Safety of D‐allulose as a novel food pursuant to Regulation (EU) 2015/2283

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 D‐allulose as a novel food (NF) pursuant to Regulation (EU) 2015/2283. During the risk assessment, the Panel identified a number of data gaps, which needed to be addressed by the applicant. Therefore, an EFSA request for additional information was sent to the applicant. The requested data concerned the identity, the production process, proposed uses and use levels, genotoxicity and human data of the NF. Despite being contacted several times, the applicant did not reply to EFSA's requests for additional data. Based on the available data, the Panel concludes that the safety of the NF, i.e. D‐allulose, cannot be established.

INTRODUCTION

Background and terms of reference as provided by the requestor

On 04 August 2018, the company Petiva Europe SA submitted a request to the European Commission in accordance with Article 10 of the Novel Food Regulation (EU) 2015/2283, to place D-allulose on the Union market as a novel food (NF).

In accordance with Article 10 (3) of Regulation (EU) 178/2283, the European Commission asks the European Food Safety Authority to provide a scientific opinion by carrying out the assessment for D-allulose as a novel food.

DATA AND METHODOLOGIES

Data

The safety assessment of this NF is based on the data provided in the original application, as well as on additional information submitted by the applicant in response to the request for supplementary data issued by EFSA in February 2020¹ and on multiple subsequent occasions. However, the applicant did not submit the additional information requested by the Panel.

Administrative and scientific requirements for NF applications referred to in Article 10 of Regulation (EU) 2015/2283 are listed in Commission Implementing Regulation (EU) 2017/2469.²

A common and structured format on the presentation of NF applications is described in the EFSA guidance on the preparation and presentation of an NF application (EFSA NDA Panel, 2016). As indicated in this guidance, it is the duty of the applicant to provide all of 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.

Methodologies

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.

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

Introduction

The NF that is the subject of the application is D-allulose. The NF is produced by enzymatic conversion of D-fructose to D-allulose in a reaction catalysed by D-psicose 3-epimerase (EC 5.1.3.30); it consists of an epimer of D-fructose, which differs from fructose only in the position of the hydroxyl group on the third carbon (C-3). The NF is proposed to be used as a sugar substitute in cereals, chewing gum, confectioneries, frostings, jams, jellies, soft drinks, candies, chocolates, cookies and low-calorie foods. The target population is the general population.

Identity of the NF

The NF D-allulose (trade name Honeytose) is produced by epimerisation of D-fructose at C-3, in a reaction catalysed by D-psicose 3-epimerase (EC 5.1.3.30) (Table 1; Figure 1).

TABLE 1 Chemical identity of the ‘D-allulose (Honeytose)’.

Chemical substance
Chemical (IUPAC) name	(3R, 4R, 5R)-2-(hydroxymethyl) oxane-2, 3, 4, 5-tetrol
Common name	D-allulose
Synonyms	D-psicose
Trade names, abbreviations	Honeytose
CAS number	551-68-8
Molecular formula	C6H12O6
Molecular weight	180.16

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The identity of the NF was confirmed through H NMR, C NMR and LC-MS analyses.

Production process

According to the information provided, the NF is produced in line with good manufacturing practice (GMP) and hazard analysis critical control point (HACCP) principles.

Production occurs via enzymatic reaction using a D-psicose 3-epimerase (EC 5.1.3.30) and an invertase (EC 5.2.1.26). As reported by the applicant, the enzyme D-psicose 3-epimerase is immobilised onto a solid matrix and used for the conversion of D-fructose into D-allulose. The product is further subjected to downstream processing to remove the process residuals, and D-allulose is separated from unconverted D-fructose by countercurrent chromatography. The D-allulose solution is filtered and condensed, through evaporation, into syrup (> 85% w/w). The syrup is concentrated with a rotary drum dryer and converted to powder and/or crystals.

The D-fructose, which is an input to the process, is produced by hydrolysing sucrose into D-glucose and D-fructose, and by enzymatically converting the remaining D-glucose into D-fructose.

Following an additional data request from EFSA, the applicant provided supplementary information clarifying aspects of the production process, i.e. immobilisation of the invertase, pH values, purification steps. Nevertheless, in order to assess the safety of the manufacturing process, the applicant was requested to perform molecular biology tests for the potential detection of the enzyme and residual DNA, i.e. ELISA and Slot-Blot, in three batches of the NF. The requested information was not submitted to EFSA; therefore, the Panel considered that the production process was insufficiently described and does not allow a proper hazard identification.

Compositional data

The NF consists of D-allulose in crystalline/powder form (≥ 98% crystalline D-allulose) and in liquid form (> 90% D-allulose).

In order to confirm that the manufacturing process is reproducible and adequate to produce on a commercial scale a product with certain characteristics, the applicant provided analytical information for two batches of the NF in crystalline/powder form (Table 2) and one batch of the NF in liquid form (Table 3).

TABLE 2 Batch to batch analysis of the NF in crystalline/powder form.

Parameter (unit)	Batch number	Method/technique of analysis
Sub 1	Sub 2
Appearance	Powder	Powder	Physical observation
Odour	No off-odour	No off-odour
Allulose (purity) g/100 g	99.85	–	HPLC-RI
Fructose, g/100 g	< 0.5	–	HPLC-RI
Optical rotation (°)	α D25 + 4.7	–
Loss on drying (moisture), %	–	3.65	AOAC 935.29
Protein, g/100 g	< 0.10	–	Kjeldahl (titrimetry)/HPLC
Fat, g/100 g	< 0.10	–	AOAC 945.48
Ash, %	–	0.51	AOAC 945.46
Lead (Pb), mg/kg	< 0.10	< 0.10	IFSH/MI022
Arsenic (As), mg/kg	< 0.10	< 0.10	IFSH/MI022
Mercury (Hg), mg/kg	< 0.10	< 0.10	IFSH/MI022
Cadmium (Cd), mg/kg	< 0.10	< 0.10	IFSH/MI022
Microbiology
Total plate count CFU/g	< 10	< 10	IS 5402:2012
Yeast & mould, CFU/g	< 10	< 10	IS 5403:1999
Coliforms, CFU/g	< 10	< 10	IS:5401(II):2012
Salmonella in 25 g	Absent	Absent	ISO:6579:2014
Shigella in 25 g	Absent	Absent	IS:5887 (VII) 1999
S. aureus/g	Absent	Absent	IS:5887 (II) 1976
E. coli/g	Absent	Absent	IS:5887 (I) 1976

TABLE 3 Batch to batch analysis of the NF in liquid form.

Parameter (unit)	Batch number	Method of analysis
Sub 1	Sub 2
Appearance	Clear yellow liquid	–	Physical observation
Odour	No odour	–
Allulose (purity), %	73.75	–	HPLC-RI
Fructose, g/100 g	2.99	–	HPLC-RI
Loss on drying (moisture), %	–	–	–
Protein, g/100 g	< 0.10	–	Kjeldahl (titrimetry)/HPLC
Fat, g/100 g	< 0.10	–	AOAC 945.48
Total carbohydrates, g/100 g	76.74	–	HPLC-RI
Ash, %	< 0.5%	–	AOAC 945.46
Lead (Pb), mg/kg	–	0.12	IFSH/MI022
Arsenic (As), mg/kg	–	0.22	IFSH/MI022
Mercury (Hg), mg/kg	–	< 0.10	IFSH/MI022
Cadmium (Cd), mg/kg	–	< 0.10	IFSH/MI022
Microbiology
Total plate count/g	–	< 10	IS 5402:2012
Yeast & mould, CFU/g	–	< 10	IS 5403:1999
Coliforms, CFU/g	–	< 10	IS:5401(II):2012
Salmonella in 25 g	–	Absent	ISO:6579:2014
Shigella in 25 g	–	Absent	IS:5887 (VII): 1999
S. aureus/g	–	Absent	IS:5887 (II) 1976
E. coli/g	–	Absent	IS:5887 (I) 1976

Information on the accreditation of the external laboratories that performed the analyses was provided. The applicant also performed in-house tests, and the analytical methods were described.

In the course of the safety assessment, the Panel requested the applicant to provide further information on specific parameters, e.g. optical rotation, protein determination; however, this information was not provided. In addition, compositional information for five independent batches of the NF, as required by EFSA, was not provided either. Hence, the Panel considers that the information provided on the composition was not sufficient to adequately characterise the NF.

Stability

The applicant performed different stability studies.

Two studies (one for the crystalline form and one for the liquid form) were performed to assess the microbiological and sensory stability of the NF. These were carried out under accelerated conditions for 21 and 22 weeks, respectively, corresponding to ‘21 and 22 months in room conditions’, according to the applicant. From the data provided on one batch, the product was stable. However, since no information was provided on the experimental conditions, the Panel could not assess the adequacy and relevance of these two stability tests.

Two additional studies were performed to assess the chemical stability, i.e. concentration of allulose in the NF food. These tests were performed over 24 h, with the NF stored in PET or glass bottles in unsterilised water and at pH 2.8, ‘under the sun’, as stated by the applicant. Different acids were used to reach pH 2.8, i.e. hydrochloric, acetic, citric and phosphoric acids. The applicant reported to have performed the study also with no acid; however, it was unclear at which pH value. Analogous tests (i.e. NF in PET or glass bottles at pH 2.8 and no acid) were performed at 25°C for 10, 20, 25 and 30 days. These tests were performed on three batches, in duplicate. The applicant reported that the NF remained stable under both conditions, i.e. ‘under the sun’ and at 25°C. However, the raw data showed that the NF degraded to some extent (concentration: ca. −2.2%). Since no information was provided on the degradation products (only the concentration of allulose was reported), the Panel could not conclude on the safety of the NF under the tested conditions.

No information was provided on the stability of the NF under the intended conditions of use, including uses at high temperature. Additional data were requested to the applicant in relation to the experimental conditions for stability testing and for stability of the NF under the intended conditions of use. However, no reply was received from the applicant.

The Panel considers that the data provided are not sufficient to assess the stability of the NF.

Specifications

The specifications of the NF as proposed by the applicant are indicated in Tables 4 and 5.

The applicant described D-allulose as a C-3 epimer of D-fructose, differing from fructose only in the position of the hydroxyl group on the third carbon. D-allulose has the same chemical formula of fructose and D-glucose, i.e. C₆H₁₂O₆. D-allulose naturally occurs in small quantities in food products. It has the physical characteristics of monosaccharides, and it is 70% as sweet as sucrose.

TABLE 4 Specifications of the NF in crystalline/powder form.

Parameter	Specification	Method
Physical appearance	White crystal or powder
Odour	No odour
Moisture	2.0%
Purity by HPLC	≥ 98%
Other saccharides	≤ 2%	HPLC-RI Glucose and fructose; LoQ < 2%
pH	3.0–7.0
Ash	< 0.5%
Heavy metals
Lead	< 1.0 mg/kg
Cadmium	< 1.0 mg/kg
Mercury	< 1.0 mg/kg
Other metals	< 10 ppm	LoQ < 1 ppm
Copper	< 3 ppm
Zinc	< 5 ppm
Microbiological
Total plate count	< 100 CFU/g
Yeast and mould	< 100 CFU/g
Coliform	< 10 CFU/g
Salmonella	Not detected in 25 g

TABLE 5 Specifications of the NF in liquid form.

Parameter	Specification	Method
Physical appearance	Liquid
Odour	No odour
Moisture/sugar content	± 1% of label (total sugar in solution is decided by final client)	HPLC LoQ < 1%
Purity by HPLC	≥ 98%	LoQ < 1%
Other saccharides	≤ 2%	HPLC Glucose and fructose; LoQ < 2%
pH	3.0–7.0
Ash	< 0.5%
Heavy metals
Lead	< 1.0 mg/kg
Cadmium	< 1.0 mg/kg
Mercury	< 1.0 mg/kg
Other metals	< 10 ppm	LoQ < 1 ppm
Copper	< 3 ppm
Zinc	< 5 ppm
Microbiological
Total plate count	< 100 CFU/g
Yeast and mould	< 100 CFU/g
Coliform	< 10 CFU/g
Salmonella	Not detected in 25 g

The Panel requested the applicant to report the moisture content (as %) of the NF in liquid form. The applicant replied that ‘Allulose is soluble in water, so moisture (water content) can vary from 99% to solubility limit pending client requirement.’

The Panel requested to clarify the identities of ‘other saccharides’ and ‘other metals’ in the proposed specifications. The applicant replied by confirming the quantitative values provided in the tables above, however, did not indicate the identity of the specific sugars and metals. In addition, there is not a clear indication on the analytical method to be used for determining ‘other metals’.

The Panel considers that the information provided on the specifications of the NF is not sufficient.

History of use of the NF and/or of its source

History of use of the source

The production of the NF involves a fermentation step in which the feedstock material is sucrose produced from sugar beet. The applicant reported that also D-glucose, from ‘corn starch liquefaction’, could be used as a feedstock.

History of use of the NF

Allulose is naturally present in small quantities in different foodstuffs, e.g., cane, beet, coffee, wheat plant, raisin (Oshima, 2006). In addition, allulose is found in processed foods, e.g. brown sugar and ketchup contain approximately 71 mg/100 g and 40 mg/100 g of allulose, respectively (Oshima, 2006).

FDA did not question the three GRAS notifications regarding the use of allulose as a sugar substitute in certain conventional foods and beverages (GRAS Notification Number (GRN) 400 (Ref. 5), GRN 498 (Ref. 6) and GRN 693 (Ref. 7)) (FDA, 2020).

Proposed uses and use levels and anticipated intake

Target population

The applicant claimed that the NF is intended to be consumed by the general population.

Proposed uses and use levels

The applicant intends to market the NF as a sugar substitute at use levels ranging from 1 to 100% in selected foods. Anticipated uses of D-allulose are shown in Table 6.

TABLE 6 Proposed uses and use levels for the NF.

Category	Use level (g NF/100 g)
Sugar substitutes	100
Chewing gum	50
Non-alcoholic beverages, low calorie, reduce calorie, sugar free	3.5
Alcoholic beverages, reduced calorie	2
Fruit juice drinks, low or reduced calorie	1
Dietetic or low-calorie bakery products (cakes, rolls, cookies, bread, muffin)	10
Hard candies (low calorie, reduced calorie or sugar free)	70
Soft candies (low calorie, reduced calorie or sugar free)	25
Chocolate, low or reduced calorie	30
Jams & jellies	10
Yoghurt (regular & frozen), low calorie, reduced calorie, sugar free	5
Frozen dairy desserts (ice cream, soft serve, sorbet), low calorie, reduced calorie, sugar free	5
Fat-based creams and fillings used in modified fat/calorie cookies, cakes, pies and pastries	10
Ready to eat cereals (< 5% sugar)	10
Coffee mix	30
Sauces & syrups, low or reduced calorie	5
Nutrition bars (meal replacement bars, protein bars and energy bars)	10

During the assessment, EFSA requested the applicant to (i) revise the initially proposed uses and use levels of the NF using the EFSA standardised FoodEx2 classification system, (ii) indicate the foods for which maximum daily intakes of the NF are proposed and (iii) provide estimates of the anticipated daily intake of the NF (per kg body weight (bw) and in absolute amounts) using the summary statistics of the EFSA Comprehensive Food Consumption Database. However, this information was not submitted by the applicant, who persisted in using the FAIM tool.

The Panel considers that the concerns on the proposed uses and use levels of the NF were not addressed.

Absorption, distribution, metabolism and excretion (ADME)

The applicant in the 2018 original submission provided a limited literature search for the assessment of ADME.

Available human data from the literature were not provided by the applicant. Following the evaluation of effects of allulose in subchronic and chronic toxicity studies, EFSA requested a human study (see also Section 3.10.4) to clarify the toxicokinetics of the NF in humans.

No reply was received from the applicant.

Nutritional information

Based on the limited information provided, the Panel cannot conclude whether or not the consumption of the NF at the proposed uses and use levels is nutritionally disadvantageous.

Toxicological information

The Panel notes that no toxicological studies with the NFs were performed by the applicant, who referred only to published data.

Genotoxicity

Genotoxicity studies performed in accordance with OECD guidelines and in compliance with good laboratory practice (GLP) using the NF were not provided.

The applicant refers to studies published in the scientific literature. The applicant has been requested to provide additional data according to the EFSA guidance (EFSA Scientific Committee, 2011).

The applicant did not reply to these data requests.

Acute and subacute toxicity

The applicant reported published data for acute toxicity comparing the LD50 of D-allulose, erythritol, beta-D-fructose, alpha-D-glucose, sucrose, maltose, table salt and alcohol.

The Panel considers that in general acute toxicity studies are not pertinent for the safety assessment of NFs.

Subchronic toxicity

No studies on the NF were performed.

Instead, in the original 2018 submission, the applicant conducted a limited literature search for toxicological studies. However, the studies identified in the literature were not performed in accordance with OECD guidelines and exhibited several limitations. Furthermore, due to the lack of data on the test substance, the representativeness of these studies for the NF could not be assessed. Consequently, while these studies may provide some insights into potential hazards, they are insufficient to support a comprehensive risk assessment, and the subchronic toxicity of the NF could not be evaluated.

Considering a risk assessment conducted by the NDA Panel based on the body of evidence provided in other allulose dossiers, the applicant was requested to provide human clinical data on long-term consumption of allulose (see Section 3.10.4).

Human data

No human data have been submitted by the applicant using the NF; however, several published studies involving human subjects are available in the literature on D-allulose.

The Panel noted that the available human trials were insufficient to address the data gaps identified during the risk assessment, especially those concerning potentially susceptible population groups, such as diabetic patients, older adults and children.

The applicant was requested to provide any additional human data that may address the safety concerns related to long-term consumption of D-allulose by humans at the proposed uses and use levels.

The applicant did not submit any additional data.

Allergenicity

The potential source of protein in the NF would be the D-psicose 3-epimerase production enzyme, and the applicant was requested to provide evidence that the production process results in the absence of proteins in the NF. However, the applicant did not reply to this request, and as a result, the Panel could not conclude on the allergenicity potential of the NF.

DISCUSSION

The NF, which is the subject of the application, is D-allulose.

The NF is intended for the general population and is proposed to be used as an ingredient in a variety of food products and as a sugar replacement.

During the risk assessment, the Panel identified a number of data gaps which needed to be addressed by the applicant.

Therefore, an EFSA request for additional information/data was sent to the applicant on 19 March 2020 and on 22 February 2021. The requested data concerned the identity, the production process, compositional data, specifications, proposed uses and use levels, genotoxicity and human data regarding the NF. Following the data submission in February 2020, the applicant did not reply to any subsequent request from EFSA for additional data.

CONCLUSIONS

Based on the available data, the Panel concludes that the safety of the NF D-allulose cannot be established.

ABBREVIATIONS

• EC
• European Commission
• EFSA
• European Food Safety Authority
• EU
• European Union
• ADME
• absorption, distribution, metabolism and excretion
• AOAC
• Association of Official Analytical Chemists
• BW
• body weight
• CFU
• colony forming unit
• FAIM
• Food Additive Intake Model
• FDA
• Food and Drug Administration
• GLP
• God Laboratory Practices
• GMP
• Good Manufacturing Practice
• GRAS
• Generally RecognisedRecognized as Safe
• HACCP
• Hazard Analysis Critical Control Points
• HPLC-RI
• High Pressure Liquid Chromatography-Refractive Index
• IFSH
• Institute for Food Safety and Health
• IS
• Internal Standard
• LC-MS
• Liquid Chromatography-Mass spectrometry
• LD50
• Lethal Dose 50
• LOQ
• Limit of Quantification
• NDA
• Nutrition, Novel Foods and Food Allergens
• NF
• novel food
• NMR
• Nuclear Magnetic Resonance
• OECD
• Organisation for Economic Co-operation and Development

REQUESTOR

European Commission

QUESTION NUMBER

EFSA-Q-2018-00756

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PANEL MEMBERS

María de la Montaña Cámara Hurtado, Dominique Turck, Torsten Bohn, Jacqueline Castenmiller, Stefaan De Henauw, Karen Ildico Hirsch-Ernst, Ángeles Mencia Jos Gallego, Alexandre Maciuk, Inge Mangelsdorf, Breige Mcnulty, Androniki Naska, Kristina Pentieva, Alfonso Siani and Frank Thies.

References

EFSA NDA Panel (Panel on Dietetic Products Nutrition and Allergies). (2016). Guidance on the preparation and presentation of an application for authorisation of a novel food in the context of regulation (EU) 2015/2283. EFSA Journal, 14(11), 4594. https://doi.org/10.2903/j.efsa.2016.4594

EFSA Scientific Committee. (2011). Scientific opinion on genotoxicity testing strategies applicable to food and feed safety assessment. EFSA Journal, 9(9), 2379. https://doi.org/10.2903/j.efsa.2011.2379

FDA (U.S. Food and Drug Administration). (2020). The declaration of allulose and calories from allulose on nutrition and supplement facts labels: Guidance for industry.

Oshima, H., Kimura, I., & Izumori, K. (2006). Psicose contents in various food products and its origin. Food Science and Technology Research, 12(2), 137–143.