. (2015). Heavy Metals
revealed a significant negative correlation between Determination in Honey Samples Using
flavonoid content, Cd and Fe, while most of the Inductively Coupled Plasma-Optical
heavy metals analyzed (Zn, Cu, Ni and Pb) are Emission Spectrometry. Journal of
non-significant. Phenolic acid content showed a Environmental Health Science and
non-significant negative correlation with (Zn, Cd, Engineering, 13(39). DOI:
Fe, Cu and Ni) while a non-significant positive 10.1186/s40201-015-0189-8
correlation was observed with Pb. Aljedani, D. M. (2022). Determination of Some
Heavy Metals and Elements in Honeybee
CONCLUSION and Honey Samples from Saudi Arabia.
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to adverse health risks when consumed and losing and Mehmet, S. (2017). Analyses of
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most of the honey samples (about 80%) from open Honey Samples from South and East
markets are adulterated to some degree and might Region of Turkey Using ICP-MS.
be through honey harvesting, processing and International Journal of Analytical
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Zamfara and Kebbi states were almost in Level of Chromium, Copper, Iron,
agreement with standard values or limits and Magnesium, Manganese, Selenium, Zinc,
therefore are assumed to be free of adulteration. Cadmium, Lead and Aluminium of Honey
However, samples obtained from Katsina, Sokoto, Varieties produced in Turkey. Food
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