Analysis of Microbiological and Chemical Hazards in Edible Insects Available to Canadian Consumers

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Journal of Food Protection, Vol. 84, No. 9, 2021, Pages 1575–1581 https://doi.org/10.4315/JFP-21-099 Published 2021 by the International Association for Food Protection Copyright Ó, Her Majesty the Queen in Right of Canada, as represented by the Canadian Food Inspection Agency. This is an open access article. Research Paper Analysis of Microbiological and Chemical Hazards in Edible Insects Available to Canadian Consumers

BEATA M. KOLAKOWSKI https://orcid.org/0000-0002-5604-6978,* KRYSTYNA JOHANIUK, HELEN ZHANG, AND ETSUKO YAMAMOTO

Canadian Food Inspection Agency, Food Safety Science Services Division, 1400 Merivale Road, Ottawa, Ontario, Canada K1A 0Y9 Downloaded from http://meridian.allenpress.com/jfp/article-pdf/84/9/1575/2893105/i0362-028x-84-9-1575.pdf by guest on 26 September 2021 (ORCID: https://orcid.org/0000-0003-4786-3535 [H.Z.]; https://orcid.org/0000-0001-5533-4540 [E.Y.]) MS 21-099: Received 10 March 2021/Accepted 5 May 2021/Published Online 6 May 2021

ABSTRACT Edible insects are a novel food in most countries; their popularity is growing because of their high-protein and low-fat content, ease of cultivation, and small environmental impact. To our knowledge, this is the first report that addresses both microbiological and chemical hazards in edible insects. Samples were collected from retail stores or purchase through e- commerce. A total of 51 samples of dried whole insects or insect powder were tested for Escherichia coli, which serves as an indicator of the overall sanitation conditions throughout the food production chain, and the bacterial pathogen Salmonella spp. Neither Salmonella spp. nor E. coli (.100 CFU/g) was found in the samples analyzed. A total of 43 samples of crickets (protein bars, powders, flour, and whole insects) and 4 samples of silkworm (whole insects) were analyzed for up to 511 pesticides. Of these, 39 samples contained up to four pesticides; 34 samples were compliant and 5 samples were noncompliant with Canadian regulations. Seven pesticide residues were detected, with glyphosate and its metabolite, aminomethylphosphonic acid, as the predominant residues. Nineteen of the samples tested for pesticides were also analyzed for arsenic, cadmium, mercury, and lead; there was insufficient material remaining to allow testing of pesticides and metals. The positive rates for arsenic, cadmium, lead, and mercury were 100, 79, 58, and 74%, respectively. The detected concentrations ranged from 0.030 to 0.34 mg/kg for arsenic, from 0.031 to 0.23 mg/kg for cadmium, 0.019 to 0.059 mg/kg for lead, and from 0.94 to 28 μg/kg for mercury. Based on the lack of detection of microbiological contamination, and the positive rates and levels of pesticides and metals observed in the products, Health Canada determined that all insect products analyzed were safe for human consumption. This is a limited study; the Canadian Food Inspection Agency will continue to monitor this novel food.

HIGHLIGHTS Edible insects were tested for potential microbiological and chemical health hazards. No Salmonella spp. or E. coli (.100 CFU/g) was detected. 89% compliance was found with Canadian pesticide regulations. Glyphosate and its metabolite, AMPA, were the most commonly detected pesticides. All insect products tested were deemed safe for human consumption.

Key words: Edible insects; Escherichia coli; Glyphosate; Pesticides; Salmonella; Toxic metals

Edible insects and food products containing insect healthier and more nutritious food. The environmental ingredients are not commonly consumed by humans in footprint of insect production is minimal compared with the North America and Europe. By contrast, people in Africa, production of beef, pork, and chicken (31). Asia, and Latin America have been consuming insects for In response, a growing number of edible insect centuries. There are 1,000 to 2,000 species of insects products have appeared on the Canadian market in recent consumed globally (22). The Food and Agriculture years, such as whole and powdered insects and processed Organization of the United Nations (10) and other insect products. Dried whole insects (roasted, smoked, and organizations have been promoting the cultivation and flavored) are intended to be consumed as is, whereas consumption of edible insects as alternative sources of powdered insects are used as ingredients in other foods. protein (food security) and as sustainable forms of Processed products, such as protein bars, chips, crackers, agriculture. Edible insects have high protein-to-fat ratios and cookies, are manufactured with insects as the principal relative to plants or traditional meats, making insects a source of protein. Edible insect products can be purchased online, at specialty stores, and at mainstream grocery stores. * Author for correspondence. Tel: 613-773-6313; Fax: 613-773-5959; To date, there are no specific regulations, standards, or E-mail: [email protected]. guidelines regarding potential microbiological or chemicals 1576 KOLAKOWSKI ET AL. J. Food Prot., Vol. 84, No. 9 hazards established in Canada for edible insects or insect- multiresidue pesticide method (505 pesticides), a phenoxy containing foods. Internationally, the European Food Safety herbicide method (2 pesticides: 2,4-dichlorophenoxyacetic acid Authority requires premarket approval of individual edible [2,4-D] and 2-methyl-4-chlorophenoxyacetic acid [MCPA]), a insect species (8). A few countries in the European Union quat screen (diquat/paraquat) method, and a glyphosate method permit the marketing and sale of certain edible insect (parent compound plus metabolite) for a total of 511 pesticides. species (25). Edible insects produced for human consump- Metal method analyses were used for up to 18 metals, but only the tion and available to Canadian consumers must meet the toxic metals (arsenic, cadmium, lead, and mercury) are discussed in this article. same safety and hygiene standards as other foods available in Canada (6). All pesticide residues in insect products are Chemical analysis: multiresidue pesticide method. The subject to the general maximum residue limit (MRL) of 0.1 multiresidue pesticide method began with soaking in water for 2 h mg/kg (7). Each pesticide is assessed separately against the (1). Then, the sample was extracted in 1% acetic acid in MRL, including cases with multiple pesticides per sample. acetonitrile in the presence of isotopically labeled internal Notable exceptions are a pesticide and its metabolite or standards. Cleanup was performed using dispersive solid-phase Downloaded from http://meridian.allenpress.com/jfp/article-pdf/84/9/1575/2893105/i0362-028x-84-9-1575.pdf by guest on 26 September 2021 metabolites or multiple forms of a pesticide (e.g., spinosyn extraction with primary-secondary amine and C18. The extract A and spinosyn D, assessed as spinosad), which are was then analyzed by gas chromatography–tandem mass spec- summed and then compared against the MRL. There are trometry (GC-MS/MS) and liquid chromatography–tandem mass no regulations regarding permissible levels of arsenic, spectrometry (LC-MS/MS). cadmium, lead, or mercury in insect products in Canada (7). GC-MS/MS was performed using electron impact ionization, In this context, this article presents baseline surveillance with helium as the carrier gas (1). The samples were also analyzed data on the prevalence and levels of chemical and using reverse-phase liquid chromatography with gradient condi- microbiological hazards of edible insects. These data are tions. Detection was performed by tandem mass spectrometry with electrospray ionization in positive or negative mode with multiple important in the assessment of potential human health risks. reaction monitoring (1). A total of 505 pesticides were analyzed MATERIALS AND METHODS by these two methods. Two transitions were monitored per pesticide residue, regardless of detection method. Supplemental Insect samples. Each sample consisted of a single or Table S1 contains a list of all pesticides screened as part of this multiple unit or units (e.g., individual consumer-sized packages) method. from a single lot with a total weight of at least 50 g for microbiological analysis or at least 200 g for chemical analysis. Chemical analysis: phenoxy herbicide method. With the All samples were acquired from online retailors or collected at phenoxy herbicide method (2), the sample was extracted in 0.007 retail establishments located in Ottawa, Canada, with the intent of M HCl, pH was adjusted to 2, and ethyl ether was added. After capturing as wide a range of retail forms (i.e., brand and product centrifugation, the ethyl ether layer was evaporated to near type) and insect types as possible. The samples included dried dryness, diluted to volume with methanol, centrifuged again, and whole insects, insects in powdered form, or insects incorporated then analyzed with LC-MS/MS. The samples were analyzed using into finished products (e.g., protein bars). The products included reverse-phase liquid chromatography with isocratic conditions. domestically cultivated and processed products, domestically Detection was performed by tandem mass spectrometry with processed products, and imported products. Some products were electrospray ionization in negative mode with single reaction labeled organic. To be included in the study, the products had to be monitoring. For MCPA, m/z 199 and 201 were monitored, whereas intended for human consumption. Therefore, insects available for for 2,4-D, m/z 219.2 and 221.2 were monitored. Table S2 contains purchase as animal or pet feed were excluded. A total of 51 a list of all pesticides screened as part of this method and their samples of dried whole insects or insect powder were collected for limits of detection (LODs). microbiological hazard testing. A total of 43 samples of crickets (protein bars, powders, flour, and whole insects) and 4 samples of Chemical analysis: quat method. With the quat method (9), silkworm (whole insects) were collected for chemical hazard samples were extracted in acidic methanol (2% HCl). After testing. incubation, two rounds of centrifugation, and filtration, the samples were ready for analysis by LC-MS/MS. Microbiological analyses. Samples were analyzed for The samples were analyzed using reverse-phase, ion pair Salmonella spp. and Escherichia coli by the Canadian Food liquid chromatography with gradient elution. Detection was Inspection Agency (CFIA) using methods published in Health performed by tandem mass spectrometry with electrospray Canada’s Compendium of Analytical Methods for the Microbio- ionization in positive mode. Two transitions were monitored for logical Analysis of Foods (15). Specifically, MFLP-29 and each analyte. For paraquat, m/z 171.0 to 155.0 and 171.0 to 103.0 MFHPB-20 were used for Salmonella spp. and MFHPB-34 was were monitored. For diquat, m/z 183.0 to 157.0 and 183.0 to 130.0 used for E. coli analysis (15). were monitored. Table S3 contains a list of all pesticides screened as part of this method and their LODs. Chemical analyses. Samples were analyzed in an external, ISO/IEC 17025–accredited laboratory under contract with the Chemical analysis: glyphosate method. With the glyph- Government of Canada. The methods have been demonstrated to osate method (26), the sample was extracted in 0.05 M KOH in the provide accurate results via internationally accredited proficiency presence of an isotopically labeled internal standard. After testing programs, as well as interlaboratory sample comparison. In centrifugation, the extract was adjusted to pH 7.0 6 0.5 with addition, when the laboratories identify pesticide residue levels 0.1 M HCl. Following addition of 5% sodium thiosulfate and within 80% of the MRL for a particular commodity, they are sodium tetraborate, the sample extract was derivatized with required to repeat the assay to verify the results. Four analytical fluorenylmethyloxycarbonyl chloride. The reaction was quenched, methods were used (and will be individually described later): a filtered, and analyzed by LC-MS/MS (26). The samples were J. Food Prot., Vol. 84, No. 9 HAZARDS IN EDIBLE INSECTS IN CANADA 1577 analyzed using reverse-phase liquid chromatography with gradient TABLE 1. Distribution of sample positives and noncompliances elution. Detection was performed by tandem mass spectrometry as a function of product type with electrospray ionization in negative mode. Two marker No. of: residues were analyzed (glyphosate and its metabolite, aminomethylphosphonic acid [AMPA]), and two transitions were Insect type Format Samples Nondetects Detects Noncompliances monitored for each analyte. For glyphosate, m/z 390.0 to 150.0 and 390.0 to 168.0 were monitored. For AMPA, m/z 110.0 to 63.0 Cricket Bar 30 2 28 0 and 110.0 to 79.0 were monitored. Table S4 contains a list of all Crackers 1 0 1 0 pesticides screened as part of this method and their LODs. Flour 2 2 0 0 Powder 7 0 5 2 Multimetal analysis. In addition, where there was sufficient Whole 3 0 1 2 quantity of sample, the samples were analyzed by inductively Silkworm Whole 4 2 1 1 coupled plasma mass spectrometry (ICP-MS) for 18 metal contaminants (28, 30). The sample was extracted with 7.5 mL Downloaded from http://meridian.allenpress.com/jfp/article-pdf/84/9/1575/2893105/i0362-028x-84-9-1575.pdf by guest on 26 September 2021 of concentrated nitric acid at room temperature for 12 h. The Zambia). Salmonella spp. and E. coli (.100 CFU/g) were extract was then heated, and hydrogen peroxide and hydrochloric not detected in the samples tested. Similar to the results of acid were added sequentially. The last step was the addition of this survey, a preliminary study in Germany did not detect water to a volume of 50 mL. The sample was thus ready for the presence of Salmonella or E. coli (.100 CFU/g) in 38 analysis by ICP-MS. samples of edible insects available at retail between 2014 For determination of arsenic, cadmium, and lead by ICP-MS, and 2015 (12). In 2015 to 2016, a Dutch study found that the digestate was introduced into the Argon plasma with a genetic material from Salmonella spp. was absent from the temperature between 6,000 to 8,000 K. Ions were filtered based on mass-to-charge ratio and detected by a discrete dynode electron processed edible insect samples from one Dutch company fi multiplier. The output from the detector was proportional to the (11). However, these studies identi ed Bacillus cereus, concentration of the element in the sample. The reporting limits Pseudomonas spp., (12), Listeria spp., and Staphylococcus were 0.02 mg/kg for arsenic, 0.01 mg/kg for cadmium, and 0.015 spp. (11) in the samples analyzed, respectively. mg/kg for lead. Table S5 contains a list of all metals screened as Several studies have indicated that the total bacterial part of this method and their LODs. load of unprocessed raw insects was higher than that found Mercury (29) was analyzed by cold vapor atomic fluores- in raw ground meat and that an effective heat treatment cence spectroscopy. Mercury atoms were excited by a UV light (sterilization) is needed to reduce the total bacterial load (3, source at a wavelength of 254 nm; the fluorescence was detected 13, 24). The Federal Agency for the Safety of the Food by photomultiplier tube or UV photodiode. Argon was used as the Chain of Belgium requires all insects for human consump- μ carrier gas. The reporting limit was 0.5 g/kg. tion to undergo heat inactivation or sterilization to ensure the safety of the food products (25). Statistical analysis. The sample population consisted of all Because the samples we tested were picked up at retail, units of the targeted commodities available at retail to Canadian consumers as per the sampling design. Because all units of the no information is available on whether a heat-inactivation population could not be assumed to have an equal probability of step was used or on the conditions from breeding stage to selection, the commodity units at a store or online were drawn as final product. Our preliminary bacterial testing results randomly as possible to be reasonably representative of the indicate that the edible insects analyzed in this study appear population. The sampling method used for this study was a to have been produced under sanitary conditions. nonprobability sampling method, which does not allow standard statistical inferential methods to be invoked. Nevertheless, the Chemical testing. A total of 47 samples were tested for random selection approach provide a snapshot on the presence of up to 511 pesticides. The residue levels of the pesticides microbiological and chemical hazards in the selected foods. were assessed against Canadian MRLs for pesticide residues in food. The MRL is the maximum amount of RESULTS AND DISCUSSION residue that is expected to remain in or on food products A total of 51 samples of dried whole insects or insect when a pesticide is used according to label instructions. powder were tested for microbiological hazards. A total of Product compliance was determined by comparison of the 43 samples of crickets (protein bars, powders, flour, and reported level of a pesticide and metabolites to the MRLs whole insects) and 4 samples of silkworm (whole insects) published in Health Canada’s Pesticide MRL database (14). were tested for chemical hazard testing. The results are Many food-pesticide combinations do not have specific discussed below in the appropriate sections “Microbiolog- MRLs in Canada. In those cases, a default MRL (0.1 mg/kg) ical testing” and “Chemical testing.” is used for assessments. There are no MRLs in place for insect products, so the default MRL was used for Microbiological testing. All 51 products sampled as assessment. part of this study were analyzed for the presence of Table 1 shows a summary of the number of samples Salmonella spp. and E. coli. Table S6 presents the species of tested, the number of nondetects, the number of detects (the whole and powdered insects tested as part of this study. Of concentration is at or below the default MRL of 0.1 mg/kg), the 51 samples, 18 (35%) were domestically produced and and the number of noncompliances (the concentration 33 (65%) were imported from five countries (France, exceeds 0.1 mg/kg). The detection rate for pesticides was Thailand, the United Kingdom, the United States, and 50% in silkworms and 89% in cricket-based products (with 1578 KOLAKOWSKI ET AL. J. Food Prot., Vol. 84, No. 9

Few studies in the literature examine the levels of pesticides in retail-level insect products. One study tested locusts for pesticides (23). In that study, five pesticides were detected (β-hexachlorocyclohexane; lindane; aldrin; feni- trothion, also known as sumithion; and malathion) at concentrations ranging from 2.20 to 740 μg/kg. These pesticides are included in the scope of the multiresidue analysis used in this study but were not detected in the samples. Another survey examined 393 pesticides in fly larvae and found only chlorpyrifos (n ¼ 1) and piperonyl butoxide (n ¼ 1) (4). Chlorpyrifos was also detected in this study in a cricket-based protein bar, but piperonyl butoxide was not detected. A recent study of edible insects detected FIGURE 1. Frequency of detection of pesticide residues per Downloaded from http://meridian.allenpress.com/jfp/article-pdf/84/9/1575/2893105/i0362-028x-84-9-1575.pdf by guest on 26 September 2021 nine pesticides in one or more of the samples, but none of sample. these coincided with the pesticides detected in this study (21). Another study suggests that 99.9% of a pesticide variation depending on product format). Of the 47 samples, application is absorbed by the environment and may 5 samples (4 cricket products and 1 silkworm product, 11%) accumulate in the feed materials for insects (20). Articles had one or more noncompliant pesticide residues. have suggested that the degree of pesticide accumulation As demonstrated in Table 1 and Figure 1, most samples % depends on the type of insect, its growth stage, the (64 ) contained a single pesticide residue per sample. Eight concentration of the pesticide in the feed or environment, samples contained no detectable pesticide residues per and whether the insects were caught from the wild or sample, and seven samples contained two pesticide residues farmed. All articles, including the present study, are in per sample (one sample was associated with two noncom- agreement that pesticides in insect products do not pose a pliances). Only one sample contained three pesticide human health risk. residues (all compliant), and one sample contained four In the final part of this study, where sufficient sample pesticide residues (one compliant and three noncompliant was collected, multimetal analysis was performed. This residues). subset included 15 samples of cricket-based products and 4 Phenoxy herbicides, diquat, and paraquat were not silkworm-based products. The concentrations of the metals detected in the samples. Table 2 presents the seven in the insect products depend on numerous factors, pesticides and one metabolite that were detected in the including the levels in the environment (soil, water, and insect products. Four of the pesticides were detected only in air), the type of insect, its growth phase at harvest, its one sample each. Residues of glyphosate (including its lifespan, and the types of chemicals used on the feed metabolite, AMPA, observed singly or in combination) materials (e.g., arsenic-containing pesticides). Tables S11 were the most frequently detected pesticide residues. through S22 contain summaries of the metal levels as a Glyphosate and AMPA co-occurred in eight samples. function of metal, insect, and product type. Antimony, AMPA alone was detected in two samples. All detections beryllium, and tin were not detected in the insect samples. of glyphosate and/or AMPA occurred in cricket-based Only the results for the toxic metals (arsenic, cadmium, products. lead, and mercury) are discussed in this article. The levels of all pesticides observed were quite low, Arsenic, cadmium, lead, and mercury are of concern suggesting that the pesticides are not directly applied to the because they are associated with harmful human health insects being raised for human consumption but rather effects, ranging from affecting the brains of infants and originate from the materials used for feeding the insects children to causing cancer or death from exposure to high (e.g., grains, seeds, and grass) or other ingredients in the levels (27). The health effects vary according to the metal, products. Because the samples are being picked up at retail, its form, and the concentration. These metals typically are it is not possible to determine whether, when, or how the not deliberately added to foods (except as components of pesticides came to be in the products tested. pesticides licensed for use in Canada). They may be present

TABLE 2. Pesticides and metabolites detected in insect samples

No. of Minimum Maximum Avg Compound Function detects (mg/kg) (mg/kg) (mg/kg)

Glyphosate Herbicide 36 0.0064 0.15 0.027 AMPA Metabolite of glyphosate 9 0.007 0.45 0.16 Chlorfenapyr Insecticide 2 0.027 0.20 0.11 Chlorpyrifos Insecticide 1 0.014 Ethoxyquin Antioxidant 1 0.055 Trifloxystrobin Fungicide 1 0.0088 Tris(chloropropyl) phosphate Pesticide 1 0.18 J. Food Prot., Vol. 84, No. 9 HAZARDS IN EDIBLE INSECTS IN CANADA 1579

TABLE 3. Occurrence of arsenic in insect products

Insect type Format No. of samples No. of positives Minimum (mg/kg) Maximum (mg/kg) Avg (mg/kg)

Cricket Bar 10 10 0.035 0.16 0.073 Flour 2 2 0.052 0.059 0.056 Powder 1 1 0.34 Whole 2 2 0.12 0.28 0.2 Silkworm Whole 4 4 0.03 0.065 0.049

TABLE 4. Occurrence of cadmium in insect products

Insect type Format No. of samples No. of positives Minimum (mg/kg) Maximum (mg/kg) Avg (mg/kg)

Cricket Bar 10 10 0.031 0.23 0.087 Downloaded from http://meridian.allenpress.com/jfp/article-pdf/84/9/1575/2893105/i0362-028x-84-9-1575.pdf by guest on 26 September 2021 Flour 2 2 0.072 0.099 0.086 Powder 1 1 0.058 Whole 2 2 0.053 0.094 0.074 Silkworm Whole 4 0

TABLE 5. Occurrence of lead in insect products

Insect type Format No. of samples No. of positives Minimum (mg/kg) Maximum (mg/kg) Avg (mg/kg)

Cricket Bar 10 5 0.042 0.059 0.052 Flour 2 2 0.019 0.024 0.022 Powder 1 0 Whole 2 2 0.023 0.032 0.028 Silkworm Whole 4 1 0.02

TABLE 6. Occurrence of mercury in insect products

Insect type Format No. of samples No. of positives Minimum (mg/kg) Maximum (mg/kg) Avg (mg/kg)

Cricket Bar 10 9 0.00094 0.0056 0.0028 Flour 2 2 0.001 0.0011 0.0011 Powder 1 1 0.028 Whole 2 2 0.014 0.018 0.016 Silkworm Whole 4 0 as environmental contaminants from natural or industrial The positive rate for lead was 58% overall, 25% for sources. The laboratory cannot distinguish the source of the silkworm pupae–based products, and 67% for cricket-based metal. products (Table 5). The concentrations of lead ranged from As shown in Table 3, all insect products tested 0.019 to 0.059 mg/kg, with an average concentration of contained a detectable level of arsenic. The concentrations 0.034 mg/kg. The detected level in silkworm pupae–based detected ranged from 0.030 mg/kg (whole silkworm pupae) samples was 0.020 mg/kg versus an average level of 0.033 to 0.34 mg/kg (cricket powder), with an average concen- mg/kg in cricket products. These values do not concur with tration of 0.094 mg/kg. The average level of arsenic was the reported result of no more than 0.03 mg/kg for cricket 0.12 mg/kg in cricket products and 0.049 mg/kg in samples (21). Table S9 provides a listing of the lead content silkworm pupae. These values do not concur with the of each of the 19 samples tested. reported result of no more than 0.03 mg/kg for cricket As with cadmium, none of the silkworm pupae–based samples (18, 21). Table S7 provides a listing of the arsenic products and all cricket-based samples contained detectable content of each of the 19 samples tested. levels of mercury (Table 6). The levels of detected mercury Table 4 indicates that none of the silkworm pupae– ranged from 0.94 to 28 μg/kg, with an average level of 6.2 based products and all cricket-based samples contained μg/kg. These levels of mercury in crickets are low relative detectable levels of cadmium. The levels of detected to the 125 6 62 and 109 6 73 μg/kg observed in one study cadmium ranged from 0.031 to 0.23 mg/kg, with an average (19). Table S10 provides a listing of the mercury content of level of 0.083 mg/kg. This does not concur with previously each of the 19 samples tested. reported cadmium levels of no more than 0.03 mg/kg (21). The lack of concurrence with literature data for some Table S8 provides a listing of the cadmium content of each metal content may relate to the type of insect product of the 19 samples tested. sampled, the degree of processing, the growth phase of the 1580 KOLAKOWSKI ET AL. J. Food Prot., Vol. 84, No. 9 insect, the concentrations of heavy metals in the cultivation Available at: http://www.fao.org/fileadmin/user_upload/agns/pdf/ or harvest environment, and whether the insects are wild CXS_193e.pdf. Accessed 28 April 2021. 6. Department of Justice Canada. 2014. Food and drugs act. Available caught or farmed. This information is not available, because at: http://laws-lois.justice.gc.ca/eng/acts/F-27/. Accessed 10 March the samples are picked up at the retail level, rather than the 2021. producer level. The data provided in this article, in 7. Department of Justice Canada. 2019. Food and drug regulations— agreement with all other articles cited, does not indicate Section B.15.002(1). Available at: https://laws-lois.justice.gc.ca/eng/ # that there is a human health risk associated with the levels regulations/C.R.C.,_c._870/page-74.html h-573261. Accessed 10 March 2021. of toxic metals detected in insect products. 8. European Food Safety Authority Scientific Committee. 2015. Risk Canada has not established maximum levels (MLs) for profile related to production and consumption of insects as food and heavy metals in insect products. However, Canada has feed. EFSA 13:4257–4317. established MLs for arsenic, lead, and mercury in 9. European Union (EU) Reference Laboratories for Residues of commodities with higher consumption rates than insect Pesticides. 2013. Quick method for the analysis of residues of numerous highly polar pesticides in foods of plant origin involving

fi Downloaded from http://meridian.allenpress.com/jfp/article-pdf/84/9/1575/2893105/i0362-028x-84-9-1575.pdf by guest on 26 September 2021 products. Speci cally, none of the insect samples exceeded simultaneous extraction with methanol and LC-MS/MS determina- the Canadian MLs of 0.35 mg/kg for inorganic arsenic in tion (QuPPE-method) version 7.1. Available at: https://www.eurl- brown rice or 0.2 mg/kg for lead in beverages (16). The pesticides.eu/userfiles/file/EurlSRM/meth_QuPPe-PO_EurlSRM. highest level of mercury detected in insect products was pdf. Accessed 10 March 2021. 10. Food and Agriculture Organization of the United Nations. 2013. almost 18 times lower than the Canadian ML of 0.5 mg/kg Edible insects—future prospects for food and feed security. Available for mercury in fish (17). There are no Canadian regulations at: http://www.fao.org/docrep/018/i3253e/i3253e.pdf. 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