Safety of grain and flour from perennial intermediate wheatgrass (Thinopyrum intermedium) as a novel food pursuant to Regulation (EU) 2015/2283

TL;DR

This paper evaluates the safety of grain and flour from a new type of grass as a novel food in the EU and finds potential microbiological and allergenic concerns.

Contribution

The paper provides a safety assessment of perennial intermediate wheatgrass as a novel food under EU regulations.

Findings

The novel food has high microbial counts, indicating poor hygiene during production.

It can trigger allergic reactions in individuals with cereal allergies or celiac disease.

The provided data do not support a 12-month shelf-life for the product.

Abstract

Following a request from the European Commission, the EFSA Panel on Nutrition, Novel Foods and Food Allergens (NDA) was asked to deliver an opinion on grain and flour from perennial intermediate wheatgrass (IWG) (Thinopyrum intermedium) as a novel food (NF) pursuant to Regulation (EU) 2015/2283. The NF, composed primarily of carbohydrates, proteins and water, is produced by cultivating the IWG plant and subsequently processing it to yield either grains or flour. The target population proposed by the applicant is the general population and the NF is proposed as a food ingredient in several food products. Considering the nutritional profile of the NF and the proposed conditions of use, its consumption is considered not nutritionally disadvantageous, and the Panel considers that no toxicological studies are required on this NF. The microbiological analyses for several batches of the NF revealed high microbial counts regarding total aerobic microbial count, Enterobacteriaceae, yeasts and moulds, highlighting excessive variability in the hygiene conditions along the production process. The NF has the same potential as wheat, barley and rye to trigger IgE‐mediated allergic reactions in cereal‐allergic individuals and/or adverse reactions in individuals with coeliac disease under the proposed conditions of use. The Panel notes that, due to the high microbial loads observed in both IWG grain and flour batches, the microbiological quality of the NF is compromised. The potential presence of Enterobacteriaceae and opportunistic pathogen Pantoea species, such as P. agglomerans, and further released compounds could pose safety concerns. Moreover, the provided stability data do not support a 12‐month shelf‐life for the NF. Based on the available data, the Panel concludes that the safety of the NF, grain and flour from perennial intermediate wheatgrass (Thinopyrum intermedium) cannot be established.

Tables

TABLE 1. Proximate analysis of the NF.

Parameter (unit)	B#1	B#2	B#3	B#4	B#5	B#6	B#7	B#8	Methods of analysis
Moisture (g/100 g)	10.4	10.4	11.0	10.9	9.6	9.5	9.7	10.0	AACC 44‐15.02
Proteins (Nx6.25) (g/100 g)	19.4	21.5	15.5	15.6	19.2	21.7	15.2	16.1	AACC 46‐30.01
Total carbohydrates (g/100 g)	65.3	62.9	68.3	68.6	66.1	63.3	70.3	68.8	Calculation (by difference)
Total dietary fibre 7 (g/100 g)	23.9	23.3	21.1	21.9	20.4	20.6	17.8	22.0	AACC 32‐07.01/AOAC 991.43
Insoluble dietary fibre (g/100 g)	17.9	17.1	15.3	16.1	15.2	15.6	12.8	15.7	AACC 32‐07.01/AOAC 991.43
Soluble dietary fibre (g/100 g)	6.0	6.1	5.7	5.8	5.2	5.0	5.0	6.3	AACC 32‐07.01/AOAC 991.43
Fat (g/100 g)	2.9	3.0	2.9	3.0	2.9	3.1	2.8	3.0	AOAC 996.06
Ash (g/100 g)	2.0	2.2	2.3	1.9	2.2	2.4	2.0	2.1	AACC 08‐01.01

TABLE 2. Batch‐to‐batch microbiological analyses of the NF.

Parameter (unit)	B#9	B#10	B#11	B#12	B#13	B#14	B#15	B#16	B#17	B#18	Methods of analysis
Total aerobic microbial count (CFU/g)	8.6 × 106	4.3 × 105	2.1 × 104	5.3 × 106	7.4 × 106	8.7 × 105	8.6 × 104	1.1 × 104	1.1 × 106	1.1 × 106	ISO 4833‐1:2013
Coliforms (CFU/g)	< 10	< 10	< 10	< 10	< 10	< 10	< 10	< 10	< 10	< 10	NMKL 44:2004
Enterobacteriaceae (CFU/g)	> 1.5 × 104	3.9 × 103	6.9 × 102	> 1.5 × 104	> 1.5 × 104	> 1.5 × 104	9.2 × 103	1.3 × 103	> 1.5 × 104	> 1.5 × 104	NMKL 144:2005
E. coli (CFU/g)	< 10	< 10	< 10	< 10	< 10	< 10	< 10	< 10	< 10	< 10	NMKL 125:2005
Presumptive Bacillus cereus (CFU/g)	< 10	< 10	< 10	< 10	< 10	< 10	< 10	< 10	< 10	< 10	ISO 7932:2005
Coagulase‐positive staphylococci (CFU/g)	< 10	< 10	< 10	< 10	< 10	< 10	< 10	< 10	< 10	< 10	ISO 6888‐2:1999/Amd 1:2003
Salmonella spp. (per 25 g)	ND	ND	ND	ND	ND	ND	ND	ND	ND	ND	NMKL 71:1999
Yeasts (CFU/g)	20	1.2 × 102	10	2.7 × 102	6.0 × 103	1.9 × 102	30	< 10	6.4 × 103	< 10	NMKL 98:2005
Moulds (CFU/g)	1.9 × 104	1.7 × 103	9.5 × 102	8.2 × 104	4.2 × 103	1.6 × 103	5.1 × 102	5.7 × 102	3.9 × 104	4.9 × 102	NMKL 98:2005

TABLE 3. Specifications of the NF.

Description: Grain and flour from perennial IWG
Source: Thinopyrum intermedium
Parameter	Specification
Crude proteins	12–25 g/100 g
Total carbohydrates	60–70 g/100 g
Total dietary fibre	14–22 g/100 g
Fat	2–4 g/100 g
Ash	1–4 g/100 g
Moisture	≤ 13 g/100 g
Heavy metals
Lead	≤ 0.2 mg/kg
Arsenic	≤ 0.1 mg/kg
Cadmium 14	≤ 0.2 mg/kg
Mercury	≤ 0.05 mg/kg
Antinutritional factors
Phytic acid	< 2 g/100 g
Alpha‐amylase inhibitor activity	< 15,000 AAIU/g
Trypsin inhibitor activity	< 1000 TIU/g
Microbiological
TAMC	≤ 5 × 104 CFU/g
TYMC	≤ 100 CFU/g
Total coliforms	≤ 100 CFU/g
Presumptive Bacillus cereus	< 100 CFU/g
E. coli	< 10 CFU/g
Salmonella spp.	ND in 25 g
Coagulase‐positive staphylococci	< 10 CFU/g
Mycotoxins 14
Sum of aflatoxins (B1, B2, G1, G2)	< 0.004 mg/kg
Aflatoxins B1	< 0.002 mg/kg
Deoxynivalenol	< 1.25 mg/kg
Fumonisins (B1 and B2)	< 2 mg/kg
T2 and HT2 toxins	< 0.025 mg/kg
Zearalenone	< 0.1 mg/kg
Ochratoxin A	< 0.005 mg/kg

TABLE 4. Food categories, proposed uses and maximum use levels intended by the applicant.

Food category	Proposed food uses	Max use level (g NF/100 g)
Flour products	Cereal flour	100
Breads & similar products	All types of bread & rolls	85
	Crackers, crispbreads, rusks, breadsticks	100
Fine bakery wares	Baked goods, e.g. biscuits, cakes, pastry, etc.	70
Breakfast cereals	Ready‐to‐eat cereal	100
	Cereal bars	70
Snack foods	Savoury snacks (e.g. chips, pretzels, snack mix)	100
Pasta & noodles	Pasta	100
	Noodles	100
	Stuffed/filled pasta	20
Soups	Ready‐to‐eat soups	30
Beverages	Cereal‐based beverages	70
Alcoholic beverages	Beer	100
	Vodka, whiskey, other distilled beverages	100

Full text

INTRODUCTION

1

Background and Terms of Reference as provided by the requestor

1.1

On 18 December 2019, the company Patagonia Provisions, Inc.1 submitted a request to the European Commission in accordance with Article 10 of Regulation (EU) 2015/22832 to authorise placing on the Union market of the grain3 from perennial intermediate wheatgrass (Thinopyrum intermedium) as a novel food.

The application requests to authorise use of the grain from perennial intermediate wheatgrass (Thinopyrum intermedium) in a number of foods.

In accordance with Article 10(3) of Regulation (EU) 2015/2283, the European Commission asks the European Food Safety Authority to provide a scientific opinion on the grain from perennial intermediate wheatgrass (Thinopyrum intermedium) as a novel food.

DATA AND METHODOLOGIES

2

Data

2.1

The safety assessment of this NF is based on data supplied in the application and information submitted by the applicant following EFSA's requests for supplementary information.4

Administrative and scientific requirements for NF applications referred to in Article 10 of Regulation (EU) 2015/2283 are listed in the Commission Implementing Regulation (EU) 2017/2469.5

A common and structured format on the presentation of NF applications is described in the EFSA guidance on the preparation and presentation of a NF application (EFSA NDA Panel, 2016). As indicated in this guidance, it is the duty of the applicant to provide all the available (proprietary, confidential and published) scientific data (including both data in favour and not in favour) that are pertinent to the safety of the NF.

This NF application does not include a request for the protection of proprietary data.

Methodologies

2.2

The assessment follows the methodology set out in the EFSA guidance on NF applications (EFSA NDA Panel, 2016) and the principles described in the relevant existing guidance documents from the EFSA Scientific Committee. The legal provisions for the assessment are laid down in Article 11 of Regulation (EU) 2015/2283 and in Article 7 of Commission Implementing Regulation (EU) 2017/2469.

In the context of this opinion, EFSA's definition of dietary fibre (i.e. non‐digestible carbohydrates plus lignin; EFSA NDA Panel, 2010a, 2010b) does not reflect the additional requirement of having a beneficial physiological effect demonstrated by generally accepted scientific evidence laid down in Annex I of Regulation (EC) 1169/20116 for:

• edible carbohydrate polymers which have been obtained from food raw material by physical, enzymatic or chemical means,
• and edible synthetic carbohydrate polymers.

It is out of the scope of this opinion to establish whether the fraction of non‐digestible carbohydrates present in the NF meets the legal definition of dietary fibre in the EU or not.

This assessment concerns only the risks that might be associated with consumption of the NF under the proposed conditions of use and is not an assessment of the efficacy of the NF with regard to any claimed benefit.

ASSESSMENT

3

Introduction

3.1

The NF, in accordance with article 3 of the NF Regulation (EU) 2015/2283, falls under the category (iv) ‘food consisting of, isolated from or produced from plants and their parts’.

The NF, which is the subject of the application, is a domesticated perennial intermediate wheatgrass (Thinopyrum intermedium) belonging to the Triticeae tribe and Poaceae family. The NF is produced using agricultural practices (growing, harvesting and post‐harvest processing) similar to those used for wheat, and mainly consists of carbohydrates, proteins and water. The NF is proposed to be used as whole grain or wholegrain flour in a number of foods in the same way as wheat, barley, oats or other cereals. The target population is the general population.

Identity of the NF

3.2

The NF is the grain and flour from perennial intermediate wheatgrass (IWG), Thinopyrum intermedium (Host) Barkworth & D.R. Dewey. Synonyms for Thinopyrum intermedium are Agropyron intermedium, Triticum repens subsp. intermedium, Elytrigia intermedia, Trichopyrum intermedium, Triticum intermedium or the common name Wild triga. The species Thinopyrum intermedium is an allohexaploid intermediate wheatgrass (2n = 6x = 42) (Mahelka et al., 2011) in the wheatgrass genus Thinopyrum (Á. Löve) within the Triticeae tribe of the grass family (Poaceae). Wheat, rye and barley also belong to the Triticeae tribe.

The NF is obtained from cultivations in the US. Perennial IWG from Europe and Asia was introduced to the US in 1932, followed by the introduction of the pubescent subspecies of wheatgrass in 1934 (Ogle, 2011). Since 1980, IWG has been subjected to seed selection for breeding domestication. Subsequently, selection breeding methods were used to improve seed size, grain yield and free threshing for sustainable production as food and fuel (Zhang et al., 2014). No GMO techniques have been used in the development of IWG so far.

In response to a request from EFSA, the applicant, which is also the tradename licensor of the NF, specified that there is a list of approved varieties and seed sources for the NF. The ‘MN‐Clearwater’ variety is among those currently used in the production of the NF and is registered with the United States Department of Agriculture (USDA), as indicated by Bajgain et al. (2020). However, according to the applicant, the list of approved varieties is regularly updated, incorporating new releases and retiring older varieties.

The applicant also reported that purity analyses of IWG seeds are performed by official seed certification agencies of the different US states. The Panel notes that a plant variety protection certificate for IWG is not available in the US.

Production process

3.3

The applicant licences growers, distributors and food and beverage manufacturers to produce and sell IWG‐based products. IWG growers implement individual food safety systems, while processing activities such as cleaning, de‐hulling and milling are conducted in accordance with BRC Global Standard principles.

IWG is sown in autumn, either in monoculture or intercropped with plants such as clover and alfalfa. Conventional growers may use non‐organic fertilisers, while organic growers follow USDA National Organic Program (NOP) guidelines, using composted manure and potentially allowing cattle grazing. Harvest occurs in summer/autumn using a combine harvester. Unlike annual crops, the soil is not tilled, and any plant residue is left on the field to decompose. The field is kept at rest throughout winter and fertilised again in spring. This cycle can last up to 5 years before tilling and reseeding. According to the applicant, the use of composted manure would minimise microbiological contamination, ideally applied in autumn for better decomposition. IWG growers aim to limit soil contact during harvest, using techniques such as direct combining or stripper headers. Swathing and combining with a pick‐up header may be used in wetter climates.

Direct cutting is preferred in stands with minimal weed pressure, ideally when 99% of kernels are brown and firm. Higher cutting reduces straw intake, and harvested IWG should be aerated immediately if moisture exceeds 13%. According to the applicant, minimising green stem parts in the combine hopper is crucial to prevent moisture issues in stored grain. The applicant also clarified that, when intercropped, IWG's taller spikes allow selective harvesting, keeping the combine header above the intercrop.

Harvested IWG is stored in clean vessels and transported using trucks. Storage can last for months or years, with immediate drying (humidity < 14%) using drying floors or grain dryers. According to the applicant, it is essential to avoid the presence of green weeds in the field, as they can introduce wet, leafy material into the bin, potentially leading to contamination. If such contamination occurs, IWG growers should implement cleaning procedures prior to storage. Processing encompasses grain receiving, cleaning, de‐hulling, milling (for flour production), storage and packaging. Particularly, cleaning and de‐hulling are essential operations in producing high‐quality IWG grains and resulting flours. Since IWG varieties are currently not free‐threshing, de‐hulling is necessary before consumption. While the applicant recommends de‐hulling soon after harvest to minimise microbial load, processors may delay this step. The hull offers protection during storage, limiting damage from pests, slowing rancidity and reducing microbial contamination. De‐hulling can be done through abrasion, friction or impact. According to the applicant, impact dehullers better preserve nutritional quality by minimising broken kernels, with around 90% hull removed via aspiration.

The Panel considers that the production process is sufficiently described.

Compositional data

3.4

The applicant provided qualitative and quantitative data on chemical and microbiological parameters for different batches of the NF (grain and flour). For all parameters, at least five independently produced batches were analysed. Certificates of accreditation for the laboratories that conducted the analyses were provided by the applicant.

The results of the proximate analysis of the NF are presented in Table 1.

Following an additional request from EFSA, the carotenoid content (lutein, beta‐carotene, alpha‐carotene, lycopene, citranaxanthin and zeaxanthin) of the NF was determined in eight batches (five of grain from conventionally or organically grown IWG and three of flour from organically grown IWG). Results were either below reporting limits or at levels that do not raise safety concerns (data not shown).

Concentrations of heavy metals (lead, cadmium, mercury and arsenic) and mycotoxins (aflatoxin B1, B2, G1 and G2, T‐2 and HT‐2 toxins, ochratoxin A, deoxynivalenol, fumonisins (B1 and B2) and zearalenone) were measured in several batches of the NF (grain and flour from conventionally or organically grown IWG). Results were below maximum amounts set for cereals and processed cereal products in Commission Regulation (EU) 2023/9158 or were found to be at levels not raising safety concerns (data not shown).

The Panel notes that pyrrolizidine alkaloids have been reported as contaminants in several plants, including wheat, corn, millet, oat, rice, sorghum and alfalfa, which may be grown in intercrops with the NF. However, analyses of 18 NF batches (ten of grain from conventionally or organically grown IWG and eight of flour from organically grown IWG) showed that concentrations of both pyrrolizidine alkaloids (detection limit: 5 μg/kg) and tropane alkaloids (atropine and scopolamine; detection limit: 1 μg/kg) were below their respective detection limits.

Furthermore, several contaminants, including polycyclic aromatic hydrocarbons (PAHs), dioxin, dioxin‐like polychlorinated biphenyls (DL‐PCBs) and non‐dioxin‐like polychlorinated biphenyls (NDL‐PCBs), were analysed in eight batches of the NF (five of grain from conventionally or organically grown IWG and three of flour from organically grown IWG) and found to be at levels that do not raise safety concerns (data not shown).

A multi‐residue screening analysis of pesticides was also conducted on eight batches of the NF (five of grain from conventionally or organically grown IWG and three of flour from organically grown IWG), and results for the majority of pesticides9 were below the maximum residue levels (MRLs) set for similar commodities (e.g. cereals) in Regulation (EC) No 396/2005.10

The Panel considers that the information provided on the chemical composition is sufficient for characterising the NF.

The applicant submitted multiple microbiological analyses for several batches of the NF (grain and flour). Results revealed high microbial counts regarding total aerobic microbial count, coliforms, Enterobacteriaceae, yeasts and moulds (total aerobic microbial count, 2.0 × 10^3^ est. – 2.7 × 10^6^ est. CFU/g; coliforms, 2.0 × 10 est. – 1.5 × 10^5^ CFU/g; Enterobacteriaceae, 2.0 × 10^3^ – 6.5 × 10^4^; yeasts, 4.0 × 10 est. – 3.4 × 10^4^ est. CFU/g; moulds, 1.4 × 10^2^ – 2.9 × 10^4^ CFU/g), highlighting excessive variability in the hygiene conditions along the production process. Following a recommendation from the Panel to improve process hygiene conditions and lower microbial loads in the NF to meet safety levels, the applicant reviewed the production process, specifically focusing on improved cleaning and de‐hulling procedures (see Section 3.3). Consequently, the applicant submitted additional microbiological analysis for 10 batches of the NF (Table 2). Upon a request from EFSA, information on the nature of the batches (grain or flour, organic or conventional), production country, harvesting date and storage conditions before analysis were provided by the applicant. However, no information on the storage temperature and moisture of the analysed NF batches was provided. The Panel noted that these NF batches were stored in polypropylene bags for up to 23 months prior to analysis. Enterobacteriaceae were detected at high levels in all analysed batches (6.9 × 10^2^ – > 1.5 × 10^4^ CFU/g). The total aerobic microbial count exceeded the limits established in the NF specifications in eight batches out of 10. Total yeast and mould counts exceeded the limits established in the NF specifications in all analysed batches.

The applicant provided analytical data on indole‐3‐acetic acid (IAA) and indole‐3‐butyric acid (IBA) for five batches of the NF (IWG flour from either conventional or organic production) upon EFSA's request. IAA levels ranged from 0.11 to 0.34 mg/kg, while IBA levels were below the detection limit (0.01 mg/kg) in all five batches. It should be noted that both IAA and IBA are regulated as pesticides in the EU under Regulation (EC) No 396/2005, which sets MRLs for pesticides in food and feed. Considering a standard measurement uncertainty of 50%,11 the Panel noted that one out of the five NF batches exceeded the MRL of 0.1 mg/kg established in Regulation (EC) No 396/2005 for IAA. The applicant stated, upon a request from EFSA, that IAA and IBA are not intentionally added to the NF but occur naturally in IWG.

Following a further request from EFSA, the applicant provided information on the Enterobacteriaceae genera in the NF, including the potential presence of Pantoea species to occur in IWG plants. The Panel expressed concern about the possible presence of P. agglomerans in the NF. According to the literature, P. agglomerans has been identified as a causative agent of work‐related diseases associated with exposure to grain dust and other agricultural dusts (Dutkiewicz et al., 2016). Since the production of IAA by Pantoea strains, including P. agglomerans, has been reported in the literature (Herrera et al., 2016; Luziatelli et al., 2020), EFSA asked the applicant to provide further identification analysis to investigate the potential presence of P. agglomerans and other opportunistic pathogen Pantoea species in the NF (Guevarra et al., 2021). However, the requested data were not provided. Given the natural presence of Pantoea species in IWG plants and the positive IAA analysis results, the presence of opportunistic pathogen Pantoea species, such as P. agglomerans, in the NF cannot be excluded.

The Panel considers the high variability and extremely high values for microbiological parameters (total aerobic microbial count, Enterobacteriaceae, total yeast and mould counts), along with the potential presence of opportunistic pathogen Pantoea species, including P. agglomerans, in the NF to be a safety concern under the proposed uses.

Stability

3.5

According to the applicant, cleaned and de‐hulled IWG grains and resulting flour can remain stable for up to 12 months.

The stability of three samples of IWG grain was monitored during storage. Samples were stored under accelerated conditions (35–39°C and 55%–65% relative humidity) for a period of 12 weeks and under ambient conditions (20–25°C and 35%–45% relative humidity) for a period of 12 months. The following analyses were performed: proximate composition (moisture, proteins, total carbohydrates, total dietary fibre, fat and ash), mycotoxin content (aflatoxins B1, B2, G1, G2; fumonisins B1 and B2; and ochratoxin A) and microbiological parameters (total aerobic microbial count, coliforms, E. coli, Bacillus cereus, coagulase‐positive staphylococci, Salmonella spp., yeasts and moulds). The Panel noted high variability in certain microbiological parameters (total aerobic microbial count, coliforms, yeasts and moulds) during the conducted stability studies (at both accelerated and ambient temperatures), which was consistent with the variability reported for the microbiological analysis presented in Section 3.4. Furthermore, both accelerated and ambient stability studies revealed that total aerobic microbial count, coliforms, yeasts and mould levels in various batches of the NF exceeded the limits set in the NF specifications at different storage times. Therefore, the Panel concludes that the provided data do not support a 12‐month shelf‐life for the NF.

The applicant also investigated the stability of the NF proposed as an ingredient in several food matrices. Particularly, the applicant examined processing contaminants in muffins with the NF, microbiological stability in pasta, bread and beer made with the NF. Analytical values were compared to those from foods processed without the NF (control). The Panel notes that there are no significant differences concerning potential formation of processing contaminants and microbiological counts when using the NF as an ingredient in the evaluated intended‐for‐use matrices, in comparison to the control. Nevertheless, the Panel notes that the foods containing the NF have to comply with existing legislative limits, such as microbiological limits established by Regulation (EC) 2073/200512 and the benchmark concentrations of acrylamide in bakery products established by Regulation (EU) 2017/2158.13

Specifications

3.6

The specifications of the NF are indicated in Table 3.

The Panel notes that, for TAMC, total coliforms and TYMC several batches of the NF tested for compositional analysis and/or stability studies were above specification limits.

History of use of the NF and of its source

3.7

History of use of the source

3.7.1

Plants from the genera Agropyron of the tribe Triticeae, to which the NF belongs, were used as a cereal crop in ancient China (Yang & Perry, 2013). More recently, perennial IWG was introduced to the US in 1932 from Europe and Asia (Ogle, 2011). In the US and Canada, IWG has been extensively used as a source of forage for livestock and wildlife (e.g. cattle, sheep, horse, deer, antelope and elk) (Barton et al., 1992; Hess et al., 1994; Ogle, 2011; Park et al., 1994) and to limit erosion and provide habitat for wildlife (DeHaan & Ismail, 2017; Ogle, 2011).

History of use of the NF

3.7.2

The use of IWG in foods and alcoholic beverages is still limited, primarily within niche markets in the US, where the NF has entered the commercial supply chain in 2016. Upon a request from EFSA, the applicant provided further information on the sale of NF‐based products. According to the applicant, the IWG flour is mostly mixed with wheat (20%–50%) to make breads or can be used at 100% for baked products that do not require rising, such as muffins and pancakes, while the IWG grain is used for beer and distilled beverages.

Proposed uses and use levels

3.8

Target population

3.8.1

The target population proposed by the applicant is the general population.

Proposed uses and use levels

3.8.2

The NF is intended to be used as either whole grain or wholegrain flour in a manner similar to wheat, barley, oats, or other cereals (Table 4).

Absorption, distribution, metabolism and excretion (ADME)

3.9

No ADME data have been provided for the NF.

Nutritional information

3.10

The applicant provided analytical data on total carbohydrates, proteins, and fat (see Section 3.4).

In addition, analytical data on minerals, vitamins, amino acids, fatty acids and antinutritional factors in the NF were also provided on several NF batches (grain and flour). These concentrations were compared by the applicant to those reported in various databases and reports for common wheat and other cereals, including barley, rye, spelt, buckwheat, oat and millet.15 The Panel considers that, apart from noticeably higher concentrations of calcium and total phenolic concentrations, and higher alpha‐amylase‐inhibitor activity in the NF, the nutritional composition of the NF is similar to wheat and other commonly consumed cereals.

After a request from EFSA, the applicant conducted a comparative digestion study using the harmonised INFOGEST static digestion protocol (Brodkorb et al., 2019). The simulated digestion was applied to compare the digestibility of five IWG flour samples (two conventional and three organic) with a reference flour (soft whole wheat flour, type 1). The protein digestibility was found to be higher for the IWG flour (86%–91%) compared to the reference wheat flour (82%), indicating that proteins of IWG are more bio‐accessible than proteins of soft whole wheat.

Following the recommendation by FAO (2013), the protein quality was determined by the Digestible Indispensable Amino Acid Score (DIAAS). The average DIAAS score was calculated by taking into account the amino acid composition of the NF proteins (see Appendix A) and protein digestibility using the INFOGEST protocol. The average DIAAS score for IWG conventional flour samples corresponded to 25% (infant, birth to 6 months), 30% (child, 6 months to 3 years) and 35% (older child, adolescent and adult). For IWG organic flour samples, the average DIAAS score corresponded to 29% (infant), 35% (child) and 41% (older child, adolescent and adult). The soft whole wheat flour gave a DIAAS score of 33% (infant), 40% (child) and 48% (older child, adolescent and adult). Lysine was the limiting amino acid both for IWG flours (conventional and organic samples) and soft whole wheat flour. The calculated values of DIAAS for IWG and reference flour samples are comparable to the DIAAS values for wheat as per the literature (infant: 37%; child: 20%–45%; older child, adolescent and adult: 43%–54%) (Cervantes‐Pahm et al., 2014; Han et al., 2019; Mathai et al., 2017). However, the Panel noted that sulfur amino acids, along with tryptophan amino acid, were not determined and consequently excluded from the DIAAS analysis. According to the applicant, tryptophan and sulfur‐containing amino acids (cysteine and methionine) underwent degradation during acidic hydrolysis. Regarding the method employed for the simulated gastrointestinal digestion, the Panel noted that in the harmonised INFOGEST digestion protocol, the intestinal phase is treated as a single phase, neglecting the distinct conditions present in the sequential duodenal, jejunal and ileal phases (including variations in dilutions, mineral content, pH, enzyme activities, microbial content, etc.).

The Panel considers that, based on the nutritional profile and proposed conditions of use, the consumption of the NF is not nutritionally disadvantageous.

Toxicological information

3.11

No toxicological studies with the NF were provided.

Taking into account the nature and intended conditions of use of the NF, the Panel considers that no toxicological studies are required on the NF.

Allergenicity

3.12

The applicant collected data on the allergenicity of the NF from literature and also provided bioinformatic data upon a request from EFSA.

As the NF (IWG, Thinopyrum intermedium) is a member of the Triticeae tribe and therefore closely related to conventional wheat, it is expected to trigger similar IgE‐mediated allergic reactions in cereal‐allergic individuals and/or adverse reactions in individuals with coeliac disease.

The applicant highlighted that the major allergens in conventional wheat and IWG are the same and include the low‐MW glutenins, alpha gliadins, gamma gliadins (Maruyama et al., 1998), omega‐5 gliadin (Palosuo et al., 2001) and nonspecific lipid transfer protein (Tri a 14) (Inomata, 2009). According to Tyl and Ismail (2019), gliadins (particularly α‐ and γ‐type) were prominently represented in the 13 samples of IWG tested, while bands in the range characteristic for high‐MW glutenin subunits were either absent or much weaker than those in hard red winter wheat.

The applicant evaluated sequence homology between NF proteins and known food allergens using bioinformatic analysis (AllergenOnline.org version 21 and the NCBI Entrez Protein databases). The results showed homologies exceeding 86% between most of the proteins from the NF and allergenic proteins from wheat and close wheat relatives (barley, rye and Aegilops sp.).

Since sequence homologies were also found between protein sequences from the NF and those reported in the FARRP AllergenOnline Coeliac Disease database as toxic or immunogenic, the applicant also analysed the gluten content of both IWG grain and flour by ELISA tests. Results showed a gluten content in the analysed samples higher than 80 mg/kg using the method ‘R‐Biopharm 7002 RIDASCREEN™ ELISA’ (R5 antibodies raised against omega‐Secalin from rye). The gluten content ranged from 13,525 to 24,975 mg/kg in IWG grain and from 54,225 to 69,150 mg/kg in IWG flour using the method ‘AgraQuant ELISA’ (G12 antibodies raised against alpha 2‐gliadin 33‐mer from wheat).

The Panel considers that the NF has the same potential as wheat, barley and rye to trigger IgE‐mediated allergic reactions in cereal‐allergic individuals and/or adverse reactions in individuals with coeliac disease under the proposed conditions of use.

DISCUSSION

4

The NF, which is the subject of the application, is grain and flour from perennial intermediate wheatgrass (Thinopyrum intermedium). The NF is proposed to be used by the general population as either whole grain or wholegrain flour in a manner similar to wheat, barley, oats or other cereals. The NF is produced by cultivating the IWG plant and subsequently processing it to yield either grains or flour. The Panel notes safety concerns related to the high variability and extremely high values observed for certain microbiological parameters (i.e. total aerobic microbial count, Enterobacteriaceae, yeasts and moulds) in different batches of the NF. Furthermore, total aerobic microbial count, coliform and total yeast and mould count values in several batches of the NF analysed for compositional and/or stability purposes exceeded the limits set in the NF specifications. The potential presence of the opportunistic pathogen Pantoea species, such as P. agglomerans, represents an additional concern. The Panel further notes that the stability data provided on the NF do not support the proposed 12‐month shelf‐life.

CONCLUSIONS

5

Based on the available data, the Panel concludes that the safety of the NF, grain and flour from perennial intermediate wheatgrass (Thinopyrum intermedium) cannot be established.ABBREVIATIONSAACCAmerican Association of Cereal ChemistsAAIUAlpha‐amylase inhibitor activity unitsADMEAbsorption, distribution, metabolism and excretionAOACAssociation of Official Analytical CollaborationBRCBritish Retail ConsortiumCFUColony Forming UnitDIAASDigestible Indispensable Amino Acid ScoreDL‐PCBsDioxin‐like polychlorinated biphenylsELISAEnzyme Linked Immunosorbent AssayFAOFood and Agriculture OrganizationFARRPAbout Food Allergy Research & Resource ProgramGCGas ChromatographyGMOGenetically Modified OrganismIAAIndole‐3‐acetic acidIBAIndole‐3‐butyric acidIECIon‐Exclusion ChromatographyISOInternational Organization for StandardizationIWGIntermediate WheatgrassLC–MS/MSLiquid Chromatography with Tandem Mass SpectrometryMRLsMaximum residue levelsMWMolecular weightNANot availableNCBINational Center for Biotechnology InformationNDNot detectedNDANutrition, Novel Foods and Food AllergensNDL‐PCBsNon‐dioxin‐like polychlorinated biphenylsNFNovel foodNMKLNordic Committee on Food AnalysisNOPNational Organic ProgrammeOECDOrganisation for Economic Co‐operation and DevelopmentPAHspolycyclic aromatic hydrocarbonsTAMCTotal aerobic microbial countTYMCTotal yeast and mould countsUSUnited StatesUSAUnited States of AmericaUSDAUnited States Department of Agriculture

ACKNOWLEDGMENTS

The Panel wishes to thank Carmen Peláez and Rosangela Marchelli for the support provided to this scientific output.

REQUESTOR

European Commission

QUESTION NUMBER

EFSA‐Q‐2020‐00073

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PANEL MEMBERS

Dominique Turck, Torsten Bohn, Montaña Cámara, Jacqueline Castenmiller, Stefaan De Henauw, Karen Ildico Hirsch‐Ernst, Ángeles Jos, Alexandre Maciuk, Inge Mangelsdorf, Breige McNulty, Androniki Naska, Kristina Pentieva, Alfonso Siani and Frank Thies.