Trends in Food Science & Technology 114 (2021) 490–509

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Trends in Food Science & Technology

journal homepage: www.elsevier.com/locate/tifs

Food safety hazards of bee pollen – A review

Rita Végh a, Mariann Csóka a, Csilla Sörös b, László Sipos c, *
a
Hungarian University of Agriculture and Life Sciences, Institute of Food Science and Technology, Department of Nutrition, 1118 Budapest, Somlói út 14-16., Hungary
b
Hungarian University of Agriculture and Life Sciences, Institute of Food Science and Technology, Department of Food Chemistry and Analytical Chemistry, 1118
Budapest, Villányi út 29-43., Hungary
c
Hungarian University of Agriculture and Life Sciences, Institute of Food Science and Technology, Department of Postharvest, Commercial and Sensory Science, 1118
Budapest, Villányi út 29-43., Hungary

A R T I C L E I N F O A B S T R A C T

Keywords: Background: Bee pollen is a natural apicultural product that is becoming popular among health-conscious con­
Bee pollen sumers due to its wide variety of nutrients and bioactive substances. However, only a limited number of countries
Apiculture have established requirements for the quality and safety of the product so far.
Pesticide residue
Scope and approach: In this review, recent findings on the food safety risks of bee pollen and data about the
Heavy metal
Fungi
concentration of toxic substances detected in the products are summarized. Pollen loads may become contam­
Mycotoxin inated from the environment with pesticides, heavy metals, metalloids and mycotoxin-producing molds. In
Pyrrolizidine alkaloid addition, pollen of certain plant species initially contain hepatotoxic pyrrolizidine alkaloids in relatively large
Allergenic concentrations. Allergens and pollen grains from genetically modified plants may also be present in these
Risk assessment products. Based on literature data, a risk assessment was conducted for the most common pesticide active
Food safety substances (chlorpyrifos, fluvalinate, carbendazim, thiacloprid), toxicologically important elements (arsenic,
Margin of exposure cadmium, mercury, lead), common mycotoxins (aflatoxin-B1, ochratoxin-A, fumonisins, zearalenone, deoxy­
nivalenol, T-2 toxin) and pyrrolizidine alkaloids.
Key Findings and Conclusions: Our results suggest that pesticide residues usually do not pose a chronic risk to
consumers, but the estimated acute exposure values can be close to the acute reference dose (ARfD). Arsenic,
cadmium, lead and pyrrolizidine alkaloid content of bee pollen potentially pose a health risk to consumers,
therefore it is recommended to set a maximum limit for these substances and monitor their concentration in
commercially available products. Since scientific data regarding the mercury and mycotoxin content of bee
pollen is incomplete, further studies are needed in order to summarize the food safety hazards of bee pollen
pollutants.

acceptable techniques for preservation (Thakur & Nanda, 2020).

1. Introduction
As a result of the growing importance of health- and environmentally
conscious lifestyle, there is an increasing demand for low-processed
foods of natural origin, which contain bioactive ingredients. There­
fore, apicultural products have come into prominence in recent decades,
of which bee pollen is produced in relatively large quantities (Kieliszek
et al., 2018). Pollen is mixed with nectar and glanular secretions by
forager bees and compressed into their pollen baskets. When transferred
to the hive, the product can be harvested by the beekeeper with the use
of a device called pollen trap (Campos et al., 2008). Pollen is usually
marketed after drying, but freezing and lyophilization are also
Different bee pollens possess diverse chemical compositions and thera­
peutic effects as these properties are mainly determined by the species of
the source plants and are also influenced by the place of origin, the
weather conditions and the processing and storage technology. By
knowing the botanical origin of the products, important information can
be acquired about their quality (Bayram et al., 2021; Thakur & Nanda,
2020). A commonly used technique for determining the botanical origin
of pollen loads is the microscopic pollen analysis since, the size, shape
and surface properties of pollen grains are characteristic to particular
plant species (Kieliszek et al., 2018).
Bee pollen contains essential nutrients in exceptionally high con­
centrations, therefore it can be used as a dietary supplement. According

* Corresponding author.
E-mail addresses: vegh.rita@phd.uni-szie.hu (R. Végh), Csoka.Mariann@uni-mate.hu (M. Csóka), Marczika.Andrasne.Soros.Csilla@uni-mate.hu (C. Sörös), Sipos.
Laszlo@uni-mate.hu (L. Sipos).

https://doi.org/10.1016/j.tifs.2021.06.016
Received 1 February 2021; Received in revised form 4 June 2021; Accepted 5 June 2021
Available online 12 June 2021
0924-2244/© 2021 Published by Elsevier Ltd.
R. Végh et al. Trends in Food Science & Technology 114 (2021) 490–509

Abbreviations ICP-MS Inductively Coupled Plasma - Mass Spectrometry

ICP-OES Inductively Coupled Plasma - Optical Emission
3-AcDON 3-Acetyldeoxynivalenol Spectrometry
AAS Atomic Absorption Spectroscopy JMPR The Joint FAO/WHO Meeting on Pesticide Residues
ADI Acceptable Daily Intake LC50 Lethal Concentration
AFL Aflatoxins LC-MS Liquid Chromatography – Mass Spectrometry
ALARA As Low As Reasonably Achievable LC-MS/MS Liquid Chromatography – Tandem Mass Spectrometry
ARfD Acute Reference Dose LD50 Lethal Dosage
aw water activity LOAEL Lowest Observed Adverse Effect Level
BMDL Benchmark Dose Limit LOD Limit of Detection
CIE International Commission on Illumination LOQ Limit of Quantitation
CONTAM Panel on Contaminants in the Food Chain MOE Margin of Exposure
DAS Diacetoxyscirpenol MRL Maximum Residue Level
DNA Deoxyribonucleic acid NEO Neosolaniol
DON Deoxynivalenol NIV Nivalenol
EFSA European Food Safety Authority NOAEL No Observed Adverse Effect Level
ELISA Enzyme-linked Immunosorbent Assay OTA Ochratoxin-A
FAO Food and Agriculture Organization of the United Nations PA Pyrrolizidine alkaloid
FUM Fumonisins PANO Pyrrolizidine alkaloid N-Oxide
FUS-X Fusarenone-X PMGF pollen-mediated gene flow
GC-MS Gas Chromatography – Mass Spectrometry ppb part per billion
GC-MS/MS Gas Chromatography – Tandem Mass Spectrometry ppm part per million
GM Genetically modified QuEChERS an acronym of quick, easy, cheap, effective, rugged and
HBGVs Health-Based Guidance Values safe
HGT Horizontal gene transfer TDI Tolerable Daily Intake
HMPC Committee on Herbal Medicinal Products TMI Tolerable Monthly Intake
HPLC/FD High-Performance Liquid Chromatography with TWI Tolerable Weekly Intake
Fluorescence Detection WHO World Health Organization
IARC International Agency for Research on Cancer ZEA Zearalenone

to Campos et al. (2008), carbohydrates (13–55%), proteins (10–40%), 2. Research methodology

lipids (1–13%) and fibers (0.3–20%), all contribute to the composition of
pollen. In addition, they are rich in biologically active micronutrients
like minerals, polyphenols and vitamins. Based on a report by Habryka
et al. (2016), the product is used in apitherapy mainly for its antioxidant,
anti-inflammatory, antibiotic and antiallergic effects. Bee pollen im­
proves blood supply to the nerve tissue, thereby increasing mental
performance and eliminating the state of fatigue. Research works have
also shown a positive effect on some diseases of the liver, heart and
prostate. The main consumers of bee pollen are the followers of the
health- and environmentally conscious lifestyle, as well as the elderly,
who use it due to its antioxidant and other therapeutic effects.
Only a few countries possess legislation on bee pollen, such as
Poland, Bulgaria, Switzerland, Brazil and Argentina (Shahali, 2015;
Thakur & Nanda, 2020). Currently, there is no European standard
document for beekeeping products. The Technical Committee for Food
Products of the International Organization for Standardization estab­
lished a subcommittee on beekeeping products (ISO/TC34/SC19),
which has recently begun standardizing the product. At present, how­
ever, due to the lack of regulations, bee pollen of different origin can be
characterized by heterogeneous nutritional properties and food safety
risks.
According to international publications, the most important food
safety risks associated with bee pollen are pesticides, toxic metals and
metalloids, mycotoxins, pyrrolizidine alkaloids, allergen proteins and
pollen grains of GM plants (Campos et al., 2008; Thakur & Nanda,
2020). The aims of our work are to present new research data on the
food safety risks of bee pollen, to summarize general conclusions and to
perform a risk assessment on the most commonly found toxic com­
pounds and elements, based on scientific data.
The literature review was conducted between June 2020 and March
2021, based on studies dealing with the food safety risks of bee pollen.
The sources of the published data are academic journals from electronic
databases (ScienceDirect, Scopus, SpringerLink, Web of Science, Wiley
Online Library), doctoral dissertations and chapters of scientific books
as well. The keywords used in the research were “bee pollen”, “apicul­
tural product”, “food safety”, “pesticide residue”, “heavy metal”, “toxic
element”, “mycotoxin”, “pyrrolizidine alkaloids”, “allergenic”, “case
study” and “risk assessment”. The collected studies were grouped by
topics for easier handling, while excluding irrelevant ones and those
older than 20 years (except for case studies). In this review, mainly the
results of experimental studies are presented, but 12 literature reviews
and 8 medical case studies are also included in the references.
3. Hazards indicating food safety risks
3.1. Pesticide residues
Sörös (2019) defines plant protection products as preparations of
natural, synthetic or chemical origin that are capable of reducing,
attracting or alerting pests or regulating the life processes of pests and
plants. According to Ambrus et al. (2020), the use of pesticides is
essential for the food supply of the Earth’s population, so residues are
generally present in plant foods. Alternatives of these products are
grown by organic farming. Main principles of organic farming are sus­
tainability, rural development, animal welfare and food safety. Due to
the benefits of organic agriculture, the global interest is constantly
growing for these products (Bosta et al., 2019). The production and
labelling of organic foods are regulated at European level (Regulation
2018/848/EU). This document summarizes the requirements for
organic beekeeping practices on bee nutrition, veterinary drug use,

491
R. Végh et al.
animal welfare and hive placement. Under the regulation, apiaries
should be located at areas which ensure the availability of pesticide-free
nectar and pollen sources. Under favorable environmental conditions,
bees typically forage in a radious of 2 km from the hive (Garbuzov et al.,
2015), therefore a distance of at least three km must be provided from
bee plants treated with conventional pesticides (Regulation
2018/848/EU). However, since honeybees are capable to forage at
distances of up to 10 km (Garbuzov et al., 2015), the pesticide
contamination of organic beekeeping products can not be completely
excluded.
The toxicity of active substances varies widely, and is most often
characterized by LD50, LC50, NOAEL, LOAEL, ADI and ARfD values.
Prior to the authorization of plant protection products, a number of
toxicological studies are carried out to determine the effect of active
substances to human health and the environment. However, the
continuous development of analytical test methods and the improve­
ment of their sensitivity often lead to new research outcomes that make
it necessary for regular review of results in this field (Ambrus et al.,
2020). There are currently about 1500 substances in the European
Union’s Pesticides database with nearly one third being approved.
Maximum Residue Level (MRL) values for honey vary between 0.01 and
1 mg/kg, but for other apicultural products no MRLs are applicable until
individual products have been identified and listed (EU Pesticides
Database, 2021).
Pesticide active substances are generally classified on the basis of the
affected taxonomic groups. The attainment of adequate selectivity is
usually difficult in organisms belonging to closely related taxonomic
groups, like useful and harmful insects, due to the similar nature of
biochemical processes (Sörös, 2019). Determining the pesticide expo­
sure of bee species is a key issue of pollinator protection. According to
Gierer et al. (2019), pollen plays an important role in estimating bee
exposure to pesticides, because it is their main source of nutrients be­
sides nectar. To date, researchers have investigated pesticide residue
content of various pollen samples resulting in a number of available
data. The results vary widely, as they are influenced by a number of
factors including the botanical origin of the sample, the method and
time of application of the pesticides, the doses used, the
physico-chemical properties of the active substances and environmental
conditions like temperature, humidity, water activity and soil
characteristics.
Table 1 summarizes the results of 20 studies from different parts of
the world, which aimed to determine pesticide residue contents of bee
pollen. Relatively little data are available from Asia, Africa and South
America (Toselli & Sgolastra, 2020). In the selected studies, several
active substances were detected from pollen samples simultaneously
using multiresidue methods. The table shows the most common active
substances and their average and maximum concentrations. Studies
reported the widespread occurrence of chlorpyrifos (8), fluvalinate (7),
carbendazim (7) and thiacloprid (7) in pollen samples. The chemical
structures of these compounds are shown in Fig. 1. The concentrations of
the active substances vary in the ppb range in most studies, however, in
certain products; residues were detected in ppm levels. The list of pes­
ticides present in outstanding amounts are diverse.
Chlorpyrifos was a widely used organic phosphorus insecticide that
poses a potential risk to honeybees (Roszko et al., 2016) and also to
human health (Burke et al., 2017). The Committee proposed the with­
drawal of this pesticide, which was accepted by the member states
(Commission Implementing Regulation 2020/18/EU). Fluvalinate
(tau-fluvalinate) is an acaricide used in apiaries and therefore it is often
found in bee products. Carbendazim is a fungicide that inhibits cell di­
vision. This active substance has been banned in the European Union
(Commission Implementing Regulation 542/2011/EU), but it is an
active metabolite of several other components, like fuberidazole, thia­
bendazole and thiophanate-methyl (Sörös, 2019). Thiacloprid is a
human carcinogen and reproductive toxicant that has recently been
banned by the European Commission (Commission Implementing
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Trends in Food Science & Technology 114 (2021) 490–509
Regulation 2020/23/EU).
One of the most comprehensive studies on the topic was conducted
by Johnston et al. (2014), who examined the pesticide residue content of
107 bee pollen samples and 25 bee bread samples from 12 European
countries. They determined the concentration of 300 active substances.
Two-thirds of the products were contaminated with at least one active
agent and a total of 53 pesticides were identified including insecticides,
acaricides, fungicides and herbicides. The most common substances
were chlorpyrifos-ethyl, boscalid and thiacloprid. In a few cases, con­
centrations of some fungicides exceeded 1 ppm, for example boscalid
(Sweden), dimethomorph (Italy), fenhexamide (Germany) and cypro­
dinil (Switzerland). Böhme et al. (2018) conducted studies on the
pesticide residue content of bee pollen over five years in Germany,
involving 281 samples. Approximately four-fifths of the samples were
contaminated with at least one substance. A total of 73 active substances
were detected, mostly fungicides. Most pesticides were revealed in
samples from hives located near orchards, followed by products from
cereal plantations and meadows. The most common active substance in
bee pollen was thiacloprid with the highest concentration of 470 μg/kg.
Relatively few studies were conducted on the pesticide contamina­
tion of bee pollen in countries outside Europe. Nai et al. (2017) screened
155 pollens for 232 pesticides in Taiwan. It was concluded that at least
one pesticide was detectable in three-quarter of the samples. A total of
56 active substances were identified, of which fluvalinate and chlor­
pyrifos showed the highest detection frequency and also high residue
levels. Tong et al. (2018) examined the pesticide contamination of 189
pollen samples originated from major beekeeping areas of China. There
were insecticides (imidacloprid, thiametoxam, fenpropathrin, bifen­
thrin, chlorpyrifos), acaricides (coumaphos, fluvalinate) and fungicides
(carbendazim, triadimefon) among the most common active substances
in these samples. Mullin et al. (2010) conducted a broad survey of
pesticide contamination of apicultural products across 23 states of the
USA and 1 Canadian province. Most pollen samples contained at least
one systemic pesticide and nearly half of the samples contained both
in-hive acaricides (fluvalinate, coumaphos) and fungicides (chlor­
othalonil). Authors emphasized that the scientific knowledge is incom­
plete on the biological effects of pesticide combinations.
Bees from hives deployed near mass flowering crops tend to collect
pollen from other species if it is possible. It is generally assumed, that the
pesticide residue content of bee pollen is determined by the pesticide
treatment of crops. However, Botías et al. (2015) suggested that the
chronic exposure of bees to neonicotinoids came from residues in
non-focal crop pollen. Wildflowers, for example, can become contami­
nated due to the fact that some pesticides are persistent in the soil.
Böhme et al. (2018) pointed out that pesticide residues in bee pollen
pose a risk not only to bee health, but also raise safety concerns in
human nutrition. In light of this fact, a human risk assessment was
performed for the most common pesticide residues (chlorpyrifos, flu­
valinate, carbendazim and thiacloprid) and pesticides detected in
exceptionally high concentrations in bee pollen. The results are reported
in sections 4.1. and 4.2.
3.2. Toxic metals and metalloids
As a result of increased industrial pollutant emissions and inadequate
waste management, more and more heavy metals are accumulating in
the soil, water and air. The term heavy metal is commonly used for el­
ements with atomic number above 20 and a density greater than 5 g/
cm3. Many of them are essential in small amounts for the enzymatic and
metabolic processes of biological organisms, but in higher concentra­
tions they are harmful. From a toxicological point of view, lead (Pb),
cadmium (Cd) and mercury (Hg) are the most important heavy metals
and certain forms of arsenic (As) can also be toxic at very low concen­
trations. Accumulation of heavy metals in the human body can cause
severe health consequences including carcinogenesis, mental illnesses,
and various disorders affecting growth, development, metabolism and
R. Végh et al. Trends in Food Science & Technology 114 (2021) 490–509

Table 1
Summary of the literature data on pesticide contamination of bee pollen.
Number of Country of Technique Number of screened active Widely occurring active ingredients Concentration Reference
samples origin ingredients (%>LOD) (μg/kg)

Mean Max.

53 Poland GC-MS 161 tebuconazole (21) 30 65 Roszko et al. (2016)

LC-MS/MS thiacloprid (19) 61 136
chlorpyrifos (13) 16 40
205 Poland LC-MS/MS 5 thiacloprid (62) 89 1002 Pohorecka et al. (2012)
acetampirid (45) 4 26
thiametoxam (37) 4 10
281 Germany GC-MS/ 282 thiacloprid (52) n. d. 470 Böhme et al. (2018)
MS protiokonazol-destio (36) n. d. 79
LC-MS/MS
102 Germany LC-MS/MS 262 trifloxystrobin (63) 41 707 Friedle et al. (2021)
thiacloprid (46) 14 258
myclobutanil (38) 12 334
boscalid (35) 8 201
154 Luxembourg GC-MS/ 112 thiacloprid (29) n. d. 133 Beyer et al. (2018)
MS permethrin-cis (12) n. d. 40
LC-MS/MS
554 Italy LC-MS/MS 66 dimethoate (30) 10 179 Tosi et al. (2018)
mandipropamid (20) 9 261
45 Spain LC-MS/MS 63 coumaphos (89) 56 374 Calatayud-Vernich et al.
fluvalinate (47) 11 72 (2018)
amitraz (38) 18 246
chlorpyrifos (31) 10 100
41 Spain GC-MS/ 253 coumaphos (73) 301 1833 Vázquez et al. (2015)
MS fluvalinate (59) 31 71
LC-MS/MS chlorfenvinphos (44) 10 73
chlorpyrifos (39) 9 69
carbendazim (29) 3 6
thiabendazole (29) 2 4
thiacloprid (29) 1 2
128 France GC-MS/ 80 carbendazim (34) 24 2595 Lambert et al. (2013)
MS amitraz II (15) 7 129
LC-MS/MS
107 Europe GC-MS/ 300 chlorpyrifos-etil (17) 23 705 Johnston et al. (2014)
MS boscalid (13) 19 1081
LC-MS/MS thiacloprid (13) 10 250
189 China GC-MS 66 carbendazim (76) 169 4516 Tong et al. (2018)
LC-MS/MS fluvalinate (53) 66 531
chlorpyrifos (51) 39 327
fenpropathrin (45) 57 1323
bifenithrin (36) 37 790
36 Thailand GC-MS 178 chlorpyrifos (31) n. d. 244 Chaimanee et al. (2019)
LC-MS/MS fluvalinate (19) n. d. 47
155 Taiwan GC-MS/ 232 fluvalinate (48) 915 16260 Nai et al. (2017)
MS chlorpyrifos (42) 128 956
LC-MS/MS carbaryl (11) 83 220
32 USA (Maine) LC-MS/MS 190 carbendazim (34) 2 6 Drummond et al. (2018)
atrazine (31) 2 14
propioconazole (28) 3 17
313 USA (New LC-MS/MS n.d. coumaphos (47) 6 163 Stoner & Eitzer (2013)
York) carbaryl (41) 28 227
phosmet (33) 227 16556
carbendazim (29) 50 1800
atrazine (27) 3 88
350 North America GC-MS 200 fluvalinate (88) 95 2670 Mullin et al. (2010)
LC-MS/MS coumaphos (75) 180 5828
chlorpyrifos (44) 53 830
280 chlorothalonil (53) 3015 98900
160 USA (Virginia) GC-MS 11 fluvalinate (29) n.d. 25 Fulton et al. (2019)
coumaphos (27) n.d. 338
chlorothalonil (15) n.d. 130
21 Brazil GC-MS/ 26 permethrin (19) n.d. 70 de Oliveira et al. (2016)
MS aldrin (14) n.d. 102
12 Uruguay LC-MS/MS 19 azoxystrobin (100) 1 5,5 Niell et al. (2015)
carbendazim (92) 3 9
carbaryl (75) <1 1
30 Seychelles LC-MS/MS 108 carbendazim (100) n.d. 3 Muli et al. (2018)
imidacloprid (100) n.d. 1
chlorpyrifos (100) n.d. 1
metalaxyl (100) n.d. 3
azoxystrobin (100) n.d. 2
diazinon (100) n.d. <1
carbosulfan (100) n.d. 5
hexaconazole (100) n.d. 1

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R. Végh et al.
Key: n.d.: no data, GC: gas chromatography, LC: liquid chromatography, MS: mass spectrometry, LOD: limit of detection.
Fig. 1. Chemical structures of pesticides commonly found in bee pollen (Pubchem, 2021).
nervous system (Mishra et al., 2018). In biological systems, inorganic
contaminants compete with essential trace elements for free binding
sites, thereby negatively impacting on the maintenance of homeostasis
(Sattler et al., 2016).
Numerous studies demonstrate that bees and beekeeping products
can be used as biological indicators of the heavy metal contamination of
a given area (Bargánska et al., 2016; Murashova et al., 2020; Roman
et al., 2009). According to Bargánska et al. (2016), bees forage pollen,
nectar and propolis within an approximate 7 km2 area from early spring
to late fall. As bees consume those products during winter, which were
stored in summer, they can be exposed to environmental contaminants
all year round. Investigation of inorganic contaminants of bee products
is also a food safety issue (Roman et al., 2016). Morgano et al. (2010)
estimated the extent of contribution of bee pollen consumption to the
provisional tolerable weekly intake (PTWI) of toxic elements. The most
significant contributions are shown by Al (27%) and As (8%). Campos
et al. (2008) developed a comprehensive set of quality criteria for
commercially available bee pollen, which was published on the website
of the International Honey Commission and is often quoted in scientific
research. Maximum limits have been proposed for Pb (500 μg/kg), Cd
(30 μg/kg) and Hg (10 μg/kg) as well.
Several studies support that toxic metal concentrations of bee pollen
significantly depends on the degree of environmental pollution (Conti &
Botré, 2001; Kostić et al., 2015; Lambert et al., 2012; Morgano et al.,
2010; Roman et al., 2009). Campos et al. (2008) suggest beekeepers to
collect pollen at least three km distant from pollution sources. However,
bees can forage up to 10 km from the hive (Garbuzov et al., 2015), thus
pollen can become contaminated from more distant areas. According to
Roman (2009), the concentrations of Cd, Hg and As were significantly
higher in bee pollen originating from a former military airfield than in
those from agricultural and woodland regions. For Cd, Pb and Hg, 96, 60
and 8% of all samples, respectively, exceeded the limits recommended
by Campos et al. (2008). Cadmium also accumulated to a large extent in
bee pollen collected in agricultural and woodland regions, presumably
due to its presence in mineral fertilizers and certain pesticides. Conti and
Botré (2001) examined beekeeping products that originated from the
downtown and outskirts of Rome and found that intense vehicular traffic
in the city center increased heavy metal content of bee pollen, propolis
and beeswax significantly, but it was not true for honey. In bee pollen
from the midtown, large amounts of cadmium and lead had been
accumulated compared to other products. In contrast, their chromium
content was relatively low and independent of the provenance. The Pb
content of pollen samples was below the limit recommended by Campos
et al. (2008) in all cases, but the Cd concentration exceeded the limit of
30 μg/kg in 43% of the samples (100% of the samples from the city
center). According to Sattler et al. (2016), pollen can easily be
contaminated with cadmium, as it is present in many industrial pro­
cesses and can be found in phosphate-based fertilizers as well.
Kostić et al. (2015) determined the botanical composition and
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Trends in Food Science & Technology 114 (2021) 490–509
element content of bee pollen samples from six regions of Serbia. They
concluded that the element content of bee pollen is much more influ­
enced by botanical composition than by geograpichal origin. They
observed high concentrations of aluminium, iron, chromium, cobalt and
zinc in samples with high proportion of certain taxonomic groups
(Helianthuus, Rosaceae, Raxinus, Sophora). On the other hand, it has also
been shown that urban pollutants had a significant effect on the toxic
element content of samples from Belgrade. The authors suggested ana­
lysing elements only in monofloral pollen samples of the same plant
species that originated from different areas, in order to determine the
environmental impact on the product.
Some research has been conducted around the world on the variation
of heavy metals in bee pollen at different periods of the year. Morgano
et al. (2010) examined bee pollen samples collected from three different
locations of Brazil on a monthly basis for a year. The concentrations of
aluminium, cadmium, cobalt and lead were higher in the dry season
(from August to October), as this period is more proned to air pollution.
Theoretically, other inorganic pollutants may also be disseminated by
air even though their concentration in bee pollen has not shown a def­
inite pattern during the studied period. In case of aluminium, another
explanation may be that soil leaching in the rainy season promotes the
accumulation of this element in plants. The accumulated Al will appear
in the next blossoms that will occur during the dry period.
Álvarez-Ayuso & Abad-Valle (2017) examined Spanish bee pollen
samples collected in three different periods. They found that the con­
centrations of As and Zn were the highest during the driest period (June)
and lowest in the period with the highest pluviometric precipitation
(September). The reason for this is that dry weather favours the
contamination of pollen with particulate matter. Lambert et al. (2012)
obtained similar results by examining the lead content of bee pollen
samples collected in 2008 and 2009 from four locations of France. The
samples were collected during four different months in both years. It was
found that the lead content of products collected in the dry months (end
of June/beginning of July, end of September/beginning of October)
were significantly higher. The patterns obtained for the four products of
different origin showed great similarity during the studied period. In
contrast, Al Naggar et al., 2013 concluded that the reason for seasonal
differences of Cu, Zn, Cd, Pb and Fe content in bee pollen samples from
Egypt collected in spring and summer might be the variation of botanical
composition.
Table 2 summarizes the literature data on the concentration of
toxicologically important elements (As, Cd, Hg, Pb) in bee pollen. The
arithmetic mean of the measured concentration values was given for the
four elements. Mean values exceeding the limits recommended by
Campos et al. (2008) are marked with an asterisk (*). Arsenic content of
pollens are usually between LOD and 500 μg/kg, but monofloral samples
from different parts of China are strongly contaminated with this
element, regardless of plant origin. The mean values for Cd concentra­
tion of bee pollen samples exceeded 30 μg/kg in most studies. The data
R. Végh et al. Trends in Food Science & Technology 114 (2021) 490–509

Table 2
Summary of the literature data on toxic metal contamination of bee pollen.
Number of Year of Botanical origin Country of origin (Type of Technique Mean concentration (μg/kg) Reference
samples harvest area)
As Cd Hg Pb

23 1998 n.d. Italy (Outskirts of Rome) AAS n.a. 30 n.a. 112 Conti & Botré (2001)
6 1998 n.d. Italy (Center of Rome) AAS n.a. 67* n.a. 272
10 2007–2009 Multifloral Italy (50 km from post AAS n.a. 310* n.a. 750* Satta et al. (2012)
mining sites)
22 2007–2009 Multifloral Italy (post-mining sites) AAS n.a. 2965* n.a. 3620*
18 2008–2009 n.d. France (hedgerow) AAS n.a. n.a. n.a. 237 Lambert et al. (2012)
72 2005–2006 n.d. Poland (agricultural- ICP-OES 31 253* 4 648* Roman (2009)
woodland)
72 2005–2006 n.d. Poland (former military ICP-OES 78 360* 6 771*
airfield)
25 2012 n.d. Poland (area of copper ICP-OES 290 33* n.a. 147 Roman et al. (2016)
industry)
80 2009 n.d. Poland FAAS n.a. 51* n.a. 1673* Formicki et al. (2013)
3 2014 Multifloral Spain (mine area) ICP-MS 340 97* n.a. n.a. Álvarez-Ayuso & Abad-Valle
3 2014 Multifloral Spain (3 km from a mine ICP-MS 133 157* n.a. n.a. (2017)
area)
25 2011 n.d. Serbia ICP-OES n.a. 67* n.a. n.a. Kostić et al. (2015)
17 2010 n.d. Bulgaria AAS n.a. 155* n.a. 900* Zhelyazkova & Atanassova
(2011)
1 2007 Papaver somniferum Slovakia AAS n.a. 250* 5 <100 Kacániová et al., 2010
1 2013 Betula verrucosa Slovakia AAS <300 46* 5 240 Shevtsova et al. (2014)
3 2011 Betula verrucosa Ukraine AAS <300 291* 4 387
15 2018 n.d. Europe ICP-MS n.a. 88* n.a. 247 Adaškevičiütė et al. (2019)
9 2016 n.d. Turkey ICP-AES n.a. 68* n.a. 347 Özcan et al. (2019)
7 2016 Multifloral Turkey ICP-MS n.a. <LOD n.a. 96 Temizer et al. (2018)
1 2020 Rhododendron Turkey ICP-MS <LOD n.a. <LOD <LOD Bayram (2021)
ponticum
20 n.d. n.d. Turkey ICP-OES 391 69* n.a. 193 Altunatmaz et al. (2017)
1 n.d. n.d. Canada ICP-OES 8 77* n.a. <LOD
2 n.d. n.d. China ICP-OES 474 64* n.a. 2
1 n.d. n.d. Chile ICP-OES 1054 44* n.a. 339
1 n.d. Escallonia rubra Chile (near Llaima Volcano) ICP-OES n.a. 20 n.a. 20 Mejías et al. (2018)
Lotus uliginosus
1 n.d. n.d. Chile (distant Llaima ICP-OES n.a. 30 n.a. 30
Volcano)
43 2007–2008 n.d. Brazil (mostly rural area) ICP-OES 400 34* 1 120 Morgano et al. (2010)
5 2011 n.d. Brazil ICP-OES n.a. 9 n.a. 15 Sattler et al. (2016)
5 2016 n.d. Brazil ICP-MS 169 2 n.a. 148 Oliveira et al. (2017)
3 2013 n.d. Brazil (mine areas) ICP-OES 237 <LOQ n.a. 121 Nascimento et al. (2018)
ICP-MS
1 2013 n.d. Brazil (natural area) ICP-OES 24 <LOQ n.a. <LOQ
ICP-MS
8 n.d. n.d. Brasil (site exposed to Voltammetry n.a. 228* n.a. 27 Silveira et al. (2013)
pollution)
6 2010 n.d. Egypt (near traffic) AAS n.a. 333* n.a. 9715* Al Naggar et al., 2013
2 2010 n.d. Egypt (agricultural) AAS n.a. 705* n.a. 1750*
10 2017 n.d. Jordan (agricultural area) ICP-OES 10 3 n.a. 326 Aldgini et al. (2019)
10 2017 n.d. Jordan (industrial area) ICP-OES <20 2 n.a. 358
2 2017 n.d. Jordan (desert) ICP-OES <20 4 n.a. 20
1 2013 n.d. Russia (near a highway) AAS 340 90* n.a. 480 Murashova et al. (2020)
8 2016–2017 Multifloral Philipphines ICP-MS 21 83* n.a. 61 Belina-Aldemita et al. (2020)
1 2010 Brassica napus China ICP-OES 4895 215* <LOD <LOD Yang et al., 2012
1 2010 Citrullus lanatus China ICP-OES 6729 224* <LOD 506*
1 2010 Camellia japonica China ICP-OES 10890 251* <LOD 112
1 2010 Dendranthema China ICP-OES 13604 453* <LOD 980*
indicum
1 2010 Fagopyrum China ICP-OES 6436 335* <LOD 220
esculentum
1 2010 Helianthus annuus China ICP-OES 11595 296* <LOD 534*
1 2010 Nelumbo nucifera China ICP-OES 13135 348* <LOD 47
Gaertn
1 2010 Papaver rhoeas China ICP-OES 2029 232* <LOD 93
1 2010 Rosa rugosa China ICP-OES 2150 206* <LOD 628*
1 2010 Schisandra chinensis China ICP-OES 2980 149* <LOD 891*
1 2010 Vicia faba China ICP-OES 13279 246* <LOD 1298*
1 2010 Zea mays China ICP-OES 2285 142* <LOD 123

n.d.: no data.
n.a.: not analysed.
*: mean values exceeding the limit proposed by Campos et al. (2008).

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R. Végh et al.
are close to the threshold in several cases, but the average values of some
products exceed the limit more than ten times. Satta et al. (2012) re­
ported exceptionally high Cd content in pollen originated from Italian
post-mining sites. Only a few data are available for Hg, but their mean
value does not exceed 10 μg/kg in any studies. Average values of Pb
concentration show a large standard deviation. In a product from Can­
ada, the Pb concentration was below the detection limit, while in some
pollen samples from Egypt the average concentration exceeded the 500
μg/kg limit nearly twenty times. In Poland, threshold limits were
established for As (200 μg/kg), Cd (50 μg/kg), Hg (33 μg/kg) and Pb
(500 μg/kg) content of bee pollen (Roman et al., 2016). Although Polish
standards for Cd and Hg are slightly higher than the limits recommended
by Campos et al. (2008), even in this case can be concluded that most
products are heavily contaminated with cadmium. Applying the data of
Table 2, a risk assessment for the heavy metal and arsenic content of bee
pollen was performed in sections 4.1. and 4.3.
3.3. Molds and mycotoxins
Mycotoxins are secondary metabolites produced by filamentous
fungi that evoke a toxic response even in low concentrations to higher
organisms. They occur mainly in foods of plant origin, but some my­
cotoxins can also be detected in milk and meat of animals that consumed
contaminated feed. The growth of mycotoxin-producing molds is
affected by a number of factors, like temperature, humidity, oxygen
content of the air, physical damage of crops and the presence of fungal
spores as well (Alarcón et al., 2019). Mycotoxin contamination is a
complex and multifactorial phenomenon that poses a challenge to food
safety. The spread of molds is significantly affected by climate change
scenarios being that every mold species have their own optimum envi­
ronmental condition for growth and secondary metabolite production.
Consequently, the presence of mycotoxins in foods is considered to be
unpredictable (Pereira et al., 2019). Global warming extensively in­
fluences the climate of temperate zones, resulting in aflatoxins becoming
more common in these areas (Kostić et al., 2019). Most mycotoxins are
difficult to eliminate during processing due to their stability against
heat, physical and chemical treatments. To date, more than 300 myco­
toxins have been identified, but in food and animal feedstuffs only a few
occur frequently. These are aflatoxins (AFL), ochratoxins (OTA), fumo­
nisins (FUM), patulin, zearalenone (ZEA) and trichothecenes including
deoxynivalenol (DON) and T-2 toxin (T-2) (Alshannaq & Yu, 2017). The
International Agency for Research on Cancer (IARC) classifies myco­
toxins based on sufficient evidence for carcinogenicity. Aflatoxin-B1
belongs to Group 1 (carcinogenic to humans), because it significantly
increases the risk of developing a malignant tumour in the liver.
Ochratoxin-A and fumonisins had been classified to Group 2B (possibly
carcinogenic to humans), while patulin, zearalenon and trichothecenes
to Group 3 (not classifiable as to its carcinogenicity to humans).
According to Kostić et al. (2019), the physico-chemical properties of
bee pollen are optimal for the growth of several microorganisms
including mycotoxin-producing molds. The European Commission has
set maximum tolerable levels for a number of mycotoxins in various
foods (Commission Regulation 1881/2006/EC), but there are no limits
for beekeeping products. Campos et al. (2008) proposed limit values for
aflatoxins (B1: max 2 μg/kg; B1+B2+G1+G2: max 4 μg/kg) and for the
total amount of yeasts and molds (max 50,000 CFU/g).
Bee pollen can easily be contaminated with aflatoxins, in which
climatic conditions, processing methods and human hygiene play a
prominent role. The hive is another possible source of contamination,
but propolis effectively protects the bees against aflatoxin-producing
molds (Kostić et al., 2019). According to recent studies, propolis ex­
tracts have inhibitory effect on the growth of Aspergillus parasiticus and
they reduce the expression of aflR, nor-1, ver-1 and omtA genes that are
involved in the biosythesis of aflatoxins (Hosseini et al., 2020; Pour
et al., 2020). The presence of fumonisins in bee pollen usually indicates
that the product contains pollen from maize (Zea mays), but the
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Trends in Food Science & Technology 114 (2021) 490–509
contamination can also occur under certain weather conditions, espe­
cially in case of damages caused by insects (Kostić et al., 2019).
The moisture content plays a key role in the safety of bee pollen and
is the main determinant of its shelf-life. The water content of freshly
collected products vary between 20 and 30% before drying. Adequate
microbiological stability is usually provided by drying, but alternative
methods like freezing and lyophilisation are also used for preservation to
protect heat-labile components like vitamin C or provitamin A. If the
moisture content of dried pollen exceeds 6%, the pollen may be
contaminated with microorganisms, but moisture content below 5% can
lead to objectionable organoleptic properties (Campos et al., 2008).
National regulations require a maximum moisture content of 6% for bee
pollen in Poland and Switzerland, but different requirements have been
set it Bulgaria (max 10%), Argentina (max 8%) and Brazil (max 4%).
Differences in regulations may be due to the fact that the obtained
moisture value is influenced by the analytical method applied for the
determination. By using the conventional drying method at 105 ◦ C,
decomposition reactions and loss of volatile compounds can occur. Thus,
the obtained results can be twice as high as values measured by alter­
native methods like infrared radiation, lyophilization, vacuum drying or
Karl-Fischer titration (Melo & Almedia-Muradian, 2011).
Mycotoxin accumulation usually occurs when pollen harvest is not
carried out frequently, high humidity of the open air favours the spread
of molds. It is therefore critical to empty the pollen traps daily, then
store the product in sterile environment and process it as soon as
possible (Kačániová et al., 2011). Garcia-Villanova et al. (2004)
concluded with similar suggestions to avoid biotic hazards both from
natural habitat and from human handling. They recommended to collect
pollen at least every 48 h and to reduce water activity (aw) under 0.60
within 24 h of collection. According to Arruda et al. (2017), collecting
bee pollen on rainy or wet days is not advised due to its hygroscopic
properties. To protect the product against high humidity, the most
appropriate packaging materials for storing are glass and hard plastic.
Over the past decades, several studies have been conducted in
Europe on the mycotoxin content of bee pollen samples, but very little
data are available from other continents. The results of the reviewed
studies are summarized in Table 3. Information of pollen collection and
preservation methods was not available in most cases suggesting that
researchers used fresh bee pollen samples for the determination. How­
ever, commercial (Rodríguez-Carrasco et al., 2013) and dried samples
(Garcia-Villanova et al., 2004; Nuvoloni et al., 2021) were also listed in
the table.
The mean values of aflatoxin contamination exceeded the limits of 2
μg/kg (AFL-B1) and 4 μg/kg (AFL B1+B2+B3+B4) recommended by
Campos et al. (2008) in many cases. Two studies reported that the
average concentrations of AFL-B1 exceeded the limit more than four
times in multifloral samples from Serbia (Kostić et al., 2017; Petrović
et al., 2014). The mean concentrations of total aflatoxins were relatively
low in products from rape (Brassica napus), sunflower (Helianthus
annuus) and silver birch (Betula verrucosa), but it reached 16 μg/kg in
poppy (Papaver somniferum) pollen. The amount of aflatoxins and OTA
were below the limit of quantification in dried bee pollen samples from
Spain (Garcia-Villanova et al., 2004). The authors emphasized that re­
sults may be related to rapid drying and optimal moisture content. Ac­
cording to Nuvoloni et al. (2021), the mean concentration of aflatoxins
can exceed 20 μg/kg in both fresh and dried pollen samples. Rodrí­
guez-Carrasco et al. (2013) reported very low mycotoxin contamination
in bee pollen, although, they tested only Fusarium toxins. Of the 15
samples examined, only 2 contained mycotoxins, namely neosolaniol
and nivalenol, above the limit of detection. The analysed samples were
randomly purchased in supermarkets, so it can be assumed, that their
moisture content was reduced to the optimal level.
Several genera of molds have been isolated from bee pollen but
Alternaria, Aspergillus, Fusarium and Penicillium are the most common. It
is important to note that not all fungi can be isolated from mycotoxin-
containing bee pollen and mycotoxins are not always detectable from
R. Végh et al. Trends in Food Science & Technology 114 (2021) 490–509

Table 3
Summary of the literature data on mycotoxin contamination of bee pollen.
Bee pollen sample Country of origin Technique Detected toxins Mean concentration (μg/kg) Isolated mold genera Reference

Multifloral, dried (n = 18) Italy ELISA AFL (total) 26 n.a. Nuvoloni et al. (2021)
DON 29
Multifloral, fresh (n = 11) Italy ELISA AFL (total) 21
DON 45
Multifloral Lithuania ELISA ZEA 70 Alternaria Sinkevičienė et al. (2019)
(n = 12) DON 50 Fusarium
T-2 <5 Mucor
Penicillium
Multifloral Serbia ELISA AFL-B1 8 Alternaria Kostić et al. (2017)
(n = 26) Aspergillus
Fusarium
Mucor
Penicillium
Rhizopus
Multifloral Serbia ELISA AFL-B1 9 Acremonium Petrović et al., 2014
(n = 33) Alternaria
Aspergillus
Cladosporium
Epiccocum
Fusarium
Mucor
Penicillium
Rhizopus
Betula verrucosa Ukraine. Slovakia HPLC/FD AFL (total) <4 Alternaria Shevtsova et al. (2014)
(n = 4) OTA <0.5 Aspergillus
FUM-B1 <100 Cladosporium
FUM-B2 <100 Penicillium
ZEA <10 Rhizopus
DON <0.1
T-2 <10
Multifloral China LC-MS/MS AFL-B1 <LOQ n.a. Xue et al. (2014)
(n = 20) AFL-B2 <LOQ
AFL-G1 <LOQ
AFL-G2 <LOQ
OTA <LOQ
Multifloral Spain GC -MS/MS DON <LOD n.a. Rodríguez-Carrasco et al. (2013)
(n = 15) 3-AcDON <LOD
FUS-X <LOD
DAS <LOD
NIV <LOD
NEO 3.47
HT-2 <LOD
T-2 <LOD
Papaver somniferum Slovakia ELISA AFL (total) 16 Alternaria Kačániová et al. (2011)
(n = 5) OTA 6 Aspergillus
FUM (total) <LOD Cladosporium
ZEA 362 Fusarium
DON 183 Mucor
T-2 299 Paecilomyces
Brassica napus Slovakia ELISA AFL (total) 4 Penicillium
(n = 5) OTA 10 Rhizopus
FUM (total) <LOD
ZEA 182
DON 215
T-2 265
Helianthus annuus Slovakia ELISA AFL (total) 1
(n = 5) OTA 7
FUM (total) 10
ZEA 147
DON 203
T-2 365
Multifloral Spain HPLC/FD AFL-B1 <LOQ n.a. Garcia-Villanova et al. (2004)
(n = 20) AFL-B2 <LOQ
AFL-G1 <LOQ
AFL-G2 <LOQ
OTA <LOQ

n.a.: not analysed.

mold-contaminated products. Therefore, microbiological and toxin
analytical tests are both required for food safety monitoring of bee
pollen (Kostić et al., 2019).
Campos et al. (2008) concluded that in general, microbiological
characteristics of bee pollen seems to be of greater importance for food
safety than pesticide and heavy metal contamination. Based on the
available data of human exposure and tolerable intake values, a risk
assessment was performed for AFL-B1, OTA, FUM, ZEA, DON and T-2
content of bee pollen. Results are shown in sections 4.1. and 4.3.

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3.4. Pyrrolizidine alkaloids
According to Mädge et al. (2020), pyrrolizidine alkaloids (PAs) are
secondary metabolites that contribute the survival of certain plant
species as a part of their natural defence mechanism against herbivores.
To date, more than 660 PA compounds have been identified that have a
structure characteristic of the source plant. Their common feature is the
necine base composed of two fused five-membered rings with a nitrogen
atom at the bridgehead. Necine base is esterified with one or two necic
acids. These are mono- or dicarboxylic acids with branched carbon
chains containing hydroxy-, alkoxy-, epoxy- and carboxyester groups.
The schematic structure of pyrrolizidine alkaloids is represented in
Fig. 2. Chemical modifications of both necine base and necic acids are
responsible for the structural diversity of PAs. In plants, PAs are natu­
rally found in the form of tertiary bases and/or their corresponding
N-oxides (PANOs), where the nitrogen is oxidised.
Globally, more than 6000 plant species contain pyrrolizidine alka­
loids, which mainly belong to Boraginaceae (all genera), Asteraceae
(tribes Eupatoriae and Senecioneae) and Fabaceae (Crotalaria) plant
families. The PA content of a product is influenced by botanical and
geographical origin, plant organ, growth stage and environmental fac­
tors. Humans are mainly exposed to PAs through traditional medicine
and foods contaminated by PA-producing plants like tea, herbal infu­
sion, honey, dietary supplements and spices. According to Moreira et al.
(2018), the toxicity of different PA metabolites varies according to their
chemical structures. The liver is the main target organ of toxicity, but
carcinogenic, genotoxic, teratogenic and neurotoxic effects as well as
lung lesions have also been reported.
In 2011, The EFSA Panel on Contaminants in the Food Chain
(CONTAM) concluded that no tolerable daily intake could be established
for pyrrolizidine alkaloids. Instead, they applied the Margin of Exposure
(MoE) approach. A benchmark dose lower confidence limit for a 10%
excess cancer risk (BMDL10) of 70 μg/kgbw/day was calculated as the
reference point for comparison with the estimated dietary exposure. The
panel concluded that exposure to PAs from pollen, tea, herbal infusions
and herbal dietary supplements could potentially pose a risk to the
consumer. Therefore, chronic and acute dietary exposures of PAs were
estimated in the European population in 2016. Chronic exposure to PAs
for consumers of pollen-based products ranged between 0.7 and 12 ng/
kgbw/day, while acute exposure was between 2.8 and 44 ng/kgbw/day.
Based on a reassessment of rodent carcinogenicity studies and revised
guidelines for benchmark dose modelling, a new reference point of 237
μg/kgbw/day was established (EFSA, 2017).
The Joint FAO/WHO Expert Committee on Food Additives also used
the MoE approach. The Committee used BMDL10 of 182 μg/kgbw/day
and estimated the chronic exposure for children and adults from con­
sumption of honey and tea, separately. It was concluded that the
calculated MOEs for honey and tea for high consumers may be a concern
(World Health Organization, 2016).
The Committee on Herbal Medicinal Products (HMPC) of the Euro­
pean Medicines Agency issued a Public Statement in 2014, which set an
adult limit of 0.35 μg toxic unsaturated pyrrolizidine alkaloid per day
Fig. 2. Schematic representation of the chemical structure of pyrrolizidine al­
kaloids (Mädge et al., 2020).
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Trends in Food Science & Technology 114 (2021) 490–509
over a maximum of 14 days of consumption. Two years later, a transi­
tional limit of 1.0 μg was recommended by the HMPC in order to
implement measures to reduce PA contamination. Most European au­
thorities accepted the limit. The transitional period was extended until
31 May 2021 (Steinhoff, 2019).
Bees collect pollen and nectar from a wide range of wildflowers
(Botías et al., 2015) so consequently, these toxins can accumulate in
apicultural products. The PA content of honeys has been extensively
investigated, but a number of studies on pollen are also conducted
mainly in Switzerland and Germany. The PA content of bee pollen de­
pends primarily on its botanical composition. According to European
studies, bee pollen containing PAs can be characterized by a relatively
high proportion of pollen from plant species belonging to the genera
Echium, Senecio and Eupatorium (Boppré et al., 2008; Dübecke et al.,
2011; Kast et al., 2018; Kempf et al., 2010).
Kempf et al. (2010) examined the PA content of 55 commercial bee
pollens. It was concluded that PA-positive pollen are more frequent
(31%) compared with honey (9%). In addition, the average PA content
of pollen is 100 times higher than those found in honey. It was also
demonstrated by palinological studies that 94% of PA-positive samples
contain a significant proportion of pollen from plants belonging to the
genus Echium. On the other hand, species from Senecio and Eupatorium
occurred rarely. Samples with increased PA concentrations (>10 μg/g)
had the highest proportions of Echium pollen (75–83%). However, in
some cases low PA concentrations were detected despite a great amount
of Echium pollens. Researchers agree that microscopic analysis without
examining the PA concentration is not sufficient for monitoring
apicultural products (Kast et al., 2019; Kempf et al., 2010).
Several studies confirm that pollen from Echium vulgare is the main
contributor to PA contamination of European bee pollens (Kast et al.,
2019; Kempf et al., 2010; Lucchetti, 2017). According to Kast et al.
(2019), the main blooming season of this plant is June and July, but the
flowering can start as early as April and last until autumn. The PA
content of 262 bee pollen samples collected in three different years was
determined and palynological analysis was also performed. The all-years
sum of Echium-type pollen did not reach 0.02%, however, the PA content
of samples collected in July exceeded 4000 μg/kg. Boppré et al. (2008)
examined the pyrrolizidine alkaloid content of pure pollens and mono­
floral pollen pellets of Echium vulgare, Eupatorium cannabinum, Senecio
jacobaea, Senecio ovatus and Echium plantagineum. Their results indicated
that the PA content of pollen directly collected from flowers was higher
than bee-collected pollen loads although, pollen pellets also had a very
high PA content of between 6000 and 350,000 μg/kg. These findings
support the suggestion that the presence of pollen grains of certain plant
species contributes significantly to the PA contamination of honeys. In
addition, authors investigated the effect of heating on the total pyrro­
lizidine alkaloid level of pollen loads. Gentle treatment (35 ◦ C, 72 h)
resulted in approximately 30% reduction of total PAs, but when higher
temperatures were applied for a longer period (40 ◦ C, 84 h and 60 ◦ C, 84
h), a decrease of 65% and 75%, respectively, were observed.
Kast et al. (2019) investigated the amount and thermal stability of
pyrrolizidine alkaloids in Swiss bee breads. The samples contained PAs
between 9 and 2538 μg/kg. There was no significant change in samples
stored at 15 ◦ C, whereas the amount of PAs in the products stored at
30 ◦ C decreased by approximately one third after 6 months. The
decrease was mainly due to the heat sensitivity of N-oxides, but many PA
compounds remained stable at high temperatures. Only a few studies
have been published on the PA content of royal jelly and propolis, but
those studies showed that they had a lower concentration of PAs
compared to honey and bee pollen (Mulder et al., 2018; Lucchetti,
2017). Lucchetti (2017) found that adult bees have a high pyrrolizidine
alkaloid tolerance, but larvae are sensitive to pollen even with low
concentrations of this toxin. However, it has also been observed that
royal jelly contains orders of magnitude less PA than pollen. The author
concluded that nurse bees play an important role in the survival of the
colony by filtering PAs during royal jelly secretion.
R. Végh et al. Trends in Food Science & Technology 114 (2021) 490–509

Scientific data on the pyrrolizidine alkaloid content of apicultural
products are summarized in Table 4, with particular attention to bee
pollen. Each product is likely to contain these compounds above the
limit of detection. In general, the PA content of pollens is higher
compared to honey, royal jelly and propolis samples. The average PA
content of multifloral bee pollens range between 100 and 1597 μg/kg,
but the maximum concentration reaches 37,855 μg/kg. Based on the
data of multifloral bee pollen, a risk assessment was performed in sec­
tion 4.3. Boppré et al. (2008) used monofloral pollen pellets of Echium,
Eupatorium and Senecio species for their experiments, which showed
extraordinarily high PA content. As PA-producing plants are widespread
in many environments (Lucchetti, 2017), it is suggested to monitor
hepatotoxic pyrrolizidine alkaloid levels in pollen supplies intended for
human consumption.
3.5. Allergens
Allergy is a specific reaction of the immune system to certain sub­
stances, which is manifested in various clinical symptoms. The number
of allergic patients is constantly increasing, and half of the next gener­
ation of Europe may be affected (Nanotte-Varly, 2016). The most com­
mon allergens are pollen from anemophilous (wind-pollinated) plants,
like birch (Betulaceae), ragweed (Ambrosia artemisiifolia) and mugwort
(Artemisia vulgaris), but pollen from entomophilous (insect-pollinated)
plants can also cause allergic symptoms (Yang et al., 2019). According to
Pitsios et al. (2006), bee pollen contains 400,000–6,400,000 pollen
grains per gram, which can cause serious allergic side effects, even
anaphylaxis. Nanotte-Varly (2016) found that the occurrence of bee
pollen allergy in the Polish population is 0.5%, however, cases of mild
reaction are rarely reported and this value is presumably
underestimated.
Table 5 summarizes 10 case studies from the last 40 years on the
allergic symptoms caused by bee pollen consumption. Important
information of patients and data on the botanical origin of pollen that
triggered reactions are also included.
Several research groups have found that people with pollen allergy,
atopy or asthma have an increased risk to have an allergic reaction after
ingesting bee pollen. Patients were between 4 and 56 years of age, with
an average of 31 years. The most common symptoms are swelling of the
mouth and tongue, swallowing difficulties, sore throat, itching, rushes,
gastrointestinal symptoms, and general malaise. The botanical origin of
pollens presumed to trigger reactions shows great variety, but dandelion
(Taraxacum officinale) and other Asteraceae species were mentioned in
most cases. Greenberger and Flais (2001) suggested that allergic reac­
tion can also be induced due to the presence of fungi.
According to Shahali (2015), it is a common misconception that bee
pollen contains pollen grains only from entomophilous plants as
airborne pollen can easily incorporated into the product. Furthermore,
bees collect pollen from anemophilous plants under certain conditions.
In early spring, it is particularly common that bees collect pollen from
ash (Fraxinus), willow (Salix) and oak (Quercus) in large quantities.
Pollen loads collected in mid-summer often contains large amounts of
grains from maize (Zea mays) and species of Chenopodioideae. According
to Sebestyén (2014), bees tend to collect pollen from highly allergenic
ragweed (Ambrosia artemisiifolia) at certain times, therefore it can
potentially get into different apicultural products. However, a study by
Basista et al. (2012) suggests that bee pollen allergy is associated with
the occurrence of proteins from insect saliva.
Cross-reactions can also develop due to structural similarities of
proteins in different pollens. An example of this is the pollen of birch
(Betulaceae), which shows cross-reaction with the pollen of many plant
species belonging to different taxonomic groups like apple (Malus),
celery (Apium graveolens) and soybean (Glycine max). Pollen of mugwort
(Artemisia vulgaris) cross-reacts with pollen from several plants preferred
by bees like sunflower (Helianthus annuus), dandelion (Taraxacum offi­
cinale) and species of goldenrod (Solidago). Panallergens (e.g. profilin)

Table 4
Summary of the literature data on pyrrolizidine alkaloid content of apicultural products.
Samples Country of origin Technique %> PA mean (μg/ PA max. (μg/ Reference
LOD kg) kg)

Honeys of various botanical origins (n = 381) Europe LC-MS/ 65 17 225 Dübecke et al.
MS (2011)
Honeys of various botanical origins (n = 2839) Central- and South-America LC-MS/ 68 46 1087 Dübecke et al.
MS (2011)
Honeys of various botanical origins (n = 60) China LC-MS/ 40 3 16 Zhu et al. (2018)
MS
Honeys of various botanical origins (n = 60) Marketed in China LC-MS/ 75 31 289 Zhu et al. (2018)
MS
Bee pollens of various botanical origins (n = 119) n.d. LC-MS/ 60 1108 37855 Dübecke et al-,
MS 2011
Bee pollens of various botanical origins (n = 55) Europe, Asia, USA, Mexico, New- GC-MS 31 1597 16350 Kempf et al. (2010)
Zealand
Bee pollens of various botanical origins (n = 32) Marketed in Switzerland LC-MS/ 31 100 1185 Kast et al. (2018)
MS
Bee pollens of various botanical origins (n = 5) Switzerland LC-MS/ 80 240 586 Kast et al. (2018)
MS
Bee pollens of various botanical origins (n = 5) Marketed in Belgium LC-MS/ 80 555 1672 Picron et al. (2019)
MS
Bee pollens of various botanical origins (n = 44) Marketed in Italy and online stores LC-MS/ 77 402 3356 Inacio et al. (2020)
MS
Bee pollen of Echium vulgare (n = 4) Germany LC-MS 100 350,000 n.d. Boppré et al. (2008)
Bee pollen of Eupatorium cannabinum (n = 10) Germany 100 120,000 n.d.
Bee pollen of Senecio jacobaea (n = 17) Germany 100 100,000 n.d.
Bee pollen of Senecio ovatus (n = 14) Germany 100 70,000 n.d.
Bee pollen of Echium plantagineum (n = 2) Australia 100 6000 n.d.
Supplements containing bee pollen (n = 12) Marketed in Europe LC-MS/ 100 576 1911 Mulder et al., 2018
MS
Bee breads of various botanical origins (n = 8) Switzerland LC-MS/ 100 n.d. 2538 Kast et al. (2019)
MS
Supplements containing propolis and royal jelly (n Marketed in Europe LC-MS/ 47 8 48 Mulder et al., 2018
= 17) MS

n.d.: no data.

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Table 5 packaging of commercially available bee pollen should include infor­

Summary of case studies on bee pollen-induced allergic reactions. mation about potential allergenicity (Yang et al., 2019). Several re­
Age/Gender Symptoms Presumable allergens Reference searchers (Martín-Muñoz et al., 2010; Pitsios et al., 2006; Shahali, 2015)
known allergic attracted attention to the lack of regulation that contributes to mis­
disease conceptions about the product.
35/M Nausea, dizziness, Pollen from Kanneppady
Seasonal cough, wheezing, dandelion or et al. (2018) 3.6. Pollens of genetically modified plants
allergic chest tightness, chrysanthemum
rhinitis itchiness, redness,
(summer) swelling of lips and
Due to agricultural innovations, it become possible to supply billions
gums of people with food, fuel and clothes even with limited land availability
40/M Nasal congestion, Pollen from black McNamara & and fewer farmers (Turnbull et al., 2021). Biotechnological innovations,
Seasonal shortness of breath, willow, cocklebur, Pien (2018) especially genetic modification (GM) techniques are effectively
allergic hives, tongue and marsh elder or
contributing to the success of agriculture as they help to reduce the
rhinitis hand, swelling ragweed
(autumn) during a strenuous environmental footprint, improve food quality and increase productivity
outdoor run (Giraldo et al., 2019). According to the definition of the Cartagena
40/M Nausea, vomiting, Pollen from Choi et al. Protocol on Biosafety (2000), a plant is genetically modified if it con­
Seasonal abdominal pain, dandelion or (2015)
tains a novel combination of genetic material, which was introduced by
allergic diarrhea, chrysanthemum
rhinitis generalized
modern technology. There are two types of modern biotechnological
(autumn) urticaria, facial methods: the fusion of cells beyond the taxonomic family and in vitro
edema, dyspnea nucleic acid techniques. The latter includes recombinant DNA and direct
30/F Swelling of eyelids, Pollen from timothy Jagdis & injection of nucleic acid into cells or organelles (Turnbull et al., 2021).
Seasonal lips and throat, grass Sussman (2012)
Currently, transgenic crops are cultivated on almost 200 million
allergic difficulty of
rhinitis swallowing, hives, hectares, the vast majority of which is concentrated in seven countries:
(summer) shortness of breath, USA, Canada, Brazil, Argentina, Paraguay, India and China. The wide­
paleness, feel of spread global occurrence of transgenic plants has raised concerns about
faint and weak
their impacts on human and animal health (Delaney et al., 2018;
4/M Intense itching in Pollens from speaces Martín-Muñoz
Seasonal the mouth, tongue of the family et al. (2010)
Domingo, 2016; Giraldo et al., 2019), on non-target organisms (Arpaia
allergic and pharynx with Asteraceae et al., 2021; Pinheiro and Faria, 2021), and on the environment (Brookes
rhinitis uvula edema, & Barfoot, 2018; Tsatsakis et al., 2017a). There are no persuasive data
(spring, swallowing that GM foods pose higher health risks compared to those encountered
autumn) difficulties, isolated
daily (Hilbeck et al., 2020), however, according to Domingo (2016), it is
hives on the thorax.
56/F Palmar pruritus, Pollens (rageweed, Greenberger & necessary to conduct studies on the long-term health effects of GM
Perennial generalized grasses, tree pollens) Flais (2001) plants, including tests of mutagenicity, teratogenicity, carcinogenicity
allergic urticaria, acute and/or fungi and allergenicity. The literature provides contrasting evidence of the
rhinitis, dyspnea, throat (Alternaria, effects of GM crops on non-target insects. Most studies show neutral or
atopy tightness, fatigue Cladosporium)
19/M Facial itch and Pollen from Prichard &
“negligible” effects, some reported negative effects, while a few others
Asthma swelling, difficulty eucalyptus Turner (1985) reported positive effects (Pinheiro & Faira, 2020; Abrol, 2012). Hon­
in breathing, eybees (Apis mellifera) are suitable organisms for non-target risk as­
stridor, wheeze, sessments, because they consume pollen. Pollen produced by
edema
insect-resistant, genetically engineered plants contains insecticidal Cry
32/F Throat tickle, Cross- reactive Cohen et al.
Allergic itching of the face Asteraceae antigens (1979) proteins from the bacterium Bacillus thuringiensis (Bt), which have been
rhinitis and eyes, facial (e.g. from dandelion) widely adopted. According to a recent study, chronic oral exposure of Bt
(mainly in urticaria, breathing Cry78Ba1 rice pollen causes negative effects neither to worker bees nor
summer) and swallowing to larvae at the maximum tested concentration (Han et al., 2021).
difficulties,
anxiety, paleness
Extensive cultivation of GM plants is a controversial issue amongst
27/F Watery eyes, sore Cross- reactive Cohen et al. policy makers, scientists and the consumers (Tsatsakis et al., 2017b).
Allergic throat, swelling of Ascteraceae antigens (1979) Labeling policies can include a ban on labeling, voluntary labeling
rhinitis the tongue and lips, (e.g. from dandelion) indicating that a product is GM free, or mandatory labeling indicating
(mainly in generalized pruritis
that a product contains at least one ingredient that is coming from a
Aug–Sept) and urticaria
25/M Swelling and Cross- reactive Cohen et al. plant with an altered genetic composition. In the latter case, a legal
Allergic itching in the Asteraceae antigens (1979) tolerance threshold is set, which varies between countries: in the United
rhinitis mouth, tightness in (e.g. from dandelion) States, Canada and Japan, conventional products may contain up to 5%
(mainly in the throat. GM material before they must be labeled. In Australia, New Zealand,
Aug–Oct) cutaneous itching,
flushing, profuse
South Africa, Brazil and China, the tolerance threshold is set at 1%,
sweating, rapid, while in the EU, a 0.9% limit is applied to approved products and zero
weak pulse, tolerance to unapproved products (Giraldo et al., 2019; Ramessar et al.,
hypotension 2010). To date, mandatory GM labeling laws are not applied to products
n.d.: no data. from animals fed with GM feed as well as GM processing aids and en­
zymes (Van-Eenennaam & Young, 2017). However, some EU member
are responsible for cross-reactions between different allergen sources countries have established regulations and guidelines to label animal
(Pitsios et al., 2006; Shahali, 2015). Some molds (Aspergillus, Cladospo­ products voluntarily as non-GM (Venus et al., 2018).
rium) commonly found in bee pollen can also cause a severe allergic Gene flow is a major evolutionary force, which causes changes in
reaction (Choi et al., 2015; Greenberger & Flais, 2001). gene frequencies along with mutation, genetic drift and selection
To date, only seven countries (Switzerland, Poland, Bulgaria, Brazil, (Tsatsakis et al., 2017a). Natural gene flow occurs through vertical gene
China, Australia and New-Zealand) have requirements that the transfer and is generally subjected to strict taxonomic boundaries.
Horizontal gene transfer (HGT) refers to the movement of a genetic

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material between unrelated organisms, e.g. plants to microorganisms,
humans or pollinators (Lal et al., 2020, pp. 335–394). In the case of GM
organisms, movement of transgene(s) into other species have been
concerned relating to health and natural environments (Giraldo et al.,
2019).
Because many GM crops are self-pollinated, it is generally assumed
that they are not visited by pollinators. However, honeybees as well as
wild bees tend to forage GM crops like canola, maize, alfalfa, soybean
and cotton. Consequently, there is a possibility that transgene DNA and/
or novel proteins may be present in the plant tissues and secretions
collected by bees (Abrol, 2012; Seide et al., 2018). Pollen is considered
the most likely source of GM material in bee products (Malone, 2002).
Compared to leaves, which contain about 2% protein, pollen have a
protein content between 8% and 40%, and for this reason transgenes
may be expected to express novel proteins at reasonable levels in this
tissue. Higher levels of novel proteins may be present in pollen if the
transgene includes a pollen specific promoter. The actual concentration
of GM material in pollen loads depends on the relative amount of GM
pollen to pollen from other plants and the degree of transgene DNA/­
novel protein production in pollen grains (Abrol, 2012).
Honeybees commonly forage up to 2 km from the hive, but among
others, oilseed rape is such an attractive plant source that bees may
travel more than 5 km for it. In fact, under certain conditions, they can
fly distances of more than 10 km (Abrol, 2012). With this in mind,
strategies should be applied to minimise the presence of GM material in
apicultural products. These include separating GM and non-GM crops,
using bee attractants such as a sugar syrup spray on non-GM crops, using
bee repellents such as some pesticides on GM crops or using GM plants
where the transgene is not expressed in pollen (Malone, 2002). Campos
et al. (2008) drew attention to the lack of research on the health impact
of bee pollen containing GM materials. Authors suggest labeling bee
pollen that contain pollen grains of GM plants according to the current
European legislation (Regulation 1829/2003/EC).
4. Risk assessment of toxicologically important contaminants
and components of bee pollen
EFSA defined risk assessment as “a specialised field of applied sci­
ence, that involves reviewing scientific data and studies in order to
evaluate risks associated with certain hazards. It involves four steps:
hazard identification, hazard characterisation, exposure assessment and
risk characterisation.” This task is typically performed by comparing
statistical exposure data with toxicological threshold levels or other
reference points. The results are usually expressed as percent contribu­
tion to a threshold level or a margin of exposure value (EFSA, 2019).
Risk assessment of bee pollen is essential for decision-making, as safety
is a public health priority and an important requirement for trade. In the
following sections, exposure assessments are presented for the common
contaminants and toxic substances of pollen, based on literature data.
The exposure values were compared to official reference values in order
to draw conclusions about the extent of risks for the consumers of the
product.
The recommended daily portion of bee pollen is up to 25 g. Many
authors (Altunatmaz et al., 2017; Melo & Almedia-Muradian, 2011;
Morgano et al., 2010; Sattler et al., 2016) performed calculations with
this intake value. In our study, exposure was also estimated assuming a
daily pollen consumption of 25 g and an average body weight of 70 kg.
4.1. Risk assessment of non-genotoxic carcinogen contaminants of bee
pollen
There are several toxic contaminants in bee pollen, for which
acceptable daily intake (ADI) or tolerable daily, weekly or monthly
intake (TDI, TWI, TMI) values have been established (Table 6). Conse­
quently, four pesticide active substances (chlorpyrifos, fluvalinate, car­
bendazim and thiacloprid), two heavy metals (cadmium and mercury)
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Trends in Food Science & Technology 114 (2021) 490–509
and five mycotoxins (OTA, FUM, ZEA, DON, T-2) are listed. Table 6
shows the estimated exposure as well as the acceptable or tolerable daily
intake values of these compounds. The daily contribution of 25 g pollen
consumption to the acceptable or tolerable daily intake was also
calculated. Contribution data above 10% are marked with an asterisk
(*). For the estimation of exposure, average concentrations of above
mentioned compounds and elements were used based on literature data.
The sources of reference values are the EU Pesticides Database and a
working document of Codex Alimentarius Commission (World Health
Organization, 2019). Of the mentioned pesticides, only fluvalinate is
authorized in the European Union currently. Carbendazim has been
banned for several years (Commission Implementing Regulation
542/2011/EU), however, it can be detected in bee pollens as a degra­
dation product of other pesticides. Chlorpyrifos was banned in early
2020 (Commission Implementing Regulation 2020/18/EU), after it was
proven based on scientific studies that it can not be excluded that
chlorpyrifos has a genotoxic potential and adverse effect on neural
development and reproduction. Thiacloprid was also banned recently
(Commission Implementing Regulation 2020/23/EU) due to its poten­
tial carcinogenicity and reproductive toxicity.
According to the European Union’s Pesticide Database, the available
data is insufficient to set ADI value for chlorpyrifos. Consequently, risk
assessment can not be performed for this substance. The ADI values of
the other pesticides exceeded the daily exposures 15–28,000 times. The
consumption of bee pollen contributed to less than 1% to the acceptable
daily intake of the three pesticides in most cases. Cadmium was present
in significant amounts in many products, and in a few cases, its intake
with pollen surpassed 10% of the tolerable exposure. Satta et al. (2012)
reported exceptionally high levels of Cd contamination in pollens orig­
inated from Italian post-mining sites. Consuming of 25 g of this product
contributes 92% to the established TMI. The mercury content of bee
pollen is included only in six studies, which show low levels of
contamination. Therefore, their significance in the mercury load of the
human body is virtually negligible. Our present knowledge of mycotoxin
contamination of pollen is also insufficient to draw far-reaching con­
clusions. On the other hand, it can be stated that mycotoxins can be
accummulated in significant amounts in the product. Based on the data
published by Kačániová et al. (2011), the exposure of T-2 toxin from
everyday consumption of 25 g pollen exceeds its tolerable daily intake.
The same study reported an average ochratoxin-A concentration of 8
μg/kg, which contributes 20.43% to the tolerable intake of this toxin in
case of a pollen consumption of the same amount. Regarding zear­
alenone and deoxynivalenol, the relevant data are 230 and 207 μg/kg,
with the contributions of 16.43 and 7.39%. According to these results,
Aspergillus and Fusarium contamination may be a serious problem
regarding the food safety of pollen pellets.
4.2. Acute risk assessment of pesticides detected in high concentrations
from bee pollen
According to EFSA (2019), acute exposure assessments are used to
estimate short-term exposure to a substance and are typically compared
to acute reference doses (ARfD). For calculation of acute exposure, in­
dependent daily intakes are considered. The ARfD of a chemical is
defined as “an estimate of the amount of a substance in food and/or
drinking water, normally expressed on a body weight basis, that can be
ingested in a period of 24 h or less without appreciable health risk to the
consumer on the basis of all known facts at the time of the evaluation”.
Concentrations of pesticides are usually below the detection limit in
bee pollens, but sometimes they are present at concentrations of ppm
range. Due to the large variance, acute risk for some pesticides was also
investigated, which is presented in Table 7. Exposure estimation was
performed for samples containing pesticides at concentrations above 2
ppm, assuming a daily intake of 25 g and an average bodyweight of 70
kg. Only those pesticides are listed, for which applicable ARfD values
have been established (EU Pesticides Database, 2021). The contribution
R. Végh et al. Trends in Food Science & Technology 114 (2021) 490–509

Table 6
Risk assessment of non-genotoxic carcinogens of bee pollen.
Compound/ Mean conc. Reference Estimated daily ADIb/TDIc/ Acceptable/Tolerable daily Contribution of pollen to the
element (μg/kg) intakea (ng/kgbw/day) TWId/TMIe intake (ng/kgbw/day) acceptable/tolerable daily intake
(%)

chlorpyrifos 16 Roszko et al. (2016) 5.71 Not set due to insufficient data Can not be determined
10 Calatayud-Vernich et al. 3.57
(2018)
9 Vázquez et al. (2015) 3.21

39 Tong et al. (2018) 13.93

128 Nai et al. (2017) 45.71
53 Mullin et al. (2010) 18.93
fluvalinate 11 Calatayud-Vernich et al. 3.93 ADI: 5000 5000 0.08
(2018)
31 Vázquez et al. (2015) 11.07 0.22
33 Johnston et al. (2014) 11.79 0.24
66 Tong et al. (2018) 23.57 0.47
915 Nai et al. (2017) 326.79 6.54
95 Mullin et al. (2010) 33.93 0.68
carbendazim 3 Vázquez et al. (2015) 1.07 ADI: 20000 20000 <0.01
24 Lambert et al. (2013) 8.57 0.04
169 Tong et al. (2018) 60.36 0.30
71 Nai et al. (2017) 25.36 0.13
2 Drummond et al. (2018) 0.71 <0.01
50 Stoner & Eitzer (2013) 17.86 0.09
3 Niell et al. (2015) 1.07 <0.01
thiacloprid 61 Roszko et al. (2016) 21.79 ADI: 10000 10000 0.22
89 Pohorecka et al. (2012) 31.79 0.32
14 Friedle et al. (2021) 5.00 0.05
1 Vázquez et al. (2015) 0.36 <0.01
10 Johnston et al. (2014) 3.57 0.04
Cadmium 38 Conti &Botré, 2001 13.57 TMI: 25000 833 1.62
(total) 2135 Satta et al. (2012) 763 91.55*
307 Roman (2009) 109.64 13.16*
33 Roman et al. (2016) 11.79 1.41
51 Formicki et al. (2013) 18.21 2.19
127 Álvarez-Ayuso & 45.36 5.45
Abad-Valle (2017)
67 Kostić et al. (2015) 23.93 2.87
155 Zhelyazkova & Atanassova 55.36 6.65
(2011)
250 Kacániová et al., 2010 89.29 10.72*
230 Shevtsova et al. (2014) 82.14 9.86
88 Adaškevičiütė et al. (2019) 31.43 3.77
68 Özcan et al. (2019) 24.29 2.92
<LOD Temizer et al. (2018) Low Low
68 Altunatmaz et al. (2017) 24.29 2.92
25 Mejías et al. (2018) 8.93 1.07
34 Morgano et al. (2010) 12.14 1.46
9 Sattler et al. (2016) 3.21 0.39
2 Oliveira et al. (2017) 0.71 0.09
<LOQ Nascimento et al. (2018) Low Low
228 Silveira et al. (2013) 81.43 9.78
426 Al Naggar et al., 2013 152.14 18.26*
3 Aldgini et al. (2019) 1.07 0.13
340 Murashova et al. (2020) 121.43 14.58*
83 Belina-Aldemita et al. 29.64 3.56
(2020)
258 Yang et al., 2012 92.14 11.06*
Mercury (total) 5 Roman (2009) 1.79 TWI: 4000 571 0.31
5 Kacániová et al., 2010 1.79 0.31
4 Shevtsova et al. (2014) 1.43 0.25
<LOD Bayram (2021) Low Low
1 Morgano (2010) 0.36 0.06
<LOD Yang et al., 2012 Low Low
Ochratoxin-A 8 Kačániová et al. (2011) 2.86 TWI: 100 14 20.43*
<0,5 Shevtsova et al. (2014) <0.18 <1.29
<LOQ Garcia-Villanova et al. Low Low
(2004)
<LOQ Xue et al. (2014) Low Low
Fumonisins 3 Kačániová et al. (2011) 1.07 TDI: 2000 2000 0.05
(B1+B2) <100 Shevtsova et al. (2014) <35.71 <1.79
Zearalenone 230 Kačániová et al. (2011) 82.14 TDI: 500 500 16.43*
70 Sinkevičienė et al. (2019) 25.00 5.00
<10 Shevtsova et al. (2014) <3.57 <0.71
Deoxynivale nol 29 Nuvoloni et al. (2021) 10.36 TDI: 1000 1000 1.04
(continued on next page)

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Table 6 (continued )
Compound/ Mean conc. Reference Estimated daily ADIb/TDIc/ Acceptable/Tolerable daily Contribution of pollen to the
element (μg/kg) intakea (ng/kgbw/day) TWId/TMIe intake (ng/kgbw/day) acceptable/tolerable daily intake
(%)

45 Nuvoloni et al. (2021) 16.07 1.61

207 Kačániová et al. (2011) 73.93 7.39
50 Sinkevičienė et al. (2019) 17.86 1.79
<0,1 Shevtsova et al. (2014) <0.04 <0.01
<LOD Rodríguez-Carrasco et al. Low Low
(2013)
T-2 toxin 201 Kačániová et al. (2011) 71.79 TDI: 60 60 119.65*
<10 Shevtsova et al. (2014) <3.57 <5.95
<5 Sinkevičienė et al. (2019) <1.79 <2.98
<LOD Rodríguez-Carrasco et al. Low Low
(2013)
a
Assuming a consumption of 25 g/day and average bodyweight of 70 kg.

b
Acceptable Daily Intake, ng/kgbw/day.

c
Tolerable Daily Intake, ng/kgbw/day.

d
Tolerable Weekly Intake, ng/kgbw/week.

e
Tolerable Monthly Intake, ng/kgbw/month.

of pesticide contaminated pollens to the ARfD was calculated and
expressed as a percentage. Contribution values above 50% are marked
with an asterisk (*).
The estimated acute exposure was below the ARfD in all cases. Only
two substances showed a contribution of more than 50% to the ARfD.
Mullin et al. (2010) reported that a product contained chlorothalonil at
an extremely high concentration (nearly 100 ppm), while Tong et al.
(2018) detected a great amount of lambda-cyhalothrin content, for
which respectively low ARfD value has been established. The con­
sumption of these products contribute in 70.64 and 89.20% to the acute
reference dose. Other active substances showed a contribution of
0.12–13.25%, which do not pose an acute risk to consumers.
4.3. Risk assesment of genotoxic carcinogen compounds and elements of
bee pollen
For genotoxic carcinogen substances, no Health-Based Guidance
Values (HBGVs) can be established. In these cases, the use of the Margin
of Exposure (MoE) approach is recommended and an adequate MoE is
the precondition to exclude a concern for public health (EFSA, 2019).
The basis of the MoE approach is a reference point (RP) or a point of
departure (PoD) calculated from a dose-response modeling of toxico­
logical data. MoE is calculated by dividing this value with the estimated
human exposure (Cunnigham et al., 2011). MoE is a dimensionless
number, the acceptability of which depends on its magnitude. As a point
of departure, benchmark dose (BMD) is typically used. BMDL is the
lower one-sided confidence limit of the benchmark dose for a pre­
determined level of response, such as a 1.5 or 10% incidence of an effect
(World Health Organization, 2019). The lower the MoE of the genotoxic
carcinogen, the greater the public health concern is. MoEs of 10,000 or
higher are generally not a concern and do not require intervention. This
value is derived by the multiplication of two uncertainty factors. A
100-fold difference between the reference point and human exposure is
necessary to account for species differences and human variability. A
further 100-fold difference is also needed because of interindividual
variability in cell cycle control and DNA repair (Cunnigham et al.,
2011).
Currently, there is no international consensus and terminology on
what MoE values represent negligible, low and high risks, but the rela­
tive magnitude of MoE values for different substances provides useful
information. It is important to note that even with a MoE value of 10,000
and above, risk managers can make a decision that requires the intro­
duction of a measure to reduce human exposure (EFSA, 2012). For
genotoxic carcinogenic substances, applying the “as low as reasonably
achievable” (ALARA) approach is recommended irrespective of the
calculated MoE value (Cunnigham et al., 2011).
More benchmark dose lower confidence limit (BMDL) values are
established for toxic elements and compounds, which can cause
different lesions. For inorganic arsenic, a range of BMDL01 values be­
tween 0.3 and 8 μg/kgbw/day were identified for cancers of the lung,
skin and bladder, as well as skin lesions (EFSA, 2009). For lead, refer­
ence values were established for developmental neurotoxicity (BMDL01
= 0.50), effects on systolic blood pressure (BMDL01 = 1.50) and chronic
kidney disease (BMDL10 = 0.63) (EFSA, 2010).
Table 8 lists the average concentrations of genotoxic carcinogen
substances detected in bee pollens. Based on these data, human expo­
sures were calculated assuming a daily consumption of 25 g pollen and
an average bodyweight of 70 kg. The lowest BMDL values are presented
for inorganic arsenic (EFSA, 2009), lead (EFSA, 2010), AFL-B1 (EFSA,
2020) and pyrrolizidine alkaloids (EFSA, 2017). Based on the BMDL
values and the minimum margin of exposures sufficient to ensure that
there is no appreciable human risk, maximum daily intake values were
calculated. For AFL-B1 and pyrrolizidine alkaloids, a MoE of 10,000
(EFSA, 2017; EFSA, 2020), for lead a MoE of 10 (EFSA, 2010) and for
arsenic a MoE of 1 (Menon et al., 2020) were taken into account. Esti­
mated human intake and maximum daily intake are expressed in
ng/kgbw/day, the ratio of which indicates the contribution of pollen
consumption to the assumed maximum daily intake of the given
contaminant.
According to Vahter & Concha (2001), the degree of toxicity of As
depends on its chemical form. Elemental arsenic is generally more toxic
than organic forms, because the methylation of arsenic may be consid­
ered a detoxification mechanism. The resulting metabolites are less
reactive with tissue constituents and excreted in the urine to a greater
extent, while elemental arsenic, especially AsIII (arsenite) is highly
reactive with tissue constituents due to its strong binding affinity for
sulfhydryl groups. There is no information on the chemical form of
arsenic in bee pollen, therefore using the BMDL01 value of 0.3
μg/kgbw/day established for inorganic arsenic represents the worst-case
scenario. Based on literature data, the ingestion of 25 g pollen contrib­
utes in 1–49% to the assumed maximum daily intake of this metalloid in
all but one case, supposing that the arsenic content of the product is in

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Table 7 evidence that pollens from randomly selected apiaries can contain lead
Acute risk assesment of pesticides detected in high concentrations from bee above 1 ppm (Formicki et al., 2013; Yang et al., 2013; Roman, 2009).
pollen. Based on our estimates, consuming pollen even with relatively low Pb
Active Max. Reference Estimated ARfDb Contribution content also contribute greatly to the assumed daily intake. Data shown
substance c. (μg/ acute (μg/ of pollen to in the table do not necessarily represent the arsenic and lead content of
kg) exposurea kgbw) the acute commercial products, because most of them focus on demonstrating the
(μg/kgbw) reference dose
impacts of pollution. However, the interest of beekeepers is to protect
(%)
bees from pollutants, therefore hives are usually placed in natural en­
chlorothalonil 98900 Mullin 35.32 50.00 70.64*
vironments. Moreover, the lowest BMDL values established for these
et al.
(2010) elements were taken into account in the risk assessment, so the results
phosmet 16556 Stoner & 5.91 45.00 13.14 show the worst scenario.
Eitzer For AFL-B1, the assumed maximum daily intake is only 0.04 ng/
(2013) kgbw/day because of the low BMDL10 value and the uncertainty factor
fluvalinate 16260 Nai et al. 5.81 50.00 11.62
(2017)
of 10,000. Although products contained these toxins in concentrations
lambda- 12476 Tong 4.46 5.00 89.20* below 10 μg/kg, the estimated daily intakes were 70 and 80 times higher
cyhalothrin et al. than the assumed maximum daily intakes. Further research is needed to
(2018) draw appropriate conclusions on potential health risk of AFL-B1
captan 10000 Mullin 3.57 300 1.19
contamination of pollen loads.
et al.
(2010) Pyrrolizidine alkaloids were present in a concentration range of
chlorothalonil 6481 Tong 2.31 50.00 4.63 119–1597 μg/kg in bee pollen samples from various botanical origins.
et al. According to our calculations, the consumption of these pollens con­
(2018) tributes in 179–2407% to the assumed maximum daily intake. The
tebuconazole 4530 Friedle 1.62 30.00 5.39
Committee on Herbal Medicinal Products (HMPC) of the European
et al.
(2021) Medicines Agency recommended a limit of 1.0 μg/day for PAs. Based on
carbendazim 4516 Tong 1.61 20.00 8.06 literature data, PA intake from consuming 25 g of bee pollen corre­
et al. sponds to 3–40 μg/day for PAs. The results of both approaches show that
(2018)
the PA content of bee pollens may pose a risk to its consumers. Mono­
myclobutanil 4190 Stoner & 1.50 310.00 0.48
Eitzer floral pollens from Echium, Senecio and Eupatorium species can contain
(2013) up to 350,000 μg/kg of pyrrolizidine alkaloids (Boppré et al., 2008).
fluopyram 4050 Friedle 1.45 500.00 0.29 Risk assessment was not performed for these samples, because pollen
et al. loads with such a high PA content are generally not present or scattered
(2021)
in commercially available products. However, based on available data,
deltamethrin 3710 Tong 1.33 10.00 13.25
et al. the above mentioned genera may play a significant role in the PA
(2018) contamination of apicultural products.
thiophanate- 3674 Lambert 1.31 200.00 0.66
methyl et al.
5. Future challenges and trends
(2013)
dimethomorph 2678 Böhme 0.95 600.00 0.16
et al. In line with the rising demand for foods of natural origin, the number
(2018) of scientific research on bee pollen has significantly increased. These
fluvalinate 2670 Mullin 0.95 50.00 1.91 surveys focus mainly on its nutritional benefits (Benedek et al., 2021),
et al.
(2010)
but several studies highlight the food safety risks as well. Fig. 3 shows
carbendazim 2595 Lambert 0.92 20.00 4.58 the temporal trend of scientific articles published on the topic of bee
et al. pollen and its food safety over the past 30 years, based on ScienceDirect
(2013) search results. In the 1990s, only a few publications dealt with the topic,
metalaxyl 2463 Tosi et al. 0.88 500.00 0.18
but after the turn of the millennium, a continuous increase can be
(2018)
dimethomorph 2045 Johnston 0.73 600.00 0.12 observed. This trend is expected to continue in the future as many
et al. distinct issues relate to these topics.
(2014) As bees forage from a wide range of plant species, bee pollen can be
e Tolerable Monthly Intake, ng/kgbw/month. very diverse regarding their nutritional value, therapeutic effects and
a
Assuming a consumption of 25 g/day and average bodyweight of 70 kg. food safety risks. Consequently, authentic samples should be evaluated
by researchers, the application of which allows the results to be
b
Acute Reference Dose, μg/kgbw repeated. In this sense, it will also be necessary to develop inexpensive
and rapid methods for authenticity assessment (Sipos et al., 2020). The
complexity of pollen composition and the large number of potential
inorganic form. The arsenic content of commercial, monofloral pollens contaminants are also major challenges for future research.
originated from China was exceptionally high. The exposure calculated Another emerging research area is the applicability of the product as
from the mean concentration of these products is nearly nine times a functional food ingredient due to its nutrients and phytometabolites
higher than the assumed maximum daily intake value. (Kostić et al., 2020). For this purpose, it is crucially important to
Estimated daily intake of lead was higher than the assumed examine food safety aspects extensively and to establish an international
maximum daily intake in 64% of the cases. The mean Pb content was standard. In order to ensure the homogeneity of functional foods, it is
extremely high in pollen samples examined by Al Naggar et al., 2013 and recommended to use monofloral bee pollen for fortification. This re­
Satta et al. (2012). These experiments were designed to demonstrate the quires research on the optimal environmental conditions of pollen
impact of environmental pollution on the metal content of apicultural production and also on the economical and ecological aspects of harvest.
products. Most samples used in these experiments are originated from Nowdays environmental protection is one of the most significant
places near busy roads or post-mining sites. Although, studies provide public concern and is also a focus area of science. If the hives are located
in highly polluted areas, bees and their products can accumulate heavy

504
R. Végh et al. Trends in Food Science & Technology 114 (2021) 490–509

Table 8
Risk assessment of genotoxic carcinogens of bee pollen.
Compound/ Mean conc. Reference Estimated daily BMDL values Assumed maximum daily Contribution of pollen to the
Element (μg/kg) intakea (ng/kgbw/ (BMDL10a/ intakeb (ng/kgbw/day) assumed maximum daily intake
day) BMDL01b) (%)

Arsenic 55 Roman (2009) 19.64 BMDL01: 300 6.55

290 Roman et al. (2016) 103.57 0.3 34.52
237 Álvarez-Ayuso & 84.64 28.21
Abad-Valle (2017)
<300 Shevtsova et al. (2014) <107.14 <35.71
<LOD Bayram (2021) Low low
410 Altunatmaz et al. (2017) 146.43 48.81
400 Morgano et al. (2010) 142.86 47.62
169 Oliveira et al. (2017) 60.36 20.12
184 Nascimento et al. (2018) 65.71 21.90
10 Aldgini et al. (2019) 3.57 1.19
340 Murashova et al. (2020) 121.43 40.48
21 Belina-Aldemita et al. 7.50 2.50
(2020)
7501 Yang et al., 2012 2678.93 892.98
Lead 145 Conti & Botré (2001) 51.82 BMDL01: 0.50 50 103.65
2723 Satta et al. (2012) 972.50 1945.00
237 Lambert et al. (2012) 84.64 169.29
710 Roman (2009) 253.39 506.79
147 Roman et al. (2016) 52.50 105.00
1673 Formicki et al. (2013) 597.50 1195.00
900 Zhelyazkova & 321.43 642.86
Atanassova (2011)
<100 Kacániová et al., 2010 <35.71 <71.43
350 Shevtsova et al. (2014) 125.00 250.00
247 Adaškevičiütė et al., 88.21 176.43
2019
347 Özcan et al. (2019) 123.93 247.86
96 Temizer et al. (2018) 34.29 68.57
<LOD Bayram (2021) Low low
175 Altunatmaz et al. (2017) 62.50 125.00
25 Mejías et al. (2018) 8.93 17.86
120 Morgano et al. (2010) 42.86 85.72
15 Sattler et al. (2016) 5.36 10.71
148 Oliveira et al. (2017) 52.86 105.71
91 Nascimento et al. (2018) 32.50 65.00
27 Silveira et al. (2013) 9.64 19.29
7724 Al Naggar et al., 2013 2758.57 5517.14
313 Aldgini et al. (2019) 111.69 223.37
480 Murashova et al. (2020) 171.43 342.86
61 Belina-Aldemita et al. 21.79 43.57
(2020)
453 Yang et al., 2012 161.79 323.57
Aflatoxin B1 8 Kostić et al. (2017) 2.86 BMDL10: 0.04 7150.00
9 Petrović et al., 2014 3.21 0.4 8025.00
<LOQ Xue et al. (2014) Low low
<LOQ Garcia-Villanova et al. Low low
(2004)
Pyrrolizidine 1108 Dübecke et al. (2011) 395.71 BMDL10: 237 23.7 1669.66
alkaloids 1597 Kempf et al. (2010) 570.35 2406.54
119 Kast et al. (2018) 42.50 179.32
555 Picron et al. (2019) 198.21 836.33
402 Inacio et al. (2020) 143.57 605.79
576 Mulder et al., 2018 205.71 867.97

* Assuming a consumption of 25 g/day and average bodyweight of 70 kg.

** Based on BMDL values and the minimum margin of exposures sufficient to ensure that there is no appreciable human risk.
a
Benchmark Dose Lower Bound (10%): the lowest dose, which induce tumor in 10% of the experimental animals (μg/kgbw/day).

b
Benchmark Dose Lower Bound (1%): the lowest dose, which induce tumor in 1% of the experimental animals (μg/kgbw/day).

metals and other contaminants (Bargánska et al., 2016; Murashova
et al., 2020), so they are expected to be widely used as indicators of
environmental pollution. Contaminants of apicultural products are
important for researches on pollinator decline. Many factors play a role
in bee mortality, but plant protection products, especially neon­
icotinoids stay at the focus of scientific researches and media as well
(Gierer et al., 2019), because they are proved to affect the foraging and
orientation abilities of honeybees negatively (Sörös, 2019). Due to bee
toxicity or adverse effects on human health, several pesticides have
recently been banned, necessitating the on-going evaluation of alter­
native substances.
6. Conclusions
Bee pollen is a popular dietary supplement, however, there are no
regulations for the product in most countries. Consequently, these
products can be characterized by heterogenous food safety risks. Com­
mon contaminants of bee pollen are pesticides, heavy metals, metalloids

505
R. Végh et al.
Fig. 3. Temporal trend of scientific articles published on the topic of bee pollen and its food safety between 1990 and 2020 (ScienceDirect, 2021).
and mycotoxins. Pyrrolizidine alkaloids, allergenic proteins and pollen
grains from genetically modified plants may also be present in these
products. In this review, recent findings on the above-mentioned sub­
stances and data on concentrations determined in bee pollen are sum­
marized. Furthermore, a risk assessment was also performed for the
toxicologically important contaminants and constituents of bee pollen
based on literature data. Our results suggest that pesticides commonly
found in pollen do not pose a risk to human health. On the other hand,
pollen loads may be contaminated with metals, metalloids and myco­
toxins to such an extent that represents a potential risk to the consumers.
Certain plant species contain hepatotoxic pyrrolizidine alkaloids in
extremely large quantities, therefore it is suggested to monitor bee
pollen intended for human consumption.
The number of scientific researches on the topic of bee pollen asso­
ciated food safety risks is continuously increased in the last two decades,
but the information is incomplete in some areas. In Europe, many studies
are conducted, but little data is available from other continents. Based
on our findings, further research is needed on the alternatives of pesti­
cides that are common in bee pollen due to their ban. Currently, liter­
ature data is also insufficient to draw far-reaching conclusions on
mycotoxin and Hg contamination of pollen.
Funding
This paper was supported by the János Bolyai Research Scholarship
of the Hungarian Academy of Sciences. This work was partly supported
by the European Union and co-financed by the European Social Fund
(grant agreement no. EFOP-3.6.3-VEKOP-16-2017-00005). This
research was funded by National Research, Development and Innovation
Office of Hungary (OTKA, contracts No. 135700). This research was
supported by the Ministry for Innovation and Technology within the
framework of the Thematic Excellence Programme 2020- Institutional
Excellence Subpro-gram (TKP2020-IKA-12) for research on plant
breeding and plant protection.
Declaration of competing interest
There is no conflict of interest in this article.
Acknowledgements
This research was supported by the Ph.D. School of Food Science of
Szent István University. The authors are grateful for John-Lewis Zinia
506
Trends in Food Science & Technology 114 (2021) 490–509
Zaukuu for proofreading the manuscript.
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