foods

Article
Multi-Elemental Analysis of Edible Insects, Scorpions, and
Tarantulas from French (Online) Market and Human Health Risk
Assessment Due to Their Consumption: A Pilot Study
Yulianna Holowaty 1 , Axelle Leufroy 2 , Clément Mazurais 2 , Diane Beauchemin 1 and Petru Jitaru 2, *

1 Department of Chemistry, Queen’s University, 90 Bader Lane, Kingston, ON K7L 3N6, Canada;
[email protected] (Y.H.); [email protected] (D.B.)
2 Laboratory for Food Safety, University Paris East Creteil, Anses, F-94700 Maisons-Alfort, France;
[email protected] (A.L.); [email protected] (C.M.)
* Correspondence: [email protected]

Abstract: Edible insects are becoming increasingly popular as protein alternatives to traditional
animal-based products. As such, information on their elemental composition is important to ensure
they are safe for human consumption. This article describes the development and validation of a
rapid, reliable method for the simultaneous determination of 19 elements (Al, As, B, Ba, Ca, Cd, Co,
Cr, Cu, Fe, K, Mg, Mn, Mo, Na, Pb, Se, Sr, and Zn) in edible insects by inductively coupled plasma
mass spectrometry (ICP-MS) following closed vessel microwave digestion. The method was validated
using three insect certified reference materials, namely black soldier fly larvae meal (BFLY-1), cricket
flour (KRIK-1), and mealworm powder (VORM-1). The method was applied to analyze twelve
different (whole) insect species. The maximum amount of each sample was calculated for As, Cd,
and Pb with respect to their provisional tolerable daily intake values established by the Food and
Agricultural Organization/World Health Organization. Most of the samples, except for scorpions
and tarantulas, were safe to consume at large doses (1000–10,000 insects per day). Furthermore, most
of the samples contained high levels of Fe, K, Na, and Zn, providing a preliminary overview of the
Citation: Holowaty, Y.; Leufroy, A.;
nutritional profile of these novel protein alternatives.
Mazurais, C.; Beauchemin, D.; Jitaru,
P. Multi-Elemental Analysis of Edible
Keywords: trace elements; ICP-MS; edible insects; France; risk assessment
Insects, Scorpions, and Tarantulas
from French (Online) Market and
Human Health Risk Assessment Due
to Their Consumption: A Pilot Study.
Foods 2024, 13, 2353. https://doi.org/
1. Introduction
10.3390/foods13152353 Entomophagy is a term used to describe the practice of eating insects [1]. While
common in some countries, Western regions are still overcoming psychological and cultural
Academic Editors: Elisa Keating and
barriers [2]. Recent efforts have been made to eliminate these barriers and promote the
Cristina Couto
consumption of insects as an alternative protein source, given that the world’s population is
Received: 19 June 2024 estimated to reach approximately 9.7 billion by 2050 and a shortage of current animal-based
Revised: 24 July 2024 proteins is expected [3–5]. An increasing number of edible insect products are emerging on
Accepted: 24 July 2024 the market, providing consumers with environmentally friendly and nutritious options,
Published: 26 July 2024 taking into account that insects consume less water and land, emit less greenhouse gases
compared to livestock, and they contain proteins, essential vitamins, and minerals [6–10].
However, information on the total elemental composition of these commercially available
edible insects is required to ensure their safety for human consumption.
Copyright: © 2024 by the authors.
Trace elements in the human body, which are generally present at levels < 250 µg g−1
Licensee MDPI, Basel, Switzerland.
are classified as essential, nonessential, and potentially toxic, depending on the dose and
This article is an open access article
distributed under the terms and
duration of intake [11–13]. Essential trace elements play a vital role in human health and
conditions of the Creative Commons
functions. They include Cu and Fe, which participate in energy metabolism via oxidation-
Attribution (CC BY) license (https:// reduction reactions, and Fe enables transport of oxygen throughout the body via the
creativecommons.org/licenses/by/ formation of hemoglobin [11,13]. Meanwhile, potentially toxic elements (PTEs) include As,
4.0/). Cd, Hg, and Pb, which can lead to cancer and damage to the nervous system and organs

Foods 2024, 13, 2353. https://doi.org/10.3390/foods13152353 https://www.mdpi.com/journal/foods

Foods 2024, 13, 2353 2 of 13

when consumed at high doses for long periods of time [11,13–16]. There is a possibility that
insects, such as the black soldier fly larvae (BSFL, Hermetia illucens) that convert organic
matter into biomass, are bioaccumulating these PTEs from their feed, posing health risks for
humans if ingested [17,18]. To minimize these risks, trace element levels in edible insects
must be monitored.
As the emergence of insect-based proteins is relatively new, there is little information
on their elemental composition in the current literature. Although techniques such as
particle-induced X-ray emission (micro PIXE), instrumental neutron activation analysis
(INAA), and wavelength dispersive X-ray fluorescence (WDXRF) spectrometry [19–24]
have been previously used for the determination of elements in various insect samples,
inductively coupled plasma optical emission spectrometry (ICP-OES) and inductively
coupled plasma mass spectrometry (ICP-MS) allow for simultaneous multi-elemental anal-
ysis [17,18,25–29]. Notably, ICP-MS provides lower detection limits and greater sensitivity,
making it the suitable choice for this type of research [30,31].
The aim of this study was to develop and validate a rapid and reliable method for
the determination of minor, major, potentially toxic, and essential elements in a selection
of commercially available edible insects using ICP-MS. For this purpose, nineteen trace
elements were studied: Al, As, B, Ba, Ca, Cd, Co, Cr, Cu, Fe, K, Mg, Mn, Mo, Na, Pb, Se, Sr,
and Zn. As the proposed method has not previously been applied to such matrices, it was
validated through the analysis of several insect certified reference materials (CRMs) before
being applied to the analysis of real-life insect samples.

2. Materials and Methods

2.1. Instrumentation
The samples were digested using a closed microwave system (Mutiwave 7000, Anton
Paar, Courtaboeuf, France).
An Agilent 8900 ICP tandem mass spectrometry (ICP-MS/MS) instrument (Agilent
Technologies, Courtaboeuf, France) equipped with a MicroMist concentric nebulizer and
Scott-type double-pass quartz spray chamber was used for analysis. Sample digests were
introduced directly into the instrument via a peristaltic pump from tubes connected to
an SPS 4 autosampler (Agilent Technologies, Courtaboeuf, France). The operating con-
ditions are detailed in Table 1. These parameters were optimized daily by performing
short-term stability tests with the tuning solution (1 mg L−1 ) in both detection modes
(standard and collision mode) to ensure maximum sensitivity and minimal interference
from oxide (CeO+ /Ce+ < 1.2%) and doubly charged ions (Ce2+ /Ce+ < 2%). The signals
were obtained using Scientific MassHunter software (Agilent Technologies) and the raw
data were processed using Excel.

Table 1. Operating conditions of the ICP-MS/MS instrument (Agilent 8900).

Parameter Setting
RF power 1550 W
Plasma gas flow rate (Ar) 15.0 L min−1
Auxiliary gas flow rate (Ar) 0.9 L min−1
Nebulizer gas flow rate (Ar) ~1.0 L min−1
Collision gas flow rate (He) ~5.0 mL min−1
Sampling/skimmer cones Nickel
No gas mode: 11 B, 75 As, 111 Cd, 137 Ba, 208 Pb
Monitored isotopes Collision mode (He): 23 Na, 24 Mg, 27 Al, 39 K, 44 Ca, 52 Cr, 55 Mn,
56 Fe, 59 Co, 63 Cu, 66 Zn, 77 Se, 88 Sr, 95 Mo

Internal standards: 89 Y, 185 Re, 209 Bi

2.2. Chemicals and Reagents

Ultrapure water (18.2 MΩ cm, Millipore Milli-Q™, Merck Millipore, Saint Quentin
en Yvelines, France) and HNO3 (67% v/v, Suprapur, VWR, Fontenay sous-Bois, France)
Foods 2024, 13, 2353 3 of 13

were used throughout the study. High purity argon (99.996%, Linde Gas, Montereau-Fault-
Yonne, France) was used for the plasma, auxiliary, and nebulizer gases.
Stock solutions containing 1000 mg L−1 of each analyte (Al, As, B, Ba, Ca, Cd, Co,
Cr, Cu, Fe, K, Mg, Mn, Mo, Na, Pb, Se, Sr, and Zn) were purchased from LGC Standards
(Molsheim, France) and were used to prepare calibration standard solutions daily in 6%
(v/v) HNO3 .
Internal standard solutions were prepared using 1000 mg L−1 standard stock solutions
of Y, Re, and Bi that were purchased from LGC Standards. These solutions were added to
all samples, calibration standards, and blanks to compensate for drift.
A tuning solution and a Factor P/A solution were prepared separately using multi-
element solutions (Agilent Technologies) to ensure optimal instrument sensitivity over
a wide range of masses and linear response of the detector between pulse and analog
detection modes, respectively.
Solutions of HNO3 at 6% (v/v) and 10% (v/v) were used for rinsing the ICP-MS/MS
system between analyses.

2.3. Reference Materials and Samples

Three insect-based CRMs provided by the National Research Council Canada (Ottawa,
ON, Canada) were used throughout the study to validate the method, namely: BFLY-1
(black soldier fly larvae meal), KRIK-1 (cricket flour), and VORM-1 (mealworm powder).
The mussel tissue ERM-CE278k CRM from the Institute for Reference Materials and Mea-
surements (LGC Standards) was also used throughout the study as an internal quality
control sample.
Fourteen dry samples including authorized edible crickets (Acheta domesticus), worms
(Tenebrio molitor), ants (Atta laevigata), water bugs (Nepidae), locusts (Locusta migratoria), scor-
pions (Heterometrus longimanus), and tarantulas (Haplopeima albostriatum) were purchased
from the French market and online. To ensure sufficient quantities for analysis, multiple
packages of some samples were purchased. Although these packages originated from the
same batch, they were each treated as a single sample to distinguish any variations between
packages, which led to the analysis of 27 samples (see Table 2 for details). Information on
the life stage and origin of the samples is not available.

Table 2. List of edible insects, scorpions, and tarantulas samples used in the study with corresponding
code number, sample name, ingredients, and origin (if available).

Sample Name Insect Species Information Origin

Ants 1 Columbia ants Atta laevigata With salt Columbia
Ants 2 Columbia ants Atta laevigata With salt Columbia
Whole dehydrated
Crickets 1 Crickets Acheta domesticus Toulouse, France
crickets
Whole dehydrated
Crickets 2 Crickets Acheta domesticus Toulouse, France
crickets
Whole dehydrated
Crickets 3 Crickets Acheta domesticus Toulouse, France
crickets
Crickets 4 Crickets Gryllus bimaculatus - Thailand
Whole dehydrated
Crickets 5 Crickets N/A N/A
crickets
Whole dehydrated
Crickets 6 Crickets N/A N/A
crickets
Whole dehydrated
Crickets 7 Crickets N/A Netherlands
crickets
Whole dehydrated
Crickets 8 Crickets N/A Netherlands
crickets
Foods 2024, 13, 2353 4 of 13

Table 2. Cont.

Sample Name Insect Species Information Origin

Locusts 1 Locusts Locusta migratoria With salt N/A
Locusts 2 Locusts Locusta migratoria With salt N/A
Heterometrus
Scorpions 1 Black scorpions With salt Thailand
longimanus
Black Asian forest Heterometrus
Scorpions 2 With salt Thailand
scorpions longimanus
Black Asian forest Heterometrus
Scorpions 3 With salt Thailand
scorpions longimanus
Black Asian forest Heterometrus
Scorpions 4 With salt Thailand
scorpions longimanus
Tarantulas 1 Tarantula Haplopeima albostriatum - Thailand
Tarantulas 2 Tarantula Haplopeima albostriatum - Thailand
Tarantulas 3 Tarantula Haplopeima albostriatum - Thailand
Water bug 1 Giant water bug Nepidae With oil and salt N/A
Water bug 2 Giant water bug Nepidae With oil and salt N/A
Worms 1 Mealworms Tenebrio molitor - EU
Worms 2 Silk worms Bombyx mori - Thailand
Worms 3 Morio worms Zophobas morios With salt Thailand
Whole dehydrated
Worms 4 Mealworms Tenebrio molitor Toulouse, France
mealworm
Whole dehydrated
Worms 5 Mealworms Tenebrio molitor Toulouse, France
mealworm
Whole dehydrated
Worms 6 Mealworms Tenebrio molitor Toulouse, France
mealworm

2.4. Analytical Procedures

2.4.1. Sample Preparation
Each sample was removed from its original packaging and stored at room temperature
in a closed polyethylene tube while awaiting preparation for analysis. As most packages
contained multiple insects, the total weight (in g) was measured. From this, the weight per
insect (in g) was approximated by weighing 20 whole insects.
Each sample was then homogenized using a BM500 benchtop laboratory ball mill
(Anton Paar, Courtabœuf, France) and three 10 mm agate beads. The grinding frequency
and time varied depending on the type of insect. Those with hard exoskeletons (scorpions,
water bugs, and tarantulas) were ground at 30 Hz for 2 min, whereas others were ground
at 25 Hz for 45 s.

2.4.2. Multi-Elemental Analysis by ICP-MS/MS

Approximately 0.3 g of homogenous sample was weighed in a quartz digestion vial,
to which 3 mL of HNO3 (67%, v/v) and 3 mL of ultrapure water were added. The samples
were then completely digested using a closed microwave digestion system for 30 min at
250 ◦ C and a pressure of 140 bar. Once cooled to room temperature, the sample solutions
were transferred into 50 mL polyethylene flasks and 100 µL of a mixture of internal standard
solution (Y, Re, and Bi) were added before final dilution to 50 mL with ultrapure water and
subsequent analysis by ICP-MS.

2.4.3. Data Processing

Concentrations were expressed in mg kg−1 . Calibration curves were produced for
each analysis series to verify linearity (R2 ≥ 0.995). Humidity was measured in all three
insect CRMs to correct the result. A Grubbs test at the 95% confidence level was performed
to identify and remove any outliers in the experimentally obtained data.
Foods 2024, 13, x FOR PEER REVIEW 5 of 14

Foods 2024, 13, 2353 5 of 13

insect CRMs to correct the result. A Grubbs test at the 95% confidence level was performed
to identify and remove any outliers in the experimentally obtained data.
To determine the method’s accuracy when analyzing the three insect CRMs, the dif-
To determine the method’s accuracy when analyzing the three insect CRMs, the differ-
ference between the certified and measured values (Δm) were compared to their combined
ence between the certified and measured values (∆m ) were compared to their combined
uncertainties (UΔ) using the Equations (1)–(3) [32,33]:
uncertainties (U∆ ) using the Equations (1)–(3) [32,33]:
∆ = |𝑐 − 𝑐 | (1)
∆m = |cm − cCRM | (1)
𝑢∆ = 𝑠 + 𝑠
q (2)
u∆ = s2m + s2CRM (2)
𝑈 =𝑘∙𝑢 (3)
U∆∆ = k·u∆∆ (3)
where cm is average measured concentration, cCRM is certified concentration, sm is standard
where cm is average measured concentration, cCRM is certified concentration, sm is standard
deviation of the measured value, and sCRM is uncertainty of the certified value. Note, a
deviation of the measured value, and sCRM is uncertainty of the certified value. Note, a
coverage factor (k) of 2 was used, corresponding to 95% confidence level.
coverage factor (k) of 2 was used, corresponding to 95% confidence level.
3.3.Results
Resultsand
andDiscussion
Discussion
3.1.
3.1.Method
MethodValidation
Validation
To
To ensure
ensure that
that the
the proposed
proposed method
method is is applicable
applicable toto insect
insect matrices,
matrices, three
three CRMs
CRMs
(BFLY-1,
(BFLY-1, KRIK-1, and VORM-1) were were analyzed
analyzed in intriplicates
triplicatesononthree
threedifferent
differentdaysdays(n(n= =9,
9,unless
unlessotherwise
otherwisespecified)
specified)over
overaaperiod
periodofofseven
seven weeks.
weeks. The comparison of of the
theaverage
average
totalconcentration
total concentrationand
andstandard
standarddeviation
deviationdetermined
determinedfor foreach
eachofofthe
the19
19elements
elementsin inthe
the
threeCRMs
three CRMsand
andthe
thecorresponding
corresponding certified
certified value
value is
is provided
provided inin Figure
Figure 1.
1.

180
BFLY-1 KRIK-1 VORM-1
160
Recovery factor (%)

140

120

100

80

60

40

20

0
Al As B Ba Ca Cd Co Cr Cu Fe K Mg Mn Mo Na Pb Se Sr Zn

Figure 1. Average recovery factors (%) and the relative standard deviation (n = 8, patterned bars or
Figure 1. Average recovery factors (%) and the relative standard deviation (n = 8, patterned bars or
n = 9) obtained for the analysis of BFLY-1, KRIK-1, and VORM-1 CRMs by ICP-MS.
n = 9) obtained for the analysis of BFLY-1, KRIK-1, and VORM-1 CRMs by ICP-MS.
Overall, most of the elements had average percent recoveries ranging from 90 to
Overall, most of the elements had average percent recoveries ranging from 90 to 110%
110% for all three CRMs, hence confirming the method’s fit for the analysis of insects.
for all three CRMs, hence confirming the method’s fit for the analysis of insects. Further-
Furthermore, upon comparing ∆m with U∆ (Table S1), all the elements except for Co, K,
more, upon comparing Δm with UΔ (Table S1), all the elements except for Co, K, and Se in
and Se in BFLY-1, were found to have ∆m values lesser than U∆ , indicating that there were
BFLY-1, were found
no statistically to have
significant Δm valuesbetween
differences lesser than
theUmeasured
Δ, indicating that there were no statis-
and certified concentrations.
tically significant differences between the measured and certified concentrations.
3.2. Analysis of Commercially Available Edible Insects
The levels of 19 elements measured in commercially available edible insects are pre-
sented as the following: PTEs, major elements, essential elements, and others (see Tables 3–6,
respectively). It is worth to note that amongst the species analyzed in this study, only four
are currently authorized as novel food in the European Union (EU), namely the meal-
Foods 2024, 13, 2353 6 of 13

worm (Tenebrio molitor), the lesser mealworm (Alphitobius diaperionus), the locust (Locusta
migratoria), and the cricket (Acheta domesticus) [34].
The Food and Agricultural Organization/World Health Organization established pro-
visional tolerable daily intake (PTDI) values for As, Cd, and Pb of 2.10 µg kg−1 , 0.82 µg kg−1 ,
and 3.57 µg kg−1 bodyweight per day, respectively, in this type of foodstuffs [35,36]. The
maximum amount of whole edible insects that can be safely consumed daily for a 60 kg
adult and an 18 kg child was calculated using the approximated weight per insect (in g)
and is summarized in Table 3.
Based on the measured concentrations, it appears that most of the samples, notably
the crickets, locusts, and worms, can be safely consumed at rather large quantities (from
1000 to 100,000 individuals) by both children and adults. However, the maximum number
of scorpions that can be consumed in terms of Cd were significantly lower than the other
samples, with 12–32 recommended for adults and 4–10 for children.
Moreover, in the case of tarantula samples, the consumption of only 1 individual per
day is recommended for children and 2–3 for adults. However, this finding should not
pose too much concern considering the context in which insects are being considered as a
protein alternative. For instance, in Europe, insects are marketed as a food ingredient, to be
added in crackers, pasta, etc. [34].
With regard to the samples originating from the same lot, similar contamination pat-
terns were observed for the ants, crickets 1–3 and 7–8, and worms 4–6 samples. Contrarily,
measured results varied significantly between the following samples (see Tables 3–5 and 7):
crickets 5–6, locusts 1–2, scorpions 3–4, tarantulas 1–3, water bug 1–2, and worms 1–3.
Larger variances observed in scorpions, tarantulas, and water bugs can be explained by
the number of individuals in each lot. In fact, especially for these species, one lot was
composed of only one or two individuals.
Table 4 summarizes the levels of major (naturally abundant) elements (Na, K, Ca, and
Mg) in each sample. Overall, high levels of K were observed for all the samples (from 4- to
10-fold compared to Ca, Mg, and Na), suggesting that insects are rich in this element. This
is beneficial as K plays a role in proper kidney and heart regulation, muscle contraction,
and nerve transmission [37].

Table 3. Concentrations ± standard deviation (n = 2; µg kg−1 ) of potentially toxic elements in

edible insects, scorpions, and tarantulas (with corresponding sample name) by ICP-MS as well as the
number of individual of each type that can be safely consumed (per day) in relation to provisional
tolerable daily intake (PTDI) a for human adults b and children c .

As Cd Pb
Maximum
Maximum Recommended Maximum Recommended
Recommended
Sample Consumption Consumption
Measured Measured Consumption Measured
(Number per Day) (Number per Day)
(Number per Day)
Adults Children Adults Children Adults Children
Ants
1 43.86 ± 0.66 401 120 85.4 ± 1.7 80 24 106.8 ± 5.4 280 84
2 44.13 ± 0.77 429 129 79.60 ± 0.15 93 28 104.1 ± 1.2 309 93
Crickets
1 7.70 ± 0.40 145,196 43,559 36.89 ± 0.39 11,834 3550 18.296 ± 0.018 103,882 31,165
2 6.97 ± 0.18 178,103 53,431 38.95 ± 0.72 12,445 3733 15.18 ± 0.51 139,021 41,706
3 6.449 ± 0.084 182,939 54,882 29.35 ± 0.92 15,696 4709 16.7 ± 2.9 120,097 36,029
4 12.42 ± 0.65 41,399 12,420 13.75 ± 0.11 14,602 4381 36.62 ± 0.81 23,870 7161
5 3.6 ± 2.7 271,389 81,417 73 ± 10 5182 1555 53.6 ± 2.4 30,729 9219
6 2.622 ± 0.086 423,019 126,906 65.90 ± 0.49 6572 1972 33.4 ± 3.8 56,454 16,936
7 19.57 ± 0.73 13,757 4127 76.07 ± 0.92 1382 415 96.9 ± 3.0 4723 1417
8 21.07 ± 0.67 12,726 3818 76.6 ± 3.8 1367 410 62.0 ± 5.6 7352 2206
Locusts
1 35.0 ± 1.0 8137 2441 31.78 ± 0.34 3499 1050 50.194 ± 0.060 9645 2894
2 47.44 ± 0.12 4912 1474 40.9 ± 2.3 2225 667 73.4 ± 1.9 5397 1619
Foods 2024, 13, 2353 7 of 13

Table 3. Cont.

As Cd Pb
Maximum
Maximum Recommended Maximum Recommended
Recommended
Sample Consumption Consumption
Measured Measured Consumption Measured
(Number per Day) (Number per Day)
(Number per Day)
Adults Children Adults Children Adults Children
Scorpions
1 725 ± 36 62 19 539 ± 22 32 10 235 ± 14 324 97
2 383.5 ± 2.4 200 60 2272 ± 19 13 4 123.6 ± 3.1 1053 316
3 233.1 ± 3.3 226 68 1001 ± 38 21 6 162 ± 17 554 166
4 192.20 ± 0.23 127 38 771.406 ± 0.046 12 4 64.6 ± 9.7 643 193
Tarantulas
1 93.78 ± 0.51 65 19 13,200 ± 97 2 1 633.3 ± 6.3 163 49
2 191.8 ± 4.2 326 98 9650 ± 190 3 1 464.79 ± 0.89 229 69
3 120.8 ± 1.3 476 143 8582 ± 48 3 1 1064 ± 27 92 28
Water bug
1 59.31 ± 0.64 681 204 3.46 ± 0.15 4556 1367 60.96 ± 0.98 1126 338
2 86.4 ± 2.6 416 125 6.98 ± 0.14 2009 603 104.9 ± 1.4 582 175
Worms
1 39.73 ± 0.76 75,962 22,789 82.6 ± 2.1 14,267 4280 10.8 ± 1.7 475,050 142,515
2 470 ± 15 1483 445 53.9 ± 4.2 5049 1515 87.3 ± 5.2 13,571 4071
3 21.25 ± 0.62 20,567 6170 16.777 ± 0.061 10,172 3052 7.425 ± 0.011 100,064 30,019
4 25.65 ± 0.79 119,520 35,856 79.25 ± 0.76 15,105 4532 7.5 ± 2.5 694,891 208,467
5 26.50 ± 0.53 138,018 41,405 83.5 ± 2.9 17,104 5131 6.03 ± 0.28 1,031,129 309,339
6 25.66 ± 0.12 103,376 31,013 77.7 ± 2.8 13,331 3999 6.32 ± 0.39 713,524 214,057
a PTDI levels for As, Cd, and Pb are 2.10, 0.82, and 3.57 µg kg−1 bw−1 day−1 , respectively. b weight of 60 kg.
c weight of 18 kg.

Although natural samples (free from additives or other ingredients besides the insect)
were purchased for this study, some were found to have salt added to them (see ingredients
listed in Table 2). This was reflected by high levels of Na being reported in the scorpion
and tarantula samples (up to 10,597 and 40,454 mg kg−1 , respectively). Sodium, being an
essential nutrient, helps maintain plasma volume, acid-base balance, nerve transmissions,
and healthy cell functions [38].

Table 4. Concentrations ± standard deviation (n = 2; mg kg−1 ) of major elements in edible insects,

scorpions, and tarantulas (with corresponding sample name) measured by ICP-MS.

Sample Ca K Mg Na
Ants
1 539 ± 62 681 ± 2 187 ± 2 1135 ± 5
2 458 ± 87 677 ± 5 184 ± 3 1117 ± 22
Crickets
1 1268 ± 13 7567 ± 150 1215 ± 11 2577 ± 9
2 1399 ± 16 8308 ± 120 1316 ± 34 2835 ± 66
3 1397 ± 21 8538 ± 79 1314 ± 43 2874 ± 33
4 1798 ± 12 5263 ± 61 727 ± 1 1771 ± 12
5 1053 ± 66 9116 ± 330 746 ± 12 2424 ± 130
6 1034 ± 53 9078 ± 220 801 ± 10 2448 ± 15
7 496 ± 2 7017 ± 26 867 ± 13 1259 ± 12
8 411 ± 8 6559 ± 1 777 ± 2 1162 ± 6
Locusts
1 559 ± 24 6510 ± 150 749 ± 34 937 ± 57
2 481 ± 21 6350 ± 460 658 ± 4 860 ± 19
Scorpions
1 1149 ± 52 5946 ± 510 430 ± 7 10,597 ± 1009
2 1792 ± 14 5101 ± 37 542 ± 7 4152 ± 23
3 2102 ± 42 6903 ± 4 784 ± 28 6150 ± 7
4 1163 ± 4 4883 ± 53 417 ± 4 4495 ± 28
Foods 2024, 13, 2353 8 of 13

Table 4. Cont.

Sample Ca K Mg Na
Tarantulas
1 5372 ± 14 1298 ± 10 1944 ± 43 32,232 ± 320
2 2518 ± 74 1794 ± 38 1984 ± 12 27,079 ± 620
3 2748 ± 26 2220 ± 12 1765 ± 40 40,454 ± 260
Water bug
1 632 ± 2 3950 ± 4 695 ± 5 3150 ± 2
2 865 ± 18 4041 ± 18 614 ± 10 5113 ± 49
Worms
1 401 ± 10 4474 ± 250 1894 ± 69 521 ± 8
2 1132 ± 1 7462 ± 130 2565 ± 11 126 ± 1
3 788 ± 67 6040 ± 29 1343 ± 17 670 ± 6
4 499 ± 1 8335 ± 300 3074 ± 6 980 ± 10
5 492 ± 6 8533 ± 400 2896 ± 70 982 ± 38
6 501 ± 15 8294 ± 50 2959 ± 3 976 ± 10

Table 7 compares the results obtained in previous studies with those obtained in this
work for potential toxic and major elements. Many results are comparable despite the
samples being from different origins, prepared differently, etc. For example, two out of
three samples of cricket in the previous study had up to 10–38% of seasonings, whereas
only whole dehydrated crickets were analyzed in this work. Yet, except for As, the levels
of Cd, Pb, Ca, Mg, K, and Na are fairly similar. The largest differences were observed for
water bugs, which were from India in the previous study.
As can be seen in Table 5, high Cu levels were measured in the scorpion (81–130 mg kg−1 )
and tarantula samples (80–121 mg kg−1 ). Copper is involved in energy production, iron
metabolism, brain development, and immune system functioning [39]. Most of the samples
(crickets, locusts, worms, scorpions, ants, tarantulas, and water bugs) also contained high
levels of Fe, ranging from 33 to 474 mg kg−1 . This element is necessary for human growth
and development, with it circulating oxygen throughout the body [40]. Lastly, high levels
of Zn were observed for all the samples (from 100 mg kg−1 to 1000 mg kg−1 ). This is
noteworthy, as zinc is involved in cellular metabolism (catalytic activities of enzymes,
protein and DNA synthesis), immune health, and growth and development [41]. Table 8
shows that the levels of several essential elements are consistent with those previously
obtained in similar samples. Differences may be attributed to different sample origin,
sample preparation, etc. The systematically lower concentrations in water bugs reported
previously might be a result of the legs and antennas having been removed prior to analysis,
whereas the whole insect was analyzed in this work.

Table 5. Concentrations ± standard deviation (n = 2; mg kg−1 ) of essential elements in edible insects,

scorpions, and tarantulas (with corresponding sample name) by ICP-MS.

Sample Co Cr Cu Fe Mn Mo Se Zn
Ants
1 0.05241 ± 0.00074 0.12074 ± 0.00089 19.463 ± 0.042 147 ± 3 36.39 ± 0.39 0.5309 ± 0.0012 1.01 ± 0.00 113 ± 2
2 0.0519 ± 0.0032 0.114 ± 0.011 19.34 ± 0.35 146 ± 5 35.32 ± 0.41 0.5188 ± 0.0037 1.001 ± 0.012 113 ± 3
Crickets
1 0.0615 ± 0.0053 0.0561 ± 0.0096 21.50 ± 0.60 55.80 ± 0.23 55.2 ± 1.6 0.5318 ± 0.0026 0.616 ± 0.023 191 ± 2
2 0.0611 ± 0.0017 0.0390 ± 0.0088 19.54 ± 0.90 50.5 ± 2.5 59.3 ± 1.2 0.5382 ± 0.0015 0.631 ± 0.024 197 ± 2
3 0.0557 ± 0.0012 0.0406 ± 0.0042 20.891 ± 0.048 55.2 ± 2.5 58.2 ± 1.3 0.560 ± 0.015 0.6384 ± 0.0027 202 ± 8
4 0.03363 ± 0.00092 0.0915 ± 0.0031 19.98 ± 0.33 50.9 ± 1.8 63.5 ± 1.3 0.6071 ± 0.0051 0.3087 ± 0.0035 155 ± 2
5 0.0108 ± 0.0013 0.02712 ± 0.00031 24.2 ± 1.5 54.69 ± 0.029 30.262 ± 0.052 0.723 ± 0.025 0.1234 ± 0.0064 239 ± 24
6 0.0102 ± 0.0021 0.0164 ± 0.0026 26.0 ± 2.4 49.3 ± 3.3 30.1 ± 2.9 0.755 ± 0.028 0.1311 ± 0.0017 256 ± 9
7 0.0835 ± 0.0013 0.1380 ± 0.0081 53.09 ± 0.42 83.58 ± 0.17 7.69 ± 0.20 1.4969 ± 0.0053 0.19229 ± 0.00045 150 ± 37
8 0.0880 ± 0.0070 0.122 ± 0.017 49.31 ± 0.40 68.7 ± 1.4 4.937 ± 0.048 1.531 ± 0.030 0.200 ± 0.011 135 ± 15
Foods 2024, 13, 2353 9 of 13

Table 5. Cont.

Sample Co Cr Cu Fe Mn Mo Se Zn
Locusts
1 0.1218 ± 0.0049 0.0836 ± 0.0077 60.300 ± 0.030 90.33 ± 0.55 6.77 ± 0.18 0.678 ± 0.018 0.3341 ± 0.0071 177 ± 25
2 0.1246 ± 0.0047 0.2066 ± 0.0093 92.7 ± 7.7 164 ± 23 6.46 ± 0.18 0.574 ± 0.021 0.2160 ± 0.0068 173 ± 17
Scorpions
1 0.683 ± 0.030 0.1658 ± 0.0023 81.8 ± 7.4 139 ± 11 166 ± 12 0.206 ± 0.014 2.136 ± 0.081 381 ± 25
2 1.0358 ± 0.0058 0.1609 ± 0.0029 130.64 ± 0.60 199 ± 4 203.0 ± 4.0 0.3230 ± 0.0040 2.62 ± 0.14 624 ± 10
3 0.4908 ± 0.0089 0.2082 ± 0.0068 128.0 ± 1.1 107 ± 1 269.1 ± 2.0 0.28131 ± 0.00035 1.512 ± 0.011 609 ± 1
4 0.1637 ± 0.0023 0.24 ± 0.16 102.87 ± 0.40 62.07 ± 0.73 68.9 ± 2.7 0.14280 ± 0.00099 1.083 ± 0.012 552 ± 2
Tarantulas
1 0.4964 ± 0.0021 0.1479 ± 0.0029 112.33 ± 0.28 157 ± 1 735.9 ± 7.6 0.07640 ± 0.00025 1.079 ± 0.014 1011 ± 10
2 0.2461 ± 0.0042 0.3393 ± 0.0016 121.0 ± 2.8 218 ± 1 351.3 ± 1.1 0.1178 ± 0.0020 1.179 ± 0.020 821 ± 2
3 0.5862 ± 0.0049 0.2992 ± 0.0031 80.3 ± 1.2 224.2 ± 0.4 275.6 ± 4.9 0.1194 ± 0.0076 2.013 ± 0.028 734 ± 8
Water bug
1 0.10819 ± 0.00033 0.179 ± 0.031 12.69 ± 0.36 129 ± 1 10.03 ± 0.14 0.15052 ± 0.00042 0.469 ± 0.010 147.2 ± 0.4
2 0.2217 ± 0.0054 0.211 ± 0.025 11.199 ± 0.080 474 ± 340 14.674 ± 0.053 0.2358 ± 0.0086 0.5047 ± 0.0076 119 ± 1
Worms
1 0.04598 ± 0.00043 0.0231 ± 0.0045 16.6 ± 1.3 46.6 ± 5.0 10.72 ± 0.12 0.960 ± 0.010 0.819 ± 0.012 106 ± 6
2 0.001978 ± 0.000015 0.0203 ± 0.0019 9.12 ± 0.11 32.85 ± 0.97 19.390 ± 0.055 0.3557 ± 0.0035 0.6344 ± 0.0096 175 ± 4
3 0.048699 ± 0.000032 0.0504 ± 0.0074 9.714 ± 0.096 51.8 ± 2.4 17.7 ± 1.6 0.699 ± 0.011 0.2437 ± 0.0022 73.5 ± 3.0
4 0.01844 ± 0.00023 0.0237 ± 0.0043 19.08 ± 0.20 56.7 ± 9.7 11.762 ± 0.024 0.55998 ± 0.00063 0.10654 ± 0.00047 105 ± 1
5 0.01959 ± 0.00073 0.0798 ± 0.0096 20.02 ± 0.73 51.3 ± 1.8 11.187 ± 0.057 0.573 ± 0.014 0.1073 ± 0.0067 107 ± 3
6 0.0198 ± 0.0024 0.0223 ± 0.0067 18.45 ± 0.12 49.34 ± 0.49 11.10 ± 0.15 0.5681 ± 0.0089 0.10155 ± 0.00046 101 ± 1

Table 6. Concentrations ± standard deviation (n = 2; mg kg−1 ) of remaining elements in edible

insects, scorpions, and tarantulas (with corresponding sample name) measured by ICP-MS.

Sample Al B Ba Sr
Ants
1 99.5 ± 2.5 0.742 ± 0.024 17.54 ± 0.37 4.828 ± 0.091
2 105 ± 10 0.715 ± 0.031 14.753 ± 0.029 4.24 ± 0.23
Crickets
1 5.49 ± 0.74 0.757 ± 0.073 0.5008 ± 0.0019 2.794 ± 0.077
2 3.06 ± 0.43 0.5543 ± 0.0056 0.464 ± 0.013 2.75 ± 0.10
3 4.3 ± 1.7 0.553 ± 0.011 0.489 ± 0.038 2.9160 ± 0.0040
4 20.4 ± 1.2 0.398 ± 0.012 0.786 ± 0.043 2.472 ± 0.048
5 12.3 ± 7.6 0.690 ± 0.043 1.06 ± 0.16 2.54 ± 0.13
6 3.79 ± 0.72 0.314 ± 0.021 0.773 ± 0.067 2.40 ± 0.33
7 6.29 ± 0.32 0.771 ± 0.018 1.4569 ± 0.0065 1.861 ± 0.039
8 4.07 ± 0.60 0.714 ± 0.041 1.171 ± 0.048 1.636 ± 0.043
Locusts
1 55.3 ± 2.4 0.387 ± 0.031 0.9767 ± 0.0055 1.895 ± 0.028
2 119 ± 15 0.400 ± 0.020 1.298 ± 0.049 1.0888 ± 0.0059
Scorpions
1 283 ± 17 0.659 ± 0.020 8.23 ± 0.64 4.80 ± 0.45
2 64.25 ± 0.54 0.3993 ± 0.0042 11.071 ± 0.018 10.58 ± 0.11
3 69.0 ± 2.0 0.763 ± 0.034 7.078 ± 0.062 17.99 ± 0.11
4 103.7 ± 0.5 0.429 ± 0.017 4.269 ± 0.028 5.436 ± 0.055
Tarantulas
1 132.8 ± 0.05 0.898 ± 0.040 28.83 ± 0.17 31.22 ± 0.14
2 169 ± 3 1.0023 ± 0.0036 23.78 ± 0.41 31.08 ± 0.93
3 167 ± 1 1.068 ± 0.049 28.57 ± 0.12 21.73 ± 0.21
Water bug
1 81.1 ± 1.4 nd 1.0085 ± 0.0063 1.2820 ± 0.0020
2 144 ± 3 nd 1.492 ± 0.016 1.424 ± 0.036
Worms
1 14.2 ± 2.3 0.758 ± 0.012 2.238 ± 0.053 3.71 ± 0.20
2 1.406 ± 0.029 nd 0.857 ± 0.050 0.901 ± 0.016
3 14.86 ± 0.96 1.184 ± 0.028 2.22 ± 0.14 3.222 ± 0.069
4 0.490 ± 0.070 2.28 ± 0.23 3.974 ± 0.071 3.619 ± 0.064
5 1.5 ± 1.2 2.128 ± 0.070 3.83 ± 0.20 3.57 ± 0.11
6 0.4421 ± 0.0020 2.49 ± 0.36 4.003 ± 0.043 3.5216 ± 0.0024
nd: not detected.
Foods 2024, 13, 2353 10 of 13

Table 7. Comparison of concentrations previously reported for potentially toxic and major elements
with those obtained in this work.

As Cd Pb Ca K Mg Na
Insect Reference
(µg kg−1 ) (µg kg−1 ) (µg kg−1 ) (mg kg−1 ) (mg kg−1 ) (mg kg−1 ) (mg kg−1 )
Cricket
300–400 22–28 n.d.-19 612–2630 8100–13,500 740–1880 1130–15,900 [27]
(n = 3)
Cricket
3.6–21.1 13.8–76.6 16.7–96.9 411–1798 5263–9116 727–1316 1162–2874 This work
(n = 8)
Locust
300 n.d. 16 864 6060 490 2160 [27]
(n = 1)
Locust
35.0–47.4 31.8–40.9 50.2–73.4 481–559 6350–6510 658–749 860–937 This work
(n = 2)
Mealworm
200–620 30–200 n.d.-100,000 941–2380 7560–10,000 870–1450 1290–1510 [27]
(n = 3)
Mealworm
25.7–39.7 79.3–83.8 6.0–10.8 401–788 4474–8533 1343–3074 521–982 This work
(n = 5)
Silkworm
200 n.d. 9 622 1090 480 3650 [27]
(n = 1)
Silkworm
470 53.9 47.3 1132 7462 2565 126 This work
(n = 1)
Water bug
320.9–561.5 220–346 336–420 197.4–286.2 [23]
(n = 5)
Water bug
632–855 3950–4041 614–695 3150–5113 This work
(n = 2)

Table 8. Comparison of concentrations (mg kg−1 ) previously reported for essential elements with
those obtained in this work.

Insect Co Cr Cu Fe Mn Mo Se Zn Reference
Cricket
n.d.-0.045 0.174–0.267 16.3–23.9 40.9–80.7 9.62–45.8 0.327–17.2 0.474–0.899 125–173 [27]
(n = 3)
Cricket
0.0102–0.0880 0.0164–0.1380 19.54–53.09 49.3–83.6 7.7–63.5 0.560–1.531 0.123–0.638 135–256 This work
(n = 8)
Locust
n.d. 0.140 35.7 42.4 4.44 0.734 0.307 140 [27]
(n = 1)
Locust
0.1218–0.1246 0.0836–0.2066 60.3–92.7 90.3–164 6.46–6.77 0.574–0.678 0.216–0.334 173–177 This work
(n = 2)
Mealworm
0.018–0.033 0.164–0.624 11.3–28.2 36.2–253 5.36–410 0.37–0.897 0.474–0.547 93.8–126 [27]
(n = 3)
Mealworm
0.01844–0.04870 0.0223–0.0798 9.71–20.02 46.6–56.7 10.7–17.7 0.560–0.960 0.102–0.819 73.5–107 This work
(n = 5)
Silkworm
n.d. 0.054 3.90 11.7 5.71 0.118 0.357 38.6 [27]
(n = 1)
Silkworm
0.001978 0.0203 9.12 32.85 19.390 0.3557 0.6344 175 This work
(n = 1)
Water bug
22.2–420 253–1121 19.8–42.2 49.8–72.2 [23]
(n = 5)
Water bug
11.2–12.7 129–474 10.03–14.67 119–147 This work
(n = 2)

4. Conclusions
This study addresses the assessment of potentially toxic, major, and essential elements
(19 trace elements in total) in a variety of edible insects, scorpions, and tarantulas by ICP-MS.
The method accuracy was verified by analyzing several insect certified reference materials.
Based on the provisional tolerable daily intake (PTDI) values for As, Cd, and Pb, it was
concluded that most of the samples were safe to consume for both children and adults
at relatively high amounts (number of individuals ranging from 1000 to 100,000 insects
per day). However, the maximum number of scorpions that can be safely consumed is
much lower (up to ~30 for adults and up to 10 for children), while in the case of tarantulas,
consumption of only 1 individual per day is recommended for children and 2–3 for adults.
Foods 2024, 13, 2353 11 of 13

Lastly, high levels of Cu were found in the tarantula and scorpion samples as well
as K, Na, Fe, and Zn in most of the insect samples, suggesting that edible insects have a
nutritional value.
A limitation of this study is that it included no information available on the insects’ de-
velopmental stage, environmental exposition, bioaccumulation of pesticides and chemicals,
which parts of the insects were used in previous studies, and the origin of the following
samples: crickets 5–6, locusts 1–2, and water bug 1–2.
The work will be pursued by increasing the replicate size in order to assess the samples’
heterogeneity and also extending the study to other edible insect species in order to provide
a more extensive database regarding the benefit–risk balance related to trace elements via
consumption of this type of foodstuffs.

Supplementary Materials: The following supporting information can be downloaded at: https:
//www.mdpi.com/article/10.3390/foods13152353/s1, Table S1: Statistical comparison of measured
concentrations in insect CRMs with their certified values.
Author Contributions: Conceptualization: Y.H., A.L., D.B. and P.J.; data curation: Y.H., A.L., D.B. and
P.J.; formal analysis: Y.H. and C.M.; funding acquisition: D.B. and P.J.; investigation: Y.H., A.L., D.B.
and P.J.; methodology: Y.H., A.L., C.M., D.B. and P.J.; project administration: D.B. and P.J.; resources:
D.B. and P.J.; supervision: A.L., D.B. and P.J.; validation: Y.H., C.M. and A.L.; writing—original draft
preparation: Y.H.; writing—review and editing: D.B., A.L. and P.J. All authors have read and agreed
to the published version of the manuscript.
Funding: This work was supported by a Mitacs Globalink Research Award (Canada) (funding
number FR101710).
Institutional Review Board Statement: Not applicable.
Informed Consent Statement: Not applicable.
Data Availability Statement: The original contributions presented in the study are included in the
article/Supplementary Materials; further inquiries can be directed to the corresponding author.
Acknowledgments: The authors gratefully acknowledge the National Research Council Canada
for providing the certified reference materials. They appreciate the support from the Metallic Trace
Elements and Minerals Unit at ANSES as the research was carried out in their laboratory.
Conflicts of Interest: The authors declare no conflicts of interest.

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