Safety of oil from Schizochytrium limacinum (strain FCC‐3204) for use in infant and follow‐on formula as a novel food pursuant to Regulation (EU) 2015/2283
2021; Wiley; Volume: 19; Issue: 1 Linguagem: Inglês
10.2903/j.efsa.2021.6344
ISSN1831-4732
AutoresDominique Turck, Jacqueline Castenmiller, Stefaan De Henauw, Karen Ildico Hirsch‐Ernst, John Kearney, Alexandre Maciuk, Inge Mangelsdorf, Harry J McArdle, Androniki Naska, Carmen Peláez, Kristina Pentieva, Alfonso Siani, Frank Thiès, Sophia Tsabouri, Marco Vinceti, Francesco Cubadda, Thomas Frenzel, Marina Heinonen, Rosangela Marchelli, Monika Neuhäuser‐Berthold, Morten Poulsen, Miguel Prieto Maradona, Josef Schlatter, Henk Van Loveren, Emanuela Turla, Helle Katrine Knutsen,
Tópico(s)Food Allergy and Anaphylaxis Research
ResumoEFSA JournalVolume 19, Issue 1 e06344 Scientific OpinionOpen Access Safety of oil from Schizochytrium limacinum (strain FCC-3204) for use in infant and follow-on formula as a novel food pursuant to Regulation (EU) 2015/2283 EFSA Panel on Nutrition, Novel Foods and Food Allergens (NDA), Corresponding Author EFSA Panel on Nutrition, Novel Foods and Food Allergens (NDA) nda@efsa.europa.eu Correspondence:nda@efsa.europa.euSearch for more papers by this authorDominique Turck, Dominique TurckSearch for more papers by this authorJacqueline Castenmiller, Jacqueline CastenmillerSearch for more papers by this authorStefaan De Henauw, Stefaan De HenauwSearch for more papers by this authorKaren Ildico Hirsch-Ernst, Karen Ildico Hirsch-ErnstSearch for more papers by this authorJohn Kearney, John KearneySearch for more papers by this authorAlexandre Maciuk, Alexandre MaciukSearch for more papers by this authorInge Mangelsdorf, Inge MangelsdorfSearch for more papers by this authorHarry J McArdle, Harry J McArdleSearch for more papers by this authorAndroniki Naska, Androniki NaskaSearch for more papers by this authorCarmen Pelaez, Carmen PelaezSearch for more papers by this authorKristina Pentieva, Kristina PentievaSearch for more papers by this authorAlfonso Siani, Alfonso SianiSearch for more papers by this authorFrank Thies, Frank ThiesSearch for more papers by this authorSophia Tsabouri, Sophia TsabouriSearch for more papers by this authorMarco Vinceti, Marco VincetiSearch for more papers by this authorFrancesco Cubadda, Francesco CubaddaSearch for more papers by this authorThomas Frenzel, Thomas FrenzelSearch for more papers by this authorMarina Heinonen, Marina HeinonenSearch for more papers by this authorRosangela Marchelli, Rosangela MarchelliSearch for more papers by this authorMonika Neuhäuser-Berthold, Monika Neuhäuser-BertholdSearch for more papers by this authorMorten Poulsen, Morten PoulsenSearch for more papers by this authorMiguel Prieto Maradona, Miguel Prieto MaradonaSearch for more papers by this authorJosef Rudolf Schlatter, Josef Rudolf SchlatterSearch for more papers by this authorHenk van Loveren, Henk van LoverenSearch for more papers by this authorEmanuela Turla, Emanuela TurlaSearch for more papers by this authorHelle Katrine Knutsen, Helle Katrine KnutsenSearch for more papers by this author EFSA Panel on Nutrition, Novel Foods and Food Allergens (NDA), Corresponding Author EFSA Panel on Nutrition, Novel Foods and Food Allergens (NDA) nda@efsa.europa.eu Correspondence:nda@efsa.europa.euSearch for more papers by this authorDominique Turck, Dominique TurckSearch for more papers by this authorJacqueline Castenmiller, Jacqueline CastenmillerSearch for more papers by this authorStefaan De Henauw, Stefaan De HenauwSearch for more papers by this authorKaren Ildico Hirsch-Ernst, Karen Ildico Hirsch-ErnstSearch for more papers by this authorJohn Kearney, John KearneySearch for more papers by this authorAlexandre Maciuk, Alexandre MaciukSearch for more papers by this authorInge Mangelsdorf, Inge MangelsdorfSearch for more papers by this authorHarry J McArdle, Harry J McArdleSearch for more papers by this authorAndroniki Naska, Androniki NaskaSearch for more papers by this authorCarmen Pelaez, Carmen PelaezSearch for more papers by this authorKristina Pentieva, Kristina PentievaSearch for more papers by this authorAlfonso Siani, Alfonso SianiSearch for more papers by this authorFrank Thies, Frank ThiesSearch for more papers by this authorSophia Tsabouri, Sophia TsabouriSearch for more papers by this authorMarco Vinceti, Marco VincetiSearch for more papers by this authorFrancesco Cubadda, Francesco CubaddaSearch for more papers by this authorThomas Frenzel, Thomas FrenzelSearch for more papers by this authorMarina Heinonen, Marina HeinonenSearch for more papers by this authorRosangela Marchelli, Rosangela MarchelliSearch for more papers by this authorMonika Neuhäuser-Berthold, Monika Neuhäuser-BertholdSearch for more papers by this authorMorten Poulsen, Morten PoulsenSearch for more papers by this authorMiguel Prieto Maradona, Miguel Prieto MaradonaSearch for more papers by this authorJosef Rudolf Schlatter, Josef Rudolf SchlatterSearch for more papers by this authorHenk van Loveren, Henk van LoverenSearch for more papers by this authorEmanuela Turla, Emanuela TurlaSearch for more papers by this authorHelle Katrine Knutsen, Helle Katrine KnutsenSearch for more papers by this author First published: 19 January 2021 https://doi.org/10.2903/j.efsa.2021.6344 Requestor: European Commission Question number: EFSA-Q-2019-00187 Panel members: Dominique Turck, Jacqueline Castenmiller, Stefaan De Henauw, Karen Ildico Hirsch-Ernst, John Kearney, Helle Katrine Knutsen, Alexandre Maciuk, Inge Mangelsdorf, Harry J McArdle, Androniki Naska, Carmen Pelaez, Kristina Pentieva, Alfonso Siani, Frank Thies, Sophia Tsabouri and Marco Vinceti. Adopted: 24 November 2020 AboutSectionsPDF ToolsExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat 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 the safety of Schizochytrium sp. oil as a novel food (NF) pursuant to Regulation (EU) 2015/2283. Schizochytrium sp. is a single-cell microalga. The strain FCC- 3204, used by the applicant (Fermentalg), belongs to the species Schizochytrium limacinum. The NF, an oil rich in docosahexaenoic acid (DHA), is obtained from microalgae after enzymatic lysis. The applicant proposed to use the NF in infant formulae (IF) and follow-on formulae (FOF). The use level defined by the applicant was derived from Regulation (EU) 2016/127, which states the mandatory addition of DHA to IF and FOF at the level of 20–50 mg/100 kcal. The intake of DHA resulting from the use of the NF in IF and FOF is not expected to pose safety concerns. S. limacinum was attributed the qualified presumption of safety (QPS) status with the qualification 'for production purposes only'. Data provided by the applicant demonstrated the absence of viable cells in the NF. No toxicological studies were performed with the NF. However, based on the available toxicological data on oils derived from Schizochytrium sp., the QPS status of the source of the NF, the production process, the composition of the NF and the absence of viable cells in the NF, the Panel considers there are no concerns with regard to toxicity of the NF. The Panel concludes that the NF is safe under the proposed conditions of use. 1 Introduction 1.1 Background and Terms of Reference as provided by the requestor Schizochytrium sp. oil is authorised, in accordance with Regulation (EC) No 258/9711 Regulation (EC) No 258/97 of the European Parliament and of the Council of 27 January 1997 concerning novel foods and novel foods ingredients. OJ L 43, 14.2.1997, p. 1., as a novel food for a number of uses as listed in Commission Implementing Regulation (EU) 2017/247022 Commission Implementing Regulation (EU) 2017/2470 of 20 December 2017 establishing the Union list of novel foods in accordance with Regulation (EU) 2015/2283 of the European Parliament and of the Council on novel foods. OJ L 351, 30.12.2017, p. 72. establishing the Union list of novel foods in accordance with Regulation (EU) 2015/228333 Regulation (EU) 2015/2283 of the European Parliament and of the Council of 25 November 2015 on novel foods, amending Regulation (EU) No 1169/2011 of the European Parliament and of the Council and repealing Regulation (EC) No 258/97 of the European Parliament and of the Council and Commission Regulation (EC) No 1852/2001. OJ L 327, 11.12.2015, p. 1.. On 23 January 2019, the company Fermentalg submitted a request to the European Commission in accordance with Article 10 of Regulation (EU) 2015/2283 for an extension of use of Schizochytrium sp. oil as a novel food. The applicant requests to extend the use of Schizochytrium sp. oil to additional food categories, namely, infant and follow-on formulae. In accordance with Article 10(3) of Regulation (EU) 2015/2283, the European Commission asks the European Food Safety Authority to provide a scientific opinion on Schizochytrium sp. oil. 1.2 Information on existing evaluations and authorisations Three existing evaluations of the NDA Panel of EFSA need to be mentioned: In the Scientific Opinion on Dietary Reference Values for fats (EFSA NDA Panel, 2010), the Panel set an adequate intake (AI) of 250 mg for eicosapentaenoic acid (EPA) plus docosahexaenoic acid (DHA) for adults; an AI of 100 mg DHA for infants (> 6 months) and young children < 24 months; and an increase of 100–200 mg preformed DHA in addition to the AI for adults as an adequate supply of n-3 long chain polyunsaturated fatty acids (PUFA) during pregnancy and lactation. In the Scientific opinion on nutrient requirements and dietary intakes of infants and young children in the European Union (EFSA NDA Panel, 2013), the Panel concluded on the levels of nutrient and energy intakes that are considered adequate for the majority of infants and young children. In particular, the AI for DHA of 100 mg/day was confirmed for infants and young children between 6 and 24 months and was also applied to infants of 0–6 months, taking into account the concentration of essential fatty acids (FAs; including DHA) in human breast milk. It is noted that EFSA has not set AI for DHA for children after 24 months. In the Scientific Opinion on the essential composition of infant and follow-on formulae (EFSA NDA Panel, 2014), the Panel concluded that DHA should be added to IF and FOF due to its structural role in the nervous system and the retina and its involvement in normal brain and visual development. A range for the recommended concentration of DHA in IF and FOF was derived: from 20 mg/100 kcal (4.8 mg/100 kJ), based on the AI of DHA (100 mg/day) and an average energy intake of 500 kcal/day, to 50 mg/100 kcal (12 mg/100 kJ) based on the highest observed DHA concentration in human milk (1% DHA in FAs) and the amount of FA in human milk. 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 requests for supplementary information. In addition, information provided by the EFSA Panel on Biological Hazards has also been considered (EFSA BIOHAZ Panel, 2020). Administrative and scientific requirements for NF applications referred to in Article 10 of Regulation (EU) 2015/2283 are listed in Commission Implementing Regulation (EU) 2017/246944 Commission Implementing Regulation (EU) 2017/2469 of 20 December 2017 laying down administrative and scientific requirements for applications referred to in Article 10 of Regulation (EU) 2015/2283 of the European Parliament and of the Council on novel foods. OJ L 351, 30.12.2017, pp. 64–71.. 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.55 EFSA NDA Panel (EFSA Panel on Dietetic Products, Nutrition and Allergies), Turck D, Bresson J-L, Burlingame B, Dean T, Fairweather-Tait S, Heinonen M, Hirsch-Ernst KI, Mangelsdorf I, McArdle H, Naska A, Neuhäuser-Berthold M, Nowicka G, Pentieva K, Sanz Y, Siani A, Sjödin A, Stern M, Tomé D, Vinceti M, Willatts P, Engel K-H, Marchelli R, Pöting A, Poulsen M, Salminen S, Schlatter J, Arcella D, Gelbmann W, de Sesmaisons-Lecarré A, Verhagen H and van Loveren H, 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 2016;14(11):4594, 24 pp. https://doi.org/10.2903/j.efsa.2016.4594 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 of supporting the safety of the proposed NF. This NF application does not include a request for the protection of proprietary data. 2.2 Methodologies The assessment follows the methodology set out in the EFSA guidance on NF applications5 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 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 the 'DHA-rich oil from Schizochytrium sp. (strain FCC-3204)'. It is produced by the microalgae Schizochytrium sp. (strain FCC-3204). With reference to article 3 of the NF Regulation 2015/2283, the NF falls under the category 2(a)(ii): 'food consisting of, isolated from or produced from microorganisms, fungi or algae'. The production process involves the controlled growth of these algae followed by extraction and refinement of the oil produced by the algae. The oil is a mixture of triglycerides composed of PUFA in which DHA represents more than 55%. The NF is proposed to be used as an ingredient in infant formulae (IF) and follow-on formulae (FOF). 3.2 Identity of the NF The NF under assessment in the present application is an oil rich in docosahexaenoic acid (DHA). Common names to define this NF are 'DHA-rich oil from Schizochytrium sp.' or 'DHA-rich algal oil'. The oil produced by Fermentalg contains more than 55% DHA. The NF is isolated from marine microalgae belonging to the genus Schizochytrium. The taxonomic classification of the microalgae is commonly defined as follows: Kingdom: Chromista; Phylum: Bigyra; Class: Labyrinthula; Order: Thraustochytriida; Family: Thraustochytriaceae; Genus: Schizochytrium. Some databases refer to the taxonomy Eukaryota, stramenopiles instead of mentioning the Kingdom (Chromista) and the Phylum (Bigyra). Nevertheless, this is still leading to the class of Labyrinthula (https://www.uniprot.org/taxonomy/2163902). Furthermore, the taxonomic classification of the genus Schizochytrium has been subject to discussions in 2007 (Yokohama and Honda, 2007). Based on genetic and phenotypic analysis, the authors proposed changes in the classification. The genus Schizochytrium was amended and new genera such as Aurantiochytrium and Oblongichytrium were defined. Therefore, the genus Schizochytrium can now also be referred to as Aurantiochytrium. The applicant specified that the strain used to produce the NF is Schizochytrium sp. FCC-3204. This strain was obtained without use of mutagenic agents or genetic modifications. 'FCC 3204' has been deposited in the Culture Collection of Algae and Protozoa (CCAP), Scottish Marine Institute (SAMS), under the reference CCAP 4062/7. The strain FCC-3204 is a natural variant of the strain FCC-1324, which was found by the applicant after analysing the FA profiles of a number of different isolates and retained because of its unusually high DHA levels. The strain FCC-1324 was assessed by the Food Safety Authority of Ireland (FSAI, 2014) and recognised as a valid source to produce the NF 'Schizochytrium sp. oil' which is currently authorised on the Union list following the initial assessment by the UK Advisory Committee in 2002 (United Kingdom, 2002). Subsequently, following an assessment by the French Agency for Food, Environmental and Occupational Health and Safety (Anses, 2018), the DHA-rich oil produced from strain FCC-3204 was considered to be substantially equivalent to the oil from the strain FCC-1324. Therefore, it is considered that the strain FCC-3204, subject of the current application, is a valid source for the NF 'Schizochytrium sp. oil' currently authorised on the Union list. A phylogenetic tree of the Thraustochytriceae family has been reported by the applicant. According to the applicant, this classification is based on the comparison of sequences of the 18S small subunit of ribosomal DNA (18S SSU-rDNA). This analysis shows that the strain FCC-1324 (parent strain of FCC-3204) is close to strain ATCC 20888, which is the source of the currently authorised 'Schizochytrium sp. oil'. Upon EFSA's request for information, the identity of FCC-3204 at species level was addressed by the applicant based on the comparison of the genome sequence of FCC-1324, which is the parent strain of FCC-3204, with the genome sequence of the type strain Aurantiochytrium limacinum ATCC MYA-1381 (equivalent to Schizochytrium limacinum ATCC MYA-1381). According to this analysis, average nucleotide identity (ANI) between the genomes of ATCC MYA-1381 and FCC-1324 was 98.89%, and the two also have a high degree of collinearity. These data show that the strain FCC-3204 is a member of the species S. limacinum, which is a synonym of A. limacinum (Morabito et al., 2020). 3.3 Production process The unicellular microalgae Schizochytrium sp. (FCC-3204) are grown under controlled conditions (time, temperature, pH and aeration) in a liquid culture medium containing the necessary nutrients. The cultivation process starts in the laboratory. The production method and the control and verification processes ensure that the algae used in the production are pure cultures. The water used for the controlled growth of the microalgae and throughout the production process is food-grade and conforms to the requirements set out by EU Directive 98/83/CE. The entire process is carried out under inert atmosphere or vacuum conditions. Quality control evaluations are performed at each stage of production in accordance with a Hazard Analysis Critical Control Point (HACCP) system and Good Manufacturing Practice (GMP). The microalgal biomass is lysed directly via an enzymatic hydrolysis. This lysis involves a food-grade and non-genetically modified organism (GMO) enzyme. Characteristics of this enzymatic preparation are provided by the applicant (purity, pH range, temperature activity and temperature of inactivation). The manufacturer certifies that the production strain of this enzyme is not present in the enzymatic preparation and that it does not produce toxins. The enzyme was not detected in three batches of the NF. The crude oil is recovered from the lysed biomass by centrifugation. A clarification step by filtration is performed to remove solid matter. The crude oil is subsequently refined using standard techniques (neutralization, decolouration and deodorisation at high temperature). At different steps of the process, EU-authorised antioxidants are added to ensure stability. The NF is finally packaged in airtight and light-proof containers and stored at a temperature of –20°C. The applicant provided data demonstrating that the algal strain Schizochytrium sp. (FCC-3204) does not produce toxins and data on the absence of viable cells in the NF. The Panel considers that the production process is sufficiently described and does not raise safety concerns. The NF produced by the applicant is an oil which may undergo further processing steps (e.g. powdering) to be used as an ingredient of IF and FOF. However, these steps are not carried out by the applicant, but by manufacturers of IF and FOF (see also Section 3.4.1 Stability of the NF under the intended conditions of use). Therefore, the description of the production process ends with the packaging and storing of the NF in its liquid/oily form. 3.4 Compositional data The NF consists of triglycerides composed of PUFA in which DHA is the predominant one, making up together with docosapentaenoic (DPA; n-6) and palmitic acid more than 92% of the total FAs. In order to confirm that the manufacturing process is reproducible and adequate to produce a product with the required characteristics on a commercial scale, the applicant provided analytical information for 5 independent batches of the NF (Table 1). Information was provided on the accreditation of the laboratories that conducted the analyses presented in the application. The proximate analytical results show that the NF is almost entirely composed of crude fat (at least 94%). The applicant provided a new analysis of five independent batches of the NF which reported contents of proteins below the limit of quantification (LOQ) (0.25%). The batch-to-batch analysis indicates a good reproducibility of the production process. A slight variability of certain nutrients (vitamin E, β-carotene, iron) was observed. This was explained by the use of food additives and processing aids in the refining step. The concentration of total sterols varied between 9,087 and 13,375 mg/kg, which is in a similar range to that observed in different types of oils (Yang et al., 2019). The composition of the NF which is under assessment can be compared to the composition of the two authorised NFs based on DHA-rich oil from Schizochytrium currently and authorised for uses in IF and FOF: 'Schizochytrium sp. (ATCC PTA-9695) oil' and 'Schizochytrium sp. (T18) oil'. Compositional data for these two authorised NFs are available in the dossiers previously assessed by national authorities and submitted by DSM (2013) and Mara Renewable (2016). A comparison of the main long-chain (n-3)-PUFA relevant in marine oils (DHA, EPA and DPA) was made. In the present NF, the average DHA content (65% of FA) is higher compared to the two authorised NFs, in which the DHA content ranged between 37% and 44% FA. The average EPA content in the present NF (0.74% FA) is lower compared to the two authorised NFs (6% FA and 1.3% FA). Regarding DPA (n-3) concentration is low in the present NF (0.24% FA), which is also the case in the two authorised NFs (< 1% FA). In terms of chemical contaminants, the concentrations of metals (< LOQs), PCBs, dioxins and polycyclic aromatic hydrocarbons found in this batch-to-batch analysis are within the EU limits established in the respective regulations and do not present concerns from a safety point of view. Four batches of the NF were tested for the process contaminants glycidyl fatty acid esters (expressed as glycidol) and total 3-monochloro-propanol-1,2-diol (MCPD) (free and fatty acid esters). The Panel notes that for glycidyl fatty acid esters (expressed as glycidol), the maximum level (ML) in IF and FOF (liquid) is set at 6 μg/kg. For 3-MCPD (sum of 3-MCPD and 3-MCPD fatty acid esters, expressed as 3-MCPD), a ML in IF and FOF (liquid) is set at 15 μg/kg. The MLs for these contaminants are expected to be respected in the final product (IF and FOF) considering the maximum concentration of the NF in IF and FOF (63 mg NF/100 mL). The analyses on microbiological contaminants in the NF are presented in Table 1. Upon EFSA's request for information, the applicant provided analyses on Enterobacter sakazakii, which was not detected in five batches of the NF. The results of analyses in three batches of the NF showed that common marine biotoxins were below the respective LOQs (diarrhetic shellfish poisoning toxins: LOQ = 20 μg/kg; pectenotoxins: LOQ = 20 μg/kg; paralytic shellfish poisoning toxins, azaspiracids, yessotoxins, saxitoxin, okadaic acid: LOQ = 5 μg/kg; domoic acid: LOQ = 1 mg/kg). These data confirm that the NF, produced from Schizochytrium sp. (strain FCC-3204), is not expected to contain marine biotoxins, neither produced by the source organism (for which a QPS status was concluded) nor from external contamination (considering the description of the production process in Section 3.3). The Panel considers that the information provided on the composition of the NF is sufficient and does not raise safety concerns. Table 1. Batch-to-batch analysis of the NF Parameter (Unit) Batch number 0403019 0413022 0418028 0419022 0419028 Proximate analysis Energy (kcal/100 g) 900 900 851 987 851 Fat (g/100 g) > 99.22 > 99.22 > 93.78 > 99.4 > 93.78 Proteins (g/100 g)aa The applicant provided an additional analysis on total nitrogen in 5 independent batches of the NF which contained proteins below LOQ (0.25%). < 0.5ff LOQ. < 0.5ff LOQ. < 0.5ff LOQ. 0.6 < 0.5ff LOQ. Carbohydrates (g/100 g) < 0.1 < 0.1 < 0.1 < 0.1 < 0.1 Crude ash (g/100 g) < 0.25 < 0.25 < 0.25 < 0.25 < 0.25 Salt (g/100 g) 0.0135 ± 0.0131 0.0390 ± 0.0136 0.0163 ± 0.0131 < 0.01 0.0250 ± 0.0132 Sodium (g/100 g) 0.005 ± 0.005 0.016 ± 0.005 0.007 ± 0.005 < 0.005 0.011 ± 0.005 Physico-chemical parameters p-Anisidine value 6.1 2.0 7.3 1.6 5.6 Moisture and volatiles (%) < 0.1gg LOD. < 0.1gg LOD. < 0.1gg LOD. < 0.1gg LOD. < 0.1gg LOD. Unsaponifiable matter (%) 1.5 1.2 1.4 1.4 1.1 Acid value (mg KOH/g) 0.4 0.2 0.2 0.4 0.5 Peroxide value (meq/kg) 1.7 0.5 1.1 2.1 1.1 Density (kg/L) NA 0.94700 0.9452 ± 0.0008 0.9493 ± 0.0008 0.9452 ± 0.0008 Relative density NA 0.9487 0.9469 ± 0.0008 0.9480 ± 0.0008 0.9469 ± 0.0008 Tocopherols (mg/100 g) α-Tocopherol 26.2 26.2 13.9 26.8 14.1 β-Tocopherol 4.06 4.01 2.05 3.12 2.13 δ-Tocopherol 85.8 74.2 39.3 69.5 38.7 γ-Tocopherol 195 171 87.5 162 86.3 Sum of tocopherols 311 276 143 262 141 Carotenoids β-Carotene (μg/100 g) NA < 5 49.1 ± 13.7 < 5 < 5 Lutein and zeaxanthin (mg/100 g) NA < 2 < 2 < 2 < 2 Astaxanthin (mg/100 g) NA < 2 < 2 < 2 < 2 Canthaxanthin (mg/100 g) NA < 2 < 2 < 2 < 2 Fatty acids (% total FA) Myristic – 14:0 0.4 0.4 0.6 0.4 0.6 Palmitic – 16:0 13.9 13.3 15.8 12.8 15.8 Margaric – 17:0 NA 0.1 NA NA NA Heptadecenoic – 17:1 NA 0.2 NA NA NA Hexadecenoic – 16:1 TOTAL 0.1 0.1 0.1 0.1 0.1 Stearic – 18:0 0.7 0.6 0.7 0.6 0.7 Oleic – 18:1(n-9) 0.3 0.2 0.1 0.1 0.1 cis-Vaccenic – 18:1(n-7) 0.1 0.1 0.1 0.1 0.1 Linoleic – 18:2(n-6) 0.5 0.3 NA 0.1 NA γ-Linolenic – 18:3(n-6) 0.2 0.2 0.1 0.2 0.1 α-Linolenic – 18(n-3) 0.4 0.4 0.2 0.4 0.2 Stearidonic –18:4(n-3) 0.5 0.5 0.3 0.5 0.3 Arachidic – 20:0 0.1 0.1 0.1 0.1 0.1 Homo-γ-linolenic – 20:3(n-6) 0.2 0.2 0.2 0.2 0.2 Arachidonic – 20:4(n-6) 0.4 0.4 0.4 NA 0.4 Eicosatetraenoic – 20:4(n-3) 0.6 0.6 0.6 0.6 0.6 Eicosapentaenoic (EPA) – 20:5(n-3) 1.1 0.8 0.5 0.8 0.5 Behenic – 22:0 0.1 0.1 0.1 0.1 0.1 Docosapentaenoic (DPA) – 22:5(n-6) 13.4 13.3 13.8 13.4 13.9 Docosapentaenoic (DPA) – 22:5(n-3) 0.3 0.3 0.2 0.2 0.2 Docosahexaenoic (DHA) – 22:6(n-3) 65.1 66.9 64.2 67.7 64.3 Others unidentified 1.6 0.9 1.9 1.6 1.7 trans-Fatty acid < 0.05gg LOD. < 0.05gg LOD. < 0.05gg LOD. < 0.05gg LOD. < 0.05gg LOD. Sterols Total sterol content (mg/kg) 13375 9087 NA NA 9581 Metals (mg/kg) Mercury < 0.005ff LOQ. < 0.005ff LOQ. < 0.005ff LOQ. < 0.005ff LOQ. < 0.005ff LOQ. Cadmium < 0.005ff LOQ. < 0.005ff LOQ. < 0.005ff
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