Safety of L‐alpha‐glycerylphosphorylcholine (L‐alpha‐GPC) from soya phospholipids (lecithin) 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 L‐alpha‐glycerylphosphorylcholine (L‐alpha‐GPC) as a novel food (NF) pursuant to Regulation (EU) 2015/2283. The NF subject of the application is L‐alpha‐GPC produced by chemical means starting from phosphatidylcholine (PC) enriched soya lecithin and is available in two forms: a viscous liquid GPC 85% and a powder Alpha Size 100P. The identity, production process, composition and specifications of the NF do not raise safety concerns. The NF is proposed to be used in food supplements at a maximum use level of 203.7 mg/day, corresponding to 82.5 mg/day of choline. The target population comprises individuals above 3 years of age, including pregnant and lactating women. L‐alpha‐GPC is an endogenous metabolite of phosphatidylcholine, is rapidly absorbed and serves as source of choline. The Panel considers that consumption of the NF is not nutritionally disadvantageous. The safety data in laboratory animals submitted by the applicant show several limitations; however, a no observed adverse effect level (NOAEL) of 1000 mg/kg bw per day for a 90‐day study and for maternal toxicity in a teratogenicity study could be derived from published studies. Based on the nature and absorption, distribution, metabolism and excretion (ADME) of the NF, the proposed use levels, the human data provided and the margin of Exposure (MoE), derived from the 90‐day study and the teratogenicity study, for children > 3 years of age (111) and for adults (345), which the Panel considers as sufficient in this case, the Panel concludes that the NF does not raise safety concerns under the proposed conditions of use.

1. INTRODUCTION

1.1. Background and Terms of Reference as provided by the requestor

On 30 June 2023, the company Chemi S.p.A submitted an application to the European Commission in accordance with Article 10 of Regulation (EU) 2015/2283 to authorise the placing on the Union market of L‐alpha‐glycerylphosphorylcholine (L‐alpha‐GPC) from soya phospholipids (lecithin) as a novel food.

The applicant requests to authorise use of L‐alpha‐glycerylphosphorylcholine (L‐alpha‐GPC) from soya phospholipids (lecithin) as a novel food in food supplements as defined in Directive 2002/46/EC, excluding food supplements for infants and young children.

The applicant has also requested data protection under Article 26 of Regulation (EU) 2015/2283.

In accordance with Article 29(l)(a) of Regulation (EC) No 178/2002, the European Commission asks EFSA to provide a scientific opinion on L‐alpha‐glycerylphosphorylcholine (L‐alpha‐GPC) from soya phospholipids (lecithin) as a novel food in accordance with Article 10(3) of Regulation (EU) 2015/2283.

The Commission also asks EFSA to evaluate and inform the Commission as to whether and if so, to what extent, the requirements of Article 26(2)(c) of Regulation (EU) 2015/2283 are fulfilled in elaborating its opinion on L‐alpha‐glycerylphosphorylcholine (L‐alpha‐GPC) from soya phospholipids (lecithin) as a novel food regarding the proprietary data for which the applicant is requesting data protection.

1.2. Additional information

L‐alpha‐GPC (or choline alfoscerate) is the main constituent of Gliatilin® (or Delecit®), a medicinal product (authorised at the national level in some EU and not EU‐countries) that primarily acts on the central nervous system, and it is used to treat cognitive, memory and attentional disorders in elderly patients. In Europe, it is sold in Italy and Poland as capsules or as a solution for injection at the maximum dose of 1000 mg. ¹

1.3. Information on existing evaluations and authorisations

In 2016, the EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA) established Adequate Intake (AI) for choline based on observed intakes in healthy EU populations and data on the amounts needed to reverse deficiency‐related organs dysfunction, given that no suitable biomarkers allowed derivation of more precise requirements. The Panel set the following AI: 400 mg/day for adults and adolescents aged 15–17 years, 140–340 mg/day for children aged 1–14 years, 160 mg/day for infants aged 7–11 months, 480 mg/day for pregnant women and 520 mg/day for lactating women (EFSA NDA Panel, 2016).

Additionally, in 2013 the NDA Panel assessed the safety of the NF citicoline (EFSA NDA Panel, 2013), a dietary source of choline, intended to be used in food supplements for middle‐aged to elderly adults and in foods for particular nutritional uses (specifically foods for special medical purposes) with a maximum daily intake of 1000 mg/day. The Panel concluded that the consumption of the NF was not nutritionally disadvantageous and that the NF was safe under the proposed uses and use levels.

2. DATA AND METHODOLOGIES

2.1. Data

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. ²

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. ³

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, 2021a). 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.

The applicant has submitted a confidential and a non‐confidential version of a dossier following the ‘EFSA guidelines on the preparation and presentation of a NF application’ (EFSA NDA Panel, 2021a) and the ‘Administrative guidance for the preparation of applications on novel foods pursuant to Article 10 of Regulation (EU) 2015/2283’ (EFSA, 2021).

In accordance with Article 38 of Regulation (EC) No 178/2002 ⁴ and taking into account the protection of confidential information and of personal data in accordance with Articles 39 to 39e of the same Regulation, and of the Decision of EFSA's Executive Director laying down practical arrangements concerning transparency and confidentiality, ⁵ the non‐confidential version of the dossier has been published on Open EFSA. ⁶

According to Art 32c(2) of Regulation (EC) No 178/2002 and to the Decision of EFSA's Executive Director laying down the practical arrangements on pre‐submission phase and public consultations, EFSA carried out a public consultation on the non‐confidential version of the technical dossier from 12 February to 05 March 2025 for which no comments were received.

This NF application includes a request for protection of proprietary data in accordance with Article 26 of Regulation (EU) 2015/2283. The data requested by the applicant to be protected are listed in Appendix A.

2.2. Methodologies

The assessment follows the methodology set out in the EFSA guidance on NF applications (EFSA NDA Panel, 2021a) 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.

The assessment follows the methodology set out in the EFSA guidance on NF applications (EFSA NDA Panel, 2021a) and the principles described in the relevant existing guidance documents from the EFSA Scientific Committee. The legal provisions for the assessment of L‐alpha‐GPC 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.

3. ASSESSMENT

3.1. Introduction

The NF which is the subject of the application is L‐alpha‐glycerylphosphorylcholine (L‐alpha‐GPC).

According to Regulation (EU) 2015/2283, Article 3, the NF falls under the following categories: ‘Foods consisting of, isolated from or produced from plants or their parts’ and ‘food with a new or intentionally modified molecular structure, where that structure was not used as, or in, a food within the Union before 15 May 1997’.

The NF is produced by base‐catalysed transesterification of phosphatidylcholine (PC)‐enriched soya lecithin and subsequent physicochemical purification processes, and it is available in two forms: GPC 85% and Alpha Size 100P. GPC 85% is a highly viscous, transparent limpid semi‐solid mass containing 85% of the NF in water. In contrast, Alpha Size 100P is a white to off‐white crystalline powder, highly soluble in water and composed of 100% of the NF.

The NF is proposed to be used as an ingredient in food supplements. The proposed target population is individuals above 3 years of age, including pregnant and lactating women.

3.2. Identity of the NF

The NF is a water‐soluble substance obtained from the PC‐enriched fraction of soybean lecithin (to about 35% phosphorylcholine) sourced from non‐genetically modified soy plant cultivars originating from North America (USA) and South America (Argentina and Brazil). Being derived from soybean lecithin, the NF only presents the ‘L’ configuration.

The plant species ■■■■■. Common names for the plant include soybean, soya bean, soy, soya, as indicated by the applicant. The part of the plant used for the production of the NF are whole seeds from which lecithin is derived. According to the applicant, the identity of the plant material was confirmed by a deoxyribonucleic Acid (DNA) based method (■■■■■); however, no experimental evidence has been provided. The applicant provided a non‐GMO certificate on the soybean seeds which confirmed that the NF is produced from non‐genetically modified cultivars.

The NF, L‐alpha‐GPC, is a chiral chemical compound with a purity > 98% on a dry weight (dw) basis (as determined by assay). Table 1 lists the main chemical identifiers of the NF while Figure 1 shows its chemical structure.

TABLE 1.

Chemical identity of the NF, L‐alpha‐GPC.

Chemical substance
Chemical (IUPAC) name	Ethanaminium, 2‐[[[(2R)‐2,3‐dihydroxypropoxy]hydroxyphosphinyl]oxy]‐N,N,N‐trimethyl‐, inner salt
Common name	L‐alpha‐glycerylphosphorylcholine
Synonyms, trade names, abbreviations	L‐alpha‐glyceryl phosphorylcholine, D‐choline hydroxide 2,3‐dihydroxypropylhydrogen phosphate inner salt, sn‐glycero‐3‐phosphorylcholine, sn‐glycero‐3‐phosphocholine, L‐α‐GPC, L‐alpha‐glycerylphosphorylcholine, L‐α‐glycerylprosphorylcholine, glyceryl phosphoryl choline, glycerophosphorylcholine, glycerophosphocholine, choline glycerophosphate, a‐Glycerylphosphorylcholine, AGPC, A‐GPC GPC, GPC 85% (contains 85% of L‐alpha‐GPC), Alpha Size 100P (contains 100% of L‐alpha‐GPC), AlphaSize® CholineAid®
CAS Number:	28319‐77‐9
Molecular formula	C8H20NO6P
Molecular weight	257.2 Da

FIGURE 1.

Structural formula of the NF, L‐alpha‐GPC.

The applicant provided a characterisation of the identity of the NF through comparison of ¹H‐NMR and IR spectra, from each of the two NF forms, with the corresponding spectra of an authentic reference standard.

The solubility of the NF was tested, with results indicating a high solubility exceeding 1000 g/L. Despite some technical limitations in the measurements the Panel considers that there are no concerns regarding consumer exposure to small particles.

3.3. Production process

According to the information provided, the NF is produced in line to Good Manufacturing Practice (GMP) and Hazard Analysis Critical Control Points (HACCP) principles.

The NF is obtained by base‐catalysed transesterification of the starting material, a soya lecithin that has undergone an enrichment step to yield a PC‐enriched fraction. After several purification steps, a partially crystallised intermediate is obtained. From this intermediate, two distinct physicochemical purification pathways are applied: one process applies further processing and concentration steps to produce the liquid GPC 85% form, while the other involves additional crystallisation step yielding an intermediate crystalline fraction, which is then further refined to obtain the Alpha Size 100P form.

The Panel considers that the production process is sufficiently described and does not raise safety concerns.

3.4. Compositional data

The NF consists of two different forms of L‐alpha‐GPC, namely transparent limpid, viscous liquid GPC 85% and white to off‐white powder Alpha Size 100P, containing 85% and 100% of the NF, respectively.

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 five independently produced batches of the NF both in form of GPC 85% (Table 2) and Alpha Size 100P (Table 3).

TABLE 2.

Batch‐to‐batch analysis of relevant parameters of the NF form GPC 85%.

Parameters	GPC 85%					Method
	Batches
	1	2	3	4	5
Solubility	Highly soluble in water and ethyl alcohol	Highly soluble in water and ethyl alcohol	Highly soluble in water and ethyl alcohol	Highly soluble in water and ethyl alcohol	Highly soluble in water and ethyl alcohol	Visual evaluation
Specific optical rotation (°) (20°C, 589.3 nm) expressed on dw	■■■■■	■■■■■	■■■■■	■■■■■	■■■■■	USP <781S>
Water content (%)	■■■■■	■■■■■	■■■■■	■■■■■	■■■■■	USP <291> Karl Fisher titration
Assay (%) (HClO4 0.1 N) on dw	99.9	100.1	98.9	99.9	100.0	USP <541>
pH (10% w/v solution in water)	■■■■■	■■■■■	■■■■■	■■■■■	■■■■■	USP <791> Titration
Impurities (%)
Sucrose	■■■■■	■■■■■	■■■■■	■■■■■	■■■■■	In‐house HPLC–ELSD, USP <621>
Glycerylphosphorylethanolamine (GPE)	■■■■■	■■■■■	■■■■■	■■■■■	■■■■■
Pinitol	■■■■■	■■■■■	■■■■■	■■■■■	■■■■■
β‐GPC	■■■■■	■■■■■	■■■■■	■■■■■	■■■■■
Total impurities	< 0.05 b	< 0.03 a	< 0.03 a	< 0.05 b	< 0.05 b
Glycerol	■■■■■	■■■■■	■■■■■	■■■■■	■■■■■	In‐house HPLC–RID, USP <621>
Anions (mg/kg)
Sulphate	< 200 a	< 200 a	< 200 a	< 200 a	< 200 a	In‐house clarity limit test
Chloride	< 200 a	< 200 a	< 200 a	< 200 a	< 200 a	In‐house clarity limit test
Free phosphate	< 50 a	< 50 a	< 50 a	< 50 a	< 50 a	In‐house colorimetric limit test
Acetates	N/A	N/A	N/A	N/A	N/A
Residual solvents (mg/kg)
Methanol	< 10 a	< 10 a	< 10 a	< 10 a	< 10 a	In‐house HSGC–FID
Butanol	< 10 a	< 10 a	< 10 a	< 10 a	< 10 a
Microbiological parameters
TAMC (CFU/g)	< 10	< 10	< 10	< 10	< 10	Ph. Eur. monograph 2.6.12 and 2.6.13 harmonised with the USP (ref. 2021 and 2022)
TYMC (CFU/g)	< 10	< 10	< 10	< 10	< 10
Escherichia coli (in 1 g)	ND	ND	ND	ND	ND
Salmonella spp. (in 10 g)	ND	ND	ND	ND	ND

	Batches
	6	7	8	9	10
Coliforms (CFU/g)	< 10	< 10	< 10	< 10	< 10	ISO 4832
Escherichia coli (CFU/g)	< 10	< 10	< 10	< 10	< 10	ISO 166649‐2
Coagulase positive Staphylococci (CFU/g)	< 10	< 10	< 10	< 10	< 10	ISO 6888‐1
Salmonella spp. (in 25 g)	ND	ND	ND	ND	ND	ISO 6579‐1
Metals and other elements (mg/kg)
Inorganic arsenic (iAs)	< 0.1 b	< 0.1 b	< 0.1 b	< 0.1 b	< 0.1 b	ASU L 25.06‐1 (2008‐12), mod., CON‐PV 01288 (2020‐05) based on HG‐AAS
Total arsenic (As)	< 0.1 b	< 0.1 b	< 0.1 b	< 0.1 b	< 0.1 b	DIN EN 15763:2010 (2010‐04), mod. based on ICP–MS
Cadmium (Cd)	< 0.01 b	< 0.01 b	< 0.01 b	< 0.01 b	< 0.01 b
Lead (Pb)	< 0.05 b	< 0.05 b	< 0.05 b	< 0.05 b	< 0.05 b
Mercury (Hg)	< 0.005 b	< 0.005 b	< 0.005 b	< 0.005 b	< 0.005 b

Abbreviations: ASU, Official collection of analytical methods according to the German Food, Commodities and Feed Code; CFU, colony forming unit; DIN, German Institute for Standardisation; dw, dry weight; EN, European standard; GPE, glycerylphosphorylethanolamine; HG‐AAS, hydride generation‐atomic absorption spectrometry; HPLC–ELSD, high‐performance liquid chromatography with evaporative light scattering detector; HPLC–RID, high‐performance liquid chromatography with refractive index detector; HSGC–FID, headspace gas chromatography with a flame ionisation detector; ICP–MS, inductively coupled plasma mass spectrometry; ISO, International Organization for Standardization; N/A, not applicable; ND, not detected; Ph.Eur, European Pharmacopoeia; TAMC, total aerobic microbial count; TYMC, total yeast and mould count; USP, United States Pharmacopeia; w/v, weight per volume.

^a LOD.

^b LOQ.

TABLE 3.

Batch‐to‐batch analysis of relevant parameters of the NF form Alpha Size 100P.

Parameters	Alpha size 100P					Method
	Batches
	1	2	3	4	5
Specific optical rotation (°) (20°C, 589.3 nm) expressed on dw	■■■■■	■■■■■	■■■■■	■■■■■	■■■■■	USP <781S>
Water content (%)	■■■■■	■■■■■	■■■■■	■■■■■	■■■■■	USP <291> Karl Fisher titration
Assay (%) (HClO4 0.1N) on dw	100.8	99.8	100.7	99.0	99.3	USP <541>
pH (8.5% w/v water solution)	■■■■■	■■■■■	■■■■■	■■■■■	■■■■■
Impurities (%)
Sucrose	■■■■■	■■■■■	■■■■■	■■■■■	■■■■■	In‐house HPLC–ELSD, USP <621>
Glycerylphosphorylethanolamine (GPE)	■■■■■	■■■■■	■■■■■	■■■■■	■■■■■
Pinitol	■■■■■	■■■■■	■■■■■	■■■■■	■■■■■
β‐GPC	■■■■■	■■■■■	■■■■■	■■■■■	■■■■■
Total impurities	< 0.03 a	< 0.03 a	< 0.03 a	< 0.03 a	< 0.03 a
Glycerol	■■■■■	■■■■■	■■■■■	■■■■■	■■■■■	In‐house HPLC–CRID, USP <621>
Anions (%)
Sulphate	< 0.005 a	< 0.005 a	< 0.005 a	< 0.005 a	< 0.005 a	USP, IC
Chloride	< 0.005 a	< 0.005 a	< 0.005 a	< 0.005 a	< 0.005 a
Phosphate	< 0.004	< 0.001 a	<0.0015 b	< 0.0025	< 0.003
Acetates	< 0.03 a	< 0.03 a	< 0.03 a	< 0.03 a	< 0.03 a
Residual solvents (mg/kg)
Methanol	< 10 a	< 10 a	< 10 a	< 10 a	< 10 a	HSGC–FID, in‐house
Butanol	< 10 a	< 10 a	< 10 a	< 10 a	< 10 a
Ethanol	< 50 a	< 50 a	< 50 a	< 50 a	< 50 a
Hexane	< 5 a	< 5 a	< 5 a	< 5 a	< 5 a
Microbiological parameters
TAMC (CFU/g)	< 10	< 10	< 10	< 10	< 10	Ph. Eur. monograph 2.6.12 and 2.6.13 harmonised with the USP (ref. 2021 and 2022)
TYMC (CFU/g)	< 10	< 10	< 10	< 10	< 10
Escherichia coli (in 1 g)	ND	ND	ND	ND	ND
Coliforms (in 1 g)	ND	ND	ND	ND	ND
Staphylococcus aureus (in 1 g)	ND	ND	ND	ND	ND
Salmonella spp. (in 10 g)	ND	ND	ND	ND	ND

	Batches
	6	7	8	9	10
Coliforms (CFU/g)	< 10	< 10	N/A	N/A	N/A	ISO 4832
Escherichia coli (CFU/g)	< 10	< 10	N/A	N/A	N/A	ISO 166649‐2
Coagulase positive Staphylococcus (CFU/g)	< 10	< 10	N/A	N/A	N/A	ISO 6888‐1
Salmonella spp. (in 25 g)	ND	ND	N/A	N/A	N/A	ISO 6579‐1
Metals and other elements (mg/kg)
Inorganic arsenic (iAs)	< 0.1 b	< 0.1 b	< 0.1 b	< 0.1 b	< 0.1 b	ASU L 25.06‐1 (2008‐12), mod., CON‐PV 01288 (2020‐05) HG‐AAS
Total arsenic (As)	< 0.1 b	< 0.1 b	< 0.1 b	< 0.1 b	< 0.1 b	DIN EN 15763:2010 (2010‐04), mod. based on ICP–MS
Cadmium (Cd)	< 0.01 b	< 0.01 b	< 0.01 b	< 0.01 b	< 0.01 b
Lead (Pb)	< 0.05 b	< 0.05 b	< 0.05 b	< 0.05 b	< 0.05 b
Mercury (Hg)	< 0.005 b	< 0.005 b	< 0.005 b	< 0.005 b	< 0.005 b

Abbreviations: ASU, Official collection of analytical methods according to the German Food, Commodities and Feed Code; CFU, colony forming unit; DIN, German Institute for Standardisation; dw, dry weight; EN, European standard; GPC, glycerylphosphorylcholine; GPE, glycerylphosphorylethanolamine; HG‐AAS, hydride generation‐atomic absorption spectrometry; HPLC–ELSD, high‐performance liquid chromatography with evaporative light scattering detector; HPLC–RID, high‐performance liquid chromatography with refractive index detector; HSGC–FID, headspace gas chromatography with a flame ionisation detector; IC, ion chromatography; ICP–MS, inductively coupled plasma mass spectrometry; N/A, not applicable. ISO, International Organization for Standardization; ND, not detected; Ph.Eur., European Pharmacopoeia; TAMC, total aerobic microbial count; TYMC, total yeast and mould count; USP, United States Pharmacopeia; w/v, weight per volume.

^a LOD.

^b LOQ.

Analyses for both NF forms were performed in‐house according to the United States Pharmacopeia (USP) monographs or validated internal test methods. Additionally, metals and other elements (inorganic arsenic, total arsenic, cadmium, lead and mercury), aflatoxins, pesticides, proteins (soya allergens), microbial parameters and process contaminants were analysed by an accredited external laboratory using official test methods.

According to the data submitted by the applicant, the NF content for GPC 85% and Alpha Size 100P forms was determined by titration method on dry basis. The values ranged from 98.9% to 100.1% for GPC 85% and from 99% to 100.8% for Alpha Size 100P.

Specific optical rotation was determined in accordance with the relevant USP procedure in an aqueous solution, and results were expressed on a dry weight basis as required when water content is specified. For GPC 85% it was consistently −2.6°, whereas for Alpha Size 100P it ranged between −2.6° and −2.7°.

The water content of GPC 85% was reported to be between 14.7% and 15.4%, while the one of Alpha size 100P was substantially lower, ranging from 0.6% to 0.9%.

Specified impurities (e.g. sucrose, glycerylphosphorylethanolamine (GPE) pinitol, β‐GPC) were analysed using an internal high‐performance liquid chromatography with evaporative light scattering detector (HPLC–ELSD) method and were not detected in either form of the NF. Glycerol, analysed with an in‐house HPLC method using a refractive index detector (RID) was found at levels between 0.10% and 0.28% in the GPC 85% form, whereas it was below the limit of detection (LOD) in any Alpha Size 100P batch. According to the applicant, total impurities were also determined using the HPLC–ELSD method.

Residual solvents were analysed by internal headspace gas chromatography with flame ionisation detection (HSGC–FID). In GPC 85%, methanol and butanol were below LOD (10 mg/kg) while in Alpha Size 100P, which was additionally tested for hexane and ethanol, all solvents were also below LOD (5 mg/kg for hexane, 10 mg/kg for methanol and n‐butanol, and 50 mg/kg for ethanol).

Metals and other elements such as total arsenic, cadmium, lead, mercury were analysed by an internal inductively coupled plasma mass spectrometry (ICP–MS) method. All results were below the respective limit of quantification (LOQs) in both forms. Inorganic arsenic, determined by hydride generation‐atomic absorption spectroscopy (HG‐AAS), was below LOQ in both forms. All analyses complied with Regulation (EC) 2023/915, where applicable.

Microbiological analyses were conducted according to the validated procedures of the European Pharmacopoeia methods. For both NF forms, total aerobe counts and total yeast and mould counts were below 10 CFU/g, while Escherichia coli and Salmonella spp. were not detected in 1 and 10 g, respectively. Staphylococcus aureus, analysed only for Alpha Size 100P form was also not detected. Additional 5 batches were analysed by an external laboratory under ISO methodologies. These batches showed counts below 10 CFU/g for Coliforms, E. coli and Staphylococcus aureus while Salmonella spp. was not detected in 25 g. For Alpha size 100P, the Panel considers the analysis of the above‐mentioned microbiological parameters sufficient in two batches.

Mycotoxins, specifically aflatoxins B1, B2, G1 and G2, and sum of aflatoxins, were determined by using a liquid chromatography–fluorescence detection (LC–FLD) method. All results were below 0.1 μg/kg for each individual aflatoxin and below 0.4 μg/kg for the sum of aflatoxins.

Sulphates, chlorides and free phosphates remained below their respective limit of detection (LODs) (200 mg/kg for both sulphates and chlorides, 50 mg/kg for free phosphate) for both forms. Acetate, which was analysed only in Alpha Size 100P, was also below the LOQ (0.03%). Pesticide residues complied with the applicable Regulation (EC) No 396/2005.

Process contaminants, including 3‐MCPD and the sum of 3‐MCPD and its esters, were detected in both forms at levels ranging from 6.7 to 16 μg/kg. Glycidyl esters were below the LOQ (5 μg/kg) in all batches analysed. Only two batches of the Alpha Size 100P were tested. All analyses were conducted using an internal gas chromatography–mass spectrometry (GC–MS/MS) method.

Information was provided on the accreditation of the external laboratory that conducted non routine analyses presented in the application.

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

3.4.1. Stability

The applicant performed stability tests on both forms of the NF (GPC 85% and Alpha Size 100P) and on the final formulation of the NF‐containing food supplement. Six batches of GPC 85% were tested for stability in accelerated condition (40 ± 2°C and 75 ± 5% relative humidity (RH)) for a 6‐month period; a long‐term stability test was also conducted on 6 batches stored at 25 ± 2°C and 60 ± 5% RH for 5 years. For the whole length of the study, GPC 85% was kept in its packaging material consisting of high‐density polyethylene (HDPE) drums with screw closures. Additionally, one batch of Alpha Size 100P was stored for 5 years at 25 ± 2°C and 60 ± 5% RH; Alpha Size 100P was kept its original packaging material consisting of a double bag placed in an HDPE drum.

Eventually, 9 independently produced batches of different formulations as soft gel capsules of the NF (including omega‐3 fatty acids, vitamins and minerals) underwent stability tests. The tests were carried out at normal storage condition at 25°C and at 60% RH, at 30°C and at 65% RH and at accelerated conditions 40°C at 75% RH. The duration of the testing varied depending on NF formulations and conditions ranging from 3 months to 24 months. The batches were analysed for physicochemical and microbiological parameters.

No chemical or physical degradation, nor microbiological growth were observed during the reported tests. The choline content remained between 66 and 91 mg/capsule and 33–45 mg/capsule, depending on the product developed for the whole length of the test.

The Panel considers that the data provided sufficient information with respect to the stability of the NF.

3.5. Specifications

The specifications of the NF forms GPC 85% and Alpha Size 100 P are indicated in Table 4.

TABLE 4.

Specifications of the two forms of the NF.

	GPC 85%	Alpha size 100P
Description	Highly viscous, transparent limpid liquid of semi‐solid mass obtained by chemical synthesis from soy lecithin	White to off‐white crystalline powder obtained by chemical synthesis from soy lecithin
Solubility	Highly soluble in water and ethyl alcohol	Highly soluble in water 7
Specific optical rotation (20°C, 589.3 nm)	−2.4° to −2.8° on dry basis	−2.4° to −2.8° on dry basis
L‐alpha GPC content by titration (assay)	98%–102% on dry basis	98%–102% on dry basis
Water content (K.F.)	14% to 16%	≤ 1%
pH	5.0–7.0 (10% w/v solution in water)	5.0–7.0 (8.5% w/v solution in water)
Metals and other elements 8
Total arsenic	< 0.1 mg/kg	< 0.1 mg/kg
Anions content
Sulphate	≤ 200 mg/kg	≤ 200 mg/kg
Chloride	≤ 200 mg/kg	≤ 200 mg/kg
Free phosphate	≤ 50 mg/kg	≤ 50 mg/kg
Acetates	≤ 1000 mg/kg	≤ 1000 mg/kg
Impurities
Sucrose	≤ 0.10%	≤ 0.10%
GPE	≤ 0.10%	≤ 0.10%
Pinitol	≤ 0.10%	≤ 0.10%
Beta‐GPC	≤ 0.10%	≤ 0.10%
Total impurities	≤ 0.5%	≤ 0.5%
Glycerol	≤ 0.5%	≤ 0.5%
Residual solvents
Methanol	≤ 10 mg/kg	≤ 10 mg/kg
Butanol	≤ 10 mg/kg	≤ 10 mg/kg
Ethanol	N/A	≤ 50 mg/kg
n‐Hexane	N/A	≤ 5 mg/kg
Microbiological parameters
TAMC	≤ 1000 CFU/g	≤ 1000 CFU/g
TYMC	≤ 100 CFU/g	≤ 100 CFU/g
Escherichia coli	Not detected in 10 g	Not detected in 10 g
Salmonella spp.	Not detected in 25 g	Not detected in 25 g
Staphylococcus aureus	Not detected in 10 g	Not detected in 10 g
Coliforms	Not detected in 10 g	Not detected in 10 g

Abbreviations: CFU, colony forming units; GPC, glycerylphosphorylcholine; GPE, glycerylphosphorylethanolamine; K.F., Karl Fischer titration; N/A, not applicable; TAMC, total aerobic microbial count; TYMC, total yeast and mould count; w/v, weight per volume.

The applicant proposes a specification limit for butanol, ethanol and n‐hexane as ≤ 100, ≤ 500 and ≤ 50 mg/kg, respectively. The Panel notes that considering the compositional analyses of the 5 batches, a lower specification limit of ≤ 10 mg/kg for butanol, ≤ 50 for ethanol and ≤ 5 for n‐hexane could be met. Furthermore, the Panel considers including arsenic with a specification limit of ≤ 0.1 mg/kg, in line with the batch‐to‐batch analyses.

The Panel considers that the specifications of the NF is sufficient and does not raise safety concerns.

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

3.6.1. History of use of the source

The starting material for producing the NF is PC‐enriched soya lecithin. Soya lecithin is used in the EU as food ingredient and is an authorised food additive (E 322) according to Annex II and Annex III to Regulation (EC) No 1333/2008 on food additives. ⁹

Lecithins were first evaluated by the Scientific Committee on Food (SCF) in 1982 ¹⁰ and re‐evaluated by the EFSA Panel on Food Additives and Nutrient Sources added to Food (ANS) in 2017. ¹¹ The ANS Panel concluded that there was no need to establish a numerical acceptable daily intake (ADI) value for lecithins (E 322) and that there was no safety concern for the general population older than 1 year of age at the refined exposure assessment for the reported uses of lecithins (E 322) as a food additive.

Lecithin has also been evaluated including by the Joint FAO/WHO Expert Committee on Food Additives (JECFA), which established an ADI of ‘not limited’, ¹² and it is considered generally recognised as safe (GRAS) by the United States Food and Drug Administration (US FDA). ¹³ In addition to food uses, lecithins are also used in pharmaceutical products as active substances and excipients.

3.6.2. History of use of the NF

As mentioned in Section 1.2, L‐alpha‐GPC is the main constituent of Gliatilin® (or Delecit®).

In the United States, the NF has been marketed since 2012 as dietary supplements in foods and beverages in different products; in the same year, the US FDA granted GRAS status to Alpha Size® L‐alpha‐GPC. ¹⁴

3.7. Proposed uses and use levels

3.7.1. Target population

The target population proposed by the applicant are individuals above 3 years of age, including pregnant and lactating women.

3.7.2. Proposed uses and use levels

The NF is proposed to be used in food supplements. As the two forms of the NF contain different amounts of L‐alpha‐GPC, the recommended daily doses proposed by the applicant are 203.7 mg for the product Alpha Size 100P, which contains 100% L‐alpha‐GPC, and 239.65 mg for the product GPC 85%, which contains 85% of L‐alpha‐GPC. Both products provide a maximum daily intake of 203.7 mg of L‐alpha‐GPC, corresponding to an intake of 82.5 mg of choline per day.

3.7.3. Combined choline intake from the NF and other sources

As both forms of L‐alpha‐GPC release choline, a conditionally essential nutrient already present in the human diet, combined intakes from the diet and the NF were estimated by the applicant.

Estimated mean and P95 chronic intakes of choline from the background diet for all population groups were extracted from Vennemann et al. (2015). The estimated mean intakes were also used by EFSA when establishing the DRVs for choline (EFSA NDA Panel, 2016). These values were used by the applicant to calculate the combined chronic intake of choline (mean and 95th percentile) from the background diet and the NF (Table 5).

TABLE 5.

Estimated intakes of choline from the background diet and combined chronic intakes from the background diet and the NF.

Population group and age (years)	Background diet (mg/day)		Combined chronic intake (mg/day)
	Mean a	P95 a	Mean	P95
Other children (3 to < 10)	177–304	263–487	259.5–386.5	345.5–569.5
Adolescents (10 to < 18)	244–373	395–572	326.5–455.5	477.5–654.5
Adults (≥ 18)	269–468	391–773	351.5–550.5	473.5–855.5
Pregnant women	356	592	438.5	674.5
Lactating women b	356	592	438.5	674.5

^a Extracted from Vennemann et al. (2015).

^b In the absence of data on mean and 95th percentile total daily choline intake for lactating women, the applicant assumed exposures to be comparable to those of pregnant women.

3.8. Absorption, distribution, metabolism and excretion (ADME)

L‐alpha‐GPC is an endogenous choline‐containing metabolite formed during the enzymatic breakdown of dietary phosphatidylcholine. During digestion, dietary phosphatidylcholine is hydrolysed by phospholipases to yield L‐alpha‐GPC, which is efficiently absorbed in the intestine and appears in plasma mostly as free choline. Water‐soluble choline compounds, such as GPC, can also enter the portal circulation of the liver intact. L‐alpha‐GPC serves as a source of choline, a conditionally essential nutrient involved in several physiological functions (see Section 3.9). The amount of dietary choline absorbed depends on the capacity of the transport system via the saturable Choline Transporter‐Like Protein 1 (CTL1) or SLC44A. Available data do not allow defining the percentage of intestinal absorption of choline in humans, or the levels of choline intake at which the intestinal absorption capacity is exceeded (EFSA NDA Panel, 2016). A significant portion of unabsorbed choline is catabolised by the intestinal microbiota to trimethylamine (TMA), which is absorbed from the gastrointestinal tract and converted to trimethylamine N‐oxide (TMAO) in the liver.

3.8.1. Animal data

The toxicokinetics of L‐alpha‐GPC was investigated by Abbiati et al. (1993) in groups of 3–5 rats following single intravenous (i.v.) (10 mg/kg bw) or oral (p.o.) (100–300 mg/kg bw) administration. Two forms of ¹⁴C‐labelled α‐GPC were administered, one labelled on the glycerol moiety ([¹⁴G]‐GPC) and one on the choline moiety ([¹⁴C]‐GPC), allowing the metabolic fate of each component to be distinguished. This design enabled comparison of absorption efficacy, systemic availability and early distribution patterns following different routes of administration.

Intestinal absorption after oral intake of L‐alpha‐GPC was high, estimated at 90%–95% based on recovered radioactivity (95% for [¹⁴G]‐GPC; 88% for [¹⁴C]‐GPC). After i.v. administration in male rats, [¹⁴G]‐GPC radioactivity declined rapidly in the bloodstream during the first hours and more slowly thereafter. Blood‐to‐plasma ratios were close to unity, indicating that [¹⁴G]‐GPC‐equivalents distribute equally between plasma and red blood cells. Five minutes after dosing, [¹⁴C]‐GPC levels were approximately 1/6 of [¹⁴G]‐GPC, demonstrating partial and rapid metabolism of L‐alpha‐GPC. Both [¹⁴G]‐GPC and [¹⁴C]‐GPC decreased over time, but between 7 and 72 h [¹⁴C]‐GPC declined more slowly than [¹⁴G]‐GPC.

Following oral administration of 100 mg/kg bw of the radiolabelled compounds in male and female rats, no sex differences were observed. However, the kinetic profiles differed between the two radiolabels: [¹⁴G]‐GPC reached peak levels at 2–4 h and then declined more slowly than after intravenous dosing, whereas [¹⁴C‐GPC] reached its maximum only at 24 h and decreased thereafter.

Tissue distribution after oral dosing showed that [¹⁴G]‐GPC radioactivity was comparable to or lower than blood levels in most tissues, except for liver and kidney. In contrast, [¹⁴C]‐GPC accumulated in liver, kidney, lungs and spleen.

Brain‐to‐blood distribution was also different between the two radiolabels. For [¹⁴G]‐GPC, brain and blood concentrations were similar up to 32 h after oral dosing. For [¹⁴C]‐GPC brain concentration increased at 24 h and remained constant thereafter, but at 50% compared to blood concentration.

Metabolite profiles in blood showed that, after i.v. administration of [¹⁴G]‐GPC, unchanged alpha‐GPC, glycerol phosphate and two unknown metabolites were detected, while following oral administration, only the same two unknown metabolites were detected. After i.v. administration of [¹⁴C]‐GPC, unchanged alpha‐GPC, choline, choline phosphate and one unknown metabolite were detected, while after oral administration choline and three unknown metabolites were identified.

In the brain, metabolites were analysed after oral administration of [¹⁴C]‐GPC 300 mg/kg bw. Small amounts of choline and two unknown metabolites, which the author hypothesised to be the same as identified in blood, were detected. Chromatographic analysis of the phosphatidylcholine fraction of brain homogenates (after phospholipase C hydrolysis) showed increasing radioactivity from 2 to 20 h, confirming that choline entering the brain was reutilised for phospholipids biosynthesis.

Renal and faecal excretion of radioactivity was low and similar for both radiolabels. Most of the administered dose was eliminated as ¹⁴CO₂, with degradation being faster and more pronounced for the [¹⁴G]‐GPC (86% p.o., 76% i.v.) than for [¹⁴C]‐GPC (59% p.o., 21% i.v.). No sex differences were observed in any parameter (Abbiati et al., 1993).

This study suggests that [¹⁴C]‐GPC after ingestion of L‐alpha‐GPC, becomes available for central nervous system metabolism and the acetylcholine biosynthesis pathway.

3.8.2. Human data

A pharmacokinetic study (Unpublished, 1986a, 1986b) was submitted by the applicant to evaluate the plasma concentrations of free choline following administration of L‐alpha‐GPC. Twelve healthy adult volunteers participated in a controlled, randomised, cross‐over trial in which each subject received a single oral dose of 1.2 g and a single intramuscular (i.m) dose of 1 g L‐alpha‐GPC. A washout period of 5 days was implemented between the two administrations. Participants were fasted and instructed to avoid foods naturally rich in choline to minimise dietary interference. Blood samples were collected at the following time points: 0, 0.25, 0.5, 0.75, 1, 1.25, 1.5, 2, 3, 4 and 6 h post‐dose. Following i.m. administration, plasma free choline concentrations increased rapidly, with a marked rise observed at 0.25 h. Concentrations began to decline from 0.5 h onward. After oral administration, plasma choline concentrations reached their maximum at approximately 1.5 h post‐dose and subsequently decreased. However, peak choline concentrations achieved via the oral route were substantially lower than those observed after i.m. administration.

Gatti et al. (1992) conducted a comparative pharmacokinetic investigation to evaluate free choline concentrations in plasma following i.m. administration of L‐alpha‐GPC and citicoline in healthy volunteers. The study enrolled 12 healthy male participants in a randomised, single‐dose, cross‐over design. Each subject received an i.m. injection of L‐alpha‐GPC (1000 mg) and citicoline (1000 mg), with a one‐week washout period between treatments. Baseline blood samples were collected at time 0, and subsequent samples were obtained over a 6‐h post‐dose period. Baseline endogenous plasma choline concentrations remained stable throughout the study period, with no significant fluctuations being observed across time points. This stability provided a reliable reference for evaluating the pharmacokinetic response to both test substances. Administration of L‐alpha‐GPC resulted in a rapid increase in plasma free choline, with notable elevations detected at 0.25 and 0.5 h post‐injection. Citicoline produced a similar temporal profile, although the increase in plasma free choline was consistently lower than that observed with L‐alpha‐GPC. Following the initial rise, plasma choline concentrations declined with an estimated half‐life ranging from 0.5 to 6.2 h. No significant differences in half‐life were reported between the two treatments. A statistically significant difference in peak plasma choline concentration (C _max) was observed between the two compounds.

L‐alpha‐GPC produced a substantially higher C _max (mean ± SE: 28.2 ± 3.1 μmol/L; range 14.5–45.3 μmol/L) compared with citicoline that has already been authorised as NF (mean ± SE: 14.0 ± 1.2 μmol/L; range 8.2–22.1 μmol/L; p < 0.01). Similarly, the area under the plasma concentration–time curve (AUC) from 0 to 4 h (AUC_0−4h) was significantly greater following L‐alpha‐GPC administration (mean ± SE: 42.7 ± 2.7 μmol/L·h; range 27.7–58.2 μmol/L·h) than after citicoline (mean ± SE: 22.0 ± 1.6 μmol/L·h; range 12.5–31.7 μmol/L·h; p < 0.001). However, when AUC values were normalised to the choline content of each compound (L‐α‐GPC: 405 mg choline; citicoline: 213 mg choline), the difference between treatments was no longer statistically significant. This suggests that the higher unadjusted AUC and C _max values observed for L‐alpha‐GPC primarily reflect its greater intrinsic choline content rather than differences in metabolic handling or bioavailability.

A controlled clinical study was conducted to assess the effect of oral L‐alpha‐GPC on plasma concentration of free choline in humans (Perruca et al., 1991). Twelve healthy male volunteers participated in a randomised, placebo‐controlled, cross‐over design. Each subject received a single oral dose of L‐alpha‐GPC (1200 mg) or placebo on two separate occasions, with a one‐week washout period between treatments. All participants fasted overnight before each administration and followed a controlled diet excluding choline‐rich foods to minimise dietary interference. Blood samples were collected at 0.25, 0.5, 0.75, 1, 1.25, 2, 4 and 6 h post‐dose, and plasma free choline concentrations were quantified. Following placebo administration, plasma free choline concentration decreased after 4 and 6 h. Oral administration of L‐alpha‐GPC induced a statistically significant increase in plasma choline AUC compared with placebo (placebo: −4.18 ± 8.99 μmol/L·h; L‐alpha‐GPC: 11.70 ± 8.87 μmol/L·h). Plasma choline concentrations began to rise significantly from 0.75 h onward. Baseline choline concentrations at time 0 averaged 10.1 ± 0.66 μM, reaching a maximum concentration of 13.6 ± 1.04 μM at 2 h post‐dose.

These findings demonstrate that oral L‐alpha‐GPC increases circulating free choline in humans under controlled dietary conditions.

3.9. Nutritional information

Both forms of the NF mainly consist of L‐alpha‐GPC (> 98% on dry basis).

The NF releases choline, a quaternary amine (2‐hydroxyethyl‐N,N,N‐trimethylammonium) present in foods in free and esterified forms, including phosphatidylcholine, free choline, phosphocholine, glycerophosphocholine and sphingomyelin. Although choline can be synthesised endogenously, this synthesis may be insufficient, making choline a conditionally essential component of the diet. Choline is an integral constituent of phospholipids involved in membrane structure and function and plays a key role in lipid and cholesterol metabolism, including the assembly and secretion of very low‐density lipoproteins by the liver. It is also a precursor of the neurotransmitter acetylcholine and of betaine, through which choline participates in one‐carbon metabolism. Dietary choline deficiency can result in hepatic steatosis and liver and muscle damage, indicating that endogenous synthesis alone may not meet physiological requirements (EFSA NDA Panel, 2016).

In 2016, EFSA established adequate intakes (AIs) for choline for all population groups (EFSA NDA Panel, 2016), including adults and adolescents 15–17 years of age (400 mg/day), infants (7–11 months: 160 mg/day), children (1–3 years: 140 mg/day; 4–6 years: 170 mg/day; 1–10 years: 250 mg/day) and adolescents 11–14 years of age (340 mg/day). For pregnant and lactating women, EFSA set AIs of 480 and 520 mg/day, respectively, noting that pregnant or lactating women may need more choline than non‐pregnant non‐lactating women (EFSA NDA Panel, 2016).

ULs for choline have been established by the US Institutes of Medicine (IoM, 1998) for all population groups (except infants) based on the amounts of choline that are associated with hypotension and fishy body odour: 1000 mg/day for children 1–8 years of age; 2000 mg/day for children 9–13 years of age; 3000 mg/day for adolescents 14–18 years of age; and 3500 mg/day for adults, including pregnant and lactating women.

The Panel considers, that taking into account the composition of the NF and the proposed conditions of use, the consumption of the NF is not nutritionally disadvantageous.

3.10. Toxicological information

The applicant provided two in vitro genotoxicity studies performed with Alpha Size 100P, as requested by EFSA, conducted in compliance with Good Laboratory Practice (GLP) and the relevant OECD Test Guidelines (TG 471 and TG 487). In addition, the applicant submitted several toxicological studies performed with GPC 85% in 1986, which were not conducted according to OECD Test Guidelines and were not GLP compliant.

3.10.1. Genotoxicity

Upon EFSA's request, the applicant provided an Ames test and in vitro micronucleus test (Table 6) performed with the NF Alpha size 100P, as it was considered the worst‐case scenario since this form of the NF may contain traces of solvents (i.e. ethanol and/or hexane) that are not used during the production of GPC 85%.

TABLE 6.

List of genotoxicity studies with the NF.

Reference	Type of study	Test system	Dose
Unpublished (2025a)	Bacterial reverse mutation test (GLP, OECD TG 471)	S. Typhimurium TA98, TA100, TA1535 and TA1537 E. coli WP2 uvrA (pKM101)	Up to 5000 μg/plate (absence and presence of S9 mix)
Unpublished (2025b)	In vitro Mammalian Micronucleus Assay (GLP, OECD TG 487)	Human Lymphocytes	Up to 2000 μg/plate (absence and presence of S9 mix)

Abbreviations: GLP, Good Laboratory Practices; OECD, Organisation for Economic Co‐operation and Development; TG, test guidelines.

The potential of the NF (L‐alpha‐GPC in form of Alpha Size 100P) (purity 98%–102%) to induce gene mutations was evaluated in S. Typhimurium strains (TA1535, TA1537, TA98, TA100) and E. coli strain WP2uvrA both in the presence and absence of metabolic activation (Unpublished, 2025a). The study was performed according to the OECD Test Guideline 471 (1997, corrected 2020) and in compliance with GLP. Alpha Size 100P, dissolved in water, was tested at six concentrations (i.e. 31.6, 100, 316, 1000, 2500 and 5000 μg/plate) selected on the basis of a preliminary cytotoxicity test. Two independent experiments were conducted applying plate incorporation and pre‐incubation methods. No precipitation or toxicity were observed under any experimental condition. No biologically relevant changes in the number of revertant colonies were induced by the test item compared to the vehicle controls at any concentration and with any tester strain. Positive and negative control values were within historical laboratory data, confirming assay validity and the functional integrity of the S9 metabolic activation system.

To evaluate the potential of the NF (L‐alpha‐GPC in form of Alpha Size 100P) (purity 98%–102%) to induce chromosomal damage, an in vitro micronucleus assay was performed in human peripheral blood lymphocytes according to the OECD Test Guideline 487 (2016, corrected 2023), in a study compliant with GLP (Unpublished, 2025b). Based on the results of a preliminary test of cytotoxicity, three concentrations (i.e. 500, 1000 and 2000 μg/mL) were selected for the analysis of the frequency of micronuclei applying a short treatment (4 + 40 h of recovery) with and without metabolic activation and a continuous treatment (44 + 0 h recovery) without metabolic activation. Mild cytotoxicity, up to 11%, was only observed at the highest concentration tested after continuous treatment. No increase in the frequency of micronuclei was observed in treated cultures compared to vehicle control cultures at any concentration and under any experimental condition. Positive and negative controls were within the historical data. The Panel notes that the applicant conducted the requested MN test only with the powdered form of the NF (i.e. Alpha Size 100P) and considered that sufficient also to evaluate genotoxicity for GPC 85%.

Taking into account the test results provided and considering the nature, source and production process of the NF, the Panel considers that there are no concerns regarding genotoxicity.

3.10.2. Subchronic toxicity

The applicant has provided subchronic toxicity studies in rats and dogs.

These studies, also summarised by Brownawell et al. (2011), presented several limitations (e.g. not performed according to GLP guidance or OECD TG, lack of functional operational battery, male and female data reported together, lack of raw and individual data). Moreover, the representativeness of the test item could not be established since limited compositional data were provided. The Panel considers that the studies submitted were not adequate for establishing a NOAEL for the safety assessment of the NF.

A GLP‐compliant 90‐day subchronic oral toxicity study performed in rats by Tian et al. (2025) was submitted. The test item was a chemically synthesised L‐alpha‐GPC with purity above 98%, which the Panel considers representative of the NF. The animals were given L‐alpha GPC by gavage at doses of 0, 500, 1000 and 2000 mg/kg bw per day for 90‐days, followed by a 4‐week recovery period. Overall, alpha‐GPC was well tolerated, with no mortality and no treatment related ophthalmologic abnormalities. Clinical observations were limited to occasional mild diarrhoea in animals treated at the highest dose.

In female rats receiving the highest dose, mean body weight and weight gain were significantly reduced compared to the control group from weeks 3 to 13 (approximately 15% lower mean body weight) accompanied by consistent decreases in weekly and total food consumption. Similar reductions in food intake were observed in the female high dose satellite group, whereas male satellite animals showed only sporadic decreases. These findings indicated a treatment related effect on food intake and growth in females at the highest dose.

Haematology and clinical chemistry revealed several statistically significant changes. The increase in creatinine observed at the mid‐dose group in females was considered non toxicologically relevant since it was not dose dependent. At the highest dose female group, statistically significant increases in alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) and prothrombin time (PT), together with decreases in total protein and albumin concentrations, suggested hepatic effects. Other altered parameters at the highest dose tested (i.e. decrease in eosinophils (EO)% and increase in haematocrit (HCT)%) were minor and not considered adverse. These effects were not observed in male rats. At the end of the recovery period, isolated decreases in glucose concentrations (female satellite group) and of the activated partial thromboplastin time (APTT) and increase in neutrophils (male satellite group) lacked toxicological significance.

Urinalysis showed dose related drops in pH in male and female rats and increases in specific gravity at the low and high dose in female and at the mid dose in male rats. These changes were not considered of toxicological concern.

In female rats, organ‐weight analysis revealed decreased empty‐stomach weight at the mid and high doses, together with increased relative weights of the brain, liver and kidneys at the high dose. No treatment related histopathological lesions were identified. Observed microscopic findings, including mild inflammatory infiltrates and hepatic steatosis, occurred with similar frequency in control and high dose groups.

Despite the applicant's claim that the study was conducted according to the OECD Test Guideline, no assessment of the functional observational battery was performed, which is a limitation of the study.

The Panel considers that the mid dose tested (1000 mg/kg bw per day) can be considered as no observed adverse effect level (NOAEL) of this study.

3.10.3. Reproductive and developmental toxicity

The applicant submitted several reproductive toxicity studies. The test items were parenterally (rather than orally) administered and not conducted following GLP or in accordance with OECD TG. Moreover, the representativeness of the test item could not be established since limited compositional data were provided. The Panel considers that these studies are not adequate for establishing a NOAEL for the safety assessment of the NF.

A teratogenicity developmental toxicity study was conducted by Tian et al. (2025), which followed the principles of OECD TG 414 including some deviations (e.g. limited dosing period (day 6 to 15 of pregnancy vs. day 6 to 20 in the OECD TG)) and included major maternal and fetal endpoints. L‐alpha‐GPC (chemically synthesised, with purity above 98%) was administered to pregnant rats at 0, 500, 1000 and 2000 mg/kg bw. Mated females were randomly assigned to groups and dosed by gavage from gestation days 6–15. Body weight was monitored throughout pregnancy, and on day 20 the uterus and foetuses were examined for implantation outcomes, fetal viability, growth and abnormalities involving external features, viscera and the skeleton.

Maternal toxicity was seen at the high dose group. Pregnant dams receiving the highest dose showed significantly reduced body weight and weight gain at several time points. Despite this maternal effect, reproductive parameters – including numbers of corpora lutea, implantations, viable fetuses, dead fetuses and resorptions – did not differ from controls and no statistically significant reduction of fetal weight was observed.

Few and sporadic external or visceral malformations were observed. Skeletal abnormalities presented with a low incidence and no dose–response pattern. The NOAEL for developmental toxicity is 2000 mg/kg bw per day under the conditions of this study (i.e. exposure duration and overall study design were not explicitly aligned with OECD test guideline specifications).

The Panel concludes that, overall, L‐alpha‐GPC did not induce embryo‐fetal toxicity or teratogenic effects at the tested doses. The NOAEL for maternal toxicity is 1000 mg/kg bw per day.

3.10.4. Human data

The applicant submitted several human intervention studies which investigated the effects of L‐alpha‐GPC administered for ≥ 7 days. In some studies, the administration of L‐alpha‐GPC was exclusively intramuscular (Abbati et al., 1991; Frattola et al., 1991; Gatti et al., 1992; Muratorio et al., 1992; Unpublished 1997a). These studies will not be discussed further, as such a route of administration is not relevant for the safety assessment of L‐alpha‐GPC as novel food. The main characteristics of the 10 remaining studies, the relevant safety endpoints assessed and the results are shown in Appendix A .

Most studies were therapeutic interventions conducted in older adults (males and females) with various forms of dementia or senile cognitive decline (n = 8) with doses of 1200 mg/day (range: 800 to 1600 mg/day) given for one to 6 months. The number of patients in the L‐alpha‐GPC arms was variable, ranging from 20 to 2058 individuals. L‐alpha‐GPC was well tolerated with few adverse events reported, which were mild and generally did not require withdrawal. In the three placebo‐controlled trials which monitored haematology, clinical chemistry and urinalysis, no adverse effects on these endpoints were reported. The two studies conducted in young healthy adults (Canal et al., 1991; Ferini Strambi et al., 1991) were small (n = 8–32), of short duration (7–10 days) and aimed to assess the effect of L‐alpha‐GPC (1200 mg/day) on sleep‐related parameters (a concern raised in relation to therapeutic interventions in older adults) and on amnesia induced by an intramuscular dose of scopolamine. No effects of any sleep‐related parameters or adverse events were reported.

The Panel notes that the human intervention studies submitted, most of which were conducted at doses of 1200 mg/day for up to 6 months in male and female older adults, support the safety and tolerability of the NF under the proposed conditions of use (203.7 mg/day).

3.11. Allergenicity

The starting material for producing the NF is PC‐enriched soya lecithin. ‘Soybeans and products thereof’ are listed under the Annex II (i.e. substances or products causing allergies or intolerances) of Regulation (EU) No 1169/2011. ¹⁵

The applicant provided an enzyme‐linked immunosorbent assay (ELISA) for soy protein (LOQ = 1.25 mg/kg) that was conducted in five batches of the NF by an accredited laboratory. Soya protein was below the LOQ in all batches. Considering the intended daily dose of 203.7 mg/day, this would lead to an intake of 0.25 mg of soya protein from the NF on a single occasion.

The Panel notes the low allergenic potential of the NF for soya‐allergic individuals. However, the Panel also notes that the NF is subject to mandatory allergen labelling under Regulation (EU) No 1169/2011, and that exemptions from mandatory allergen labelling require a specific scientific evaluation by EFSA (EFSA NDA Panel, 2021b).

4. DISCUSSION

The NF subject to the application is L‐alpha‐glycerylphosphorylcholine (L‐alpha‐GPC) provided in two forms: GPC 85% and Alpha Size 100P.

The NF is intended for use as ingredient in food supplements as source of choline. The proposed target population is the general population above 3 years of age, including pregnant and lactating women. The proposed dose of L‐alpha‐GPC is 203.7 mg/day for all groups of the target population, corresponding to an intake of choline of 82.5 mg/day.

The Panel considers that the information provided on the identity, production process, composition and the amended specifications of the NF is sufficient and does not raise safety concerns.

L‐alpha‐GPC is an endogenous metabolite of phosphatidylcholine and is naturally present in the human body. Following oral intake, it is rapidly absorbed and serves as source of choline. It is metabolised to free choline and glycerophosphate, both normal constituents of human metabolism involved in acetylcholine synthesis and phospholipid formation.

A NOAEL of 1000 mg/kg bw per day was identified for the 90‐day repeated‐dose oral toxicity study in rats and for maternal toxicity in a teratogenicity study (Tian et al., 2025). Based on the default body weight, an intake of 203.7 mg/day corresponds to an estimated exposure of approximately 9 mg/kg bw per day in children older than 3 years, resulting in a margin of exposure (MoE) of 111. For adults, the same intake equates to an exposure of 2.9 mg/kg bw per day, corresponding to an MoE of approximately 345.

The NF (L‐alpha‐GPC) is intended for use in food supplements for individuals older than 3 years of age at doses up to 203.7 mg/day, which correspond to 82.5 mg choline per day. This amount corresponds to between 16% (for lactating women) and 59% (for children of 3 years of age) of the AI for the respective groups of the target population. The Panel notes that combined intakes from the diet (P95) and the NF are below the UL for choline established by IoM for all population groups The Panel also notes that the human intervention studies submitted, most of which were therapeutic interventions conducted at doses of 1200 mg/day for up to 6 months in male and female older adults, support the safety and tolerability of the NF under the proposed conditions of use (203.7 mg/day).

Based on:

1. the nature and ADME of the NF (i.e. a metabolite that is formed in the GI tract from the breakdown of dietary phosphatidylcholine, which serves as source of choline when absorbed);

2. the proposed use levels (within the AI for choline for groups of the target population and below the UL for choline for all population groups when combined with background intake);

3. the human data provided (well tolerated, with only rare and mild adverse effects reported in older adults in therapeutic interventions using doses about 6 times higher than the proposed conditions of use for up to 6 months); and

4. the MoE derived from the 90‐day toxicological study and the teratogenicity study for children > 3 years of age (111) and for adults (345), which the Panel considers to be sufficient in this case,

the Panel concludes that the NF does not raise safety concerns under the proposed conditions of use.

5. CONCLUSIONS

The Panel concludes that the NF, L‐alpha‐GPC, is safe under the proposed conditions of use.

5.1. Protection of Proprietary data in accordance with Article 26 of Regulation (EU) 2015/2283

The Panel could not have reached the conclusion on the safety of the NF under the proposed conditions of use without the data claimed as proprietary by the applicant and reported in Appendix  B .

ABBREVIATIONS

[¹⁴C]‐GPC

¹⁴Carbon‐labelled choline moiety

[¹⁴G]‐GPC

¹⁴Carbon‐labelled glycerol moiety

1H‐NMR

Proton Nuclear Magnetic Resonance

3‐MCPD

3‐monochloropropane‐1,2‐diol esters

ADI

acceptable daily intake

ADME

absorption, distribution, metabolism and excretion

AE

adverse event

AI

adequate intake

ALP

alkaline phosphatase

ALT

Alanine aminotransferase

ANS

EFSA Panel on Food Additives and Nutrient Sources added to Food

APTT

activated partial thromboplastin time

AST

aspartate aminotransferase

AST

aspartate aminotransferase

ASU

Official collection of analytical methods according to the German Food, Commodities and Feed Code

AUC

area under the curve

bw

body weight

CAS

Chemical Abstracts Service

CDP

cytidine diphosphate

CFU

colony forming unit

C _max

peak plasma concentration

CO₂

carbon dioxide

CTL1

choline transporter‐like protein 1

DIN

German Institute for Standardisation to German Institute for Standardisation/ Deutsches Institut für Normung

DRV

dietary reference values

dw

dry weight

ELISA

enzyme‐linked immunosorbent assay

EN

European standard

EO

eosinophils

FDA

Food and Drug Administration

GC–MS/MS

Gas chromatography‐tandem mass spectrometry

GLP

Good Laboratory Practice

GMO

genetically modified organism

GMP

Good Manufacturing Practice

GPC

glycerylphosphorylcholine

GPE

glycerylphosphorylethanolamine

GRAS

generally recognise as safe

HACCP

hazard analysis critical control points

HCT

haematocrit

HDPE

High‐density polyethylene

HG‐AAS

hydride generation‐atomic absorption spectrometry

HPLC–ELSD

high‐performance liquid chromatography with evaporative light scattering detector

HPLC–RID

high‐performance liquid chromatograph refractive index detector

HSGC–FID

headspace gas chromatography with a flame ionisation detector

i.m.

intramuscular

i.v.

intravenous

IC

ion chromatography

ICP–MS

inductively coupled plasma mass spectrometry

IoM

Institute of Medicine

IR

infrared

ISO

International Organization for Standardization

IUPAC

International Union of Pure and Applied Chemistry

JECFA

Joint FAO/WHO Expert Committee on Food Additives

K.F.

Karl Fischer titration

LC–FLD

liquid chromatography–fluorescence detection

LOD

limit of detection

LOQ

limit of quantification

MN

micronucleus

MoE

margin of exposure

N/A

not applicable

ND

not detected

NDA

EFSA Panel on Nutrition, Novel Foods and Food Allergens

NF

Novel food

NOAEL

no observed adverse effect level

OECD

Organisation for Economic Co‐operation and Development

p.o.

Oral administration

P95

95th percentile

PC

phosphatidylcholine

Ph.Eur

European Pharmacopoeia

PT

prothrombin time

RH

relative humidity

RID

refractive index detector

SCF

Scientific Committee on Food

SE

standard error

TAMC

total aerobic microbial count

TG

Test guidelines

TMA

trimethylamine

TMAO

trimethylamine N‐oxide

TYMC

total yeast and mould count

UL

tolerable upper intake level

USP

United States Pharmacopeia

w/v

weight per volume

y

years

REQUESTOR

European Commission

QUESTION NUMBER

EFSA‐Q‐2023‐00492

COPYRIGHT FOR NON‐EFSA CONTENT

EFSA may include images or other content for which it does not hold copyright. In such cases, EFSA indicates the copyright holder and users should seek permission to reproduce the content from the original source.

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.

LEGAL NOTICE

Relevant information or parts of this scientific output have been blackened in accordance with the confidentiality requests formulated by the applicant pending a decision thereon by EFSA. The full output has been shared with the European Commission, EU Member States (if applicable) and the applicant. The blackening may be subject to review once the decision on the confidentiality requests is adopted by EFSA and in case it rejects some of the confidentiality requests.

APPENDIX A. Summary of human studies

Reference	Study design	Study population	Age and sex	L‐alpha‐GPC dose; route of administration; n	Comparator, route of administration dose; n	Duration of the intervention	Safety‐related endpoints	Results
Ferini Strambi et al. (1991)	Single‐blind controlled study	Healthy	21–30 y Males	1200 mg/day; oral n = 8	N/A	7 days	Sleep‐related parameters	No effect
Canal et al. (1991)	Single‐arm, uncontrolled (for L‐alpha‐GPC administration)	Healthy	19–38 y Males Females	1200 mg/day; oral n = 32	N/A	10 days	Tolerability AEs	Good tolerance No AEs
Ban et al. (1991)	Single‐arm, uncontrolled	Patients with multi‐infarct dementia, primary degenerative dementia or mixed dementia	> 60 y	1200 mg/day; oral n = 817	N/A	6 months	Tolerability AEs	Good tolerability 14 non‐severe AEs in 34 patients: agitation (13), heartburn (7), nausea (5), headache (3), insomnia (3) and orthostatic hypotension (2) Unclear if related to the intervention
Parnetti et al. (1993)	Multicenter, double‐blind, randomised, controlled trial	Patients with suspected senile dementia of Alzheimer's type	65–85 y Males Females	1200 mg/day; oral n = 65	Acetyl‐L‐carnitine; oral 1500 mg/day n = 61	6 months	Tolerability AEs	Good tolerability AEs for L‐alpha‐GPC were insomnia, gastralgia and restlessness (one patient each) of mild entity. AEs for acetyl‐L‐carnitine were agitation (n = 3), pyrosis (n = 2), nausea (n = 2) and gastralgia (n = 1). Unclear if related to the treatment, no treatment withdrawal needed
Barbagallo Sangiorgi et al. (1994)	Multicentre, single‐arm, uncontrolled	Patients with acute cerebral ischemia	45–85 y	1000 mg/day; i.m. (days 1–28) 1200 mg/day; oral (day 29–6 months) n = 2058	N/A	6 months	Tolerability AEs	Good tolerability The most frequently reported AEs were heartburn, excitation, insomnia, nausea and headache. A total of 4 patients withdrew during the oral dose phase for one of these reasons
Unpublished (1997a)	Randomised controlled trial, parallel	Patients with senile mental decline	59–80 y Males Females	800 mg/day i.m. (days 1–20) and 1600 mg/day; oral (days 21–40) n = 31 1600 mg/day; oral n = 30	Placebo; oral n = 26	40 days	AE Standard haematology and clinical chemistry Urinalysis Prothrombin time	No AE or adverse effects reported
Unpublished (1997b)	Single‐blind, randomised controlled trial	Patients with senile mental decline	62–95 y Males Females	800 mg/day; oral n = 20	Phosphatidylcholine; oral 1200 mg/day n = 20	30 days	Tolerability AE Standard haematology and clinical chemistry Urinalysis	Good tolerability No AE or adverse effects reported
Unpublished (1997c)	Single‐blind, randomised controlled trial, parallel	Patients with senile mental decline	61–89 y Males Females	800 mg/day; oral n = 20	Phosphatidylcholine; oral 1200 mg/day n = 20	20 days	Tolerability AE Standard haematology and clinical chemistry Urinalysis	Good tolerability No AE or adverse effects reported
De Jesus Moreno Moreno (2003)	Multicentre, double‐blind, randomised, placebo‐controlled trial	Patients with dementia of Alzheimer's type	60–80 y	1200 mg/day; oral n = 132	Placebo, oral n = 129	180 days	Tolerability Monthly physical examinations	Good tolerability 15 AEs (10 episodes of constipation, 5 episodes of nervousness) were reported in 11 patients of the L‐alpha‐GPC versus 6 AEs in 3 patients of the placebo group. AEs were mild

Abbreviations: AEs, adverse events, CDP, cytidine diphosphate, GPC, glycerylphosphorylcholine, i.m., intramuscular, N/A, not applicable, y, years.

APPENDIX B. List of elements of the dossier for which data protection was requested

NF section	Elements of the application dossier for which a request for data protection was filed by the applicant	File considered as pertinent for the NDA panel to reach the conclusion on the safety (yes/no)
Identity	Annex 1, Annex 2	No
Production process	Annex 3a, Annex 3b, Annex 4, Annex 5, Annex 6a	Yes
Compositional data	Annex 6b, Annex 6c, Annex 10, Annex 11, Annex 12, Annex 12b, Annex 13, Annex 14, Annex 15a, Annex 15b, Annex 16	Yes
ADME	Annex 19a, Annex 19b
Toxicological information	Annex c.1 Annex c2	Yes
	Annex 20, Annex 21 (a–g), Annex 22, Annex 23, Brownawell et al. (2011), Annex c3, Annex c4, Annex d1, Annex d2, Annex d3 Amendment to Sections 2.3, 2.9.6; Amendment to Section 2.9.5	No

EFSA NDA Panel (EFSA Panel on Nutrition, Novel Foods and Food Allergens) , Turck, D. , Bohn, T. , Cámara, M. , Castenmiller, J. , De Henauw, S. , Jos, Á. , Maciuk, A. , Mangelsdorf, I. , McNulty, B. , Naska, A. , Pentieva, K. , Siani, A. , Thies, F. , Aguilera‐Gómez, M. , Frenzel, T. , McArdle, H. J. , Moldeus, P. , Neuhäuser‐Berthold, M. , … Hirsch‐Ernst, K. I. (2026). Safety of L‐alpha‐glycerylphosphorylcholine (L‐alpha‐GPC) from soya phospholipids (lecithin) as a novel food pursuant to Regulation (EU) 2015/2283. EFSA Journal, 24(5), e10008. 10.2903/j.efsa.2026.10008