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American Journal of  
Chemistry and Pharmacy (AJCP)

Dietary Risks Assessment of  Antibiotic Residues and Microbial Safety of  
Local Honey in Adamawa State

Dowell B.F1*, Maitera O.N1, Daniel M.W1, Micah M.M1

Volume 1 Issue 1, Year 2022
ISSN: 2834-0116 (Online)

DOI: https://doi.org/10.54536/ajcp.v1i1.492
https://journals.e-palli.com/home/index.php/ajcp

Article Information ABSTRACT

Received: August 11, 2022

Accepted: September 02, 2022

Published: September 12, 2022

This study investigated the microbial safety and dietary risk assessments of  antibiotic resi-
dues in honey. The determination of  antibiotic residues in this study were carried out using 
HPLC (LC 1200 series Agilent Tech). The results of  the microbial analysis of  the honey 
samples showed that, some of  the honey samples were contaminated with micro-organ-
isms. The antibiotics with highest mean concentration in the study were chloramphenicol 
(6.468±0.03 µg/kg) and sulfonamide (5.553±0.04 µg/kg) from sample HS1 and HG2 re-
spectively. Estimated daily intake (EDIs) of  detected antibiotic residues which were com-
pared with the recommended acceptable daily intake (ADIs) for each antibiotic allowed in 
food. Chloramphenicol was observed to possess the highest risk for adults and children 
(1.08E-04 µg/kg/bw) and (4.31E-04 µg/kg/bw) respectively in sample HS1. All the EDIs 
evaluated for antibiotic residues falls below acceptable daily intake (ADI). The target hazard 
quotient (THQ) and health index (HI) for antibiotic residues in all the samples were < 1, 
this suggested no risks associated with the level of  antibiotic residues risks thus having less 
danger to human health through its consumption.

Keywords
Antibiotic Residues, Estimated 
Daily Intake, Health Index, 
Honey, Microbial Safety 

1 Chemistry department, Modibbo Adama University (MAU), P.M.B 2076, Yola, Adamawa State, Nigeria
* Corresponding author’s e-mail: dowellfarimani@yahoo.com

INTRODUCTION
Honey is the natural sweet, viscous substance produced 
by honeybees from the nectar of  flowers or from the 
secretion of  living parts of  plants or excretions of  plant 
sucking insects on the living parts of  plants, which 
honeybees collect, transform and combine with specific 
substances of  their own, store and leave in the honey 
comb to ripen and mature (Abeshu and Geleta, 2016). 
It is one of  the most widely sought products because of  
its unique nutritional and medicinal properties (James et 
al., 2009). The variety produced by honey bees (the genus 
Apis) is the most commonly sought for because it is 
collected by most beekeepers and consumed by people. 
Honey is also produced by bumble bees, stingless bees 
and other hymenopteran insects such as honey wasps, 
though the quantity is generally lower and they have 
slightly different properties compared with honey from 
the honey bees. Honey bees convert nectar into honey by 
a process of  regurgitation and evaporation: they store it 
as a primary food source in wax honeycombs inside the 
beehive (Satarupa and Subha, 2014). Honey is composed 
primarily of  the sugar: glucose and fructose; its third 
greatest component is water (Singh et al., 2012).  Honey 
also is composed of  a complex mixture of  carbohydrates 
and other less frequent substances, such as organic acids, 
amino acids, proteins, minerals, vitamins, lipids (Blasa 
et al., 2006; Ball, 2007; Zerrouk et al., 2011), aromatic 
compounds, flavonoids, vitamins, pigments, waxes, 
pollen grains, several enzymes and other phytochemicals 
(Gomes et al., 2010).
In Nigeria, domestic consumption rate of  honey was 
estimated at 380,000 tonnes, with a global price of  about 
4.5 billion dollars (Vanguard News, 2017).  Though, 
the country has the potential to produce about 800,000 

tonnes, and can generate over 10 billion dollars from 
local and international trade, her current potential honey 
output is less than 3% (Punch News, 2017). The current 
production of  honey liquid in Nigeria is 2 million liters 
but has potential to produce 20 million liters of  honey 
annually (The Guardian news, 2018). Across Nigeria, 
Net return analysis showed that honey bee production 
is profitable, encouraging gross margin and net income 
(Duruson, 2011; Igbokwe and Mbanaso, 2006; Uduma 
and Udah, 2015, Abdullahi et al., 2014, Folayan and 
Bifarin, 2014). Though, the returns generated from the 
businesses have an encouraging outlook, these figures 
are considered far beyond the estimated market potential 
endowed within Nigeria and her young populace.
Antibiotics, particularly streptomycin, sulfonamide, 
and chloramphenicol are often used by beekeepers to 
treat bee-related diseases and as growth enhancers. The 
relatively long shelf-life of  antibiotic residues in foods 
could indirectly leads to the emergence of  bacteria 
strain that can resist the antibiotics overtime. Could also 
induces allergic reactions in hypersensitive individuals, 
and could leads to the disorder of  the hemopoietin 
system (Tillotson et al., 2006).  Antibiotic residues in 
honey have become a major consumer concern. Some 
drugs have the potential to produce toxic reactions in 
consumers directly while some other is able to produce 
allergic or hypersensitivity reactions (Velicer et al., 2004). 
Considering these negative effects of  antibiotic, the 
residual level in foods from plant and animal origin are 
regulated in developed economy (Vragović et al., 2012), 
however, these limits have received poor recognition in 
developing economy like Nigeria, thus posed a serious 
public health risk burden (Mensah et al., 2014). The 
determination of  antibiotic residues and microbial safety 

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in honey and other bee products has become a growing 
concern considering the growing popularity of  honey 
in human daily diets. Its ingestion without knowing its 
source and safety might carry significant health hazards. 
Residual level of  contamination cannot be changed 
through various production techniques, hence the need to 
study the microbial safety and dietary risks of  antibiotics 
residues in the samples of  local honey.

METHODOLOGY
Materials and Chemicals/Reagents
Standard of  tetracycline, Streptomycin, Sulfonamides, 
chloramphenicol, distilled water, deionized water, 
acidified water, sodium acetate, anhydrous magnesium 
sulphate, sodium hydroxide, plastic bottles, methanol, 
acetonitrile, volumetric volumes, electric centrifuge, 
beaker, glass tubes, polyethylene bottles, petri dish, sterile 
pipette, Potato Dextrose agar, tryptone Neomycin agar, 
potassium chloride, HPLC (LC 1200 series Agilent Tech). 
All are analytical grade.

Study area and Geographical Location
Adamawa is a state in North-eastern Nigeria, with its 
capital at Yola. It is located between latitude 9°20′N and 
longitude 12°30′E (Fig 3.1). It occupied an area of  36,917 

square kilometers. The state has population of  3,178,950 
(NPC, 2006). Adamawa is one of  the largest States of  
Nigeria, the State of  Borno borders it to the Northwest, 
Gombe to the West and Taraba to the Southwest. It’s 
eastern bordered with Cameroon. It comprises of  21 
local Government area.

Sample collection, Storage and Preservation
The honey samples were collected from three (3) local 
government areas of  Adamawa state (Ganye, Song and 
Mubi North from the Southern, Central and Northern 
senatorial zones respectively). The samples were mixed to 
make a composite sample that represents each sampling 
location (Table 3.1). Six (6) composite samples were 
collected purposively from the bee-keepers farms namely: 
Sugu, Ganye I, Dirma, Shimba, Lokuwa and Yewa Hosere 
at different locations. Similarly, three (3) composite 
samples were purchased from retailers at different point 
(Ganye II, Song and Mubi) in the same geographical area. 
The samples were collected during dry season (January-
March 2021). The collected samples were properly stored 
at room temperature and preserved to avoid loss due to 
poor storage, was timely analyzed to minimize loss due to 
prolonged storage. Samples were stored in polyethylene 
bottles preserved for analysis (Japhet et al., 2018).

Figure 1: Map of  the study Area (Google map 2020).

Table 1: Sample locations of  honey and coding
L.G.A of  the senatorial zones in Adamawa State Sample Locations Code
Ganye (Southern Senatorial zone) Sugu HG1

Ganye I HG2
Ganye II HG3

Song (Central Senatorial zone) Dirma HS1

Shimba HS2
Song HS3

Mubi North (Northern Senatorial zone) Lokuwa HM1
Yewa Hosere HM2
Mubi HM3

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Figure 2: Samples collected for analysis

Microbial Contaminants analysis
Standard plate count 
Ten grams of  honey was suspended in 90 ml of  0.1% 
phosphate buffer solution.  A series of  dilutions was then 
carried out and 0.1 ml spread on Plate Count Agar (PCA) 
(OXOID).  The culture was incubated for 72 h at 37°C.

Yeast and mold count 
Isolation of  microorganisms 
One ml of  each sample was picked with the aid of  a sterile 
pipette. A plastic rack was arranged with 9 sterile test 
tubes containing 9 milliliters of  sterile distilled water. A 
ten-fold dilution adopted by (Eleazu et al., 2013) was done 
by dispersing 1 ml of  the sample into the first test tube 
(10-1) which will be well shaken. One milliliter was then 
taken again from 10-1 dilution and transferred to the next 
test tube (10-2). The dilution continued to 10-9. Each test 
tube was shaken vigorously before each transfer. 

Inoculation
Shree & Arlis (2003) used the pour plate method to plate 
all the samples. 0.1 ml from dilution 10-2 was dispersed 
into sterile petri dish with the aid of  a sterile pipette. 
Potato dextrose agars was poured into the plates (10 ml) 
and isolation of  fungi were carried out using the potato 
dextrose agar. The plates were swirled gently for easy 
mixing of  the samples and the media. All plates were 
allowed to solidify on the bench and each plated sample 
was duplicated.

Incubation
The potato dextrose agar (PDA) plates were transferred 
to an incubator at 25˚C for 3-5 days. All incubated plates 
were examined daily for mycelia and colony growth.

Subculture and purification
After the incubation period, a flamed surgical knife was 

used to subculture different color of  mycelia growth from 
potato dextrose agar (PDA) plates on newly prepared 
PDA plates. All while the PDA plates were incubated at 
25˚C for 3-5 days.

Counting of  colonies
After incubation of  all the plates, counts of  the number of  
colonies in each plate was done with a hand tally counter 
(Fawole et al., 1988). The mean counts were obtained and 
multiplied by the appropriate dilution factor. The mean 
count was calculated as:
Mean =  (Total viable count)/(Number of  plates)         
The estimation of  the viable counts in each sample was 
made in colony forming unit (CFU) and Total viable 
count = (Number  of  colonies X Dilution Factor)/
(Amount Plated (ml))

Sulfite-Reducing Anaerobic Bacteria (clostridia)
The technique of  enumeration carried out the sulfite-
reducing anaerobic bacteria analysis in a solid medium 
in tubes. To 1 ml decimal dilution (10−2), contained 
in sterile tubes, 20 ml of  the melted Sulfite Tryptone 
Neomycin agar was added and cooled in a water bath at 
45˚C. After homogenization and solidification, the tubes 
were incubated anaerobically at 37˚C for 24–48 hours. 
Colonies characteristics of  sulfite-reducing anaerobic 
bacteria appear black in the tubes (Francois et al., 2018)
Total Coliform and fecal coliform (ISO 4831: 2006)
The total coliform search was carried out according to 
ISO 4831: 2006. 1 ml of  decimal dilution (10−2) was 
poured aseptically into sterile plates. Purple crystal, bile-
lactose neutral red agar (VRBL), melted and cooled in a 
water bath at 45˚C, was added to the inoculum at a rate 
of  15 ml per dish. The mixture was then homogenized by 
rotary movements. After solidification of  the first layer, 
a second 5 ml layer of  VBRL was added. Control of  the 
sterility of  the medium was carried out in a Petri dish 

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with approximately 15 ml of  VBRL. The total coliform 
count was done directly after incubation at 30˚C for 
24–48 hours. Fecal coliforms are characterized by a small 
mass of  fluorescent colonies with a diameter of  0.5 mm 
(Francois et al., 2018).

Extraction and determination of  Antibiotic residues 
using HPLC
The extraction of  honey samples was carryout subjecting 
to deproteinizing chemical procedure using Acetonitrile 
(ACN). 2 g of  the honey sample was place into 10 ml test 
tubes and shake intensively with 3 ml acetonitrile (ACN) 
for 1minute. The mixture was centrifuge for 15 minutes 
at 5000 rpm. The supernatant was collected and dried 
under Nitrogen stream at 40°C. The residue will be re-
dissolved in methanol and filtered through 0.45µm filter 
paper describe by Pagliuca et al., 2002).  
The presence of  antibiotic residues in the honey samples 
was carried out using different mixture of  aqueous 
mobile phase (A) Acidified water and organic mobile 
phase (B) methanol/ACN with a flow rate of  1 ml/min. 
The respective antibiotic residues were quantified by a 
modified method described by (Albino et al., 2005 and 
Shafqat et al., 2012), detected at 210-240 nm. 

The health risk  assessment and hazard characterization 
of  antibiotic residue
The health risk assessment and hazard characterization 
were carryout by first estimating the daily intake (EDI) of  
the respective antibiotics in the honey. This was achieved 
by integrating the average concentration of  the antibiotics 
in the honey,  the average consumption rate of  the 
honey and the average body weight per person as described 
in equation 1 (Forkuoh et al., 2018; USEPA, 1997).
EDI=(Ch×Hir)/BW…………………… equation 1
The Ch is the antibiotic concentration (µg/kg) in the 
honey, Hir represents the average honey consumption rate 
or intake rate for an average child and adults (0.001). The 
BW is the average body weight of  children (15 kg) and 
adults (60 kg) (USEPA, 2000; Akbari et al., 2012).     

The potential non-carcinogenic risk from the 
consumption of  the antibiotics
The potential non-carcinogenic risk from the consumption 
of  the antibiotics were estimated using Target Hazard 
Quotient (THQ) and the health index (HI) as described 
by the United State Environmental Protection Agency 
(USEPA, 1997). The THQ was estimated by integrating 
the ratio of  the EDI to the acceptable daily intake (ADI) 
values for each antibiotic (FAO/WHO, 2002 and 2010; 
USEPA, 1996; Bwatanglang et al., 2019; Bwatanglang, 
2019). The expression for estimating the THQ are 
described in equation 2.
THQ=EDI/ADI…………….........…equation 2
The HI, expressed as the sum of  the THQ as described 
in equation 3 is the cumulative effect pose by the 
combination of  the individual antibiotics presents in the 
honey (Forkuoh et al., 2018; Reffstrup et al., 2010).
HI = (EDI1/ADI1) +(EDI2/ADI2) +(EDI3/ADI)+….
(EDIi/ADIi)……………………..equation 3
Were the EDIi represents the estimated daily intake dose 
of  the individual antibiotics (1, 2, 3….) in the honey 
and the ADIi is the acceptable daily intake dose for the 
individual antibiotics (1, 2, 3……)

Statistical analysis
The mean and standard deviation of  the results were done 
using Minitab 19 and data was generated in triplicate. The 
results were express as Mean± SD 

RESULTS AND DISCUSSION
Microbial analysis
The results of  the microbial analysis of  the honey 
samples showed that, some of  the honey samples are 
contaminated with micro-organisms. Microbial analysis 
results are shown in Table 2 and 3 The SPC were found in 
low numbers in all samples of  honey with a mean count 
which varies within 1.0 x 104 – 3.3 x 104 CFU/ml. The 
yeast and mold count of  honey samples were less than 10 
CFU/ml in sample in all the samples.  

Table 2: Mean results of  microbial analysis
Sample Location Sample SPC in CFU/ml Mold and Yeast 

(CFU/ml) x 10^2
Sulfite-reducing 
Clostridia

Total coliform and 
fecal Coliform

GANYE HG1 1.7 x 10^4    2 ND ND
HG2 2.0 x 10^4    ND ND ND
HG3 3.2 x 10^4    1.8 ND ND

SONG HS1 1.0 x 10^4    ND ND ND
HS2 ND    ND ND ND
HS3 3.3 x 10^4    1.6 ND ND

MUBI-NORTH HM1 1.3 x10^4    ND ND ND
HM2 ND    1 ND ND
HM3 ND    1.5 ND ND

SPC = Standard Plate Count
ND = Not detected
CFU = Colony forming units

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Table 3: Isolated micro-organisms PDA (Fungi) from local honey samples in Adamawa State
Sample 
Location

Sample Aspergillus 
fumigatus

Aspergillus 
flavus

Aspergillus 
niger

penicillum sp Trichophyton 
rumbrum

GANYE HG1     +   -   -   -    +
HG2     -   -   -   -    -
HG3     -   +   -   +    -

SONG HS1     -   -   -   -    -
HS2     -   -   -   -    -
HS3     +   -   +   -    -

MUBI-
NORTH

HM1     -   -   -   -    -
HM2     -   -   +   -    +
HM3     -   +   -   -    -

Key: + = Present
- = Absent
PDA = Potato Dextrose Agar

Microbial Contamination
The mean level of  microbial contamination of  
different honey samples were presented in Table 2. 
The contamination with standard plate count (SPC) 
for Aerobic Mesophilic Bacteria varies within 1.0 x 104 
– 3.3 x 104 CFU/ml. This result conforms with the 
result of  Ndife et al., 2014. According to Tchoumboue 
et al., 2008, the contamination with fungi and bacteria 
indicate inadequate hygienic conditions during collecting, 
manipulating, processing and storing. Microbial 
contamination during and post processing can also result 
in spoilage or persistence of  some bacteria in honey.
The yeast and mold count of  honey samples were less than 
10 CFU/ml in samples HG1, HG3, HS3, HM2 and HM3 
but was not detected in Sampling points HG2, HS1, HS2 
and HM1 (Table 3). There was total absence of  sulfite-
reducing clostridia and fecal coliforms in all the samples, 
indicating that honeys were produced in accordance with 
good hygiene practices during extraction, packaging and 
storage (Guiraud, 2003). The low microbial loads of  
honey samples could be attributed to their low pH values 
and high amount of  total soluble sugars and possibly 
phenolic compounds and their synergistic interaction 
(Alvarez-Suarez et al., 2010).
Yeast, mold and spore-forming bacteria (coliform) have 
been implicated to survive in honey and are indicative 
of  the sanitary quality of  the honey (Ezeama, 2007; 
Eleazu et al., 2013). In a study conducted by Omafuvbe 
and Okanbi 2009, observed that no mold contamination 
was detected. The micro-organisms that were isolated 
in the honey samples from all sampling points are fungi 
(Table 3). The presence of  micro-organisms in honey 
can sometimes influence the stability of  the product and 
its hygienic quality. Normal honey must lack pathogenic 
micro-organisms or micro-organisms that produce 
enteric illnesses (Popa et al., 2009).

Determination of  Antibiotic Residues
The determination of  antibiotic residues in all the 
sampling points was carried out using HPLC (LC 1200 
Series, Agilent Tech).  The Health Risks Assessment, 
Target Hazard Quotient and Health Index was estimated 

using various mathematical expressions and results 
presented in Tables.
Maximum residues level (MRLs) are yet to be established 
for honey and other bee products (Al- Waili et al., 2012). 
The EU (European Union) under the council Directive 
2001/110/EC has set a reference point for action (RPAs) 
for antibiotic residues in honey which is also used as 
provisional MRL (Forsgen, 2010; Johnson et al., 2010; 
EFSA, 2013). The RPAs are residue concentrations which 
are technically feasible for analytical considerations place 
as a bench mark to reject any bee products exceeding 
these limits (Johnson et al., 2010; ESFA, 2013; Mutinelli, 
2003). Provisional MRL in parts per billion (ppb) for 
oxytetracycline (25 ppb), chloramphenicol (0.3 ppb) and 
nitrofurans (1.0 ppb) were established by EU for honey 
(Johnson et al., 2010).

Mean concentration of  antibiotic residues
The samples’ average mean± Sd concentration of  
antibiotic residues were analyzed and the results presented 
in Table 4. All the samples contained at least two or more 
antibiotic residues detected. Streptomycin were below 
the detection level (BDL) in samples HG1, HG2, HG3, 
HS1, HS2 and HM2. Only samples from Song (HS3) 
and Mubi North (HM1 and HM3) contains the level of  
streptomycin antibiotic residues. Similarly, tetracycline 
was below the detection limit (BDL) in samples from 
HG1, HS1, HS2, HM2 and HM3 as can be seen in Table 
4.
Among the antibiotics analyzed, sulfonamide was detected 
in virtually all the samples except sample HS3 and HM1 
from Song and Mubi-North respectively. The antibiotic 
with the highest concentration detected in the honey 
samples from all locations are: Chloramphenicol and 
Sulfonamide (Table 4). The highest mean concentration 
of  tetracycline and streptomycin were found in sampling 
point HG3 from Ganye (1.513± 0.01 µg/kg) and 
sampling point HS3 from Song (1.513± 0.05 µg/kg) 
respectively. The samples HG2 and HS1 from Ganye and 
Song were observed to contain the highest concentration 
of  sulfonamide with a mean concentration of  5.553±0.04 
µg/kg and 5.386±0.07 µg/kg respectively. Similarly, highest 
mean concentration of  6.468± 0.03 µg/kg and 6.447± 
0.03 µg/kg were detected for chloramphenicol in sampling 
point HS1 and HG2 from Song and Ganye respectively.

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In general, the consumption of  honey containing 
antibiotic residues represents a real danger to human 
health, considering that the indiscriminate consumption 
of  these compounds is the main cause of  bacteria 
resistance to antibiotics, especially in the case of  
aminoglycosides (streptomycin).  Also, antibiotics can 
cause allergic reactions in people sensitive to certain 
classes of  compounds and can cause hypersensitivity 
in up to 5% of  individuals exposed to these substances 
(Brum, 2018; Ford, 2017). 
Aminoglycosides are antibiotics that act by interfering 
with the replication of  bacterial DNA by blocking protein 
synthesis. This class of  antibiotics causes serious toxic 
adverse effects on the kidneys and the auditory system 
and, when used in high concentrations, can cause a serious 
risk of  neuromuscular paralysis. Regarding tetracyclines, 
these cause adverse effects to the gastrointestinal system, 
causing irritation and may lead to deficiency disorders 
in the absorption of  some vitamins. Besides, they cause 
kidney damage and bone marrow disorders, and can 
also cross the placental barrier and deposit in the bones 
(Brum, 2018; Ford, 2017).
Sulfonamides, better known as sulfas, are antibiotics 
that act by inhibiting the metabolism of  bacterial cells 
by inhibiting folic acid, making the cells unable to grow 

and multiply. Regarding the adverse effects caused 
by these substances, these can range from itchy skin, 
hypersensitivity reactions, liver problems to problems 
related to blood cells. (Brum, 2018; Ford, 2017; Omidi 
et al., 2016).
Bwatanglang et al., (2019) reported present of  antibiotics 
residues from Uba/Uvu, Mubi/Vimtim and Gombi/
Garkida for both Raw Honey Sample (RHS) and 
Commercial Honey sample (CHS). It was found 
that tetracycline, chloramphenicol, sulfonamide and 
streptomycin were all detected in the RHS from all the 
sampling points. 
Studies conducted in other countries also reported 
the presence of  antibiotic residues in honey samples. 
Twenty nine percent of  251 honey samples produces 
across Greece were observed to contain residual level 
of  tetracycline from 0.018- 0.100 mg/kg (Saridaki-
Papakonstadinou et al., 2006). About 13 honey samples 
out of  34 imported from Asian countries into Switzerland 
were found to 0.4 and 9.0 µg/kg of  chloramphenicol with 
at least two samples containing up to 5.0 µg/kg (Orteli et 
al., 2004). In another study, streptomycin 3- 10,820 µg/
kg, sulfonamide 5- 4,592 µg/kg, chloramphenicol 0.1- 
169 µg/kg were detected in honey samples from EU 
(Diserens, 2007).

Table 4: Mean Concentrations of  Antibiotic Residues (µg/kg) in Honey samples
Sample Location Sampling 

Points
Chloramphenicol Sulfonamide Tetracycline Streptomycin

GANYE HG1 0.102±0.05 2.232±0.02 ND ND
HG2 6.447±0.03 5.553±0.04 0.031±0.03 ND
HG3 ND 0.576±0.01 1.513±0.01 ND

SONG HS1 6.468±0.03 5.386±0.07 ND ND
HS2 0.194±0.02 0.150±0.01 ND ND
HS3 1.743±0.05 ND 1.292±0.03 1.513±0.05

MUBI-NORTH HM1 ND ND 0.060±0.01 0.126±0.07
HM2 1.042±0.04 0.687±0.01 ND ND
HM3 ND 5.042±0.03 ND 0.215±0.02

ND = Not Detected

Potential health risk of  antibiotic residues associated 
with consumption of  honey samples
The results of  human health risk assessments were 
presented in Table 5. The Estimated Daily Intake (EDIs) 
of  detected antibiotic residues for both adults and children 
in all the samples were compared with the recommended 
acceptable daily intake (ADIs) for each antibiotic in all 
food. The dietary exposure assessment is very critical 
towards evaluating the risks associated with antibiotics 
in honey. Helping to determining whether a residual of  
concerns to pose a potential risk to public health.
Based on the antibiotic residues detected in all the 
sampling points, chloramphenicol was found to 
possessed the highest exposure risks to both adults and 
children (1.08E-04 µg/kg/bw) and (4.31E-04 µg/kg/bw) 
respectively in sample HS1 (see Table 5). Sulfonamide 
presents the highest EDIs for adults (9.26E-05 µg/kg/
bw) and children (3.70E-04 µg/kg/bw) from sample HG2 
(Table 5). Similarly, the highest EDIs for tetracycline was 

found in sampling point HG3 (2.52E-05 µg/kg/bw) and 
(1.00E-04 µg/kg/bw) for adults and children respectively 
while EDIs for streptomycin was only obtained in sample 
points HS3, HM1 and HM3 from Ganye and Mubi-
North for both adults and children (Table 5)
Due to the smaller body weight (15 kg) and physiological 
susceptibility in children, the EDI were observed to 
be higher in children than that of  the adults. With the 
exception of  chloramphenicol which has no define ADI, 
all the EDIs calculated for each antibiotic residue were 
found to be below their recommended ADIs (Johnson 
et al., 2010). This shows that the consumption has a 
negligible risk to human health having < 1% ADI in all 
the samples.
Estimated Daily Intake (EDI) of  2.09 ng/kg body weight 
(BW)/day and 1.83 ng/kg BW/day for tetracycline and 
penicillin residues were determined in dairy products 
resulting in 0.007% and 0.006% of  the ADI respectively 
(Kabrite et al., 2019). Other related study reported various 
exposure indices due to dietary intake of  antibiotics by 
human. Residues of  quinolones and sulfonamides were 

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found to be widely distributed in in cultured fish samples 
from the Pearl River Delta, South China. The EDI results 
showed that the consumption of  the fishes to dietary 

intakes of  quinolones and sulfonamides were far below 
the acceptable daily intake (ADI) and poses no risk to the 
public health (He et al., 2016).  

Table 5: Estimated Daily Intake (EDI) in µg/kg/bw for Antibiotic Residues
ADULTS

  GANYE SONG    MUBI NORTH
ANTIBIOTICS HG1 HG2 HG3 HS1 HS2 HS3 HM1 HM2 HM3 ADI
Chloramphenicol 1.70E-06 1.07E-04 0.00E+0 1.08E-04 3.23E-06 2.91E-05 0.00E+0 1.74E-05 0.00E+0 NA
Sulfonamide 3.72E-05 9.26E-05 9.60E-06 8.98E-05 2.50E-06 0.00E+0 0.00E+0 1.15E-05 8.40E-05 5.00E+01
Tetracycline 0.00E+0 5.17E-07 2.52E-05 0.00E+0 0.00E+0 2.15E-05 1.00E-06 0.00E+0 0.00E+0 3.00E+01
Streptomycin 0.00E+0 0.00E+0 0.00E+0 0.00E+0 0.00E+0 2.52E-05 2.10E-06 0.00E+0 3.58E-06 5.00E+01

CHILDREN
GANYE SONG MUBI NORTH

ANTIBIOTICS HG1 HG2 HG3 HS1 HS2 HS3 HM1 HM2 HM3 ADI
Chloramphenicol 6.80E-06 4.30E-04 0.00E+0 4.31E-04 1.29E-05 1.16E-04 0.00E+0 6.95E-05 0.00E+0 NA
Sulfonamide 1.49E-04 3.70E-04 3.84E-05 3.59E-04 1.00E-05 0.00E+0 0.00E+0 4.58E-05 3.36E-04 5.00E+01
Tetracycline 0.00E+0 2.07E-06 1.00E-04 0.00E+0 0.00E+0 8.61E-05 4.00E-06 0.00E+0 0.00E+0 3.00E+01
Streptomycin 0.00E+0 0.00E+0 0.00E+0 0.00E+0 0.00E+0 1.01E-04 8.40E-06 0.00E+0 1.43E-05 5.00E+01

Table 6: Target Hazard Quotient (THQ) and Health Index (HI) for Antibiotic Residues 
ADULTS

  GANYE SONG    MUBI NORTH
ANTIBIOTICS HG1 HG2 HG3 HS1 HS2 HS3 HM1 HM2 HM3
Chloramphenicol - - - - - - - - -
Sulfonamide 7.44E-07 1.86E-06 1.92E-07 1.80E-06 5.00E-08 0.00E+0 0.00E+0 2.30E-07 1.68E-06
Tetracycline 0.00E+0 1.72E-08 8.40E-07 0.00E+0 0.00E+0 7.17E-07 3.33E-08 0.00E+0 0.00E+0
Streptomycin 0.00E+0 0.00E+0 0.00E+0 0.00E+0 0.00E+0 5.04E-07 4.20E-08 0.00E+0 7.20E-08
HI 7.44E-07 1.88E-06 1.03E-06 1.80E-06 5.00E-08 1.22E-06 7.53E-08 2.30E-07 1.75E-06

CHILDREN
  GANYE SONG    MUBI NORTH

ANTIBIOTICS HG1 HG2 HG3 HS1 HS2 HS3 HM1 HM2 HM3
Chloramphenicol - - - - -         - -          -             -
Sulfonamide 3.00E-06 7.40E-06 7.68E-07 7.18E-06 2.00E-07 0.00E+0     0.00E+0 9.16E-07 6.72E-06
Tetracycline 0.00E+0 6.90E-08 3.33E-06 0.00E+0 0.00E+0 2.87E-06 1.33E-07 0.00E+0 0.00E+0
Streptomycin 0.00E+0 0.00E+0 0.00E+0 0.00E+0 0.00E+0 2.02E-06 1.68E-07 0.00E+0 2.86E-07
HI 3.00E-06 7.47E-06 4.10E-06 7.18E-06 2.00E-07 4.89E-06 3.01E-07 9.16E-07 7.01E-06

Target Hazard Quotient (THQ) and Health Index 
(HI) for antibiotic residues
The results of  THQ and HI for potential non-
Carcinogenic risks were evaluated and presented in Table 
6. Based on the results obtained for THQ and HI, shows 
that the exposure to the antibiotics through the dietary 
consumption of  honey possess no immediate effect to 
human health as a result of  non- carcinogenic related 
risks (Table 6).  The Health Index (HI) for all the samples 
analyzed falls below the level of  concern to human health, 
shows HI < 1 in all the samples as can be seen in Table 6.
The THQ and HI analysis are health-based statistical 
probability expressed as a function of  quantified 
level of  concern; a process developed to exposure to 
environmental pollutants (Bwatanglang, 2019).  The 
target hazard quotient (THQ) and health index (HI) 
obtained < 1 in all the sampling points also suggest 
no risks associated with the level of  chloramphenicol, 
sulfonamide, tetracycline and streptomycin in honey, 
thus having less danger to human health through its 
consumption. But children are more adaptable to 
encountered the risks associated with their health due 
to the high value of  THQ and HI observed more than 

the adults. Similar study on the risk assessment due to 
dietary exposure to oxytetracycline, tetracycline and 
chlortetracycline through milk consumption in India 
showed HQ < 1 (Chauhan et al., 2018).
Though ADI was not defined for chloramphenicol and 
the THQ and HI could not be evaluated in this study, the 
consumption of  the honey samples analyzed in this study 
points toward potential health risk to chloramphenicol. 
Lack of  availability of  ADI for chloramphenicol further 
suggest a zero-tolerance level to the antibiotics. An ADI 
could not be established for chloramphenicol due to lack 
of  genotoxic and toxicological data, in addition to lack of  
definable NOAEL (No Observe Adverse Effect Level) or 
LOAEL (Lowest Observed Adverse Effect Level) (EFSA, 
2013). These makes residual level of  chloramphenicol not 
allowed in the animal food-production chain. And thus, 
reported to constitute threat to public health (EFSA 
2013; Commission Regulation (EU) No 37/2010; and 
No 165/2010).

CONCLUSION
Honey is consumed on a large scale throughout the world 
and becomes mandatory to carryout microbial analysis, 
monitoring and evaluation of  the risk to the health of  

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Journal of  chemical education, 84(10), 1643.
Blasa, M., Candiracci, M., Accorsi, A., Piacentini, M. P., 

Albertini, M. C. and Piatti, E. (2006). Raw Millefiori 
honey is packed full of  antioxidants. Food Chemistry, 
97, 217-222.

Brum, L. F. S. (2018). Farmacologia Aplicada à Farmácia (1 
ed). Porto Alegre: SAGAH.

Bwatanglang I (2019). Diclorvos-Mediated Heavy Metal 
Uptake in Leafy Vegetables and Potential Health Risk 
on Consumption. London Journal of  Research in Science: 
Natural and Formal, 19(3), 15-24

Bwatanglang IB, J Bimba, ST Magili, Y Musa and SP 
Zira (2019). Dietary exposure to antibiotics residue 
in honey and the potential health risks to consumers 
in Adamawa state, Nigeria. International Journal of  
Chemical Studies, 7(4), 1672-1679. 

Bwatanglang IB, Magili ST (2019). Health Risk 
Assessment of  Insulin supporting Elements in 
Commonly Consumed Anti-Diabetic Medicinal 
Plants. London Journal of  Research in Science: Natural and 
Formal, 19(4),11-22

Chauhan SL, Priyanka, Garg SR, Jadhav VJ (2019). 
Dietary exposure assessment of  tetracycline residues 
in milk in Haryana. International Journal of  Chemical 
Studies, 7(1), 1862-1865.

Codex Alimentations (2001). Draft revised standard 
for standard for honey (at step 10 of  the Codex 
procedure). Alinorm, 1(25), 19–26

Commission regulation (EU), (2010). No 165/2010 
on amending Regulation (EC) No 1881/2006 
setting maximum levels for certain contaminants in 
foodstuffs as regards aflatoxins. Off. J. Eur. Union. 
2010, 50, 8-12.

Commission regulation (EU), (2010) on pharmacologically 
active substances and their classification regarding 
maximum residue limits in foodstuffs of  animal 
origin. Off. J Eur.  Commun, 15(1), 72.

Diserens, J. (2007). Contaminants and residues in Food. 
Strategies (if  any) to screen and analyze veterinary 
drug residues in food from animal origin. In Proceedings 
of  the 5th International Fresenius Conference Nestle Research 
Center.

Dolan, J. W (2002). Peak Tailing and Resolution. LC●GC 
Europe, (pp 34-36.).

Duruson, G. C. (2011). Profitability of  apiculture as a 
bussiness venture in Ikwuano Local Government Area 
of  Abia State. B. Sc. Project. Department of  Agricultural 
Economics and Extension, Abia State University, Umuahia 
Campus, Umuahia, Abia state, Nigeria.

EFSA Panel on Contaminants in the Food Chain 
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principles and scientific methods to be taken into 
account when establishing Reference Points for 
Action (RPAs) for nonallowed pharmacologically 
active substances present in food of  animal origin. 
EFSA Journal, 11(4), 3195.

Eleazu, C. O., Iroaganachi, M. A., Eleazu, K. C. and 
Okoronkwo, J. O. (2013). Determination of  the 

the consumers. The result of  the microbial safety show 
that the samples studied are of  very good microbiological 
and hygienic quality, though there were isolated micro-
organisms in the honey samples from some sampling 
points. This maybe as a result of  improper handling 
during the production process and also air dust from the 
environment. The presence of  micro-organisms in honey 
can sometimes influence the stability of  the product and 
its hygienic quality. The potential human health risks 
associated with exposure to antibiotic residues were 
found to be lower than each acceptable daily intake (ADI) 
but the presence of  antibiotic and pesticide residues in 
honey is a concern to health. Due to the smaller body 
weight (15 kg) and physiological susceptibility in children, 
the EDI were observed to be higher in children than that 
of  the adults.

RECOMMENDATIONS 
The following recommendations have been suggested 
to enable better understanding and improve the results 
obtained from such research:

I. There should be educational programs to the bee 
keepers within the state on beehive management, this will 
create awareness on the safety of  consumers and public 
health.

II. The presence of  some pathogenic fungus in the 
samples necessitates an urgent need to always monitor 
microbial status of  honey.

III. Microbial testing should guarantee both good 
hygienic and good marketable qualities of  this products 
and good production efficiency.

IV. There should be continual monitoring and 
evaluation of  antibiotic and residues in honey, this will 
help to assess the potential risks to human health.

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