<b>Bijsluiter</b>. De hyperlink naar het originele document werkt niet meer. Daarom laat Woogle de tekst zien die in dat document stond. Deze tekst kan vreemde foutieve woorden of zinnen bevatten en de opmaak kan verdwenen of veranderd zijn. Dit komt door het zwartlakken van vertrouwelijke informatie of doordat de tekst niet digitaal beschikbaar was en dus ingescand en vervolgens via OCR weer ingelezen is. Voor het originele document, neem contact op met de Woo-contactpersoon van het bestuursorgaan.<br><br>====================================================================== Pagina 1 ======================================================================

<pre>Lycopeen
Tweede beoordeling van de veiligheid voor de consument, volgens de
Europese verordening 258/97 betreffende nieuwe voedingsmiddelen en
nieuwe voedselingrediënten
Lycopene
Second opinion regarding consumer safety, in accordance with European
Regulation 258/97 concerning novel foods and novel ingredients
Gezondheidsraad:
Commissie Veiligheidsbeoordeling nieuwe voedingsmiddelen (VNV)
Health Council of the Netherlands
Committee on the Safety Assessment of Novel Foods
aan/to
de minister van Volksgezondheid, Welzijn en Sport/
the Minister of Health, Welfare and Sport
de minister van Landbouw, Natuur en Voedselkwaliteit/
the Minister of Agriculture, Nature and Food Quality
Nr 2004/02VNV, Den Haag, 24 juni 2004
No. 2004/02VNV, The Hague, June 24, 2004
</pre>

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<pre>De Gezondheidsraad, ingesteld in 1902, is een adviesorgaan met als taak de regering en
het parlement “voor te lichten over de stand der wetenschap ten aanzien van vraagstuk-
ken op het gebied van de volksgezondheid” (art. 21 Gezondheidswet).
     De Gezondheidsraad ontvangt de meeste adviesvragen van de bewindslieden van
Volksgezondheid, Welzijn & Sport; Volkshuisvesting, Ruimtelijke Ordening & Milieu-
beheer; Sociale Zaken & Werkgelegenheid en Landbouw, Natuur en Voedselkwaliteit.
De Raad kan ook eigener beweging adviezen uitbrengen. Het gaat dan als regel om het
signaleren van ontwikkelingen of trends die van belang kunnen zijn voor het overheids-
beleid.
     De adviezen van de Gezondheidsraad zijn openbaar en worden in bijna alle gevallen
opgesteld door multidisciplinaire commissies van – op persoonlijke titel benoemde –
Nederlandse en soms buitenlandse deskundigen.
The Health Council of the Netherlands, established in 1902, is an independent scientific
advisory body. Its remit is “to advise the government and Parliament on the current level
of knowledge with respect to public health issues...” (Section 21, Health Act).
     The Health Council receives most requests for advice from the Ministers of Health,
Welfare & Sport; Housing, Spatial Planning & the Environment; Social Affairs &
Employment, and Agriculture, Nature and Food Quality. The Council can publish advi-
sory reports on its own initiative. It usually does this in order to ask attention for devel-
opments or trends that are thought to be relevant to government policy.
     Most Health Council reports are prepared by multidisciplinary committees of Dutch
or, sometimes, foreign experts, appointed in a personal capacity. The reports are avail-
able to the public.
Deze publicatie kan als volgt worden aangehaald:
Gezondheidsraad. Commissie Veiligheidsbeoordeling nieuwe voedingsmiddelen. Lyco-
peen. Den Haag: Gezondheidsraad, 2004; publicatie nr 2004/02VNV.
Preferred citation:
Health Council of the Netherlands. Committee on the Safety Assessment of Novel
Foods. Lycopene. The Hague: Health Council of the Netherlands, 2004; publication no.
2004/02VNV.
auteursrecht voorbehouden/all rights reserved
U kunt het advies downloaden van www.gr.nl/
This report can be downloaded from www.healthcouncil.nl
</pre>

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<pre>  Inhoud/Contents
  Brief aan de Minister van Volksgezondheid, Welzijn en Sport 4
  Letter to the Dutch Minister of Health, Welfare and Sport 7
  Literatuur/Literature 10
  Bijlagen/Annexes 12
A De Adviesaanvraag/Request for advice 13
B De commissie/The committee 15
C EU-procedure/EU-procedure 17
D Samenvatting van het dossier/Executive summary of the dossier 20
E Eerste beoordeling/First assessment 27
  Inhoud/Contents                                                  3
</pre>

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<pre>Gezondheidsraad                               Vice-voorzitter
Health Council of the Netherlands
Aan de Minister van Volksgezondheid,
Welzijn en Sport
Onderwerp         : Tweede beoordeling veiligheid Lycopeen uit Blakeslea trispora
Uw kenmerk        : VGB/VL2483343
Ons kenmerk       : 2004/02VNV, U-868 /JvdW/cv/622-CX
Datum             : 24 juni 2004
Mijnheer de minister,
Dit schrijven dient ter beantwoording van de adviesaanvraag over de veiligheid van nieuwe
voedingsmiddelen en nieuwe voedselingrediënten, die door u mede namens de Minister van
Landbouw, Natuurbeheer en Voedselkwaliteit aan de Gezondheidsraad is voorgelegd. Aan de orde
is een zogenoemde tweede beoordeling, conform de Europese verordening 258/97, van het gebruik
van Lycopeen uit Blakeslea trispora. De lycopeen is bedoeld als voedingssupplement en als
ingrediënt in boter en margarine, melk en melkproducten, sauzen, soepen en smaakmakers, en in
gebak in concentraties van 2 tot 7 mg/kg. Het product zal vermarkt worden in een oliesuspensie
van 5% en een van 20% lycopeen. De aanvrager die dit product op de markt wil brengen is de
firma Vitatene. De beoordeling is verricht door de Commissie ‘Veiligheidsbeoordeling nieuwe
voedingsmiddelen’ van de Gezondheidsraad (Commissie VNV).
De eerste beoordeling van de aanvraag voor markttoelating is verricht door de Engelse
deskundigencommissie ‘Advisory Committee on Novel Foods and Processes’ (ACNFP). De
ACNFP heeft enkele aanvullende vragen gesteld bij het ingediende dossier. De ACNFP heeft
lycopeen uit Blakeslea trispora beoordeeld als veilig voor de gevraagde toepassingen. De
bevoegde autoriteit, de Engelse Food Standards Agency, heeft dit advies overgenomen.
De Commissie VNV baseert haar oordeel op het rapport van de eerste beoordeling door de
ACNFP en op de informatie in het dossier. De Commissie VNV stemt grotendeels in met de
Engelse beoordeling. De lycopeen uit de nieuwe bron, de schimmel Blakeslea trispora, is
vergelijkbaar met de lycopeen in groenten en fruit. Lycopeen is een carotenoïd waaraan anti-
oxidatieve eigenschappen worden toegeschreven. De schimmel Blakeslea trispora is al eerder door
Bezoekadres                                                            Postadres
Parnassusplein 5                                                       Postbus 16052
2511 VX    Den Haag                                                    2500 BB   Den Haag
Telefoon (070) 340 7520                                                Telefax (070) 340 7523
                                                                       www.gr.nl
                                                                                             1
</pre>

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<pre>Gezondheidsraad                             Vice-voorzitter
Health Council of the Netherlands
Onderwerp          : Tweede beoordeling veiligheid Lycopeen uit Blakeslea trispora
Ons kenmerk        : 2004/02VNV, U-868 /JvdW/cv/622-CX
Pagina             :2
Datum              : 24 juni 2004
internationale deskundigencommissies goedgekeurd als bron voor β-caroteen. De aanvrager heeft
aanvullend een 28-dagen toxiciteitsonderzoek gedaan met de lycopeenrijke biomassa. Dit leverde
geen aanwijzingen op voor nadelige effecten.
      Met geconcentreerd lycopeen uit verschillende bronnen zijn door derden uitgebreide
toxicologische onderzoeken gedaan zoals acuut, subchronisch en chronisch proefdieronderzoek.
Ook resultaten van carcinogeniteits-, mutageniteits- en reproductietoxiciteitsonderzoek door
derden zijn beschikbaar en leveren geen aanwijzingen voor nadelige invloeden van consumptie
van lycopeen. Gegevens over hoge lycopeeninname van mensen duiden niet op
gezondheidsrisico’s. De aanvrager heeft met het eigen lycopeenpreparaat van 20% lycopeen in
olie gesuspendeerd een 90-dagen onderzoek in proefdieren laten uitvoeren. Daaruit is een No
Observed Adverse Effect Level (NOAEL) afgeleid van 601 mg per kg lichaamsgewicht per dag.
Deze waarde wordt vervolgens vergeleken met de gemiddelde inname van lycopeen door de
Engelse consument als deze uitsluitend de voorgestelde voedingsmiddelen met de nieuwe
lycopeen zou nuttigen in het huidige eetpatroon. Dit leidt tot consumptieniveaus van 44,9 µg per
kg lichaamsgewicht voor kinderen en 22,6 µg per kg lichaamsgewicht voor volwassen mannen (97
percentiel). Tussen de geschatte toekomstige inname en NOAEL is de veiligheidsmarge meer dan
10.000. Als de NOAEL wordt vergeleken met de inname door het gebruik van reeds op de markt
zijnde voedingssupplementen met 20 mg lycopeen is er nog altijd een veiligheidsmarge van 2000.
      De huidige inname van lycopeen uit groenten en fruit varieert zeer sterk van persoon tot
persoon en van dag tot dag. Indicaties van gemiddelde innames zijn: 0,8 mg per persoon per dag in
Finland, 1 mg per persoon per dag in het Verenigd Koninkrijk (lycopeenrijk eetpatroon) en 5 mg
tot uitschieters van 25 mg per persoon per dag in de Verenigde Staten. De inname van lycopeen
door de Engelse consument als deze uitsluitend de voorgestelde voedingsmiddelen met de nieuwe
lycopeen zou nuttigen in het huidige eetpatroon wordt 0,65 mg per persoon per dag voor jonge
kinderen en 1,5 mg per persoon per dag voor volwassenen (97 percentiel). Het lycopeenpreparaat
bevat geen meetbare hoeveelheden eiwit waardoor de kans op allergene eigenschappen
verwaarloosbaar is.
      De Commissie VNV is het grotendeels eens met het oordeel van de ACNFP. Zij heeft de
volgende aanvullingen en kanttekeningen bij het oordeel. Er zijn de commissie twee onderzoeken
bekend die een indicatie geven van de consumptie van lycopeen door de Nederlandse bevolking.
Het ene onderzoek is een cohortonderzoek bij ouderen en betreft een berekening van de inname
Bezoekadres                                                            Postadres
Parnassusplein 5                                                       Postbus 16052
2511 VX    Den Haag                                                    2500 BB    Den Haag
Telefoon (070) 340 7520                                                Telefax (070) 340 7523
                                                                       www.gr.nl
                                                                                               2
</pre>

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<pre>Gezondheidsraad                              Vice-voorzitter
Health Council of the Netherlands
Onderwerp         : Tweede beoordeling veiligheid Lycopeen uit Blakeslea trispora
Ons kenmerk       : 2004/02VNV, U-868 /JvdW/cv/622-CX
Pagina            :3
Datum             : 24 juni 2004
van carotenoïden. De inneming van lycopeen bedroeg bij mannen 1,0 mg per persoon per dag (SD
1,56) en bij vrouwen 1,3 mg per persoon per dag (SD 1,88) (Gol98). Gegevens uit het andere
onderzoek, bij personen van 25-45 jaar, geven een mediane inname van 4,9 mg lycopeen per
persoon per dag (spreiding 2,8 – 7,5 mg) (O’Ne01). Nederlanders consumeren dus al relatief veel
lycopeen en de procentuele toename door het op de markt komen van het nieuwe preparaat is naar
verwachting klein. De Commissie VNV is van mening dat de door de aanvrager voorgestelde
toepassingen en gehalten de suggestie wekken van gebruik als kleurstof. Ten opzichte van het
pakket aan toxicologisch onderzoek dat standaard geëist wordt voor toelating van een
voedseladditief (SCF01) schiet het dossier op enkele punten tekort. Daardoor kan er ook geen
acceptable daily intake (ADI) worden vastgesteld en slechts een no observed adverse effect level
(NOAEL). Voor stoffen die toepassing vinden als voedselingrediënt én als voedseladditief
verdient een volledig dossier en afleiding van een ADI de voorkeur. Omdat het
productenassortiment beperkt blijft en de gehaltes laag, vindt de Commissie VNV dit
lycopeendossier vooralsnog acceptabel.
De Commissie VNV maakt geen bezwaar tegen toelating van de nieuwe lycopeen als
voedselingrediënt op de Europese markt op de voorgestelde wijze.
Ik onderschrijf de conclusies en aanbevelingen van de Commissie VNV.
Hoogachtend,
prof. dr JGAJ Hautvast
Bezoekadres                                                           Postadres
Parnassusplein 5                                                      Postbus 16052
2511 VX    Den Haag                                                   2500 BB   Den Haag
Telefoon (070) 340 7520                                               Telefax (070) 340 7523
                                                                      www.gr.nl
                                                                                              3
</pre>

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<pre>Letter to the Dutch Minister of Health,
Welfare and Sport
On June 15, professor JGAJ Hautvast, Vice-president of the Health Council of the
Netherlands wrote as follows to the Minister of Health, Welfare and Sport:
This letter had been prepared in reply to your request for advice regarding the safety of
novel foods and food ingredients, also made on behalf of the Minister of Agriculture,
Nature and Food Quality. The subject in question is a so-called second opinion, in
accordance with European Regulation 258/97, concerning Lycopene from Blakeslea
trispora. The lycopene is intended for use as a dietary supplement and as an ingredient
in butter and margarine, milk and milk products, sauces, soups and seasonings, and in
confectionery in concentrations of 2–7 mg/kg. The product will be marketed in an oil
suspension of 5% and of 20% lycopene. The applicant wishing to place this product on
the market is Vitatene. This assessment was carried out by the Health Council's Com-
mittee on Safety Assessment of Novel Foods (VNV Committee).
     The initial assessment of the application for marketing authorisation was carried out
by the Advisory Committee on Novel Foods and Processes (ACNFP), in the United
Kingdom. The ACNFP raised several supplementary questions in connection with the
dossier that was submitted. The ACNFP judged lycopene from Blakeslea trispora to be
safe for the proposed uses. The Competent Authority, the UK Food Standards Agency,
has adopted this advice.
     The VNV Committee bases its opinion on the report from the initial assessment by
the ACNFP and on the information in the dossier. The VNV Committee largely agrees
with the UK assessment. The lycopene from the novel source, the fungus Blakeslea tris-
Letter to the Dutch Minister of Health, Welfare and Sport                                  7
</pre>

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<pre>pora, is comparable to the lycopene in vegetables and fruit. Lycopene is a carotenoid to
which anti-oxidative properties are ascribed. The Blakeslea trispora fungus has already
been approved by international expert committees as a source of β-carotene. The appli-
cant has additionally carried out a 28-day toxicity study with the lycopene-rich biomass.
This did not produce any indications of adverse effects.
     Extensive toxicological studies (e.g. acute, sub-chronic and chronic animal studies)
have been performed by third parties using concentrated lycopene from different sour-
ces. Results are also available from carcinogenicity, mutagenicity and reproductive toxi-
city studies conducted by third parties and these show no evidence of adverse effects
from consumption of lycopene. Data on high-level human intake of lycopene do not
suggest any health risks. The applicant has had a 90-day study performed in experimen-
tal animals using its own lycopene preparation containing 20% lycopene suspended in
oil. From this a No Observed Adverse Effect Level (NOAEL) of 601 mg per kg body
weight per day has been derived. This value is then compared with the average intake of
lycopene by the UK consumer if the proposed foods with the novel lycopene is consu-
med in accordance with current food consumption patterns. This results in consumption
levels of 44.9 µg per kg body weight for children and 22.6 µg per kg body weight for
adult males (97 percentile). There is a safety margin of more than 10,000 between esti-
mated future intake and the NOAEL. If a comparison is made between the NOAEL and
the intake through the use of dietary supplements containing 20 mg lycopene already on
the market, there is still a 2,000-fold safety margin.
     Current intake of lycopene from vegetables and fruit varies massively from person
to person and from day to day. The following are indications of average intakes: 0.8 mg
per person per day in Finland, 1 mg per person per day in the United Kingdom (lyco-
pene-rich eating patterns), and a range of between 5 mg and 25 mg per person per day
(in outliers) in the United States. The intake of lycopene by the UK consumers if they
were to consume only the proposed foods with the novel lycopene in accordance with
current eating patterns is 0.65 mg per person per day for young children and 1.5 mg per
person per day for adults (97 percentile). As the lycopene preparation contains no
measurable amounts of protein, the risk of allergenic properties is negligible.
     While the VNV Committee is largely in agreement with the ACNFP opinion, it has
the following additions and comments to make in connection with it. The Committee
knows of two studies that give an indication of the consumption of lycopene by the
Dutch population. One is a cohort study with elderly people which estimates the intake
of carotenoids. In the case of men, intake of lycopene was 1,0 mg per person per day
(SD 1,6) and for women it was 1.3 mg per person per day (SD 1,9) (Goldbohm et al,
1998). Data from the other study with people of 25-45 years of age give a median intake
of 4.9 mg lycopene per person per day (variation 2.8 – 7.5 mg) (O’Neill et al, 2001).
Thus the Dutch consume relatively large amounts of lycopene and the percentage
Letter to the Dutch Minister of Health, Welfare and Sport                                 8
</pre>

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<pre>increase resulting from the market introduction of the novel preparation is expected to
be small. The VNV Committee believes that the applications and levels proposed by the
applicant closely resemble use as a colourant. As far as the package of toxicological stu-
dies that is usually required for authorisation of a food additive (SCF 2001) is con-
cerned, the dossier is lacking in some areas. It is therefore not possible to establish an
acceptable daily intake (ADI), only a no observed adverse effect level (NOAEL). For
substances that are used as a food ingredient and as a food additive, it is preferable to
have a complete dossier and derivation of an ADI. Because the product assortment is
still limited and the levels are low, the VNV Committee finds this acceptable for the
time being.
      The VNV Committee does not object to the authorisation of the novel lycopene as a
food ingredient on the European market in the proposed way.
I endorse the conclusions and recommendations of the VNV Committee,
(signed) professor JGAJ Hautvast
Letter to the Dutch Minister of Health, Welfare and Sport                                  9
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<pre>      Literatuur/Literature
EG97  Verordening (EG) nr. 258/97 van het Europees Parlement en de Raad van 27 januari 1997 betreffende
      nieuwe voedingsmiddelen en nieuwe voedselingrediënten. Publikatieblad van de Europese
      Gemeenschappen 1997; L43: 1-6
      (Regulation (EC) No 258/97 of the European Parliament and of the Council of 27 January 1997 concerning
      novel foods and novel food ingredients. Official Journal of the European Communities 1997; L43: 1-6).
EG97a Aanbeveling (EG) nr. 97/618/EG van de Commissie van 29 juli 1997 betreffende de wetenschappelijke
      aspecten en de presentatie van de informatie die nodig is om aanvragen voor het in de handel brengen van
      nieuwe voedingsmiddelen en nieuwe voedselingrediënten te ondersteunen alsmede het opstellen van de
      verslagen van de eerste beoordeling uit hoofde van Verordening (EG) nr. 258/97 van het Europees
      Parlement en de Raad. Publikatieblad van de Europese Gemeenschappen 1997; L253: 1-36
      (97/618/EC: Commission Recommendation of 29 July 1997 concerning the scientific aspects and the
      presentation of information necessary to support applications for the placing on the market of novel foods
      and novel food ingredients and the preparation of initial assessment reports under Regulation (EC) No 258/
      97 of the European Parliament of the Council. Official Journal of the European Communities 1997; L253:
      1-36).
EG03  Verordening (EG) nr. 1829/2003 van het Europees Parlement en de Raad van 22 september 2003 inzake
      genetisch gemodificeerde levensmiddelen en diervoeders. Publikatieblad van de Europese
      Gemeenschappen 2003; L268: 1-23.
      (Regulation (EC) nr. 1829/2003 of the European Parliament and of the Council of 22 September 2003
      concerning genetically modified food and feed. Official Journal of the European Communities 2003; L268:
      1-23).
FAO96 Biotechnology and Food Safety. Report of a joint FAO/WHO Consultation. Rome, FAO 1996.
      Literatuur/Literature                                                                                      10
</pre>

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<pre>Gol98  Goldbohm RA, Brants HAM, Hulshof KFAM, van den Brandt PA. The contribution of various foods to
       intake of vitamin A and carotenoids in the Netherlands. Internat. J. Vit. Nutr. Res. 1998; 68:378-383.
GR92   Commissie Toxicologische aspecten van biotechnologisch bereide producten. Productveiligheid bij nieuwe
       biotechnologie. Den Haag, Gezondheidsraad 1992, publicatienummer 1992/03.
GR02   Commissie Veiligheidsbeoordeling nieuwe voedingsmiddelen. Veiligheidsbeoordeling van nieuwe
       voedingsmiddelen. Den Haag, Gezondheidsraad 2002, publicatienummer 2002/05VNV.
       Committee on the Safety Assessment of Novel Foods. Safety assessment of novel foods. The Hague, Health
       Council 2002, publication number 2002/05VNV.
OECD93 Safety evaluation of foods derived by modern biotechnology. Concepts and principles. Paris, OECD 1993.
OECD96 OECD Workshop on Food Safety Evaluation. Paris, OECD 1996.
O’Ne01 O’Neill ME, Caroll Y, Corridan B., e.a. A European carotenoid database to assess carotenoid intakes and
       its use in a five-country comparative study. Br. J. Nutr. 2001; 85: 499-507.
SCF01  Guidance on submissions for food additive evaluations by the Scientific Committee on Food. Brussels,
       Scientific Committee on Food, 2001, documentcode SCF/CS/ADD/GEN/26Final.
WHO91  Strategies for assessing the safety of foods produced by biotechnology. Report of a joint FAO/WHO
       Consultation. Geneva, WHO 1991.
       Literatuur/Literature                                                                                   11
</pre>

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<pre>A De adviesaanvraag/Request for advice
B De commissie/The committee
C EU-procedure/EU-procedure
D Samenvatting van het dossier/Executive summary of the dossier
E Eerste beoordeling/First assessment
  Bijlagen/Annexes
                                                                12
</pre>

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<pre>Bijlage A
        De Adviesaanvraag/Request for advice
        Op 18 augustus 1999 schreef de Minister van Volksgezondheid, Welzijn en Sport aan de
        Voorzitter van de Gezondheidsraad (brief kenmerk GZB/VVB 993428):
        Sinds mei 1997 is in de Europese Unie de Verordening (EG) 258/97 van kracht inzake nieuwe voedingsmid-
        delen en nieuwe voedselingrediënten. Daarmee werd de veiligheidsbeoordeling onderdeel van een com-
        munautaire procedure.
             Met u is reeds de mogelijkheid besproken de beoordeling door de Gezondheidsraad te laten uitvoeren.
        Ik verzoek u dan ook mede namens de Staatssecretaris van Landbouw, Natuurbeheer en Visserij, in deze
        eerste fase van uitvoering van de Europese Verordening (EG) 258/97 gedurende een aantal jaren, de veilig-
        heidsbeoordeling gestalte te geven. Voor het onderbrengen bij de Gezondheidsraad pleit het experimentele
        karakter dat de beoordeling de eerste jaren zal hebben. Dit experimentele karakter komt voort uit het feit dat
        het een nieuw soort beoordeling betreft van deels nieuwe categorieën van voedingsmiddelen of voedse-
        lingrediënten. Het is namelijk een veiligheidsbeoordeling vóór het op de markt brengen van met name voe-
        dingsmiddelen van een genetisch gemodificeerde oorsprong en zogenaamd functional foods (nutriceutica).
        Daarnaast ga ik ervan uit dat de onafhankelijke wetenschappelijke advisering door de Gezondheidsraad het
        vertrouwen van de Europese Commissie en de andere lidstaten in het Nederlandse oordeel nog versterkt.
             Mijn beleid is erop gericht een zo groot mogelijke openheid en transparantie te realiseren van de
        gevolgde procedure en de beoordeling om de consument vertrouwen te geven in de veiligheid van de
        De Adviesaanvraag/Request for advice                                                                           13
</pre>

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<pre>nieuwe voedingsmiddelen. Ik verzoek de Gezondheidsraad hieraan bij te dragen door bijvoorbeeld inzage te
geven in de dossiers waarvoor een aanvraag wordt ingediend, waarbij uiteraard bedrijfsvertrouwelijke gege-
vens worden beschermd en door de criteria, waarop de veiligheid zal worden beoordeeld, te publiceren.
De Minister van Volksgezondheid, Welzijn en Sport,
w.g. dr E Borst-Eilers
English translation
On 18 August 1999, the Minister of Health, Welfare and Sport wrote as follows to the
President of the Health Council of the Netherlands (under reference GZB/VVB
993428):
Since May 1997, Regulation (EC) 258/97 concerning novel foods and novel food ingredients has been in
force in the European Union. Under the Regulation, the safety of novel foods has to be assessed as part of a
community procedure.
      Following discussions regarding the possibility of the Health Council making such assessments, the
State Secretary for Agriculture, Nature Management and Fisheries and I wish the Council to take responsi-
bility for safety assessment for a period of several years during the fist phase of implementation of European
Regulation (EC) 258/97. It is considered appropriate that the Health Council should initially take on this
role because the assessment activities will be of an experimental nature, involving both a new form of asses-
sment (i.e. pre-marketing assessment) and, in many cases, new categories of foodstuff (primarily foodstuffs
with a genetically modified basis and functional foods or nutraceuticals). We also feel that if assessments
are made by a body with the Council's independent scientific status, this will support the validity of the
Netherlands' opinion in the eyes of the European Committee and other member states.
      My wish is to make the procedure and the assessment as open and transparent as possible, so as to
enhance consumer trust in the safety of novel foods. I would like the Health Council to support this objec-
tive by, for example, allowing perusal of the application dossier (insofar as consistent with the need to pro-
tect the confidentiality of commercially sensitive information) and publishing the criteria upon which safety
assessments are made.
The Minister of Health, Welfare and Sport,
(signed) dr E. Borst-Eilers
De Adviesaanvraag/Request for advice                                                                           14
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<pre>Bijlage B
        De commissie/The committee
        •  Prof. dr EG Schouten, voorzitter/chairman
           hoogleraar epidemiologie; Wageningen Universiteit and Researchcentrum/ profes-
           sor of epidemiology; Wageningen University and Research centre
        •  Prof. dr CAFM Bruijnzeel-Koomen
           hoogleraar dermatologie/allergologie; Academisch Ziekenhuis Utrecht/professor of
           dermatology/allergology; Academic Hospital Utrecht
        •  Ir EJ Kok
           toxicoloog; RIKILT-DLO Wageningen/toxicologist; State Institute for Quality Con-
           trol of Agricultural Products, Wageningen
        •  Dr CF van Kreijl
           moleculair-bioloog; RIVM Bilthoven/molecular biologist; National Institute of
           Public Health and the Environment, Bilthoven
        •  Prof. dr P van der Laan
           emeritus hoogleraar statistiek; Technische Universiteit Eindhoven/professor of sta-
           tistics; Technical University Eindhoven
        •  Dr FM Nagengast
           gastro-enteroloog; Academisch Ziekenhuis Nijmegen/gastro-enterologist; Acade-
           mic Hospital Nijmegen
        •  Dr ir JMA van Raaij
           voedingsfysioloog; Wageningen Universiteit and Researchcentrum/ food physiolo-
           gist; Wageningen University and Research centre
        De commissie/The committee                                                             15
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<pre>•   Prof. dr ir G Schaafsma
    hoogleraar voeding; TNO Voeding, Zeist/professor of nutrition; TNO Nutrition and
    Food Research, Zeist
•   Dr GJA Speijers
    toxicoloog; RIVM Bilthoven/toxicologist; National Institute of Public Health and
    the Environment, Bilthoven
•   Prof. dr WJ Stiekema
    hoogleraar bioinformatica; Wageningen Universiteit en Researchcentrum/ professor
    of bioinformatics; Wageningen University and Research centre
•   Ir R Top, adviseur/advisor
    Ministerie van VWS; Den Haag/Ministry of Health, Welfare and Sport; The Hague
•   Prof. dr WM de Vos
    hoogleraar microbiologie; Wageningen Universiteit en Researchcentrum/ professor
    of microbiology; Wageningen University and Research centre
•   Dr RA Woutersen
    toxicoloog, toxicologisch patholoog; TNO Voeding, Zeist/toxicologist, toxicologic
    pathologist; TNO Nutrition and Food Research, Zeist
•   Dr ir F van der Wilk, adviseur/advisor
    COGEM, Bilthoven/Committee on Genetic Modification, Bilthoven
•   Dr JAG van de Wiel, senior secretaris/senior scientific staff member
    Gezondheidsraad, Den Haag/Health Council of the Netherlands, The Hague
Administratieve ondersteuning/Administrative assistance: CL Vuijst, MSc; Gezond-
heidsraad, Den Haag/Health Council of the Netherlands, The Hague.
Layout: J van Kan; Gezondheidsraad, Den Haag/Health Council of the Netherlands, The
Hague.
De commissie/The committee                                                            16
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<pre>Bijlage C
        EU-procedure/EU-procedure
        Als een fabrikant een nieuw voedingsmiddel op de markt brengt, dient de veiligheid
        voor de consument gewaarborgd te zijn. In 1997 werd de Europese verordening van
        kracht waarin de procedure is geregeld voor de goedkeuring voor marktintroductie van
        een nieuw voedingsmiddel (EG97). Bij deze procedure zijn verschillende actoren
        betrokken. De aanvrager moet beoordelen of het product werkelijk 'nieuw' is, dat wil
        zeggen dat het nog niet eerder in de Europese Unie in substantiële mate voor menselijke
        voeding is gebruikt en ook niet wezenlijk gelijkwaardig is aan een bestaand product.
        (Voor een wezenlijk gelijkwaardig product kan worden volstaan met een kennisgeving
        van de marktintroductie.) Ook moet het niet gaan om een levensmiddelenadditief, aroma
        of extractiemiddel, omdat deze producten op een andere wijze worden beoordeeld. Voor
        een nieuw voedingsmiddel in de zin van de Europese verordening moet de aanvrager
        een veiligheidsdossier overleggen volgens aanbevelingen van de Europese Commissie
        (EG97a). Deze aanbevelingen zijn gebaseerd op rapporten van verschillende instanties
        die zich met het onderwerp nieuwe voedingsmiddelen bezighouden, te weten de OECD
        (OECD93, OECD96) en de WHO/FAO (FAO96, WHO91). Ook de Gezondheidsraad
        heeft zich al eerder over dit onderwerp gebogen (GR92). Sinds het verschijnen van de
        aanbevelingen van de EU wordt in internationaal verband gewerkt aan explicitering en
        aanpassing aan de stand van de wetenschap (GR02).
            De fabrikant levert het volgens de richtlijnen samengestelde dossier in bij het land
        waar het product het eerst op de markt zal komen. Daarop komt de nationale veiligheids-
        beoordelingsautoriteit in actie. In Nederland is dat de Minister van Volksgezondheid,
        Welzijn en Sport. Zij heeft de Gezondheidsraad verzocht haar van advies te dienen. De
        EU-procedure/EU-procedure                                                                17
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<pre>Voorzitter van de Gezondheidsraad heeft hiertoe de commissie Veiligheidsbeoordeling
nieuwe voedingsmiddelen (commissie VNV) ingesteld.
     De commissie beoordeelt op basis van de huidige stand van de wetenschap of de
door de fabrikant geleverde gegevens juist en volledig zijn en of zij het eens is met diens
conclusies. Zij maakt een verslag van haar bevindingen — ook volgens de Europese
aanbevelingen (EG97a, deel III) — en biedt dat de minister aan. De minister formuleert
het Nederlandse oordeel over een voedingsmiddel en brengt dat in bij het Europese
overleg in het Permanent Comité voor de voedselketen en de diergezondheid. Alle Euro-
pese lidstaten worden uitgenodigd hun oordeel (de zogeheten tweede beoordeling) te
geven over het dossier en over de eerste beoordeling alvorens genoemd Comité een
eindoordeel velt. Als een dossier veel vragen oproept, gaat er een adviesvraag van de
Europese Autoriteit voor voedselveiligheid (EAV). Komt men dan nog niet tot overeen-
stemming dan beslist de Europese Ministerraad. Sinds 18 april 2004 moeten veiligheids-
dossiers van voedingsmiddelen van genetisch gemodificeerde oorsprong rechtstreeks
ingediend worden bij de EAV (EG03).
English translation
When manufacturers bring novel foodstuffs onto the market, consumer safety has to be
ensured. In 1997, a European Regulation (EG97) came into force, laying down the pro-
cedure for approving the market introduction of novel foodstuffs. The procedure recog-
nizes various actors. The applicant must decide whether a product is a novel foodstuff,
i.e. a substance that has not previously been available for human consumption to any
substantial extent within the European Union and is not substantially equivalent to any
existing product. (If a foodstuff is substantially equivalent to any existing product, it is
sufficient to inform the authorities of its market introduction). Food additives, aromas
and extracts are excluded from the provisions of the directive, since they fall within the
scope of an established assessment regime. Before marketing a novel foodstuff, the
applicant must compile a safety dossier that complies with the Recommendations of the
European Commission (EG97a). These Recommendations are based on reports by a
number of bodies that have studied the issue of novel foodstuffs, in particular the OECD
(OECD93, OECD96) and the WHO/FAO (FAO96, WHO91). The Health Council of the
Netherlands has also considered the question earlier (GR92). Since publication of the
EU recommendations, international efforts have been made to clarify and adapt the lat-
est scientific knowledge in the field (GR02).
     Having compiled a dossier in line with the guidelines, the manufacturer has to sub-
mit it to the competent authority in the country where the product is to be marketed first.
This dossier is assessed by the national safety assessment authority. In the Netherlands,
this is the Minister of Health, Welfare and Sport, who is advised by the Health Council.
EU-procedure/EU-procedure                                                                    18
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<pre>The President of the Health Council has created a Committee on the Safety Assessment
of Novel Foods (VNV Committee) to advise the minister on behalf of the Council.
     On the basis of the scientific state of the art, the committee has to decide whether the
information provided by the manufacturer is accurate and complete and whether the
manufacturer's conclusions are sound. The committee then draws up a report on its find-
ings for the minister; this report must also comply with the European Recommendation
(EG97a, part III). After considering the report, the minister formulates the Netherlands’
opinion regarding the foodstuff in question, which is discussed at European level in the
Standing Committee on the Food Chain and Animal Health. All other European member
states are invited to express a ‘second opinion’ regarding the dossier and the first opin-
ion. The Standing Committee then arrives at a final judgement. If a dossier is particu-
larly contentious, the European Commission calls upon the European Food Safety
Authority (EFSA) for advice. If consensus still cannot be reached, the issue is referred to
the European Council of Ministers. From April 18th 2004 safety dossiers of food from
genetically modified origin have to be submitted to EFSA directly (EC03).
EU-procedure/EU-procedure                                                                     19
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<pre>Bijlage D
        Samenvatting van het dossier/
        Executive summary of the dossier
        Samenvatting van het dossier/ Executive summary of the dossier 20
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<pre>    APPLICATION FOR THE APPROVAL OF
  LYCOPENE FROM BLAKESLEA TRISPORA
 Regulation (EC) No 258/97 of the European Parliament
and of the Council of 27th January 1997 concerning novel
            foods and novel food ingredients
                          SUMMARY
                 Applicant:    Vitatene
                               Antibioticos S.A.U
                               Avda. de Antibioticos, 59-61
                               24080 León,
                               Spain
                               October 22, 2003
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<pre>   APPLICATION FOR THE APPROVAL OF LYCOPENE
                    FROM BLAKESLEA TRISPORA:
    USE AS A NOVEL FOOD INGREDIENT IN EUROPE
                            Table of Contents
Applicant                                     1
Novel Food Classification                     1
Application                                   2
Information Requirements                      2
Conclusion                                    4
October 22, 2003
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<pre>   APPLICATION FOR THE APPROVAL OF LYCOPENE
                      FROM BLAKESLEA TRISPORA:
     USE AS A NOVEL FOOD INGREDIENT IN EUROPE
Applicant
The application is submitted by:
        Vitatene
        Antibioticos S.A.U.PTD Center
        Avda De Antibioticos, 59-61
        24080 Leon
        Spain
Contact person:
        Mrs Carmelita Rodriguez Otero
        Telephone:      +34 987 895 825
        Fascimile:      +34 987 895 812
        E-mail:         crodriguez@antibioticos.it
Novel Food Classification
Under Regulation (EC) No 258/97 of the European Parliament and of the Council of 27th
January 1997 concerning novel foods and novel food ingredients (hereafter referred to as EC
258/97), Vitatene seeks approval to market lycopene, derived from the fungus Blakeslea
trispora, for use as a novel food ingredient in Europe. Vitatene therefore submit following an
application dossier (“Application for the approval of Lycopene from Blakeslea trispora”) pursuant
to the Commission Recommendation of 29 July 1997 concerning the scientific aspects and the
presentation of information necessary to support applications for the placing on the market of
novel foods and novel food ingredients (hereafter referred to as the Commission
Recommendation of 1997), concerning the scientific aspects of information necessary for the
placing on the market of novel foods and novel food ingredients.
Article 1(2) of EC 258/97 states that the regulation “…shall apply to the placing on the market
within the Community of foods and food ingredients which have not hitherto been used for
human consumption to a significant degree within the Community and which fall under the
following categories…(d) foods and food ingredients consisting of or isolated from
October 22, 2003                                                                                1
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<pre>microorganisms, fungi or algae”. Lycopene from B. trispora is thus considered a novel
food/food ingredient due to its source organism.
Application
This application seeks approval for the use of lycopene in the form of an oil suspension as a
nutritional food ingredient. The individual proposed food-uses and their maximum usage levels
for lycopene from B. trispora in the E.U. are summarized in the following Table.
Table 1           Summary of the Individual Proposed Food Uses and Use-Levels for
                  Lycopene in the E.U.
           Food Category                                Proposed Food-Use                      Use-Level
                                                                                                (ppm)
Fat Spreads                          Butter                                                       2.0
                                     Margarine and Margarine-Like Spreads                         5.0
Milk and Milk Products               Desserts, Puddings, and Custards                             4.0
                                     Ice Cream, Ice Milk, Frozen Yogurt, Sherbet and Novelties    6.0
                                     Processed Cheese                                             3.0
                                     Ripened Orange, Yellow, and White Cheese                     3.0
Miscellaneous                        Condiments, Seasonings, Relishes, and Pickles                6.0
                                     Mustard                                                      5.0
                                     Savoury Sauces and Gravies                                   7.0
                                     Soups and Soup Mixes                                         6.0
Sugar, Preserves, and Confectionery  Sweet Spreads, Fillings, and Icings                          5.0
Information Requirements
Section 4 of the Commission Recommendation of 1997 outlines recommendations pertaining to
the “Scientific Classification of Novel Foods for the Assessment of Wholesomeness”, which
facilitates the safety and nutritional evaluation of a given novel food/food ingredient. Lycopene
from B. trispora is classified in Class 2 as a “Complex Novel Food from non-GM source”, since
the production strains of B. trispora have been developed by conventional techniques, with no
use of genetic modification. Lycopene from B. trispora would be further allocated under Sub-
Class 2.2: “the source of the novel food has no history of food use in the Community”. Pursuant
to Table II, Part I under the Commission Recommendation of 1997, the essential information
requirements corresponding with this classification are outlined as follows, expanded upon in
the dossier (“Application for the approval of Lycopene from B. trispora”), and summarized
herein.
I.       Specification of the Novel Food
II.      Effect of the production process applied to the Novel Food
October 22, 2003                                                                                         2
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<pre>III.     History of the organism used as the source of the Novel Food
IX.      Anticipated intake/extent of use of the Novel Food
X.       Information form previous human exposure to the Novel Food or its source
XI.      Nutritional information on the Novel Food
XII.     Microbiological information on the Novel Food
XIII.    Toxicological information on the Novel Food
Executive Summary
Manufacturing/specifications
The manufacturing of lycopene is carried out in two phases. The fermentation phase refers to
the production of lycopene through a co-fermentation process using the 2 sexual mating types
(plus and minus) of the fungus B. trispora. The extraction and recovery phase involves solvent
extraction of lycopene from the biomass of the fermentation broth, and recovery of the product
as high purity lycopene crystals. The final lycopene product is formulated into an oil suspension
(5 or 20% lycopene) prior to packaging. Results of analysis for representative lots of 5 and 20%
lycopene oil suspensions demonstrated compliance with final product physical, chemical, and
microbiological specifications.
History/Safety of Lycopene and the Source Organism
Safety of the fungal source of lycopene has been confirmed in two toxicological studies
including a 28-day oral feeding study conducted with the B. trispora biomass in rats, and a 90-
day study with the extracted lycopene formulation and two mutagenicity studies including a
bacterial mutation test and a chromosome aberration test. The safety of B. trispora is further
supported by expert reviews of the literature, which concluded that the B. trispora strains are
considered to be non-toxigenic and non-pathogenic, and by the SCF and JECFA who have
verified the safety of the microorganism and the resultant production of β-carotene. In addition,
stability tests, microbiological tests and protein presence analysis have been performed on the
final lycopene product, which demonstrate that it is free of protein, mycotoxins and other toxic
metabolites.
Previous Human Exposure
Lycopene from B. trispora is chemically and nutritionally equivalent to naturally occurring
lycopene, since the biosynthetic routes from the fungal source are identical to those occurring in
nature. Lycopene is a normal constituent of the human diet due mainly to its presence in red
fruits and vegetables, including tomatoes, watermelon, pink grapefruit, apricots, and pink
guavas, as well as in algae and fungi. A number of studies have been completed where intakes
of dietary lycopene have been assessed in various populations. These have indicated intakes
as high as 25 mg/day in Canadian males while data from an elderly female study in the United
October 22, 2003                                                                                  3
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<pre>Kingdom (U.K.) has indicated levels of intake around 1 mg/day. The consumption of lycopene
containing fruits and vegetables in the human diet has also been found to coincide with
lycopene being found as a constituent of mature breast milk and a predominant carotenoid in
human plasma.
Estimated Intake of the Novel Food
Anticipated lycopene intakes in the E.U. were estimated using the U.K. food consumption data
collected as part of the U.K. Food Standards Agency’s, Dietary Survey Programme (DSP). The
results indicated minimal exposure to the fungal-derived lycopene product in the all-person and
all-user specific demographic groups in the U.K. population using consumption data and
information pertaining to the individual proposed food-uses for lycopene. On an all-user basis,
the highest mean and 97.5th percentile intakes of lycopene by the U.K. population from all
proposed food-uses in the E.U., as observed in male adults, were estimated to be 0.60
mg/person/day (8.1 µg/kg body weight/day) and 1.68 mg/person/day (22.6 µg/kg body
weight/day). On a body weight basis, children consumed the greatest amount of lycopene, with
mean and 97.5th percentile all-user intakes of 15.1 and 44.9 µg/kg body weight/day,
respectively.
Safety of the Novel Food
The safety of lycopene from B. trispora is supported by a 90-day toxicity study and two genetox
studies as well as an extensive knowledge of lycopene metabolism, a history of use due to the
natural presence of lycopene in food, and published literature on the safety (acute toxicity, sub-
chronic toxicity, reproductive toxicity, carcinogenicity, mutagenicity/genotoxicity, and clinical
trials) of lycopene derived from sources other than fungal (i.e., dietary lycopene). Based on the
average no-observed-effect level (NOEL) of 601 mg lycopene/kg body weight per day from the
rat sub-chronic study, and the maximum level of human intake from proposed food uses and
dietary supplement intake, it can be concluded that there is a large margin of safety.
Conclusion
In conclusion, this application demonstrates the safety of lycopene from B. trispora, which is
supported by the purity of lycopene from B. trispora (>95%), the conformity between
biosynthetically-derived lycopene in nature and chemically-derived lycopene from B. trispora,
the historical consumption of lycopene as a normal component of the diet (e.g., red fruits and
vegetables including tomatoes, watermelon, pink grapefruit, apricots), minimal exposure under
the conditions of intended use, safety data provided by Vitatene for the final lycopene
suspension and for the biomass, additional safety data for the biomass, and published
toxicological and clinical data conducted with lycopene (from sources other than B. trispora).
October 22, 2003                                                                                   4
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<pre>Bijlage E
        Eerste beoordeling/First assessment
        Eerste beoordeling/First assessment 27
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<pre>ADVISORY COMMITTEE FOR NOVEL FOODS AND PROCESSES
OPINION ON AN APPLICATION UNDER THE NOVEL FOODS REGULATION FOR
LYCOPENE FROM BLAKESLEA TRISPORA
Applicant                     Vitatene
Responsible personDr Rodríguez-Otero
EC Classification 2.2
INTRODUCTION
1. An application was submitted by Vitatene to the UK Competent Authority for
   authorisation of lycopene derived from the fungus Blakeslea trispora for use as a novel
   food ingredient.
2. Lycopene (C40H56) is an aliphatic branched hydrocarbon with a molecular weight of
   536.9 Daltons. It exists predominantly in the trans- form and is a red crystalline powder
   soluble in fats and organic solvents, but virtually insoluble in water, methanol or ethanol.
3. Solvent extracted lycopene from tomatoes is approved for use as an additive (E160d)
   and is used in dietary supplements and as an ingredient (food colour) in a range of foods.
   Synthetic lycopene is also used as a dietary supplement outside the EU, but is not
   permitted for use as a colour additive. Blakeslea trispora is a fungus found on a number
   of tropical plants, and strains of B. trispora are able to synthesise large quantities of
   carotenoids. Following the publication of a positive opinion from the SCF in 2001 β-
   carotene from B. trispora was approved for food additive use. Although lycopene per se
   has a history of consumption, and is produced using the same biosynthetic pathway as
   β- carotene, the organism has not hitherto been used for production of lycopene sold in
         Eerste beoordeling/First assessment                                                 28
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<pre>     the EU and the product requires authorisation under regulation (EC) 258/97 before it can
     be marketed.
I. Specification of the novel food
      Information on this aspect is provided on pages 1 – 6 of the Application dossier
4. The applicant intends to market lycopene from B. trispora as a nutritional food ingredient.
     The purified, crystalline lycopene is dissolved in high oleic sunflower oil, supplemented
     with tocopherol to minimise oxidation. Tocopherol is added at levels consistent with
     those specified in the relevant food additives directive 95/2/EC. The Novel Food (NF) will
     be available in this oil suspension form (5% and 20%) only.
5. Detailed compositional analyses of the NF are given in the Application dossier For these
     analyses the company has tested both crystalline lycopene and oil suspensions. The
     Applicant’s specification of the novel food states that it should be not less than 95%
     lycopene of which at least 90% is trans-lycopene. The remainder comprises of a number
     of low level contaminants, such as the extraction solvent, isobutyl acetate (not greater
     than 1%), sulphated ash (not greater than 1%) and subsidiary colouring matters (not
     greater than 5%). This company’s specification was exceeded in each of three non-
     consecutive, representative lots described in the application.
     Discussion. The Committee was satisfied with the specification of the novel food.
II. Effect of the production process applied to the novel food
      Information on this aspect is provided on pages 5, 7-14 of the Application dossier
6. Lycopene from B. trispora is obtained by the co-fermentation of 2 sexual mating types of
     the fungus, obtained using classical strain selection techniques to increase the efficiency
     of lycopene production. The strains used are the same as those approved for the
     production of β-carotene. The mating types are stable and are preserved and maintained
     using GLP methods and are deposited in a culture collection.
7. Fermentation of the fungi to produce lycopene is a two-stage process. Flasks are
     inoculated with each of the mating types, and grown under controlled conditions. Once
     vegetative growth is established, the contents of the flasks are individually transferred
     asceptically to larger growth tanks containing sterile medium. Once sufficient cell mass
     has accumulated the strains are transferred asceptically into another tank where co-
     fermentation commences. It is at this point that the fungi start to produce lycopene. The
            Eerste beoordeling/First assessment                                               29
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<pre>     process is further controlled by the addition of imidazole which inhibits the formation of
     carotene.
8. After completion of the fermentation process, lycopene rich biomass is subject to an
     initial purification process using isopropyl alcohol, which removes any oils and other
     lipophilic substances. The residue is evaporated to dryness, milled and extracted with
     isobutyl acetate. The resulting enriched solvent is separated and concentrated by
     vacuum distillation. The lycopene is then crystallised. Due to its susceptibility to oxidation
     the lycopene is crystallised under nitrogen. The crystalline lycopene is dissolved in high
     oleic acid sunflower oil containing tocopherol (1%) and diluted in accordance with the
     desired specification. The purification and extraction processes are identical to those
     used in the production of beta-carotene from B. trispora, which have been examined and
     cleared by the SCF. Each batch of the final product is assayed to check compliance with
     the specification Application dossier Section 1.e.
9. The applicant has supplied data indicating that in comparison with lycopene from other
     sources, lycopene from B. trispora is predominantly present in the trans- form (at least
     90%). The data also indicates that the purity of the fungal lycopene is comparable with
     synthetic lycopene (Application dossier Table II c-1).
10. The applicant has demonstrated that the NF (20% oil suspension) is stable for a period
     of at least two years when stored at 5°C. Other studies demonstrate that lycopene (5%
     and 20% oil suspension) can be stored in sealed containers for at least 6 months at a
     range of temperatures (3°C, 25°C and 40°C) with no appreciable deterioration in product
     quality. In all cases the tests took place in conditions conducive to oxidation as, although
     the NF was sealed in bottles, the applicant did not sparge with nitrogen.
     Discussion. The Committee was satisfied that the production process is controlled and
     that the in-process monitoring steps are appropriate to ensure a safe and consistent
     product, that does not deteriorate during storage. The Committee accepted clarification
     from the applicant that consumption of the novel food in a dietary supplement form did
     not raise levels of exposure to the extraction solvent, isobutyl acetate to levels that would
     be toxicologically significant.
III. History of the organism used as a source of the novel food
      Information on this aspect is provided on pages 15 – 17 of the Application dossier
11. The applicant has based previous dietary exposure to B. trispora on its use as a source
     of β-carotene, noting that the safety of the organism was assessed by the SCF (2000)
             Eerste beoordeling/First assessment                                                 30
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<pre>    and the Joint Expert Committee on Food Additives (JECFA) (2001). The SCF concluded
    that, based on the information supplied, the organism is non-pathogenic and non-
    toxigenic. A subsequent 28-day oral feeding study using Wistar rats, Jonker (2000) (see
    also Section XIII) also demonstrated that the organism was both non-toxigenic and non-
    pathogenic.
12. JECFA concluded that β-carotene from B. trispora is acceptable for food additive use,
    providing that it met the specification of its synthetic counterpart. The applicant is of the
    view that this finding is consistent with their view that the source organism is safe.
13. The applicant also carried out mycotoxin assays on each of three non-consecutive
    batches to determine whether aflatoxin B1, Mycotoxin T2, ochratoxin and zearalenone
    were present. The results, for both crystalline and oil suspended lycopene, were all
    negative.
    Discussion The Committee was reassured that the SCF assessment of the use of the
    source organism in the production of beta-carotene provided reassurance that there was
    a history of safe food use. The Committee also noted the similarity of the production
    process for production of the novel food would not give raise to any additional concerns.
IX. Anticipated intake/extent of use of the novel food
     Information on this aspect is provided on pages 21-27 of the Application dossier
14. The applicant intends to use the NF as a nutritional food ingredient. In addition to its use
    in dietary supplements, the ingredient will be used in a range of foodstuffs, including fat
    spreads, milk products and confectionery. A full list of the proposed uses is given in the
    Application dossier (Table IX a-1).
15. In order to predict the intake of the NF the applicant has used the most up to date
    information available from UK dietary surveys. The applicant has used proposed
    maximum use levels for all foods described above to predict potential intake. In order to
    compare the data over a 7-day period across a number of different surveys that target
    different sub-groups of the UK population, the applicant has applied a weighting factor.
    The UK CA sought the views of experts in the Food Standards Agency who were
    satisfied with the validity of the methodology.
16. The applicant has used dietary intake data for children (1.5–4.5), young people (4-10),
    male and female teenagers and male and female adults. Given that the proposed range
    of foodstuffs is wide, the applicant notes that the percentage of potential users was high
    amongst all age groups (>98%).
           Eerste beoordeling/First assessment                                                 31
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<pre>17. The intake estimates are summarised below. The largest consumers of the NF on an
    absolute basis are predicted to be male adults, whereas children have the highest
    predicted intakes on a body weight basis. These figures are likely to overestimate actual
    consumption, as they are based on the assumption that consumers always select foods
    that are fortified at the maximum level.
                                              ESTIMATED DAILY INTAKE
       Population Group       Mean          97th %tile     Mean       97th %tile
              (age)            (mg)            (mg)      (µg/kg bw)  (µg/kg bw)
            Children           0.22            0.65         15.1        44.9
            (1½-4½)
         Young People          0.37            0.93         14.6        36.0
              (6-11)
          Teenager (F)         0.40            1.02          7.6        20.6
             (11-18)
          Teenager (M)         0.42            1.18          7.9        23.8
             (11-18)
            Adult (F)          0.46            1.23          7.4        21.0
             (16-64)
            Adult (M)          0.60            1.68          8.1        22.6
             (16-64)
18. Based on the available intake data the applicant notes that the highest amount of
    lycopene from a food source would be obtained by consumption of fortified soups and
    soup mixes.
19. The applicant also intends to market the NF in supplement form at levels up to 20mg per
    day. Supplements containing lycopene from other sources are currently on the market in
    the EU, and it is likely that the NF would replace those already being consumed and
    overall consumption levels would not increase. In contrast, incorporation of lycopene into
    foods would result in additional intake.
20. The applicant has used the most recent adult dietary survey data available, however the
    Food Standards Agency is able to make estimates of intake based on a 2001 survey of
    British adults, which is not currently in the public domain in a form that the applicant
    could use to assess consumption of their product. Analyses of these data that confirm
    the applicant’s consumption estimated are similar to those obtained with the newer
    survey data..
           Eerste beoordeling/First assessment                                              32
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<pre>    Discussion. As the proposed levels of incorporation were low the Committee was
    content that the intended use of the product did not give any cause for concern, based on
    scientific information currently available.
X Information from previous human exposure to the novel food or its source
     Information on this aspect is provided on pages 28-30 of the Application dossier
21. Lycopene is a normal constituent of the diet in a number of red fruits and vegetables
    such as tomatoes and watermelon. Levels of lycopene in tomato are dependent both on
    the species of tomato and the degree of ripening but are generally in the range 3.1-7.7
    mg / 100g.
22. The applicant highlighted a 1996 UK study that indicated that consumption of a
    lycopene-rich diet would lead to consumption of 1.03mg/person/day lycopene. These
    results are similar to levels seen in Finland (0.70 and 0.87 mg/day for females and males
    respectively).
23. However the applicant also highlighted other studies that show that intake of lycopene
    outside the EU shows markedly varied levels of consumption. The applicant has
    summarised a number of North American dietary surveys that reinforce the European
    findings that consumption of lycopene is intrinsically varied and dependent on dietary
    preference. Consumption of lycopene in North America indicates a large variation
    dependent upon method of data collection, however in all cases mean levels were
    significantly higher than those seen for UK subjects. A USDA study showed that mean
    lycopene intake for the general US population was 4.7mg/day however a number of
    other dietary surveys indicate that consumption could be as high as 25.2mg/person/day.
24. The Applicant also notes that there are no reliable consumption figures available for the
    current consumption of lycopene in dietary supplement form despite such products being
    freely available in Europe and the North America.
    Discussion The Committee was reassured that lycopene has a history of consumption
    in the EU, albeit from a different source. The Committee noted that, in addition to its
    presence in fresh fruit and vegetables, dietary supplements containing lycopene
    extracted from tomatoes at levels in excess of 20mg were widely available in the UK.
           Eerste beoordeling/First assessment                                             33
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<pre>XI. Nutritional information on the novel food
     Information on this aspect is provided on pages 31-33 of the Application dossier
25. The applicant is of the view that, although the source of lycopene is novel, the nutritional
    value of the novel food is unchanged when compared to existing lycopene. Other
    constituents of the novel food (high oleic acid sunflower oil and tocopherol) will have a
    negligible impact on the nutritional value of the lycopene oil suspension as they are
    relatively common in the diet.
26. Lycopene is an effective antioxidant, and these antioxidant properties are perceived to
    be primarily responsible for the potential health benefits of dietary carotenes.
    Discussion The Committee was reassured that altering the source of the novel food
    would not affect its nutritional value.
XII. Microbiological information on the novel food
     Information on this aspect is provided on pages 34-35 of the Application dossier
27. Microbiological information supplied by the applicant indicates that, three non-
    consecutive batches had no detectable moulds, yeast, Salmonella or Escherichia coli.
    These findings applied to both the crystalline lycopene, and oil suspension (5% and 20%
    forms).
    Discussion The Committee was content with the microbiological data supplied, but
    requested further information from the applicant to demonstrate the absence of the
    anaerobic spore forming pathogen Clostridium botulinum. The applicant was able to
    supply this information, and the Committee was satisfied that the absence of this
    organism from the final product could be demonstrated.
XIII Toxicological Information on the Novel Food
     Information on this aspect is provided on pages 36-57 of the Application dossier
28. The applicant presented a number of toxicological studies on both the novel food and the
    source organism. The applicant has noted that the NF is chemically comparable to
    others on the market (Application dossier Table 2.c-1) and has therefore included
    toxicological studies on lycopene products from other manufacturers as supporting data.
Summary of studies
29. The applicant assessed the sub-chronic toxicity of the source of the novel food by testing
    the lycopene-rich biomass extracted from B. trispora. Supplementary information to
    demonstrate the safety of the source organism has been supplied from an independent
           Eerste beoordeling/First assessment                                                34
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<pre>    scientist, the SCF and JECFA. A 90-day oral toxicity study has been carried out on the
    NF (20% oil suspension).
30. The applicant also highlighted details of acute, sub-chronic and chronic, carcinogenicity,
    mutagenicity and genotoxicity, reproductive toxicity trials and human safety data for
    lycopene from other sources. Developmental toxicity investigations were carried out on
    two US lycopene products, whilst human safety data were mostly based on high levels of
    consumption of commonly available lycopene-rich foods.
Lycopene biomass (Application dossier p37)
31. Lycopene-rich biomass obtained under the fermentation conditions described in section
    II was used in a sub-chronic toxicity study. Four groups of 40 rats (20/sex) were
    assigned. The first formed a control group whilst the other three received lycopene
    biomass at levels of 0.1, 0.3 or 1% of the total diet. These percentages corresponded to
    daily doses of 90, 272 and 906 mg/kg body weight in males and 87, 260 and 868 mg/kg
    bodyweight in females respectively. The lycopene-enriched diet was administered for a
    period of 28 days following which the animals were sacrificed.
32. Clinical observations, neurobehavioural observations, growth, food consumption and
    food conversion efficiency were assessed throughout the study and haematology,
    clinical chemistry, organ weights and macroscopic and microscopic examinations were
    carried out at necropsy.
33. No treatment related differences were found in mean body weights and relative/absolute
    organ weights between the control and treatment groups. Food consumption and food
    conversion efficiency were also not adversely affected by the treatment. No treatment
    related clinical signs or neurotoxic indications were found as a result of the lycopene
    biomass administration. These were assessed using neurobehavioural observations and
    motor activity assessments.
34. Haematological measurements showed a statistically significant decrease in mean
    corpuscular volume and prothrombin time in the high dose male group only. However no
    significant changes were noted for other red blood cell groups, coagulation variables,
    white blood cell counts, packed cell volume or haemoglobin concentrations and the
    authors considered the decrease in mean corpuscular volume as an incidental finding
    and of no toxicological significance. The decrease in prothrombin times was found to be
    small (6%) and within the limits of historical controls.
           Eerste beoordeling/First assessment                                             35
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<pre>35. No adverse effects were noted in the clinical chemistry variables and macroscopic and
    microscopic examinations at necropsy revealed no treatment related changes except a
    statistically significant decreased incidence of increased hyaline droplet nephropathy in
    the high dose male group. Again, the authors of the study attached no toxicological
    significance to this finding.
Toxicological assessment of B. trispora (Application dossier p40)
36. The two mating strains of B. trispora are stable cultures that are preserved under
    conditions that adhere to good manufacturing practices. The strains are considered to be
    non-toxigenic and non-pathogenic on the basis of 28-day oral feeding study described
    above. The applicant also notes that B. trispora is formally classified in Germany as “risk
    group 1”, organisms that pose no risk for humans and vertebrates
37. The production of lycopene by B. trispora is an intermediary of the beta-carotene
    synthetic pathway and the SCF considered the use of B. trispora as a source of beta-
    carotene as acceptable. The Committee concluded that the “source organisms and the
    production process yielded no grounds to suppose that the final crystalline product, beta-
    carotene, differs from the chemically synthesised beta-carotene used as a food
    colourant” (SCF, 2000)
Final Product (Application dossier p38)
38. A 90-day oral toxicity study was carried out to assess the toxicity of the 20% lycopene oil
    suspension in male and female Wistar rats. Groups of 20 rats received a diet containing
    0, 0.25, 0.5, or 1.0 % lycopene in the form of a sunflower oil suspension. These
    percentages corresponded to daily doses of 0, 145 291 and 586 mg/kg bodyweight for
    males and 0, 156, 312 and 616 mg/kg bodyweight for females.
39. The animals were monitored for viability, clinical signs of toxicity, body weights and food
    consumption. Prior to necropsy, neurobehavioural testing and ophthalmoscopic
    examinations were performed and blood and urine analyses were obtained. Following
    necropsy, gross and histopathological examinations of various tissues were performed
    and organ weights recorded.
40. A pink discolouration of the fur was noted in all animals in the high dose group and many
    in the mid-dose group. This was attributed to the direct contact of the animals to the red
    staining lycopene mixture in the diet. No adverse effects were noted from the
    examinations described above and as a result the no observed effect level (NOAEL) was
    set at 1% in the diet. This was equivalent to a dose of 601mg / bodyweight per day,
    averaging the doses received by the male and female groups.
           Eerste beoordeling/First assessment                                               36
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<pre>41. The genotoxicity of a 20% cold water dispersal of lycopene from B. trispora was
    assessed using a bacterial mutation test and an in vitro chromosome aberration test. As
    a result of these studies the investigator concluded that lycopene is not genotoxic.
Margin of safety (Application dossier p39)
42. Comparing the NOEL of 601mg lycopene/kg bodyweight/day from the sub-chronic rat
    study with the anticipated maximum intake from food use of between 1 and 2 mg/day
    gives a 20000-fold safety margin. Likely intake from food supplement at a level of 20mg/
    day is associated with a 2000-fold safety margin.
Toxicological assessment of lycopene from sources other than B. trispora
43. The applicant has supplied details of additional toxicological studies with lycopene
    derived from natural tomato extracts, tomato paste and synthetically produced lycopene
    in a number of forms including cold water dispersible (CWD) and water-soluble (WS)
    beadlet formulations and dietary supplements.
²   Acute toxicity studies (Application dossier p40).
²   Sub-chronic and chronic toxicity studies (Application dossier p41).
²   Carcinogenicity studies (Application dossier p45).
²   Mutagenicity / Genotoxicity studies (Application dossier p46).
²   Reproductive toxicity studies (Application dossier p49).
²   Human safety data (Application dossier p45).
    Discussion. The Committee was satisfied with the toxicological data supplied by the
    applicant. However the Committee requested further information on the relevance of a
    significant change in the incidence of hyaline droplets in the sub-chronic toxicity study
    (Application dossier p38). The Committee also requested confirmation that the sub-
    chronic toxicity study parallel tests done using beta carotene biomass (Application
    dossier p38) did not raise any additional concerns. The applicant has responded to these
    comments highlighting that the increase in hyaline droplet nephropathy seen in male rats
    is not a toxicologically significant finding, noting that the mechanism of action, is of no
    relevance to humans. The applicant also confirmed that the parallel test with the beta
    carotene biomass revealed no additional toxicological findings. The Committee was
    content with the applicant’s responses.
           Eerste beoordeling/First assessment                                               37
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<pre>Allergenicity
     Information on this aspect is provided on page 58 of the Application dossier
44. The applicant is reported that the primary source of allergenic material, the source
    organism, is not present in the final products to any significant degree. This is borne out
    by the microbiological information (See para.30 above). Protein assays carried out on
    both the novel food (5% and 20% suspensions) and the sunflower oil were negative at
    the limit of detection (1µg protein/ml or 1µg protein in 400mg lycopene oil suspension).
    The applicant concludes that this is indicative of the absence of allergenic potential.
    Discussion The Committee was content that the final product did not give rise to any
    allergenic potential.
Labelling
     Information on this aspect is provided on page 22 of the Application dossier
45. The applicant proposes that the ingredient would be described on food labels as
    "lycopene" without identifying the source to the consumer. The applicant confirms that
    labelling of products containing the NF will comply with current EU regulations and may
    include the statement ‘contains an additional source of lycopene’.
    Discussion The Committee was of the view that the proposed labelling should be
    expanded to indicate the source of the lycopene, in order that individuals who do not
    wish to consume products derived from, or containing fungi are adequately informed.
OVERALL DISCUSSION
46. The applicant has provided a clear specification of the proposed novel food and
    indicated, on the basis of analysis from a number of non-consecutive batches, that the
    specification is achievable. The process is similar to the production of beta-carotene from
    Blakeslea trispora, which was given a positive evaluation by the SCF in 2001.
47. Given that lycopene is present in a large range of fresh fruits and vegetables, and
    lycopene extracted from tomatoes is widely available no additional nutritional concerns
    or benefits associated with consumption of the novel food have been identified. Based
    on scientific information currently available to the applicant there is sufficient
    reassurance that consumption of the novel food does not give rise to any toxicological
    concerns.
           Eerste beoordeling/First assessment                                                38
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<pre>48. The applicant has demonstrated that the novel food is stable under normal conditions
    and when subject to mild temperature abuse. The applicant has also demonstrated that
    the novel food is microbiologically safe.
49. Although the proposed labelling of the product is adequate, the applicant should comply
    with general food labelling legislation and ensure that the labelling of the products and
    the source does not mislead the consumer.
Conclusion
50. The Advisory Committee on Novel Foods and Processes is satisfied by the evidence
    provided by Vitatene that the range of uses for lycopene from Blakeslea trispora is
    acceptable subject to the applicant’s adherence to the proposed specification, and the
    production parameters described above.
                                                                                   April 2004
          Eerste beoordeling/First assessment                                              39
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