<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>Betaïne
Tweede beoordeling van de veiligheid voor de consument, volgens de
Europese verordening 258/97 betreffende nieuwe voedingsmiddelen en
nieuwe voedselingrediënten
Betaine
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 2003/03VNV, Den Haag, 23 oktober 2003
No. 2003/03VNV, The Hague, October 23, 2003
</pre>

====================================================================== Einde pagina 1 =================================================================

<br><br>====================================================================== Pagina 2 ======================================================================

<pre></pre>

====================================================================== Einde pagina 2 =================================================================

<br><br>====================================================================== Pagina 3 ======================================================================

<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.
Betaïne. Den Haag: Gezondheidsraad, 2003; publicatie nr 2003/03VNV.
Preferred citation:
Health Council of the Netherlands. Committee on the Safety Assessment of Novel
Foods. Betaine. The Hague: Health Council of the Netherlands, 2003; publication no.
2003/03VNV.
auteursrecht voorbehouden/all rights reserved
U kunt het advies downloaden van www.gr.nl/
This report can be downloaded from www.healthcouncil.nl
</pre>

====================================================================== Einde pagina 3 =================================================================

<br><br>====================================================================== Pagina 4 ======================================================================

<pre></pre>

====================================================================== Einde pagina 4 =================================================================

<br><br>====================================================================== Pagina 5 ======================================================================

<pre>  Inhoud/Contents
  Brief aan de Minister van Volksgezondheid, Welzijn en Sport 7
  Letter to the Dutch Minister of Health, Welfare and Sport 21
  Literatuur/Literature 27
  Bijlagen/Annexes 29
A De Adviesaanvraag/Request for advice 31
B De commissie/The committee 33
C EU-procedure/EU-procedure 35
D Samenvatting van het dossier/Executive summary of the dossier 39
E Eerste beoordeling/First assessment 65
  Inhoud/Contents                                                  5
</pre>

====================================================================== Einde pagina 5 =================================================================

<br><br>====================================================================== Pagina 6 ======================================================================

<pre>6 Betaïne/Betaine</pre>

====================================================================== Einde pagina 6 =================================================================

<br><br>====================================================================== Pagina 7 ======================================================================

<pre>Gezondheidsraad                                Vice-voorzitter
Health Council of the Netherlands
Aan de Minister van Volksgezondheid,
Welzijn en Sport
Onderwerp          : Tweede beoordeling veiligheid Betaïne
Uw kenmerk         : VGB/VL 2407857
Ons kenmerk        : 2003/03VNV, U-1520/MR/cv/622-CM
Datum              : 23 oktober 2003
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, Natuur en Voedselkwaliteit aan de Gezondheidsraad is voorgelegd. Aan de orde is een
zogenoemde tweede beoordeling, conform de Europese verordening 258/97, van betaïne. De
chemische naam hiervan is trimethylglycine. De aanvrager die dit nieuwe voedselingrediënt op de
markt wil brengen is de firma Finnfeeds Finland Ltd. Betaïne komt niet direct beschikbaar voor de
consument, maar het zal worden verwerkt in verschillende categorieën levensmiddelen. 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 in Finland door de Novel
Food Board (NFB). De belangrijkste conclusie van de NFB is dat, alhoewel de aanvrager veel
informatie heeft verstrekt over het effect van betaïne in bepaalde patiënten, informatie uit
wetenschappelijk onderzoek bij gezonde vrijwilligers beperkt is. De NFB stelt dat een
betrouwbare beoordeling van de veiligheid voor de consument daarom niet mogelijk is.
De Commissie VNV maakt bezwaar tegen toelating op de markt van betaïne als voedselingredient.
Zij baseert haar oordeel op het rapport van de eerste beoordeling door de NFB (zie Bijlage E), de
informatie in het dossier (voor een samenvatting zie Bijlage D) en de wetenschappelijke literatuur.
De Commissie VNV stemt ten dele in met de Finse beoordeling en maakt zelf nog enkele kritische
opmerkingen bij het dossier. Het is de Commissie VNV niet altijd duidelijk hoe zwaarwegend de
kanttekeningen die de NFB heeft geplaatst, moeten worden opgevat. De belangrijkste kritiek van
de Commissie VNV op het veiligheidsdossier is dat de veilige bovengrens van inneming
onvoldoende wordt onderbouwd. Deze kritiek is gebaseerd op het feit dat adequaat onderzoek bij
gezonde mensen ontbreekt en dat ongewenste metabole effecten zijn waargenomen bij
Bezoekadres                                                              Postadres
Parnassusplein 5                                                         Postbus 16052
2511 VX    Den Haag                                                      2500 BB   Den Haag
Telefoon (070) 340 7520                                                  Telefax (070) 340 7523
E-mail: gr@gr.nl                                                         www.gr.nl
                                                                                               7
</pre>

====================================================================== Einde pagina 7 =================================================================

<br><br>====================================================================== Pagina 8 ======================================================================

<pre>8</pre>

====================================================================== Einde pagina 8 =================================================================

<br><br>====================================================================== Pagina 9 ======================================================================

<pre>Gezondheidsraad                               Vice-voorzitter
Health Council of the Netherlands
Onderwerp          : Tweede beoordeling veiligheid Betaïne
Uw kenmerk         : VGB/VL 2407857
Ons kenmerk        : 2003/03VNV, U-1520/MR/cv/622-CM
Datum              : 23 oktober 2003
proefdieren. Gezien voornoemde onzekerheden oordeelt de commissie ook negatief over het brede
productassortiment dat de aanvrager voorstelt. In de tekst hieronder wordt het oordeel van de
Commissie VNV verder uitgewerkt.
Algemene productinformatie
De drie producten, die de aanvrager op de markt wil brengen te weten Betafin BF20, AP en
TMG20, bestaan uit bijna zuivere betaïne (minstens 99%) en bevatten geen ongewenste
bestanddelen of microbiologische verontreinigingen. De Commissie VNV is het eens met de NFB
dat de productinformatie in het dossier representatief is voor de op de markt te brengen
voedselingrediënten betaïne. Deze informatie is voldoende om de veiligheidsbeoordeling te
kunnen uitvoeren.
      In verband met de stabiliteit van betaïne merkt de Commissie VNV op dat het nieuwe
ingrediënt niet aan levensmiddelen mag worden toegevoegd die in de thuissituatie mogelijk boven
de 200 °C verhit zullen worden (bijvoorbeeld in ovengerechten) in verband met de verhoogde kans
op ongewenste pyrolytische esterificaties. Ook zou volgens de Commissie VNV betaïne niet
geschikt zijn om te worden verwerkt in levensmiddelen die een mogelijke bron van Lysteria
besmetting vormen (zoals zuivelproducten van rauwe melk), omdat bij osmotische of koude stress
deze bacteriën in voedsel worden beschermd door de aanwezigheid van betaïne.
      Betaïne wordt verkregen uit een extract van suikerbietmelasse waarbij voornamelijk
standaardtechnieken worden gebruikt. De Commissie VNV stemt in met de NFB dat de kwaliteit
van het industriële productieproces gewaarborgd is. Evenals de Finse beoordelaars accepteert de
Commissie VNV dat producten uit niet-genetisch gemodificeerde suikerbiet een lange
geschiedenis van gebruik kennen in de menselijke voeding. Suikerbietmelasse wordt beschouwd
als een veilige grondstof voor levensmiddelen.
Veiligheid en werkzaamheid
De Commissie VNV beoordeelt alleen de veiligheid en niet de werkzaamheid van het bio-actieve
ingrediënt betaïne. Volgens de aanvrager is een dagelijkse inneming van 4 gram betaïne ruim
voldoende om de serumhomocysteïnespiegel effectief te verlagen hetgeen gunstig zou zijn ter
voorkoming van hart en vaatziekten. Aangezien de aanvrager belang hecht aan de veronderstelde
gezondheidsbevorderende werking van betaïne wijst de Commissie VNV op het definitieve
voorstel van de Europese Commissie voor een Verordening inzake voedings- en
gezondheidsclaims voor levensmiddelen (EC03).
Bezoekadres                                                             Postadres
Parnassusplein 5                                                        Postbus 16052
2511 VX    Den Haag                                                     2500 BB    Den Haag
Telefoon (070) 340 7520                                                 Telefax (070) 340 7523
E-mail: gr@gr.nl                                                        www.gr.nl
                                                                                              9
</pre>

====================================================================== Einde pagina 9 =================================================================

<br><br>====================================================================== Pagina 10 ======================================================================

<pre>10</pre>

====================================================================== Einde pagina 10 =================================================================

<br><br>====================================================================== Pagina 11 ======================================================================

<pre>Gezondheidsraad                              Vice-voorzitter
Health Council of the Netherlands
Onderwerp          : Tweede beoordeling veiligheid Betaïne
Uw kenmerk         : VGB/VL 2407857
Ons kenmerk        : 2003/03VNV, U-1520/MR/cv/622-CM
Datum              : 23 oktober 2003
Veilige bovengrens van inneming
De Commissie VNV stemt in met de beoordeling van de NFB dat het dossier voldoende
voedingskundige informatie bevat over het nieuwe voedselingrediënt. De commissie wijst erop dat
betaïne onderdeel is van de normale methyleringcyclus in het menselijk lichaam; het is een
metaboliet van choline en fungeert als donor van een methylgroep. Blootstelling aan betaïne is niet
nieuw. Het is van nature aanwezig in bepaalde plantaardige en dierlijke producten en wordt ook
als technische hulpstof toegevoegd aan bepaalde levensmiddelen (voor details zie Bijlage D en E).
De dagelijkse inneming die de aanvrager voorstelt is zo’n 2 à 4 maal meer dan de hoeveelheid die
men via de gewone voeding binnenkrijgt. In de Verenigde Staten varieert dit van gemiddeld
ongeveer 1 g tot 2,5 g bij een seafood-rijke voeding.
      In de EU zijn sinds 1982 voedingsupplementen op de markt met betaïne in de vorm van het
hydrochloride zout, met een dagelijks aanbevolen hoeveelheid variërend van 7 tot 324 mg. De
NFB wijst erop dat gedocumenteerde gegevens over de gevolgen van het gebruik hiervan
ontbreken. Het is ook niet bekend in hoeverre ongewenste effecten kunnen optreden, als de
consumptie de dosering die de aanvrager voorstelt overschrijdt.
      In 1996 is betaïne is als weesgeneesmiddel∗ ter behandeling van homocystinuria, een
zeldzame stofwisselingziekte, toegelaten in Amerika door de Food and Drug Administration
(handelsnaam ’Cystadane’) en daarna is het ook in Canada en Australië geregistreerd. Als
medicijn is de dagelijks dosering gewoonlijk 6 g, maar kan oplopen tot 20 g. De Commissie VNV
ondersteunt de kritiek van de NFB dat het dossier geen informatie bevat over de
veiligheidsbeoordelingen bij deze registraties. In aanvulling op het dossier en het
beoordelingsrapport van de NFB meldt de Commissie VNV dat in de EU betaïne het predikaat
‘weesgeneesmiddel’ is toegekend in juli 2001, maar dat de autorisatieprocedure voor
markttoelating nog niet is afgerond (EU01). Op basis van productinformatie (Can03) constateert
de Commissie VNV dat de toxicologische gegevens die nodig waren voor de registraties als
weesgeneesmiddel (buiten Europa) beperkt zijn, zo hoefde er bijvoorbeeld geen
vruchtbaarheidsonderzoek en carcinogeniciteitsonderzoek bij proefdieren te worden uitgevoerd.
Ook is er geen farmacokinetisch onderzoek bij mensen gedaan.
Mensgebonden onderzoek. De Commissie VNV stelt dat de conclusie van de aanvrager dat
consumptie tot 30 g betaïne per dag extra geen gezondheidskundig of voedingskundig risico
∗
  geneesmiddelen voor zeldzame aandoeningen (zie ook Linthorst GR, Hollak CEM. Europese verordening
inzake weesgeneesmiddelen: kansen en bedreigingen. Ned Tijdschr Geneeskd 2003;147:143-145)
Bezoekadres                                                              Postadres
Parnassusplein 5                                                         Postbus 16052
2511 VX    Den Haag                                                      2500 BB    Den Haag
Telefoon (070) 340 7520                                                  Telefax (070) 340 7523
E-mail: gr@gr.nl                                                         www.gr.nl
                                                                                               11
</pre>

====================================================================== Einde pagina 11 =================================================================

<br><br>====================================================================== Pagina 12 ======================================================================

<pre>12</pre>

====================================================================== Einde pagina 12 =================================================================

<br><br>====================================================================== Pagina 13 ======================================================================

<pre>Gezondheidsraad                                Vice-voorzitter
Health Council of the Netherlands
Onderwerp          : Tweede beoordeling veiligheid Betaïne
Uw kenmerk         : VGB/VL 2407857
Ons kenmerk        : 2003/03VNV, U-1520/MR/cv/622-CM
Datum              : 23 oktober 2003
oplevert, onvoldoende is onderbouwd. De aanvrager heeft de resultaten besproken van 43
mensgebonden onderzoeken, waarvan er 36 bij verschillende typen patiënten zijn uitgevoerd. In 20
% hiervan is het effect van langdurige blootstelling (minstens één jaar) aan relatief hoge
doseringen betaïne (6 – 20 g per dag) bestudeerd. Alhoewel er geen acute klachten zijn
gerapporteerd, tekent de Commissie VNV aan dat ongewenste bijwerkingen niet systematisch in
kaart zijn gebracht in deze patiëntenonderzoeken.
     Van de zeven onderzoeken bij gezonde vrijwilligers is in de meeste gevallen het effect
onderzocht van een eenmalige blootstelling aan betaïne (2- 6 g), terwijl in twee onderzoeken een
blootstelling van 6 g per dag gedurende meerdere weken is geëvalueerd. De Commissie VNV
merkt op dat, alhoewel deze onderzoeken waren opgezet om de werkzaamheid te bestuderen, de
resultaten hiervan wel ondersteunende waarde hebben voor de veiligheidsbeoordeling van betaïne.
Bijlage 14 van het dossier bevat de resultaten van een placebo-gecontroleerd onderzoek bij 48
gezonde vrijwilligers met overgewicht, dat gerandomiseerd en dubbelblind parallel is uitgevoerd
(Sch02). Bij een blootstelling van 6 g per dag gedurende 12 weken traden er volgens de aanvrager
geen bijwerkingen op. In een ander onderzoek met 15 gezonde vrijwilligers die gedurende drie
weken dagelijks 6 g consumeerden werden eveneens geen ongewenste effecten waargenomen,
maar dit onderzoek was niet placebo-gecontroleerd uitgevoerd en heeft daarmee minder
bewijskracht.
     Alhoewel de Commissie VNV op grond van het beperkt klinisch onderzoek geen
aanwijzingen heeft dat er duidelijke klachten zullen optreden bij een dagelijkse inneming van ten
hoogste 6 g betaïne extra, is zij het niet eens met de conclusie van de aanvrager dat er geen
nadelige bijwerkingen zouden zijn. Onderzoekers hebben namelijk wel significante veranderingen
in het cholesterolmetabolisme ten gevolge van betaïne inneming waargenomen. Dit is beschreven
in verschillende onderzoeken die in het dossier zijn opgenomen, uitgevoerd met vrijwilligers
(Sch02) of met patiënten (Abd01, McG02), als ook in ander ongepubliceerd onderzoek waarvan de
commissie over de vertrouwelijke resultaten beschikt. De Commissie VNV concludeert dat de
verhouding ’totaal /HDL cholesterol’ in ongunstige zin verschuift, en beoordeelt dit als een
ongewenste bijwerking. Ook wijst zij erop dat dit het door de aanvrager beoogde positieve effect
van het nieuwe voedselingrediënt tegengaat. Om de veiligheid van betaïne voor de consument te
kunnen beoordelen, eist de Commissie VNV dat er uitgebreider onderzoek wordt gedaan bij
gezonde vrijwilligers met een zo hoog mogelijk verantwoorde dosis. Hierbij moeten, behalve de
verschillende leverenzymen, ook alle gangbare cardiovasculaire risicofactoren worden bestudeerd.
Bezoekadres                                                              Postadres
Parnassusplein 5                                                         Postbus 16052
2511 VX    Den Haag                                                      2500 BB    Den Haag
Telefoon (070) 340 7520                                                  Telefax (070) 340 7523
E-mail: gr@gr.nl                                                         www.gr.nl
                                                                                              13
</pre>

====================================================================== Einde pagina 13 =================================================================

<br><br>====================================================================== Pagina 14 ======================================================================

<pre>14</pre>

====================================================================== Einde pagina 14 =================================================================

<br><br>====================================================================== Pagina 15 ======================================================================

<pre>Gezondheidsraad                               Vice-voorzitter
Health Council of the Netherlands
Onderwerp          : Tweede beoordeling veiligheid Betaïne
Uw kenmerk         : VGB/VL 2407857
Ons kenmerk        : 2003/03VNV, U-1520/MR/cv/622-CM
Datum              : 23 oktober 2003
Proefdieronderzoek. Uit het dossier blijkt dat betaïne niet acuut toxisch is en niet mutageen in de
gebruikelijke onderzoeken naar genotoxiciteit. In subchronisch toxiciteitsonderzoek bij ratten, die
betaïne gedurende 90 dagen in het voer kregen toegediend, werden echter wel
behandelingsgerelateerde effecten waargenomen bij alle geteste doseringen waarvan de laagste 0,8
g/kg/dag en de hoogste 4,4 g/kg/dag was (overeenkomend met respectievelijk 56 g en 308 g bij
een lichaamsgewicht van 70 kg). Deze hematologische een hepatologische veranderingen zijn
volgens de aanvrager het gevolg van de onfysiologisch hoge blootstellingen waardoor de normale
evenwichten worden verstoord en zijn derhalve niet relevant bij gewone menselijke consumptie
die de aanvrager voorstelt. Uit de resultaten van een zogeheten reversibiliteitsonderzoek bij ratten,
met een gemiddelde dagelijkse dosis van ten hoogste 5,7 g/kg, concludeert de aanvrager dat de
door betaïne te weeg gebrachte effecten omkeerbaar zijn; de veranderingen in de lever zouden het
gevolg zijn van intermediair metabolisme. De Commissie VNV onderschrijft de mening van de
NFB dat, alhoewel de geïnduceerde effecten mild en reversibel lijken, het moeilijk is in te schatten
wat hiervan de betekenis zal zijn voor de gezondheid van de consument.
      Om deze metabole aspecten beter te onderzoeken zijn er op de Brandeis Universiteit in de
Verenigde Staten vervolgonderzoeken uitgevoerd met alleen vrouwelijke ratten die gedurende 28
en 90 dagen voer met betaïne kregen toegediend. Dit mechanistisch voedingsonderzoek, waarvan
de uitkomsten in het dossier zijn opgenomen, was voor wat betreft de rattenstam en het
betaïnegehalte van het voer vergelijkbaar met de voornoemde toxicologische onderzoeken. Ook
bleek dat de effecten die in de lever optraden vergelijkbaar waren, maar minder ernstig. De
onderzoekers schrijven dit verschil in effect toe aan verschillen in de samenstelling van de
gebruikte standaard rattenvoeren. De Commissie VNV acht dit een plausibele verklaring, maar
blijft van mening dat de waargenomen afwijkingen in de lever moeten worden toegeschreven aan
betaïne en als ongewenst moeten worden beschouwd. Zij stemt derhalve niet in met de wijze
waarop de aanvrager deze proefdierresultaten gebruikt om een voor de mens veilige bovengrens
van betaïne af te leiden.
      Samenvattend stelt de Commissie VNV dat er onvoldoende gegevens zijn om een veilig
niveau van inneming betrouwbaar te kunnen vaststellen. In het dossier ontbreekt bovendien een
duidelijk betoog, gebaseerd op het totaal aan resultaten van mensgebonden en
proefdieronderzoeken. De commissie meent dat vaststelling van een veilige bovengrens
noodzakelijk is gezien het feit dat betaïne een specifieke bio-actieve verbinding is en zal worden
toegevoegd aan producten die voor de algemene bevolking beschikbaar komen. Gebaseerd op het
huidige totaal aan beschikbare gegevens concludeert de Commissie VNV dat er vooralsnog
Bezoekadres                                                              Postadres
Parnassusplein 5                                                         Postbus 16052
2511 VX    Den Haag                                                      2500 BB     Den Haag
Telefoon (070) 340 7520                                                  Telefax (070) 340 7523
E-mail: gr@gr.nl                                                         www.gr.nl
                                                                                                15
</pre>

====================================================================== Einde pagina 15 =================================================================

<br><br>====================================================================== Pagina 16 ======================================================================

<pre>16</pre>

====================================================================== Einde pagina 16 =================================================================

<br><br>====================================================================== Pagina 17 ======================================================================

<pre>Gezondheidsraad                              Vice-voorzitter
Health Council of the Netherlands
Onderwerp          : Tweede beoordeling veiligheid Betaïne
Uw kenmerk         : VGB/VL 2407857
Ons kenmerk        : 2003/03VNV, U-1520/MR/cv/622-CM
Datum              : 23 oktober 2003
onvoldoende zekerheid is dat nadelige effecten op de gezondheid achterwege zullen blijven door
het gebruik van betaïne, waarbij de commissie anticipeert op een langdurige dagelijkse inneming
door voedingsbewuste consumenten.
Toepassing
In het licht van de hierboven genoemde beschouwing over een veilig niveau van betaïne inneming
concludeert de Commissie VNV dat het productassortiment, dat de aanvrager voorstelt, veel te
breed is, mede gezien het feit dat er geen resultaten beschikbaar zijn van chronisch
proefdieronderzoek die zo’n algemene toepassing rechtvaardigen. Door consumptie van meerdere
betaïne-houdende producten op één dag, overschrijdt de inneming de hoeveelheid die de aanvrager
aanbeveelt, te weten 4 g betaïne, en deze kan zelfs oplopen tot zo’n 8 à 12 g. De Commissie VNV
licht hieronder haar kantekeningen verder toe.
      Voor de levensmiddelen waaraan betaïne zou kunnen worden toegevoegd in concentraties
variërend van 2,0-6,7 %, stelt de aanvrager de volgende categorieën voor (zie paragraafnummer 29
in Bijlage E voor meer details):
     - dranken variërend van mineraalwater, frisdrank, koffie, thee tot alcoholische dranken
     - graanproducten zoals müsli, cornflakes, energierepen
     - zuivelproducten
     - snoepgoed
Voor elk van deze productcategorieën vermeldt het rapport van de eerste beoordeling het
gemiddelde en de 90 percentiel van de dagelijkse inneming. Op basis hiervan is berekend dat
mensen die op één dag producten uit alle categorieën consumeren (stapeling), respectievelijk 21 en
39 g betaïne zullen binnenkrijgen. Hiervoor zijn consumptiegegevens uit Finland, Zweden en
Denemarken gebruikt, aangevuld met informatie over inneming van zuivelproducten in het
Verenigd Koninkrijk, Duitsland, Spanje en Nederland. Het is de Commissie VNV echter niet
duidelijk hoe de verschillende sets van consumptiegegevens uit deze zeven Europese lidstaten zijn
gecombineerd om de innemingen af te leiden.
      De Commissie VNV constateert dat de consumptie van bepaalde producten, die met betaïne
verrijkt zullen gaan worden, zeer sterk uiteen kan lopen in de verschillende Europese lidstaten of
zelfs onbekend is. Zij stemt in met de NFB dat, mede ook vanwege nationale verschillen in de
samenstellingen van de geanalyseerde productcategorieën, de innemingsgegevens van betaïne niet
meer dan een globale indicatie zijn en benadrukt dat gemiddelden van de Europese innemingen
niet gerechtvaardigd zijn.
Bezoekadres                                                              Postadres
Parnassusplein 5                                                         Postbus 16052
2511 VX    Den Haag                                                      2500 BB   Den Haag
Telefoon (070) 340 7520                                                  Telefax (070) 340 7523
E-mail: gr@gr.nl                                                         www.gr.nl
                                                                                               17
</pre>

====================================================================== Einde pagina 17 =================================================================

<br><br>====================================================================== Pagina 18 ======================================================================

<pre>18</pre>

====================================================================== Einde pagina 18 =================================================================

<br><br>====================================================================== Pagina 19 ======================================================================

<pre>Gezondheidsraad                               Vice-voorzitter
Health Council of the Netherlands
Onderwerp           : Tweede beoordeling veiligheid Betaïne
Uw kenmerk          : VGB/VL 2407857
Ons kenmerk         : 2003/03VNV, U-1520/MR/cv/622-CM
Datum               : 23 oktober 2003
      De Commissie VNV meent dat dranken als categorie geheel zou moeten worden uitgesloten
gezien de zeer lastig in te schatten dagelijkse inneming. Omdat betaïne niet in gezonde kinderen is
onderzocht, vindt de Commissie VNV het niet acceptabel om snoepgoed te verrijken met betaïne.
      In aanvulling op de eerste beoordeling merkt de Commissie VNV nog op dat het de
verantwoordelijkheid is van de producent-en-aanvrager Finnfeeds om de eindfabrikanten van
betaïne verrijkte voedingsmiddelen te adviseren inzake het juiste gebruik en een goede etikettering
van het nieuwe ingrediënt. Bij onduidelijkheid hierover, of als de aanvrager dit niet accepteert, is
volgens de commissie deze firma niet de juiste aanvrager voor markttoelating van betaïne in
levensmiddelen. De Commissie VNV onderschrijft de mening van de NFB die nadere
etiketteringsvoorschriften nodig vindt, en voegt toe dat de consument geïnformeerd dient te
worden over hoeveel van de aanbevolen dagelijkse hoeveelheid betaïne één portie bevat. Dit alles
overwegende lijkt het de commissie volstrekt onduidelijk, hoe de aanvrager wordt geacht zicht te
houden op alle mogelijke toepassingen van het nieuwe ingrediënt.
      Op grond van bovenstaande argumenten maakt de commissie VNV bezwaar tegen het op de
markt brengen van betaïne als voedselingrediënt. Zij stelt twee mogelijkheden voor om een veilig
gebruik van betaïne door de consument te realiseren. De eerste mogelijkheid is een gelimiteerde
toepassing van betaïne in een beperkt en goed gedefinieerd productassortiment waarbij een veilige
bovengrens zal moeten worden vastgesteld op grond van de uitkomsten van adequaat klinisch
onderzoek dat nog uitgevoerd dient te worden. Met postmarketing onderzoek kan gecontroleerd
worden of de hoeveelheden betaïne die de consument dagelijks binnenkrijgt onder deze
bovengrens blijft. De tweede mogelijkheid is het bepalen van een ADI (acceptable daily intake of
wel ‘maximaal toegestane hoeveelheid’). Hiervoor is een zeer omvangrijk veiligheidsdossier
vereist, gelijk aan dat voor voedseladditieven, met de resultaten van onder andere onderzoek naar
reproductie, teratogeniteit, chronische toxiciteit en carcinogeniteit. Als deze ADI voldoende hoog
uitvalt zou een breder productassortiment mogelijk zijn.
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
E-mail: gr@gr.nl                                                          www.gr.nl
                                                                                               19
</pre>

====================================================================== Einde pagina 19 =================================================================

<br><br>====================================================================== Pagina 20 ======================================================================

<pre>20</pre>

====================================================================== Einde pagina 20 =================================================================

<br><br>====================================================================== Pagina 21 ======================================================================

<pre>Letter to the Dutch Minister of Health,
Welfare and Sport
On October 23, 2003, 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 has 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 second opinion, in accordance
with European Regulation 258/97, concerning betaine, the chemical name for which is
trimethylglycine. The applicant wishing to market this novel ingredient is the company
Finnfeeds Finland Ltd. Betaine will not be directly available for consumers, but will be
incorporated in foods of various categories. This assessment has been carried out by the
Committee on Safety Assessment of Novel Foods (VNV Committee) of the Health
Council of the Netherlands.
    The initial assessment of the application for market introduction was carried out in
Finland by the Novel Food Board (NFB). The major conclusion of the NFB is that, in
spite of the fact that the applicant has provided extensive information about the effect of
betaine in certain patients, data from scientific studies with healthy subjects is limited.
The NFB therefore states that a reliable assessment of consumer safety is not possible.
    The VNV Committee objects to market authorisation of betaine as a food
ingredient. The Committee bases its opinion on the report of the initial assessment by
the NFB (see Annex E), the information in the dossier (for a summary, see Annex D)
and the scientific literature. The VNV Committee agrees in part with the assessment
from Finland and has itself a number of criticisms of the dossier. It is not always clear to
Letter to the Dutch Minister of Health, Welfare and Sport                                    21
</pre>

====================================================================== Einde pagina 21 =================================================================

<br><br>====================================================================== Pagina 22 ======================================================================

<pre>   the VNV Committee when the comments made by the NFB should be considered
   essential to the assessment. The VNV Committee’s main criticism of the safety dossier
   is that the safe upper intake limit is inadequately supported. This criticism is based on
   the lack of adequate studies in healthy subjects and because adverse metabolic effects
   have been observed in laboratory animals. Considering these uncertainties, the VNV
   Committee also expresses a negative opinion on the wide range of products that the
   applicant is proposing. The text below sets out the assessment of the VNV Committee in
   greater detail.
   General product information
   The three products the applicant wishes to market, namely Betafin BF20, AP and
   TMG20, consist of pure betaine (at least 99%) and contain no undesirable ingredients or
   microbiological impurities. The VNV Committee agrees with the NFB that the product
   information in the dossier is representative for the betaine food ingredients to be marke-
   ted. This information is adequate as a basis for the safety assessment.
        Given the stability of betaine, the Committee VNV notes that the new ingredient
   should not be added to foods that may be heated at home to temperatures above 200 °C
   (oven dishes, for example) because of the increased possibility of undesirable pyrolytic
   esterification. Nor does the VNV Committee consider betaine suitable for use in foods
   that are a potential source of Lysteria contamination (such as dairy products made from
   untreated milk), because under osmotic or cold stress these bacteria in food are being
   protected by the presence of betaine.
        Betaine is obtained from an extract of sugar beet molasses, mainly using standard
   techniques. The VNV Committee agrees with the NFB that the safeguards for the qua-
   lity of the industrial production process are adequate. Like its Finnish colleagues, the
   VNV Committee accepts that products from genetically-unmodified sugar beet have a
   long history of use in human food. Sugar beet molasses is considered to be a safe raw
   material for foods.
   Safety and efficacy
   The VNV Committee has only assessed the safety, and not the efficacy, of the bio-active
   ingredient betaine. According to the applicant, a daily intake of 4 grams of betaine is
   sufficient for the effective decrease of blood homocysteine concentrations. It is claimed
   that this prevents cardiovascular disease. Given the fact that the applicant considers the
   assumed beneficial effects on health of betaine to be important, the VNV Committee
   refers to the definitive proposal from the European Commission for a Regulation on
   nutrition and health claims made on foods (EC03).
22 Betaïne/Betaine
</pre>

====================================================================== Einde pagina 22 =================================================================

<br><br>====================================================================== Pagina 23 ======================================================================

<pre>  Safe upper intake limit
  The VNV Committee agrees with the assessment of the NFB that the dossier contains
  enough nutritional information on the new food ingredient. The Committee wishes to
  point out that betaine is part of the normal methylation cycle in the human body; it is a
  metabolite of choline and serves as a methyl group donor. Exposure to betaine is not
  new. It is naturally present in certain vegetable and animal products and is also used as a
  technical additive in certain foods (for details, see Annexes D and E). The daily intake
  proposed by the applicant is approximately two to four times more than the intake from
  a normal diet. In the US, the latter ranges from an average of 1.0 g to 2.5 g for a high
  seafood diet.
       In the EU, food supplements with betaine in the form of the hydrochloride salt have
  been on the market since 1982, with a daily recommended dose level varying from 7 to
  324 mg. The NFB points out that there is no documented data about the consequences of
  using these supplements. It is also not known to what extent undesirable effects may
  occur if consumption exceeds the dose proposed by the applicant.
       In 1996, betaine was approved in America by the Food and Drug Administration as
  an orphan drug* (trade name 'Cystadane') for the treatment of homocystinuria, a rare
  metabolic disorder. It was also registered later in Canada and Australia. As a medicine,
  the daily dose is usually 6 g, but it can increase to 20 g. The VNV Committee supports
  the criticism of the NFB that the dossier does not contain information about the safety
  assessments made during these registration procedures. In addition to the information
  given in the dossier and the assessment report of the NFB, the VNV Committee reports
  that, in the EU, betaine was designated as an orphan medicine in July 2001, but the
  market authorisation procedure is still in progress (EU01). On the basis of product
  information (Can03) the VNV Committee notes that the toxicological data required for
  the registration as an orphan drug (outside Europe) is limited. For example, no
  reproduction or carcinogenicity studies were required in laboratory animals. Nor were
  there any human pharmacokinetic studies.
       Clinical research. The VNV Committee finds there is no adequate justification of
  the conclusion of the applicant that consumption of up to 30 g betaine extra a day
  involves no health or nutritional risk. The applicant discusses the results of 43 clinical
  studies, 36 of which were conducted in different types of patients. One fifth studied the
  effect of chronic exposure (lasting at least one year) to relatively high doses of betaine
  (6 – 20 g a day). Although no acute symptoms were reported, the VNV Committee notes
  that undesirable side-effects were not systematically explored in these patient studies.
       Most of the seven studies with healthy volunteers examined the effect of a single
  exposure to betaine (2 - 6 g), whereas two studies evaluated exposure of 6 g a day during
* Medicines for rare disorders
  Letter to the Dutch Minister of Health, Welfare and Sport                                   23
</pre>

====================================================================== Einde pagina 23 =================================================================

<br><br>====================================================================== Pagina 24 ======================================================================

<pre>   a period of several weeks. The VNV Committee notes that, although these studies were
   designed to test efficacy, the results can be used as supporting material for the safety
   assessment of betaine. Annex 14 of the dossier contains the results of a placebo-
   controlled, randomised and double-blind study with 48 obese subjects (Sch02).
   According to the applicant, there were no side-effects at an exposure of 6 g a day for 12
   weeks. Also in another study with 15 healthy volunteers who consumed 6 g daily for
   three weeks no adverse effects occurred. However, there was no placebo control group
   in the latter study and this undermined the power of the study.
        Although the VNV Committee has found no indications on the basis of the limited
   clinical research that clear symptoms will result from a maximum daily intake of an
   added 6 g betaine, it does not agree with the conclusion of the applicant that there are no
   harmful side-effects. Researchers do have observed significant changes in cholesterol
   metabolism as a result of betaine intake. This has been reported not only in various
   studies included in the dossier that were conducted with volunteers (Sch02) and with
   patients (Abd01, McG02), but also in another study, the unpublished results of which
   have been made available to the Committee on a confidential basis. The VNV
   Committee concludes that the ratio 'total to HDL cholesterol' is affected adversely, and
   considers this to be a harmful side-effect. It also points out that this is opposed to the
   beneficial effect that the applicant has in mind. In order to be able to assess the safety of
   betaine for consumers, the VNV Committee demands more extensive studies with
   healthy volunteers using a dose that is as high as responsibly possible. In addition,
   alongside the various liver enzymes, all the usual cardiovascular risk factors should be
   studied.
        Animal studies. The dossier shows that betaine is not acutely toxic or mutagenic in
   the usual tests for genotoxicity. In a subchronic toxicity study with rats given betaine in
   feed for 90 days, however, treatment-related effects were observed at all tested doses,
   the lowest of which was 0.8 g/kg/day and the highest of which was 4.4 g/kg/day
   (corresponding to 56 g and 308 g respectively at a body weight of 70 kg). These
   hematological and hepatological changes are, according to the applicant, the result of
   the unphysiologically high level of exposure, resulting in disturbances of normal
   balances so that they are unlikely to be relevant to normal human intake. On the basis of
   the results of a reversibility study in rats, with a maximum average daily dose of
   5.7 g/kg, the applicant concludes that the effects induced by betaine are reversible; the
   changes in the liver are thought to be caused by intermediary metabolism. The VNV
   Committee shares the opinion of the NFB that, although the induced effects appear to be
   mild and reversible, it is difficult to assess their significance for consumer health.
        In order to study these metabolic issues better, the Brandeis University in the United
   States has conducted follow-up trials in which female rats alone were given feed with
   betaine for 28 and 90 days. This mechanistic feeding study, the results of which have
24 Betaïne/Betaine
</pre>

====================================================================== Einde pagina 24 =================================================================

<br><br>====================================================================== Pagina 25 ======================================================================

<pre>been included in the dossier, was comparable in terms of the rat strain and the betaine
levels in the feed with the toxicological studies referred to above. The effects on the
liver were comparable but less severe. The researchers ascribe this variance to
differences in the composition of the standard rat feeds used. The VNV Committee
considers this a plausible explanation, but remains of the opinion that the observed
disorders in the liver must be ascribed to betaine and considered to be adverse. It does
not therefore agree with the way in which the applicant uses these animal results to
derive an upper limit for betaine that is safe for humans.
     In summary, the VNV Committee finds that there is inadequate data for determining
a safe level of intake in a reliable way. Furthermore, the dossier does not contain any
clearly argued reasoning that is based on the entire body of data from human and animal
studies. The Committee is of the opinion that a safe upper limit is necessary in view of
the fact that betaine is a specific bio-active compound and will be added to products that
will be available to the general population. On the basis of all the currently available data,
the VNV Committee concludes that, for the time being, it cannot be stated with certainty
that there will be no adverse health effects as a result of the use of betaine. Here, the
Committee anticipates chronic daily intake by nutritionally conscious consumers.
Application
Given the risk analysis referred to above with respect to a safe level of betaine intake,
the VNV Committee concludes that the product range proposed by the applicant is much
too broad, partly given the fact that no results are available from chronic animal studies
that justify a general application of this kind. The consumption of several products con-
taining betaine in a single day would result in intake exceeding the amount recommen-
ded by the applicant, in other words 4 g betaine, and even rising to approximately 8 à 12
g. The VNV Committee discusses its reservations in greater detail below.
     For the foodstuffs to which betaine would be added in concentrations varying from
2.0-6.7 %, the following categories are proposed (see paragraph 29 in Annex E for more
details):
     - beverages varying from mineral water, soft drinks, coffee and tea, to alcoholic
          beverages;
     - cereal products such as muesli, corn flakes and energy bars;
     - dairy products;
     - sweets.
For each of these product categories, the report of the first assessment states the average
and the 90 percentile for the daily intake. On this basis, it is calculated that people who
consume products from all categories in a single day, will ingest 21 and 39 g of betaine
respectively (cumulative intake). This calculation uses consumption data from Finland,
Sweden and Denmark, supplemented by information about the intake of dairy products
Letter to the Dutch Minister of Health, Welfare and Sport                                      25
</pre>

====================================================================== Einde pagina 25 =================================================================

<br><br>====================================================================== Pagina 26 ======================================================================

<pre>   in the United Kingdom, Germany, Spain and the Netherlands. However, it is unclear to
   the VNV Committee how the various sets of consumption data from these seven Euro-
   pean member states were combined to derive the intakes.
        The VNV Committee notes that the consumption of certain products that will be
   enriched with betaine can vary greatly in the various European member states or is even
   unknown. It agrees with the NFB that, in part because of national differences in the
   composition of the analysed product categories, the intake data for betaine is no more
   than a general indication and emphasises that averages for European intake levels are
   not justified.
        The VNV Committee is of the opinion that beverages should be excluded altogether
   as a category in view of the extreme difficulty of estimating daily intake. Because
   betaine has not been studied in healthy children, the VNV Committee does not consider
   the enrichment of sweets with betaine to be acceptable.
        In addition to the initial assessment, the VNV Committee notes that the producer/
   applicant Finnfeeds ought to be responsible for advising end-producers of betaine-
   enriched food about the correct use and the proper labelling of the new ingredient. If
   there are any ambiguities in this respect, or if the applicant does not accept this
   condition, the Committee believes that this company is not the right applicant for market
   authorisation for betaine in foodstuffs. The VNV Committee shares the opinion of the
   NFB that more detailed labelling conditions are necessary and adds that the consumer
   should be informed about how much of the recommended daily amount of betaine is
   contained in one portion. Given all this, it is by no means clear to the Committee how
   the applicant can be expected to monitor all possible uses of the new ingredient.
        Based on the arguments outlined above, the VNV Committee objects to introduction
   of betaine as a food ingredient on the European market. It proposes two options to
   realize a safe use of betaine by the consumer. The first option is a limited application of
   betaine in a restricted and well-defined product range, for which a safe upper limit needs
   to be determined on the basis of the results of adequate clinical research to be conducted
   in the future. Post-marketing research can be used to establish whether the daily intake
   of betaine by consumers remains below this upper limit. The second option is to
   determine an acceptable daily intake (ADI). This requires a highly detailed safety
   dossier on the lines of dossiers for food additives, containing the results of, amongst
   other, studies into reproduction, teratogenicity, chronic toxicity and carcinogenicity. If
   this ADI turns out to be sufficiently high, betaine might be incorporated in a wide range
   of products.
   I endorse the conclusions and recommendations of the VNV Committee,
   (signed) professor JGAJ Hautvast
26 Betaïne/Betaine
</pre>

====================================================================== Einde pagina 26 =================================================================

<br><br>====================================================================== Pagina 27 ======================================================================

<pre>      Literatuur/Literature
Abd01 Abdelmalek MF, Angulo P, Jorgensen RA, Sylvestre PB, Lindor KD. Betaine, a promising new agent for
      patients with nonalcoholic steatohepatitis: Results of a pilot study. Am J Gastoenterol 2001; 96: 2711-2717.
Can03 Canadian Product Monograph. Website http://www.orphan.com/Download/
      Canada_PRODUCT_MONOGRAPH_Eng.doc. Australian Consumer Medicine Information. Website http:/
      / www.orphan.com.au/Cystadane_CMI.html (consulted on 8 October 2003).
EC03  Voorstel voor een verordening van het Europese Parlement en de Raad inzake voedings- en
      gezondheidsclaims voor levensmiddelen. Website http://europa.eu.int/eur-lex/nl/com/pdf/2003/
      com2003_0424nl01.pdf, geraadpleegd op 8 oktober 2003
      (Proposal for a Regulation of the European Parliament and of the Council on nutrition and health claims
      made on foods. Website http://europa.eu.int/eur-lex/en/com/pdf/2003/com2003_0424en01.pdf, consulted
      on 8 October 2003).
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 1997; L43: 1-6)
EG97a 97/618/EG: Aanbeveling 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. Publicatieblad van de Europese Gemeenschappen 1997; L253: 1-36
      (97/618/EC: Commission Recommendation of 29 July 1997 concerning the scientific aspects and the
      Literatuur/Literature                                                                                        27
</pre>

====================================================================== Einde pagina 27 =================================================================

<br><br>====================================================================== Pagina 28 ======================================================================

<pre>       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 1997; L253: 1-36)
EU01   Designation of Betaine anhydrous as orphan drug. Website http://pharmacos.eudra.org/F2/register/
       alforphreg.htm (consulted on 8 October 2003; press release www.emea.eu.int/pdfs/human/comp/
       028601en.pdf).
FAO96  Biotechnology and Food Safety. Report of a joint FAO/WHO Consultation. Rome, FAO 1996.
FAO01  Evaluation of allergenicity of genetically modified foods. Report of a joint FAO/WHO expert consultation
       on allergenicity of foods derived from biotechnology. Rome, FAO 2001.
GR92   Commissie Toxicologische aspecten van biotechnologisch bereide producten. Productveiligheid bij nieuwe
       biotechnologie. Den Haag, Gezondheidsraad 1992, publicatienummer 1992/03.
McG02  McGregor DO, Dellow WJ, Robson RA, Lever M, George PM, Chambers ST. Betaine supplementation
       decreases post-methionine hyperhomocysteinemia in chronic renal failure. Kidney Int 2002; 61:1040-1046.
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.
OECD98 Report of the OECD workshop on the toxicological and nutritional testing of novel foods. Paris, OECD
       1998.
OECD00 Report of the task force for the safety of novel foods and feeds. Paris, OECD 2000.
SCF99  Opinion concerning the scientific basis for determining whether food products, derived from genetically
       modified maize, could be included in a list of food products which do not require labelling because they do
       not contain (detectable) traces of DNA or protein. Brussels, Scientific Committee on Food of the EU 1999.
Sch02  Schwab U, Torronen A, Toppinen L, Alfthan G, Saarinen M, Aro A, Uusitupa M. Betaine supplementation
       decreases plasma homocysteine concentrations but does not affect body weight, body composition, or
       resting energy expenditure in human subjects. Am J Clin Nutr. 2002; 76: 961-967.
SSC99  Opinion of the Scientific Steering Committee on microbial resistance, Brussels, Scientific Steering
       Committee of the EU 1999.
WHO91  Strategies for assessing the safety of foods produced by biotechnology. Report of a joint FAO/WHO
       Consultation. Geneva, WHO 1991.
WHO00  Safety aspects of genetically modified foods of plant origin. Report of a joint FAO/WHO expert
       consultation on foods derived from biotechnology. Geneva, WHO 2000.
28     Betaïne/Betaine
</pre>

====================================================================== Einde pagina 28 =================================================================

<br><br>====================================================================== Pagina 29 ======================================================================

<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
                                                                29
</pre>

====================================================================== Einde pagina 29 =================================================================

<br><br>====================================================================== Pagina 30 ======================================================================

<pre>30 Betaïne/Betaine</pre>

====================================================================== Einde pagina 30 =================================================================

<br><br>====================================================================== Pagina 31 ======================================================================

<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 voedings-
        middelen 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                                                                           31
</pre>

====================================================================== Einde pagina 31 =================================================================

<br><br>====================================================================== Pagina 32 ======================================================================

<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
32 Betaïne/Betaine
</pre>

====================================================================== Einde pagina 32 =================================================================

<br><br>====================================================================== Pagina 33 ======================================================================

<pre>Bijlage B
        De commissie/The committee
        •  Prof. dr LM Schoonhoven, voorzitter/chairman
           emeritus hoogleraar entomologie; Wageningen Universiteit en Researchcentrum/
           emeritus professor of entomology; 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 Control 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
           hoogleraar statistiek; Technische Universiteit Eindhoven/
           professor of statistics; Technical University Eindhoven
        •  Dr FM Nagengast
           gastro-enteroloog; Academisch Ziekenhuis Nijmegen/
           gastro-enterologist; Academic Hospital Nijmegen
        De commissie/The committee                                                       33
</pre>

====================================================================== Einde pagina 33 =================================================================

<br><br>====================================================================== Pagina 34 ======================================================================

<pre>   •   Dr ir JMA van Raaij
       voedingsfysioloog; Wageningen Universiteit and Researchcentrum/
       food physiologist; Wageningen University and Research centre
   •   Prof. dr ir G Schaafsma
       hoogleraar voeding; TNO Voeding, Zeist/
       professor of nutrition; TNO Nutrition and Food Research, Zeist
   •   Prof. dr EG Schouten, adviseur/advisor
       hoogleraar epidemiologie; Wageningen Universiteit and Researchcentrum/
       professor of epidemiology; Wageningen University and Research centre
   •   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, adviseur/advisor
       toxicoloog; TNO Voeding, Zeist/
       toxicologist; TNO Nutrition and Food Research, Zeist
   •   Dr ir F van der Wilk, adviseur/advisor
       COGEM, Bilthoven/
       Committee on Genetic Modification, Bilthoven
   •   Dr ir M Rutgers, secretaris/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.
34 Betaïne/Betaine
</pre>

====================================================================== Einde pagina 34 =================================================================

<br><br>====================================================================== Pagina 35 ======================================================================

<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 (FAO01, OECD98, OECD00, SCF99,
        SSC99, WHO00).
            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,
        EU-procedure/EU-procedure                                                                35
</pre>

====================================================================== Einde pagina 35 =================================================================

<br><br>====================================================================== Pagina 36 ======================================================================

<pre>   Welzijn en Sport. Zij heeft de Gezondheidsraad verzocht haar van advies te dienen. De
   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 Commissie naar het Wetenschappelijk Comité voor de menselijke voeding.
   Komt men dan nog niet tot overeenstemming dan beslist de Europese Ministerraad.
   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 (FAO01, OECD98, OECD00, SCF99, SSC99,
   WHO00).
        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.
36 Betaïne/Betaine
</pre>

====================================================================== Einde pagina 36 =================================================================

<br><br>====================================================================== Pagina 37 ======================================================================

<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 Scientific Committee on
Food for advice. If consensus still cannot be reached, the issue is referred to the Euro-
pean Council of Ministers.
EU-procedure/EU-procedure                                                                     37
</pre>

====================================================================== Einde pagina 37 =================================================================

<br><br>====================================================================== Pagina 38 ======================================================================

<pre>38 Betaïne/Betaine</pre>

====================================================================== Einde pagina 38 =================================================================

<br><br>====================================================================== Pagina 39 ======================================================================

<pre>Bijlage D
        Samenvatting van het dossier/Executive
        summary of the dossier
        Samenvatting van het dossier/Executive summary of the dossier 39
</pre>

====================================================================== Einde pagina 39 =================================================================

<br><br>====================================================================== Pagina 40 ======================================================================

<pre>40 Betaïne/Betaine</pre>

====================================================================== Einde pagina 40 =================================================================

<br><br>====================================================================== Pagina 41 ======================================================================

<pre>Finnfeeds Finland Ltd / Novel Food Application 15.1.2003
Summary by applicant

SUMMARY BY APPLICANT

(for the further circulation to the Member States of European Union)

An application to place betaine as a food ingredient
on the EU Novel Food Market
</pre>

====================================================================== Einde pagina 41 =================================================================

<br><br>====================================================================== Pagina 42 ======================================================================

<pre>Finnfeeds Finland Ltd / Novel Food Application 15.1.2003
Summary by applicant

TABLE OF CONTENTS

1 INTRODUCTION …… … uns onnnn on ennvensenssnnrnnenansenenenesensnerserssnrennenrenmensenn
2 SCIENTIFIC CLASSIFICATION noa venne nnenerenersnarensrsenserrsnssnensenmenuner
3 DESCRIPTION OF BETAINE avan oaeen nn enenerernrsnersansennsnnennnensnenmensnen
4 PRODUCTION PROCESS OF BETAINE. .......:cseceeceesseseeeneeneweueeuwaneenemenenrens
5 ANTICIPATED USE OF BETAINE … … „aon eenen enennenennnnneneerenseersenae
6 PURPOSE OF BETAINE ENRICHMENT ............... nn onsen nerenersensenenmennaerenn
7 SAFETY ASPECTS CONCERNING BETAINE. … „nanne eneen
TA General os nenenenneneverrenenssnnvenansnenennennnnnrnersnenmannnnannsnnenennnenn
7.2 Nutritional information ….….….sassssnnnenenes renee ermee rnsennnnnsrersnsensensensrsenn
7.3 Microbiological information … uus urn sans nennenensensenrsnnnnsensenssensnennsenn
7.4 Toxicological information … sussen ans aneersenssnnnensnnens eran
8 CONCLUSION … anar eneneeenene menens nervensenrmentenennensenrensnnseneernserseen

ODD ANNP A & &

APPENDICES

Appendix 1 Table of contents of the original novel food application

Appendix 2 Product specifications of Betafin BF 20, TMG 20 and Betafin
AP

Appendix 3 List of references of an original novel food application
</pre>

====================================================================== Einde pagina 42 =================================================================

<br><br>====================================================================== Pagina 43 ======================================================================

<pre>Finnfeeds Finland Ltd / Novel Food Application 15.1.2003
Summary by applicant

1 Introduction

The novel food application on which this summary is based, has been made to
place the novel food ingredient, betaine, both in anhydrous and monohydrate form,
to be used in beverages, confectionary, cereal and dairy products on the EU novel
food market.

Betaine (chemical name: 1-Carboxy-N,N,N-trimethylmethanaminium hydroxide inner salt)
is categorised as novel food ingredient because it has not previously been used for human

consumption to a significant degree within the European Community.

Betaine, also known as trimethylglycine or glycine betaine, is a quarternary amine, closely
related to the amino acid glycine. It is naturally present at varying concentrations, in most
living organisms, including sugar beet and other plants belonging to the Chenopodiaceae
family. Naturally oecurring betaine concentrations in plants and animals generally vary
along with growing and osmotic stress conditions. Betaine has been found at relatively
high concentrations in common foods and/or foodstuffs such as cereals (e.g. wheat, oats),

seafood, clams, vegetables and common edible mushrooms.

Betaine was first isolated in 1866, from sugar beet (Beta vulgaris) juice concentrate. Until
1960, betaine was used mainly in pharmaceutical applications. Today, it is widely used in
dietary supplements, dental! products, cosmetics, for fermentation purposes as well as in
animal feed. In these applications, betaine is used to serve a technical function such as

moisture control and/or for its nutritional benefits, particularly as a methyl donor.

In recent years, betaine has been actively investigated for its health-promoting potential. In
clinical studies it has been shown that betaine successfully lowers serum elevated
homocysteine levels in humans. High serum homocysteine levels have been shown to be
a risk marker to cardiovascular disease and thus, betaine-enriched foods may help to
lower the risk of cardiovascular disease. Other investigators have studied betaine
metabolism, including its role in the methionine cycle and betaine’s potential to protect the

liver from ethanol induced liver injury and non-alcoholic steatohepatitis. To date, there is a
</pre>

====================================================================== Einde pagina 43 =================================================================

<br><br>====================================================================== Pagina 44 ======================================================================

<pre>Finnfeeds Finland Ltd / Novel Food Application 15.1.2003
Summary by applicant

wealth of human clinical studies discussing the benefits of betaine supplementation at

dose levels ranging from 2 to 30 g/day; 6 g/day being the most common dose.

The table of contents of the original application is presented in appendix 1.

2 Scientific classification

According to European Commission's recommendations on the Assessment of Novel

foods (Part |), the novel food ingredient, betaine, is classified under

Class 1: Pure Chemicals or simple mixtures from non-GM sources.
Sub class 1.1: | The sources of the novel foods have a history of food use in the

Community.

According to the categorisation in the EC Novel Foods and Novel Food Ingredients
Regulation 258/97, the present novel ingredient falls into the following category:

e) foods and food ingredients consisting of or isolated from plants and food
ingredients isolated from animals, except for foods and food ingredients obtained
by traditional propagating or breeding practices and having a history of safe food
use.

3 Description of betaine

Betaine is 99 % pure compound of which chemical structure is presented below.
Specifications of betaine (Betafin BF 20, TMG 20 and Betafin AP) are presented in
appendix 2. Betaine as such does not have a traditional counterpart. The nutritional value
of betaine is comparable to amino acids, which are parts of proteins. Betaine is derived

from non-genetically modified source.
</pre>

====================================================================== Einde pagina 44 =================================================================

<br><br>====================================================================== Pagina 45 ======================================================================

<pre>Finnfeeds Finland Ltd / Novel Food Application 15.1.2003
Summary by applicant

Betaine: (CH3)aN*CH2COO' (CH3)aN*CH2C00° x H20

Anhydrous betaine Betaine monohydrate

Anhydrous betaine and betaine monohydrate have similar chemical structure except water
(H2O) molecule which is attached into the carbonyl group in monohydrate form.
Physicochemical characteristics of anhydrous betaine and betaine monohydrate are
similar. Anhydrous betaine is gradually converted to betaine monohydrate when the
relative humidity of environment is more than 7 %. Both betaine forms dissolve in water
when the relative humidity is more than 46 %. Nutritional, microbiological and toxicological
properties of anhydrous betaine and betaine monohydrate are equal. In the present

application, word ‘betaine’ means both betaine anhydrous and monohydrate form.

4 Production process of betaine

Sugar beet is containing 0.2 - 0.3 % betaine. Extraction of betaine from sugar beet can be
divided into three main stages — production of sugar beet molasses from sugar beet,
extraction of betaine rich fraction from sugar beet molasses and finally refining and
crystallisation of betaine rich fraction to crystalline betaine. Betaine is extracted from sugar
beet molasses by liquid chromatography method. This method is generally well known and
the same method is used for example in fructose or xylitol production. Food grade betaine
has been produced since 1983 to be sold in Japan and Korea by using this process.
Betaine marketed by Finnfeeds Finland Ltd is extracted at Naantali, Finland, from sugar
beet based concentrated liquids of betaine.
</pre>

====================================================================== Einde pagina 45 =================================================================

<br><br>====================================================================== Pagina 46 ======================================================================

<pre>Finnfeeds Finland Ltd / Novel Food Application 15.1.2003
Summary by applicant

5 Anticipated use of betaine

Betaine will be used as an ingredient in beverages, confectionary, cereal and dairy
products. These foods are produced by using the traditional ingredients except the
enrichment with betaine. Proposed new uses for betaine in foods and maximum
application levels are as follows:

- beverages, max 2,0 %

- confectionary, max 6,7 %

- cereal products, max 4,0 %

- dairy products, max 2,7%.

Betaine does not replace any particular ingredient of foods, but it replaces evenly all other
ingredients. Betaine enriched foods are manufactured as traditional foods. According to
study results, betaine is a stabile ingredient in food processing and no decomposition

happens.

Anticipated intake of betaine from the previous-mentioned betaine-enriched foods is 4
grams per day. The recommendations on package labelling of betaine-enriched foods will
advise to consume betaine-enriched products so that not more than 4 g betaine will be
obtained daily. This is because already 3 g of betaine has been shown to be effective to

decrease blood homocysteine concentrations.

6 Purpose of betaine enrichment

in clinical studies it has been shown that betaine successfully lowers serum elevated
homocysteine levels in humans. High serum homocysteine levels have been shown to be
a risk marker to cardiovascular disease and thus, betaine-enriched foods may help to
lower the risk of cardiovascular disease. Traditionally betaine has been used for
enhancing seafood flavourings, particularly crab meat substitutes. Betaine is also used as
a preservative for shrimps by lowering the water activity.
</pre>

====================================================================== Einde pagina 46 =================================================================

<br><br>====================================================================== Pagina 47 ======================================================================

<pre>Finnfeeds Finland Ltd / Novel Food Application 15.1.2003
Summary by applicant

7 Safety aspects concerning betaine

7.1 General

A safety evaluation of novel food ingredient betaine has been prepared according to the
European Commission’s Recommendations on the assessment of novel foods
(97/618/EY). In respect to betaine, based on the results of clinical studies presented
previously, nutritional impact of betaine and betaine-enriched foods happens mainly in
blood homocysteine concentrations, which decrease due to betaine enrichment. Based on
the results of several studies, betaine does not affect the bioavailability of nutrients from
the diet. No adverse effects except mild gastrointestinal symptoms have occurred in the
studies made with betaine or betaine-enriched foods. Microbiological tests show that
betaine does not cause any microbiological risks. Also toxicological tests of betaine show

that no safety concern is apparent. As an amino acid, betaine has no allergenic potential.

7.2 Nutritional information

Betaine is a normal constituent of human blood and urine and accumulates in kidney renal
medullary cells where it functions as a protective osmolyte. Betaine levels in the blood
appear to be homeostatically controlled and typical betaine plasma concentrations (20-60
umol/l) are relatively stable.

In the diet, betaine is obtained either directly from ingesting foods containing betaine or
indirectly via metabolism of foods containing choline, and choline derivatives such as
lecithin. Dietary betaine is absorbed via the small intestine into the enterocytes. It is then
released into the portal circulation and carried to the liver where significant first pass
extraction and metabolism of betaine occurs. Betaine is also metabolised in the kidneys. In
vivo, choline is largely converted to either acetylcholine and/or phospatidylcholine or
oxidized to betaine. According to kinetic studies, betaine is primarily catabolized in vivo,
urinary excretion being negligible.
</pre>

====================================================================== Einde pagina 47 =================================================================

<br><br>====================================================================== Pagina 48 ======================================================================

<pre>Finnfeeds Finland Ltd / Novel Food Application 15.1.2003
Summery by applicant

To date there are over 40 published human clinical studies (some of which comprise
several case studies on betaine); in total, more than 700 people have been studied. In
those studies, the effect of betaine to decrease the serum homocysteine concentrations
both in people with homocysteinemia and healthy people is reported. Reported betaine
dose levels range from 2 to 30 g per day; 6 g/day being the most common dose. The
period of betaine intake ranges from 1 day to 16 years. There has been no evidence of
organ toxicity, and adverse reactions to betaine have been minimal, including anecdotal
reports of nausea, diarrhoea and gastrointestinal distress. No other types of adverse
effects have been found.

7.3 Microbiological information

Betaine is manufactured under sanitary conditions and in conformance to health and
safety conditions established by the Finnish authorities. The applicant's facility is subjected
to inspections and monitoring by local health inspectors. HACCP and other internal quality
management programs have been established to ensure good manufacturing practice. In
addition, the facility has been ISO 9001 certified since September 1997.

The preservation tests of betaine-enriched beverages are being made and the results of

the tests will be attached into the present application later.

According to studies conducted by the applicant, betaine itself has some antimicrobial
effects when concentrated. As a hygroscopic product betaine tends to bind water
molecules from the surroundings. The antimicrobial effects are based on the fact that
betaine can lower the free water activity so that the growth of microbes is delayed or
inhibited.

7.4 Toxicological information

In the literature, the safety of betaine in rats has been reported in several studies. In

addition, the applicant has had toxicological studies made by expert research institutions:
short-term genotoxicity studies, acute toxicity study, A 14-day range finding study and a
</pre>

====================================================================== Einde pagina 48 =================================================================

<br><br>====================================================================== Pagina 49 ======================================================================

<pre>Finnfeeds Finland Ltd / Novel Food Application 15.1.2003
Summary by applicant

90-day sub-chronic toxicity study, 28-day sub-acute toxicity and reversibility study, 28-day
& 90-day metabolic feeding studies and sensitization / irritability studies. All studies have

been conducted in accordance with the Principles of Good Laboratory Practice (GLP).

To present one of the studies in which the NOAEL (no adverse effects level) was set: In
the 28-day and 90-day feeding studies in rats carried out with betaine as the test
substance, fifty 3-wk old Sprague-Dawley female rats were studied. As a result, no
significant adverse clinical effects were observed at any betaine dose level. Based on
modest perturbation of red blood cell physiology in growing animals and considerations for
protein metabolism, the NOAEL for betaine was set at 150 mg/g protein consumed by
growing rats fed a normal diet. The equivalent NOAEL for casually exposed adult humans
was determined to be 9-15 g betaine/day, based on the typical range of protein intake by
adult humans.

List of references of an original application is presented in appendix 3.

8 Conclusion

Based on literature, human clinical studies and toxicological tests, the novel food
ingredient betaine, is unlikely to give any rise to nutritional, microbiological or toxicological
problems. Therefore, it is considered safe for use at the anticipated consumption levels of
beverages, confectionary and cereal and dairy products as a part of the daily diet of
healthy people and especially people with risk of atherosclerosis.
</pre>

====================================================================== Einde pagina 49 =================================================================

<br><br>====================================================================== Pagina 50 ======================================================================

<pre>Finnfeeds Finland Oy / Novel food application summary 12/2002

summary:
APPENDIX 1

Table of contents of the original novel
food application
</pre>

====================================================================== Einde pagina 50 =================================================================

<br><br>====================================================================== Pagina 51 ======================================================================

<pre>Finnfeeds Finland Ltd / Novel food application 15.1.2003

TABLE OF CONTENTS

1 ADMINISTRATIVE DATA

vannnenurmeenenusverssenrsensenenenerensnnnnnnnnnenensernenrendsnnsnnd 5
2 SCIENTIFIC CLASSIFICATION sas sanan aren ennene nnen nnesensenenenennennensenennnenn 6
3 IDENTIFICATION OF ESSENTIAL INFORMATION FOR ASSESSMENT OF
WHOLESOMENESS … aa snaren ennenenenenenneuneennenrnennennneensenensnenenneennn 8
4 CONSULTATION OF STRUCTURED SCHEMES. … „asana one wenen 9
4.1 Scheme! : Specification of betaine … ns nnnsrronens ens ennnenrenennnensen 9
4.2 Scheme Il: Effect of the production process applied to betaine … … … 14
4.2.1 Production process of betaine ............... ssansnornsnr sense vennnrsenenneenenn 15
4.2.2 Stability of pure betaine „sns sun onenvenrsennveenvenrvaneranenenvennennnnnnne 17
4.2.3 Stability of betaine in foods ………..……ssannunensunnsesnonnrnannsnnnsrensenssnrnen 20
4.3 Scheme Ill: History of the organism used as the source of betaine .......... 21
4.4 Scheme IX: Anticipated intake of betaine ………….…...nseunrsnenssensananansenennansennn 24
4.4.1 Average intake of traditional foods … users snsanseenunnensenenenvenrnsnene 25
4.4.2 High intake of traditional foods „ns anansunnrunureunnenensersnensunsenven 26
4.4.3 Anticipated intake of betaine-enriched foods annen 27
4.4.4 Anticipated intake of betaine at risk groups ...............memanennnonnnane 29
4.4.5 Effects of betaine-enriched food intake on diet aaneen 30
4.5 Scheme X: Information from previous human exposure to betaine or it’s
SOUFCE …aanannannsnrnsrnnenrnenvensnnnsensnenvenennennnensunsvarnnennmendenerarornsnernennennrnnnnne 31
4.5.1 Betaine intake from diet ..............-.....::nerrmenunnsvunuonane nan nennnnnnrenen nos 32
4.5.2 Betaine use in dietary supplements in EU … un senn enso onnen enen aneen 33
4.5.3 Betaine use in dietary supplements and orphan drug in USA .......... 35
4.5.4 Other uses of betaine …….……...n-sne snor enunensennnen eneen samen senrsnrenennan ennen 35
4.5.5 Clinical studies with betaine-enriched foods oenen 35
4.6 Scheme XI: Nutritional information on betaine and betaine-enriched foods 37
4.6.1 General ……………….annanunennnennnnennensre enawensenrmennensenenenssenmansinnensnnnnnenen 38
4.6.2 Betaine metabolism/pharmacology … uns oasen senen ene senenenneenenven 38
4.6.3 Clinical studies with betaine in humans nn. sarennrsanensanennseerensen 41
4.6.4 Nutritional impact of betaine and betaine-enriched foods ............... 46
4.7 Scheme XII: Microbiological information on betaine …… ……….ansense uns eener 47
4.7.1 General nonnen nen reennren cn enenwennnrssnansssarnanvsnerenenrnennannnennnen 48
4.7.2 Preservation tests ……..….…...naserse enn ereennnnsenrnnenenn ennen enen nnenenseennnnen 48
4.7.3 Antimicrobial properties of betaine … … sues saseussennsenenneneneneensnsenven 48
4.8 Scheme XIII: Toxicological information on betaine .............,...nvenanunennnne 51
</pre>

====================================================================== Einde pagina 51 =================================================================

<br><br>====================================================================== Pagina 52 ======================================================================

<pre>Finnfeeds Finland Ltd / Novel food application 15.1.2003

4.8.1 Short-term genotoxicity studies ................»»»vnsnnnannnannnnnnannnannnnnnnnan 52
4.8.2 Acute toxicity in rats … ns. saar aaneen eenenrnenrennensnnsennennenenenrnenrenen 53
4.8.3 A 14-day range finding study and a 90-day sub-chronic toxicity study
in ratS „naaa nennnenenrerrenensensendenssnnnnsnrnanannrsenenmensnsvnnnsenenteneerse 54
4.8.4 28-day sub-acute toxicity and reversibility study in rats …… 56
4.8.5 28-day & 90-day metabolic feeding studies in rats ................nnnnsenvens 57
4.8.6 Sensitization /irritability studies …… … ….…..nsenssssarsenonnennennsensenensnnn enn 58
4.8.7 Allergenicity of betaine .............0:»:esnsnnnnninanannenennnnnnnanannnnnannan names 60
5 EVALUATION AND CONCLUSION BY THE APPLIGANT … „ananas nne manen eens 61
5.1 General considerations ………..….narsssunsnensrenenrernesnnrnuvensannnnnensenseneerrerenenn 61
5.2 Genetically modified organisms ass sns ars vensensasoneneanenmenannensnenrennenenn 61
5.3 Substantial equivalence … .….….…..ssarssanesunnnrnenennsnnnseereenen enen nerensnnnnennnnnennen 61
5.4 Compositional analysis … ross snnsonannnannsennennenveensennnensnnene senen nennnennenne 61
5.5 Anticipated intake ..........cccceccssccussenseueuesseneanereeneseecessasansnesaseremseananensnenie 61
5.6 Nutritional considerations affecting toxicological testing in animals .......... 62
5.7 Toxicological requirements … sens sansnnenrennennsnnnnnnnens enorm sene sannennenven nennen 62
5.8 Implications of novel foods for human nutrition … savannen verenvenenvenn 62
5.9 Novel microorganisms used in food ..............-s0evennnsnnnnensnennnennnanennnen ama nen 62
5.10 Allergenic potential vaas ss ors sen ne neenrenennenrennenenrsnnnenenennnenvensenenennnn 63
5.11 Assessment of marker genes … „sars unsnnunvsnsensenrvenenssrnsnnenrsseonrsnnnnrnssn 63
5.12 Conclusion … naren annanenunnansnenrsvensen eunnvennenveunsvernnnrnvennnsnanensnenrensnanen 63

REFERENCES

SUMMARY BY APPLICANT

APPENDICES

Appendix 1 a

Appendix 2
Appendix 3
Appendix 4
Appendix 5

Appendix 6
Appendix 7
Appendix 8
Appendix 9

Appendix 10
Appendix 11

ao

Product specification of Betafin BF 20 and certificates of
analyses

Product specification of TMG 20

Product specification of Betafin AP and certificates of analyses
Material safety data sheets of Betafin BF 20, TMG 20 and Betafin
AP

Analytical methods of betaine

Certificates of approvals ISO 14001:1996 and ISO 9001:1994
Kosher certificate

Statements and research reports: Concentrations of pesticides
and phenoxy herbisides and PCB and PAH compounds
Certificate of analysis: GC-analysis of isopropanol, ethanol and
methanol in betaine

Proposal for package labelling of betaine

Report: The stability of TMG 20

Report: Betaiinin hajoaminen kuumennettaessa (english
translation: Decomposition of betaine during heat treatment)
Certificate of analysis: Betaine content of cookies and brownies
Certificate of non-GMO
</pre>

====================================================================== Einde pagina 52 =================================================================

<br><br>====================================================================== Pagina 53 ======================================================================

<pre>Finnfeeds Finland Ltd / Novel food application

Appendix 12
Appendix 13
Appendix 14

Appendix 15

Appendix 16
Appendix 17
Appendix 18

Appendix 19

Appendix 20
Appendix 21
Appendix 22
Appendix 23
Appendix 24

Appendix 25

Betaine intake from natural sources

The list of human clinical studies with betaine

Final report: The effect of betaine supplementation on plasma
homocysteine concentrations, body weight, body composition
and resting energy expenditure in human subjects.
Amendment to final report: The effect of betaine supplementation
on plasma homocysteine concentrations, body weight, body
composition and resting energy expenditure in human subjects
Effect of betaine supplementation on hematology

- will be completed later. Study report: Dose-response study of
betaine and homocysteine

- will be completed later. Research report: preservation tests of
betaine-enriched beverages

Research report: Determination of microbiological stability of
betafin solutions

Short-term genotoxicity studies:

Bacterial reverse mutation assay

Mouse micronucleus test

Metaphase analysis of human lymphocytes

Acute oral toxicity study in the rats

Report: Validation of the determination of betaine in diet

Final report: A 14-day range finding study and a 90-day sub-
chronic toxicity study in the rat with betaine

Final report: A 28-day sub acute toxicity and reversibility study in
the rat with betaine

Study report: Metabolic aspects of betaine supplementation in
rats

Sensitization/irritability studies:

Human patch test for skin irritation

Sensitisation test in the guinea pig

Acute eye irritation study

15.71.2003
</pre>

====================================================================== Einde pagina 53 =================================================================

<br><br>====================================================================== Pagina 54 ======================================================================

<pre>Finnfeeds Finland Oy / Novel food application 12/2002
Summary by applicant

APPENDIX 2

Product specifications of Betafin BF 20,
TMG 20 and Betafin AP
</pre>

====================================================================== Einde pagina 54 =================================================================

<br><br>====================================================================== Pagina 55 ======================================================================

<pre>product specification
_BETAFIN BF 20

Betaine anhydrous food grade

C10A0

Formula
Molecular weight

Specification

Betaine (HPLC)
Moisture (Halogen drying)
Ash (conductometric)
Chloride (titration)
Sulphate (limit test)
Heavy metals (limit test)
Arsenic (ICP-MS)

Lead (ICP-MS)

Colour (ICUMSA)

pH (5 % solution)

Total viable count
Yeast

Mold

Coliforms

E-coli

Salmonella

Other properties
Appearance
Particle size
Bulk volume
Solubility at 25°C

Stability

Finnfeeds Finland Limited

Naantali Plant, Satamatie 1, FIN-21100 NAANTALI, Finland
Telephone +358 2 4393 300, Fax +358 2 4393 333
“Trade reg.no. 751.670, Domicile Helsinki, Business ID F11508198-2

(CH,),-N’-GH,-COO
117.15

min. 99 % d.s.
max. 2 % when packed
max. 0.1 %
max. 50 ppm
max. 100 ppm
max. 10 ppm .
max. 1 ppm
max. 1 ppm
max. 20

5-7

max. 100 cfu/g
max. 10 cfu/g
max. 10 cfu/g
not detected
not detected
not detected

‘DANISCO CULTOR

Free-flowing, white crystals, faint characteristic odour.

Typically 90% < 0,5 mm
Typically 0.6 - 0.8 kg/l

Water

Methanol
Ethanol

Heat stable up to 200°C

160 g/100 g HO
55 9/100 g MeOH
8.7 g/100 g EtOH

Hygroscopic, absorbs humidity from air
</pre>

====================================================================== Einde pagina 55 =================================================================

<br><br>====================================================================== Pagina 56 ======================================================================

<pre>Shelf life
Packaging

Storage

Finnfeeds Finland Limited

Naantali Plant, Satamatie 2, FIN-21 100 NAANTALI, Finland
Telephone +358 2 4393 300, Fax +358 2 4393 333

Trade reg.no. 751.670, Domicite Helsinki, Business ID F11508198-2

DANISCO CULTOR

3 years from the manufacturing date

20 kg cartons with polyethylene inner bag,
shrinkwrapped on one-way pallet. Standard pallet
1000 kg.

In unopened bags, protected from humidity
</pre>

====================================================================== Einde pagina 56 =================================================================

<br><br>====================================================================== Pagina 57 ======================================================================

<pre>- product specification

TMG 20

Betaine anhydrous food grade

C10B0

Formula
Molecular weight

Specification

Betaine (HPLC)
Moisture (Halogen drying)
Ash (conductometric)
Chloride (titration)
Sulphate (limit test)
Heavy metals (limit test)
Arsenic (ICP-MS)

Lead (ICP-MS)

Colour (ICUMSA)

pH (5 % solution)

Total viable count
Yeast

Mold

Other properties
Appearance
. Bulk volume
Solubility at 25°C

Stability
Shelf life
Packaging

(CH,),-N-CH,-COO ~
117.15

min. 99 % d.s.

max. 2 % when packed
max. 0.1 %

max. 50 ppm

max. 100 ppm

max. 10 ppm

max. 1 ppm

max. 1 ppm

max. 20

5-7

max. 100 cfu/g

max. 10 cfu/g ‘
max. 10 cfu/g

DANISCO

Free-flowing, white crystals, faint characteristic odour

0.5 - 0.6 kg/l |

Water . 160 9/100 g HO
Methanol 55 9/100 g MeOH
Ethanol 8.7 g/100 g EtOH

Hygroscopic, absorbs humidity from air

Heat stable up to 200°C

3 years from the manufacturing date

20 kg cartons with polyethylene inner bag,
shrinkwrapped on one-way pallet. Standard pallet

1000 kg.
</pre>

====================================================================== Einde pagina 57 =================================================================

<br><br>====================================================================== Pagina 58 ======================================================================

<pre>Storage In unopened bags, protected from humidity
</pre>

====================================================================== Einde pagina 58 =================================================================

<br><br>====================================================================== Pagina 59 ======================================================================

<pre>DANISCO
‘product description

BETAFIN AP

Betaine monohydrate pharmaceutical and food grade

C13N0

Formula
Molecular weight

Specitication
Betaine (HPLC)
Moisture (Halogen drying)

Ash (conductometric)

Chloride (titration)

Sulphate (limit test)

Heavy metals (limit test)

Arsenic (ICP-MS )
"Colour (IGUMSA)

pH (5 % solution)

Total viable count

Other properties
Appearance

Bulk volume

Betaine solubility at 25°C

Stability
Shelf life
Packaging

Storage

(CHa)g-N+-CHo-COO” x HzO
135.16

min. 99 % d.s.

max. 15 % when packed (including water of
crystallization)

max. 0.1 %
max. 50 ppm
max. 100 ppm
max. 10 ppm
max. 1 ppm
max. 20

5-7

max. 100/ g

Free-flowing, white crystals, faint characteristic odour
0.7 - 0.75 kol

Water 160 9/100 g H2O

Methanol 55 g/100 g MeOH
Ethanol 8.7 g/100 g EtOH

Hygroscopic, absorbs humidity from air
‘Heat stable up to 200°C
3 years from the manufacturing date

25 kg multiwall laminated white paper bags with

polyethylene inner bag, shrinkwrapped on one-way
pallet. Standard pallet 1000 kg.

In unopened bags, protected from humidity

_ Matters beyond the control of Finnfeeds Finland Limited (trading as Danisco Animal Nutrition) such as Incorrect storage and use of the product, animal
management, health and environment differences may cause variation in performance, Finnfeeds Finland Limited gives no express or implied warranty tor
individual results, Notwithstanding the disclaimer herein contained Finnfeeds Finland Limited's liability (If any) in respect of such matters shall under no
circumstances exceed the purchase of price of the product. i

Danisco Animal Nutrition Finnfeeds Finland Limited”
</pre>

====================================================================== Einde pagina 59 =================================================================

<br><br>====================================================================== Pagina 60 ======================================================================

<pre>Finnfeeds Finland Oy / Novel food application 12/2002
Summary by applicant

APPENDIX 3

List of references of an original
novel food application
</pre>

====================================================================== Einde pagina 60 =================================================================

<br><br>====================================================================== Pagina 61 ======================================================================

<pre>Finnfeeds Finland Ltd / Novel food application 15.1.2003

References

Abdelmalek MF, Angulo P, Jorgensen RA, Sylvestre PB, Lindor KD. Betaine, a promising
new agent for patients with non-alcoholic steatohepatitis: Results of a pilot study. Am J
Gastroenterol 2001;96:2711-2717.

Alfthan G & Aro A. Study plan and first study results of the betaine and homocysteine
dose-response study. Public Health Institute, Finland, 2002.

Andersen NL, Fagt S, Groth MV, Hartkopp HB, Moller A, Ovesen L, Warmin DL.
Danskernes kostvaner 1995. Hoved resultater. Publikation nr. 235. Seaborg;
Levnedsmiddelstyrelsen, 1996.

Andreeva SM (1971). Betaine content in mushrooms of the order Agaricales Il; Mikol.
Fitopatol 5(1), pp.29-32.

Barak AJ, Beckenhauer HC, Junnila M, Tuma DJ. Dietary betaine promotes generation of
hepatic S-adenosylmethionine and protects the liver from ethanol-induced fatty infiltration.
Alcohol 1993;17:552-555.

Barak AJ, Beckenhauer HC, Junnila M, Tuma DJ. The relationship of ethanol feeding to
the methyl folate trap. Alcohol 1993;10:495-7.

Barak AJ, Beckenhauer HC, Tuma DJ. S-Adenosyl methionine generation and prevention
of alcoholic fatty liver by betaine. Alcohol 1994;1 1:501-3.

Becker W. Befolkningens kostvaner och näringsintag | Sverige 1989. Uppsala;
Livsmedelsverket 1994.

Berlow S, Bachkman RP, Berry GT, Donnell GN, Grix A, Levitsky LL, Hoganson G, Levy
HL. Betaine therapy in homocystinemia. Brain Dysfunct 1989;2: 10-24.

Best CH, Huntsman ME. The effects of the components of lecithin upon deposition of fat in
the liver. J Physiol 1932;75:405-12.

Borsook H & Borsook ME. The biochemical basis of betaine-glycocyamine therapy. Ann
West Med Surg 1951;5:825-9.

Borscok H & Borsook ME. Treatment of cardiac decompensation with betaine and
glycocyamine. Ann West Med Surg 1951;5:830-55.

Bostom AG, Shemin D, Nadeau MR et al. Short term betaine therapy fails to lower
elevated fasting total plasma homocysteine concentrations in hemodialysis patients
maintained on chronic acid supplementation. Atherosclerosis 1995;113:129-132.

Brens CM, Serra JD, Tomas MLC, Gomez AMG, Monegal MR, Busca AV.

Homocystinuria: efficacy of pyridoxine, folic acid and betaine therapy. An Esp Pediatr
1993:1:37-41.

64
</pre>

====================================================================== Einde pagina 61 =================================================================

<br><br>====================================================================== Pagina 62 ======================================================================

<pre>Finnfeeds Finland Ltd / Novel food application 15.1.2003

Brouwer IA, Verhoef P, Urgert RI. Betaine supplementation and plasma homocysteine in
healthy volunteers. Arch Intern Med 2000;160:2546-2577.

Clarke WC et al. Effects of dietary betaine/amino acid additive on growth and seawater
adaptation in yearling Chinook salmon. Aquaculture 1994:121:137-148.

Dudman NPB, Guo X, Gordon RB, Dawson PA, Wilcken DEL. Human homocysteine
catabolism: three major pathways and their relevance to development of arterial occlusive
disease. J Nutr 1996;126:1295S-1300S.

Dudman NPB, Tyrell PA, Wilcken DEL. Homocysteinemia: Depressed plasma serine
levels. Metabolism 1987;36:198-201.

F.O. Licht GmbH. Christoph Berg. The world molasses market — supply, demand and
outlook to 2005. expert analysis of the current situation and the future outiook for world
molasses markets. United Kingdom, 1998.

Franken DG, Boers GHJ, Blom HJ, Trijbels FJM, Kloppenborg PWC. Treatment of mild
hyperhomocysteinemia in vascular disease patients. Arterioscler Thromb 1994;14:465-
470.

Frontiera MS, Stabler SP, Kolhouse JF, Allen RH. Regulation of methionine metabolism:
Effects of nitrous oxide and excess dietary methionine; J Nutr Biochem 1994; 5:28-38.

Gahl WA, Bernardini 1, Chen S, Kurtz D, Horvath K. The effect of oral betaine on vertebral
body bone density in pyridoxine-non-responsive homocystinuria. J Inher Metab Dis
1988:11:291-8.

Graybiel A, Patterson CA. Use of betaine and glycocyamine in the treatment of patients
with heart disease: preliminary report. Ann West Med Surg 1951;5:863-75.

Haahtela T, Hannuksela M, Terho E (toim.) Allergeenien luonne ja luokitus. Duodecim.
Helsinki, 1993:30.

Kim SK, Kim YC. Attenuation of bacterial lipopolysaccharide-induced hepatotoxicity by
betaine or taurine in rats. Food Chem Tox 2002;40:545-9.

Kim SK, Kim YK, Kim YC. Effect of betaine administration on metabolism of hepatic
gluthathione in rats. Arch Pharm Res 1998;21:790-2.

Knopman D, Pattersson M. An open-label, 24 week pilot study of the methyl donor betaine
in Alzheimer disease patients. Alzheimer Dis Assoc Disord 2001;15:162-5.

Lever M, Sizeland PCB, Bason LM, Hayman CM, Robson RA, Chambers ST. Glycine
betaine and proline betaine in human blood and urine. Biochim Biophys Acta
1994; 1200:259-264.

Lever M, Sizeland PCB, Bason LM, Hayman CM, Robson RA, Chambers ST. Abnormal
glycine betaine content of the blood and urine of diabetic and renal patients. Clin Chim
Acta 1994,230:69-79.

65
</pre>

====================================================================== Einde pagina 62 =================================================================

<br><br>====================================================================== Pagina 63 ======================================================================

<pre>Finnfeeds Finland Ltd / Novel food application 15.1.2003

Mar M & Zeisel SH (1999). Betaine in wine: answer to the French paradox? Medical
Hypotheses; 53(5) pp. 383-385.

McGregor DO, Dellow WJ, Robson RA, Lever M, George PM, Chambers ST. Betaine
supplementation decreases post-methionine hyperhomocysteinemia in chronic renal
failure. Kidney Int 2002;61 (3):1040-6.

Meyers S (1987) Aquaculture feeds & chemoattractants; Infofish Marketing Digest No. 1/87
pp.35-37.

Morrison LM. Use of betaine-lipotropic combinations in clinical practice. Geriatrics
1953;8:649-55.

Morrison LM. Results of betaine treatment of atherosclerosis. Am J Dig Dis 1952;Dec:381-
384.

National Academic Press, Washington D.C. Standing Committee on the Scientific
Evaluation of Dietary Reference Report (2000) Dietary reference intakes for thiamine,
riboflavin, niacin, vitamin B6, folate vitamin B12, pantothenic acid, biotin and choline;
Ch.12 — Choline.

National Public Health Institute. The 1997 Dietary survey of Finnish adults.
Kansanterveyslaitoksen julkaisuja B8/1998. Helsinki, 1998.

PDR Monograph for Nutritional Supplements (2001). Betaine and Betaine hydrochloride.

Roos G, Prättälä R. Disparities in food habits. Review of research in 15 European
Couniries. Publications of the National Public Health Institute B24/1999. Helsinki, 1999.
Gregory et al. 1990: National Diet and Nutrition Survey (NDNS).

Schwab U, Törrönen A, Toppinen L, Alfthan G, Saarinen M, Aro A, Uusitupa M. Betaine
supplementation decreases plasma homocysteine concentrations but does not affect body
weight, body composition, or resting energy expenditure in human subjects. Am J Clin Nutr
2002;76(5):961-7.

Schwahn B, Hafner D, Hohlfeld T, Laryea MD, Wendel Ul. Pharmacokenetics of oral
betaine in healthy subjects; J Inherit Metab Dis 2000;23(Suppl.1):66.

Steen MT, Kruger WD, Singh RH, Elsas LJ. Betaine increases methionine/homocysteine
ratios and decreases total plasma homocysteine (tHcy) in cystathionine beta-synthase
deficiency. J Inherit Metab Dis 2000;23:61.

Storch KJ, Wagner DA, Young VR. Methionine kinetics in adult men: Effects of dietary
betaine on L-(2H3-methyl-1-13C) methionine. Am J Clin Nutr 1991;54:386-394,

Suuronen J, Pitkanen |, Halttunen H, Moilanen R. Formation of the main gas compounds

during thermal analysis and pyrolysis. Betaine and betaine monohydrate. Journal of
Thermal Analysis and Calorimetry 2002;69:359-369.

66
</pre>

====================================================================== Einde pagina 63 =================================================================

<br><br>====================================================================== Pagina 64 ======================================================================

<pre>Finnfeeds Finland Ltd / Novel food application 15.1.2003

Van Guldener C, Janssen MJ, Lambert J, Wee PM, Donker AJ, Stehouwer CD. Folic acid
treatment of hyperhomocystenemia in peritoneal dialysis patients. Perit Dial Int
1998; 18:282-289.

Van Guldener C, Janssen MJ, de Meer K, Donker AJ, Stehouwer CD. Effect of folic acid
and betaine fasting and postmethionine-loading plasma homocysteine and methionine
levels in chronic hemodialysis patients. J Intern Med 1999;245:175-183.

Van Zandt V, Borsook H. New biochemical approach to the treatment of congestive heart
failure. Ann West Med Surg 1951;5:856-62.

Waggle DH et al. Extensive analysis of flours and mill fees from nine different wheat
mixes. Cereal Chemistry 1967;44:48-60.

Wilcken DEL & Wilcken B. The natural history of vascular disease in homocystinuria and
the effects of treatment. J Inher Metab Dis 1997;20:295-300.

Wilcken DEL, Wilcken B, Dudman NPB, Tyrell PA. Homocystinuria — the effects of betaine
in the treatment of patients not responsive to pyridoxine. N Engl J Med 1983;309:448-53.

Wiley VC, Dudman NPB, Wilcken DEL. Free and protein-bound homocysteine and

cysteine in cystathionine b-synthase deficiency: interrelations during short- and long-term
changes in plasma concentrations. Metabolism 1989;,38:734-9.

67
</pre>

====================================================================== Einde pagina 64 =================================================================

<br><br>====================================================================== Pagina 65 ======================================================================

<pre>Bijlage E
        Eerste beoordeling/First assessment
        Eerste beoordeling/First assessment 65
</pre>

====================================================================== Einde pagina 65 =================================================================

<br><br>====================================================================== Pagina 66 ======================================================================

<pre>66 Betaïne/Betaine</pre>

====================================================================== Einde pagina 66 =================================================================

<br><br>====================================================================== Pagina 67 ======================================================================

<pre>,. ~                 =              OF                                               (Co",    Tra"'"';..)
    "Ëi~      TRADE AND INDUSTRY
                            FINLAND                                                  Dno 1/618/2003
                    NovelFoodBoard                                                   12June2003
                                                                                                    .
    INITIAL ASS~SMENT REPORT
    onder Article 4 of Regulation (EC) No 258/97 of the European Parliament and the Councll
    conceming novel foods and novel food ingredients                                                     '.
    "AN APPLICATION TO PLACE BETAlNE                      AS A FOOD INGREDIENT              ON TUE EU
    NOVEL FOOD MARKET"
    1. On 24 January 2003, Finnfeeds Finland Ltd submitted an application to place betaine as a food
          ingredient on the EU novel food market to the National Food Agency. Betaine is intended foT
         use in drinks, cereal products and dairy products. The application was forwarded to the Novel
         Food Board on 30 .]anuary 2003. The Board basrequestedthe applicant toprovide. further        ". '"
         clarification on 25 Febmary 2003,6 May 2003, 19 May 2003 and 10 June 2003. The applicant
         bas respondedto the requestson 31 March 2003, 7 May 2003, 13 May 2003, 3 June 2003 and
         11 June 2003.
   2. The Novel Food Board bas examined the application in accordancewith the Commission
         Recommendation97/618/EC concerning the scientific aspectsand the presentation of
         infonnation necessaryto support applications foTthe placing on the market of novel foods and
         novel food ingredients and the preparation of initial assessmentreports under Regulation (EC)
         No 258/97 of theEuropean Parliament and of the Council.
   3. The application is considered to belong to class (e) in Article 1.2 of the Regulation, foods and
         rood ingredients consisting of or isolated trom plaats or animaJ.s.In accordance with the
         Commission Recommendation97/618/EC, the ingredient concemed by the application bas been.
         identified as belonging tQ class 1.1 "Pure chemicals or simpte mixtures trom non-aM sources;
         the source of the NF bas a history of food U$ein the Community". Accordingl y, the examination
         of this application proceeds in accordance with the schemesI, Il, fi, IX, X, XI, XII, XIII
        presented in the Commission Recommendation.
   OPINION OF THE NOVEL FOOD BOARD ON THE SAFETY OF TBR FOODS
   CONCERNED BY THE APPLICATION
   SUMMARY
   4. Betaine (synonyms glycine betaine and trimethyl glycine), intended for use as an
        ingredient in the roods concerned by the application, is a compound that is commonly
        found in animals and plants. The products concerned by the application (Betafin BF20,
        TMG20 and AP) consist up to 99% of betaine. In the products, betaine is either in
        anhydrous from or in form of betaine monohydrate. The roods for which the ingredient is
        intended correspond as totheir composition otherwise to roods that already are on the
        market.
  5. The information submitted by the applicant on the composition of the betaine ingredient
        bas been sufflcient. The calculations on anticipated consumption bas been adequately
   Betaine EAe 120603public
</pre>

====================================================================== Einde pagina 67 =================================================================

<br><br>====================================================================== Pagina 68 ======================================================================

<pre>                                                                                             2
   made.The Novel Food Board stateshoweverthai it is verf difficult to give estimateson
   the intake of betaine from foods to which betainebas beenadded, and thai the calculations
   have mainly beenbasedon the applicant's own estimateson the consumptionof single
   products.                                                                             .
6. The sourceof betaine,molassesfrom sugar beet, basa long history useas a safe raw
    material for foodstuffs.The betaine ingredient isolatedtrom sugar beetcan he regarded.as
    fit for humanconsumption.According to a testimonial presentedby the applicant, no
    geneticallymodified sugar heetsare usedin the production of the rood ingredient
    concerned.
7. The production processesusedin the manufacture of the foods concemedare traditional
    food processes,and the Board bas no objection to them. On the basisof what is said in the
    application, the processesof manufacture of the final foods cannot be expectedto have any
    marked effect on the content or the quantity of the betaine ingredient. Neither are any
    proble~ of microbiological nature to beexpected.
8. Foodscontaining the ingredient concemedby the application are not expectedto be
    nutritionally disadvantageouseven when replacing traditional foods ûSedtor comparative
    purposes.
9. The resultsof the safety evaluation studiesprovided by the applicant indicate that liver is
    the target organ of poaible betaine toxicity. The observedeffectson anirnalRwere minor
    and at least largely reversibie. The effectson the health of the consumercannot,however,
    be assessedwith adequatereliability on the basisof the materiaI presentedby the
    applicant. Betaine bas not beenproved to causeallergies or reactionsof
    hypersensitisation.
10. Basedon the existing knowledge presentedin the application and in the light of other        .
    existing knowledgeof betaine, it cao be assumedthat a daily intake of 20-30gra~ of the
    new ingredient in the roods concemed by the application would not be nutritionally
    disadvantageousfor consumers.
11. The Board holds the view thai roods to which betainebas beenadded should not be
    marketed to pregnant women, breastfeedingmothers or infants, becauseno data on the
    long-term etTectsof betaine on theseuser groups are available.
12. The Board considersthat rood labels should provide the COnsumerwith information on the
    betaine ingredient of the product. The applicant bas presenteda draft of a product label to
    be affixed on packagesintended tor sale to rood manufacturers. The applicant does not
    present any proposal on how information on the new ingredient should be disseminatedto
    consumerson the packageof the final product.
</pre>

====================================================================== Einde pagina 68 =================================================================

<br><br>====================================================================== Pagina 69 ======================================================================

<pre>                                                                              "" C"c."    '"
" .
 .,  ..
                                                                                                         3
    DETAll..ED EXAMINA nON OF mE APPLICA TION
    I. Specification of the Novel Food                                                                 .
    13. The information received 00 the products concemed by the applicatioo and on the hetaine
    ingredient of them bas been suflicient. On the basis of the materiaI preseoted, the products
    cao he considered suited for food use.
    14. Betaine is produced of the secondary flow of sugar beet intended for the production of sugar.
         An assessmentof the manufacWring processand its effects on the products is presentedin
         Chapterill.
     15. Betaine is planned to he marketed under three different brand names: Betafin BF20 (anhydrous
         form of edible quality, a description of the product in Annex la to the application), Betafin
         TMG20 (anhydrous form of edible quality conesponding to the above product, a description of
         the product in Annex 1b to the application) and Betafin AP (betaine monohydrate of edible and
         phannaceutical quality, a description of the product presentedin Annex 1d). A safety data sheet
         foT all the three prociuctsis presentedin Annex 1d Annex 2 contains a description of the
         method used in the determination of the quantity of betaine mentioned in the product
         specification.
     16. In the light of the information presentedin product specifications and in Annexes 5-6, it CaDbe
         stated that the products concerned are suited foTfood use as concerns residual materials.
     17. The data presentedin product specifications are obtained trom production on an industrial scale.
         Betaine bas been produced in commercial scale since 1979 andbetaine foT food grade betaine
         since 1983. Reference materials related to the products are available.
     18. Betaine is known under its chemical Damesglycine betaine or trimethyl glycine. lts chemical
         structure is simple -a glycine basis plus three methyl groups that are attached to ODenitrogen
         atom. In betaine monohydrate, ODewater molecule is attached to a group of carbonyls. The
         anhydrous form and the hydrated form of betaine are comparabie as to their physical and
         chemical properties. In the application, the concept "betaine" is used to describe both these
         forms.
     19. Betaine productsare highly hygroscopic. The stability ofthe products bas beenensured by
         packaging them in moisture-resistant, food quality material.
     20. Betaine is intended foTuse in drinks, sweets,cereal products and dairy products. The applicant
         states th'i~.' ~ainewil I be added to normal foods that are already on the market. The proposed
          maximu ~ taille content of the products concerned is as follows: drinks 2%, sweets6.7%,
         cereal-basedproducts 4% and milk-based products 2.7%.
                                                     .
    11. Effect of the Production Process Applied to the Novel Food
                                                                              fuL,",,"","'~""'i.,'1 'c     îIlMI
</pre>

====================================================================== Einde pagina 69 =================================================================

<br><br>====================================================================== Pagina 70 ======================================================================

<pre>                                                                                                   4
21. Tbe metbods used in tbe production of betaine and betaine products can be considered
    acceptable. Betaine is considered to he able to endure tbe manufacturing process of tbe
    foods to wbicb betaine is intended to headded.
22. The production methods of betaine productshave been presentedin the application. The
    production processis conducted according to GMP requirementsand by applying the Hazard
    Analysis and Critical Control Points (HACCP) principles. The quality system for production.
    and the environmental managementsystemhave beencertified in accordancewith the ISO
     9001:1994 and 14001:96 standards (Annex 2 to the Application).
23. Betaine can be crystallized from sugar molassesproduced from sugar beet either in an
     anhydrous form or as betaine monohydrate. Anhydrous betaine bas been proved to remain stabIe
     over the time set out for the products concemed, namely three years (Annex 8 to the
     application).
24. For anhydrous betaine, the degradation processor the esterification process starts first at a
     temperature exceeding 245OC,and betaine monohydrate is still more stabie than that. The core
     temperatureof foods to which betaine is going to be addeddoes not exceed 2000C during the
     manufacture, which means that degradation or esterification of betaine is not to be expected.
25. The stability of crystalline betaine and of a solution with a betaine content by weight of 50% has
     been testedduring 2-24 hours at temperaturesbetween800Cand 2000C (Annex 9 to the
     Application). The quantity of crystalline betaine remained unchanged under test conditions.
     even though a slight change of colour could be observed at 1200Cand especially at 20ü0C. Clear
     degradation was observed in the aqueoussolution of betaine at 200°C and an indefinabie odour
     alreadyat 1200C'However, the foods to which betaine win be added can be expected to endure
     the manufacturing process for such foods.
26. The method of manufacturing cookies did not have any effect on the betaine added (Annex 10).
111.History of tbe organism used as Novel Food source
27. Betaine is produced from sugar beet, Beta vulgaris sp., wbicb is used in tbe production of
      sugar and wbicb bas a long bistory as a food source. No genetically modified varieties are
     used in tbe production, tbe applicant presents a testimonial of that in Annex 11. Sugar
     molasses from wbich betaine is being isolated is well-known as a food ingredient
      tbroughout the EU.
 IX. Anticipated intake I extent of use of tbe Novel Food
28. The consumption calculations presented in tbe application are adequate. For many
     foodstuffs, tbe calculations are based on tbe applicant's own judgment. Tbe fact that tbe
      applicant int~nds to add betaine to certain groups of foodstuffs, and not to specific foods,
     makes tbe estimation of intake levels more difficult. Tbere are necessarily no data
     available on the consumption of single foodstuffs belonging to certain rood categories, and
     roods categories on tbe otber hand vary from country to country within the EU, which also
     makes it more difficult to make reliable estimates of consumption. Any specific estimates
      of consumption by children have not been presented.
</pre>

====================================================================== Einde pagina 70 =================================================================

<br><br>====================================================================== Pagina 71 ======================================================================

<pre>.;
    . ;
                                                                                                           5
    29. The applicant proposes that hetaine he used as an ingredient in the following foodstuffs:
             -heverages, such as soft drinks and mineral waters, non-diluted fruit juices, ready-to-
                 servecoffee and tea drinks, alcoholic drinks -excluding beer -with an alcoholic strength
                 by volume of less than 22% vol (such as long drink, cider, wine, bitter, sherry or liqueur)
             -cereal products, such as Müsli, breakfastcereals, Qat,rye, barley and wheat flakes and .
                 snack-bars("energy bars")
             -dairy products such as sour milk products
             -confectionary      such as hard candies (pastilles) and chewing gum
         The maximum hetaine content of the productsconcemed by the application is planned to he as
        follows: tor drinks 2%, tor cereal-basedproducts4%, tor sour milk products 2.7% and tor
         confectionary 6.7%.
    30. Data on the use of foodstuffs to which betaineis planned to he added in some EU countries
         (Finland, Sweden,Denmark, Great Britain, Germany, the Netherlands and Spain) is presented in
         the application and in the supplementary information submitted on 31 March 2003 and 3 June
        2003. The applicant presentsin the following data on very high consumptión of the roods to
         which hetaineis planned to he added at the level of the 9Othpercentile (intake g/day):
                           Drinks               Sweets                Cereal roducts      D.     roducts
    Swedenl                 1250                13                     176                224
    Denmark2                1250                15                     163                230
    'Riksmaten1997-1998.Kostvanorochnäringsintagi Sverige.Method and resultanalysis.Livsmedelsverket2000
    "Andersenet al, Publ. 235,Levnedsmiddestyrelsen
                                                  1996
   31. The table below presentsan estimate of the intake of betaine that has been made on the basis of
        the recommended daily intake of foods to which hetaine is added andon the consumption data
         on corresponding foods without hetaine (g/day):
                     Recommended Intake of           Average         Intake of     Maximum       Intake of
                     daily intake of betaineat        consumption    betaineat     consumption   betaineat
                     a foodstuff      recommended of a               average       of a          maximum
                                      dosage          foodstuff      consumption   foodstuff     consumption
   Drinks            200              4              493-739         10-15         1250          25
   Confection        60               4              4-8             0.2-0.5       13-15         1
   Cereal            100              4              27-45           1-2           163-176       6.5-7
     roducts
   Dairy             200              4              65-115          2-3          224-230        6
     roducts
   Cumulative                         16                             13-20.5                     39
   intake
   32. It appears trom the table above that the total daily intake of betaine trom all groups of foodstuffs
        containing betaine at recommended intake levels may be up to 16 grams. If the intake is
        calculated on the basis of the average intake of the foodstuffs concemed, the daily intake of
        betaine is estimated to be about 13-20 grams, and at large consumption about 39 grams.
</pre>

====================================================================== Einde pagina 71 =================================================================

<br><br>====================================================================== Pagina 72 ======================================================================

<pre>                                                                                                   6
 33. It is, however, improbable that a consumerwould eat all foodstuffs containing betaine
     continuously andin large quantities. The applicant considersthat a continuous high intake of
     hetaine could result in intestinal trouble, which would have a dampening effect on the
     consurner's desire to use the productsconcemedcontinuously. A study published in 2002 .
     (Abdelmalek et al) reveals that a continuous daily consumption of 20 grams during a year did
     not causeharm to the health of sevenpatientswho suffered from non-alcoholic fatty liver
     diseasei.e. non-alcoholic steatohepatosis.Similar results have been obtained in a study
     published in 1951 where the intake level was 20-30 grams per day (Borsook & Borsook).
34. The applicant proposes that adults, old people and particularly persons who are otherwise
     healthy but are running a risk of heart diseaseshould he target groups fot the products
     concemed. The products are not intended fot children or pregnant wamen or breast-feeding
     mothers, and fot that reason,the applicant bas not consideredit necessaryto make a safety
     evaluation especially with a view to thesegroups of consumers.Betaine is, however, proposed
     to he addedto groups of foodstuffs that children consume in larger quantities than adults in
     relation to their weight (e.g. sweetsand soft drinks).
35. Vegetarians and persons suffering from under-nourishmentconstitute a potential risk group if
     hetaine increasesthe need fot methionine. A study reveals that the need fot methionine by
     healthy men increased already after quite areasonabie increaseof betaine. The results of this
     study cannot, however, he directly applied to persons suffering from metbionine deficiency, and
     tests on animals do not support this observation.
36. Foodstuffs to which betaine bas been added will replace similar foodstuffs without hetaine, but
     tbis is not considered to he nutritionally disadvantageousfot user groups. The foodstuffs
     areequivalent, apart trom hetaine.
X. Information from previous human exposure to the NF or its source
37. The applicant presents data on the previous exposure to betaine, for instance, trom plant
     products that are a natural element of oor diet and from dietary supplements. There are
     no results from a systematic follow-up of the use of betaine available. These new foodstutTs
    containing betaine maf lead to a markedly higher intake of betaine, and therefore the
     ingredient is subjected to a safety assessmentin accordance with the Novel Food
    Regulation.
38. Betaine is a compound commonly found in plants and animais. It is accordingly also a natural
     element of OUtdiet. Studies of literature reveal that an analysis of the betaine content is
    available for more than 50 foodstuffs. The application presentsexamples of them, such as
    prawns (0.25-0.96 g/100 grams), spinach(0.41-0.91 g/100 grams), edible mushrooms (0.32-
    0.69 g/100 grams), breakfast cereals (0.05-29 mg/l00 grams), biscuits (5-34 mg/l00grams) and
    wille (about 1 mg/l00 mi). Annex 12 to the Application contains calculations on the intake of
    betaine from different natural sourcesand different diets. The intake varied between1 g and 2.5
    g per day.           .
39. Betaine hydrochloride is used also in dietary supplements.The doses vary between7 and 324
    mg/tablet. Most products have also beenavailable via Internet within the EU (a list of them is
    presented in the application). It must however be observed that betaine hydrochloride differs
    from betaine as to its physical and chemical properties.
</pre>

====================================================================== Einde pagina 72 =================================================================

<br><br>====================================================================== Pagina 73 ======================================================================

<pre>,~
     . '.
                                                                                                               7
     40. Betaine bas beenmarketed as food supplementin the USA since the 1960's and the FDA has
           approved it for medical use in 1996 (orphandrug, dose 6-20 grams per dar).
     41. Betaine is also used as flavour enhancerand as preservative for prawns. As an ingredient in-
          feeds, betaine bas beenused worldwide over 25 years.
     42. Betaine bas beenused in clinical testssameof which have contained analysis of the impact of
           betaine on the homocystein content of serum.
     43, No results of a systematic follow-up of the use of betaine after the products concemed have
           been introduced to the market are available.
     XI. Nutritional infonnation on the Novel Food
     44. The use of foodstufTs with added betaine concemed by the application is not expected to
           have disadvantageous nutritional effects. The intended group of users comprises adults
           and old people.
     45. Betaine is not available for traditional comparison. Betaine is not intended for use as such but as
           an ingredient in certain categories of foodstuffs that already are on the market.
     46. Anhydrous betaine and betaine monohydrate are physiologically similar. Already with the
          relative humidity at 7%, anhydrousbetaine tums into betaine monohydrate.
     47. Betaine occurs in human blood and the concentration of betaine in blood is usually quite stabie.
           It is found in urine and stored in renal medullar cells providing a source of osmotic protection.
     48. Intake of betaine takes place either directly from foodstuffs containing betaine or indirectly via
          foodstuffs containing choline/derivates of choline as metabolic products of them.
     49. Betaine comprises three methyl groups that participate in methylation. The application presents
          a description of the metabolic routes of betaine in man. Metabolization mainly takes place in the
           liver and kidney. It is further stated in the application that betaine is catabolized mainly in vivo
   ,      and is eliminated to a very little extent in urine.
     50. The application makes reference to many scientific publications (Annex 13 to the Application)
          where betaine bas been studied in clinical trials. On the basis of these analyses it can be stated
           that betaine bas been widely tested and is considered to be well tolerated at dosagesup to 30 g.
          The literature reveals that only few casesof nausea,diarrhoea or gastro-intestinal troubles have
           beenreported.
     51. Tests have beenconducted on adults running the risk of a heart attack, on healthy adults and on
          old people. Old people's tolerance to betaine bas been equal to that of adults in general.
     52. As to its nutritional effects, betaine mainly bas a lowering effect on the serum homocystein
          concentration,-A diet based on the recommendations on the intake of betaine does not involve
          any changescompared with anormal diet. The consumption volumes of sweets may, however,
          increase becausethe upper limit of the recommended daily intake of sweets containing betaine
          is planned to be set at 60 g per dar.
</pre>

====================================================================== Einde pagina 73 =================================================================

<br><br>====================================================================== Pagina 74 ======================================================================

<pre>                                                                                                           .,
                                                                                                              .
                                                                                                        8
  XII. MicrobiologicaI infonnation on the Novel Food
 53. The appUcant bas presented a sufficient explanation of the source of betaine and the
      manufacturing process of the betaine ingredient. The production process or the sources of
       raw materials do not give any reason to expect problems with the microbiological quality.
 xm.     Toxicological assessmentof the Novel Food
  54. Results of safetyevaluation studies provided by the appUcant indicated that Uver is the
      target organ of betaine toxicity. The observed effects were relatively mild and mainly
      reversibie, but their clinical significance is difficult to ~        only on the basis of animal
      experiments. Liver toxicity bas not been reported in human trials. However, the applicant
      bas provided only a very limited set of data on the safety and liver effects of betaine in
      patients. Therefore, the statistical power of the data is limited. Betaine did not show
      allergenic or hypersensitizing potential.
 55. The applicant bas carried out a set of toxicity studies with hetaine. In these studies tbe acute oral
      toxicity was low in rats. Betaine did not show eye irritating potential in rabbits, and it did not
      cause skin sensitization in guinea pigs. In tbe human patch test hetaine waS not a skin irritant. In
      addition, hetaine was not proved to he genotoxic in tbe bacterial reverse mutation assays, the
      metaphaseanalysis of human lymphocytes in vitro, and the mouse micronucleus test in vivo.
 56. Subchronic toxicity studies in rats revealed basically mild, but consistent and dose-dependent
     changes in liver. These changes included increased liver weight, increased serum levels of
     gamma-glutamyl transpeptidase (OOT) and alkaline phosphatase (AP), as weIl as increased
     incidence of hepatocellular microvacuolization. The vacuoles were shown to contain lipids.
     These changes were present already after a treatment period of 14 days, and they were more
     pronounced in female rats. A 28-day study with a 28-day recovery phase indicated tbat these
     changes were largely reversibIe. The liver changeswere observed at all dose levels studied, and ..
     therefore it was not possible to set a NOAEL value. The lowest doses studied were equivalent to
     ca. 800 mg/kg/day corresponding to 56 g/day in a 70 kg human. Although the liver changes
     observed in toxicity studies were relatively mild and reversible, their relevance for the human
     use of betaine is difficult to assess.Therefore. for the assessmentof human safety of betaine, a
     systematic and careful analysis of all human trials should he carried out. For this purpose
     placebo controlled double blind studies and studies that utilized high dose levels are most
     valuable.
57. Subchronic toxicity studies revealed also dose-dependentdecreasesin mean corpuscular volume
    and haemoglobin concentration, and increasesin blood fibrinogen concentration.
58. Betaine bas I       on market as a dietary supplement in the USA since the 1960's. It bas been
    used as a rne.~~11donor for controlling serum homocysteine levels. The recomrnended dosage
    bas been 0.5-1.5 g/day. Betaine bas also been approved by the FDA in 1996 as an orphan drug
    for treatment of homocysteinuria. The dose levels have typically been 6 g/day, but doses as high
    as 20 g/day have also been recomrnended.However, the applicant bas not referred to the safety
    evaluation studies used for registration, and no systematic post-marketing follow-up data on
    possible side effects is available.
59. For the safety of betaine the applicant bas referred to 43 published clinical trials with over 700
    subjects. Only a few of these studies were placebo controlled double blind studies. Most of the
</pre>

====================================================================== Einde pagina 74 =================================================================

<br><br>====================================================================== Pagina 75 ======================================================================

<pre>"-
   .~ .'.
  .9
           subjects were patients with a variety of illnesses, and there are only a few reports on healthy
           subjects. The dose-range has been 2-30 g/day (typically 6 g/day), and treatment times range
           from 1 day to 16 years. Reported side effects include isolated cases of nausea.diaIThoeaand
           gastrointestinal disorders, but there are no reports on adverseorgan toxicity.                .
       60, Safety issueshave not beenthe main objective in most of the trials, and they have not
           necessarilybeen adequatelymonitored in all the trials. In addition, it is problematic to
           demonstratepossible mild liver toxicity of betaine for patients with pre-existing liver disease.In
           the material presentedby the applicant effects of betaine on enzyme levels indicating liver
           damagehave been monitored only in a.few studies that included about thirty patients. Due to the
           low number of patients the statistical power of the data is limited. In addition, data on the safety
           of betaine in healthy subjectsis limited for a reliable consumer safety assessment.
                          ~
                                                           ,                                                 ,ol
</pre>

====================================================================== Einde pagina 75 =================================================================

<br><br>