P ocedia Food Science 1 (2011) 1828 – 1832
2211–601X © 2011 Published by Else ie B.V. Selec ion and/o pee - e iew unde esponsibili y o 11 h In e na ional Cong ess on Enginee ing
and Food (ICEF 11) Execu i e Commi ee.
doi:10.1016/j.p o oo.2011.09.268
A
ailable online a www.sciencedi ec .com
P ocedia – Food Science 00 (2011) 000–000
P ocedia
Food Science
www.else ie .com/loca e/p ocedi
a
11 h In e na ional Cong ess on Enginee ing and Food (ICEF11)
Nu i ional e ec s o olic acid con olled elease om
mesopo ous ma e ials
José Ba a a*, Édga Pé ez-Es e ea, And ea Be na dosa,b, Ramón Ma ínez-Mañezb
aUni e sidad Poli écnica de Valencia, Food Technology Depa men , Camino de Ve a S/N, Valencia,46020, Spain
b Uni e sidad Poli écnica de Valencia, Ins i u e o Molecula Recogni ion and Technological De elopmen , Camino de Ve a S/N,
Valencia,46020, Spain
Abs ac
Folic acid de iciency causes se ious diso de s in humans and supplemen a ion has nume ous heal h bene i s.
Howe e , he e is ini ial e idence ha sugges a nega i e impac o an inc eased exposu e o olic wi h espec o
ce ain de elopmen al and degene a i e diso de s. In his line, con olled elease o olic acid by using mesopo ous
silica ma e ials, MCM-41, has been s udied as an al e na i e o di ec supplemen a ion. Fo his pu pose, a ious
mesopo ous solids MCM-41 loaded wi h olic acid (S1) and unc ionalized wi h 3-[2-(2-
aminoe hylamino)e hylamino]p opyl- ime hoxysilane (S2) ac ing as "ga e" ha e been es ed. The esul s show ha
a pH 2 a s ongly hinde ed i amin elease is obse ed, whe eas a pH 7.5 a con olled deli e y is ound. Based on
he ob ained esul s o his s udy, olic acid con olled elease could be easible du ing a pe iod o 5h using a
sensi i e o pH ga e, and his migh educe adi ional o i ica ion nega i e e ec s, while nu i ional bene i s a e
main ained.
© 2011 Published by Else ie L d. Selec ion and/o pee - e iew unde esponsibili y o ICEF11
Execu i e Commi ee Membe s.
Keywo ds: Folic acid; con ol elease; mesopo ous ma e ial; bioa ailabili y; molecula ga es.
1. In oduc ion
Fola es a e a g oup o wa e -soluble i amins. Thei de iciency can cause neu al ube de ec s in
de eloping emb yos. To p e en he occu ence o hese and o he diseases, supplemen a ion o ce ain
oods wi h his i amin is equi ed by law in ce ain coun ies. Howe e , ecen s udies sugges ha olic
* Co esponding au ho . Tel.: +34 96 387 36 01; ax: +34 96 387 36 01.
E-mail add ess: jmba[email p o ec ed].es
© 2011 Published by Else ie B.V. Selec ion and/o pee - e iew unde esponsibili y o 11 h In e na ional Cong ess
on Enginee ing and Food (ICEF 11) Execu i e Commi ee.
1829
JosÈ Ba a e al. / P ocedia Food Science 1 (2011) 1828 – 1832
2 Au ho name / P ocedia – Food Science 00 (2011) 000–000
supplemen a ion could be a double-edged swo d, because i could be associa ed wi h he incidence o
some ypes o diseases when he excess o olic acid abso bed is di ec ed o al e na i e me abolic
pa hways. Fo example, p esen ly much a en ion is ocused on he ole o olic acid o i ica ion in
augmen ing colon cance isk. Also ea lie in he li e cycle, he i amin may addi ionally in luence insulin
esis ance [1]. Thus, ob aining sys ems ha allow a educ ion o he dose o olic acid while main aining
i s i amin ac i i y is a challenge o cu en nu i ional science.
One al e na i e o di ec o i ica ion, could be he use o encapsula ing sys ems o con olled elease
based on sup amolecula chemis y. This science has ecen ly allowed he design o new hyb id ma e ials
showing enhanced unc ional molecula ecogni ion and unc ionali ies. In his a ea, one appealing
concep is he de elopmen o “sup amolecula ga es”. These a e nanoscopic sys ems con aining
swi chable en i ies ha can con ol a will he s a e o he ga e (closed o open), allowing ei he he elease
o he con ined gues s o he en ance (o access) o molecula species o (o h ough) ce ain si es. Ga ing
p ope ies ha e been epo ed on su aces, in memb anes, o he design o a i icial channels, and ecen ly
in mesopo ous MCM-41- ype siliceous solids. Pe haps one o he mos appealing unc ions is hei
applica ion in ad anced sma deli e y sys ems. Fo ins ance, one possible use o pH-con olled ga e-like
sca oldings [2,3] could be he de elopmen o o ally applicable deli e y sys ems designed o ha e he
pa icula abili y o p o ec he ca go om he acidic condi ions o he s omach (acid pH, ga e closed) bu
will elease he load a he in es ine (basic pH, ga e open).
The aim o his s udy is o e alua e he po en ial use o olic acid encapsula ion in a mesopo ous
ma e ial MCM-41 o con olled elease o he i amin in biologic sys ems in o de o dec ease he
d awbacks o adi ional di ec o i ica ion associa ed o high abso p ion peaks.
2. Ma e ials and Me hods
2.1 Syn hesis o MCM-41
The mesopo ous MCM-41 suppo was i s syn hesized using he so-called “a ane ou e” in which
4.68 g o CTAB was added a 118°C o a solu ion o TEAH3 (25.79 g) con aining 0.045 mol o a sila ane
de i a i e (11 mL o TEOS). Nex , 80 mL o wa e was slowly added wi h igo ous s i ing a 70 °C.
A e a ew minu es, a whi e suspension was o med. This mix u e was aged a oom empe a u e
o e nigh . The esul ing powde was collec ed by il a ion and washed wi h wa e and e hanol. Finally,
he solid was d ied a 70 °C. To p epa e he inal po ous ma e ial, he as-syn hesized solid was calcined a
550 °C using oxidan a mosphe e o 5 h in o de o emo e he empla e phase.
2.2 Syn hesis o S1
In a ypical syn hesis, 0.35 g o MCM-41 and 0.07 g (0.066 mmol) o i amin B9 ( olic acid) we e
suspended in 50 mL o wa e pH=7.5, inside a ound-bo om lask in an ine a mosphe e. The mix u e
was s i ed o 24h a oom empe a u e wi h he aim o achie ing maximum loading in he po es o he
MCM-41 sca olding. The loaded solid was isola ed by acuum il a ion and d ied a oom empe a u e
o 36 h. Th oughou his ime, wo k is ca ied ou in da k (ambe ins umen al) and in an ine
a mosphe e o p e en oxida ion o olic acid (see igu e 1a).
2.3 Syn hesis o S2
The p ocedu e s a s as S1, bu a e 24h o s i ing an excess o 3-[2-(2-aminoe hylamino)-
e hylamino]p opyl- ime hoxysilane (4.3 mL, 15.0 mmol) was added. The inal mix u e was s i ed o 5.5
h a oom empe a u e. Finally, he solid (S2) was il e ed o , washed wi h 30 mL o wa e , and d ied a
oom empe a u e o 12 h (see igu e 1b).
1830 JosÈ Ba a e al. / P ocedia Food Science 1 (2011) 1828 – 1832
Au ho name / P ocedia – Food Science 00 (2011) 000–000 3
2.4 Solids cha ac e iza ion
XRD, TGA, Fluo escence spec oscopy and UV– isible spec oscopy echniques we e employed o
cha ac e ize he syn hesized ma e ials.
Fig. 1. (a) Design o loaded S1; (b) Desing o loaded S2
2.5 Vi amin elease s udies
Fo olic acid elease om S1 and S2 a ba ch sys em was designed using 10 mg o he s udy ca ie
ma e ial, which was placed in an ambe ial, o which is added a o al olume o 25 mL o he deli e y
solu ions (wa e a pH 2 and pH 7.5) and kep in s i ing o di e en elease imes s udied. A hese imes,
a sample was aken and passed h ough a nylon il e o 0.45. The deli e y o he i amin om he po e
oids o he aqueous solu ion was moni o ed ia ul a iole - isible spec oscopy by measu ing he
abso bance a 280 nm wi h a spec opho ome e .
3. Resul s and Discussion
3.1 Cha ac e iza ion o solids
Fig.2 shows powde X- ay pa e ns o he solids MCM-41 as-syn hesized, MCM-41 calcined and S1
and S2. PXRD o siliceous MCM-41 as-syn hesized shows ou low-angle e lec ions ypical o a
hexagonal a ay ha can be indexed as (100), (110), (200), and (210) B agg peaks wi h an a0 cell
pa ame e o 45.6 Å (d100 spacing o 39.5 Å). A signi ican shi o he (100) e lexion in he XRD
powde o he MCM-41 calcined sample is clea ly demons a ed in he cu e b, co esponding o an
app oxima e cell con ac ion ela ed o condensa ion o silanols du ing he calcina ion s ep. Fig. 2 also
shows cu e ha co esponds o S1 and S2. Fo hese solids he e lec ions (110) and (200) a e los , mos
likely ela ed o a educ ion o con as due o he illing o he po e oids wi h he i amin (see igu e3).
Fig. 2. Powde X- ay pa e ns o he solids (a) MCM-41 as-syn hesized, (b) MCM-41calcined, and he inal solids con aining he
i amin B9 ( olic acid) (c) S1 and (d) S2
1831
JosÈ Ba a e al. / P ocedia Food Science 1 (2011) 1828 – 1832
4 Au ho name / P ocedia – Food Science 00 (2011) 000–000
3.2 Release om S1
In he s udy o S1, olic acid elease was in ended unde di e en condi ions; i.e. pH 2 and pH 7.5.
Folic acid has low solubili y a acidic pH, so as shown in Figu e 3 i was no possible o elease mo e han
15% o he load a e 400 minu es when he deli e y is done in wa e a pH 2. When elease is pe o med
in an aqueous medium a pH 7.5, olic acid molecules a e in he o m o sal , inc easing he solubili y. In
his molecula con igu a ion, 80% o he ca go is eleased o he i s 15 minu es and a 100% elease is
achie ed a e 400 minu es.
Consequen ly, we can say ha olic acid encapsula ion in MCM-41 ma e ial, is a echnique which
accoun s o egula ing he elease o olic acid as a unc ion o pH. Wi h his beha io , i olic acid we e
in oduced o human body h ough diges i e sys em in he o m o S1, olic acid molecules would no be
deli e ed du ing passage h ough he s omach and will be eleased a he en ance o he in es ine, whe e
pH inc eases due o he p esence o bile sal s.
Fig. 3. Release p o iles o olic acid om he po es o solid S1 a pH 2 and pH 7.5
3.3 Release om S2
The e ec o polyamines as molecula ga es was s udied in S2 elease assays, using wo aqueous
solu ions (pH 2 and pH 7.5). Figu e 4 shows how a pH 2 he e is negligible elease. Howe e he e a e
no iceable di e ences om he deli e y o S2 when compa ed wi h S1.
Fig. 4. Release p o iles o olic acid om he po es o solid S2 a pH 2 and pH 7.5
1832 JosÈ Ba a e al. / P ocedia Food Science 1 (2011) 1828 – 1832
Au ho name / P ocedia – Food Science 00 (2011) 000–000 5
The i s di e ence is ha S2 shows a s onge hind ance o i amin elease a pH 2, because he e ec
o low solubili y is added o he s e ic hind ance a he exi o he po es imposed by he ancho ed
polyamine de i a i e. The second di e ence is ha a pH 7.5, a which he deli e y om S1 is e y quick,
when using solid S2 a mo e sus ained deli e y was obse ed. This beha io shows ha he use o
polyamines as pH- esponsi e ga e-like allows olic acid con olled elease o long imes (up o 7 hou s).
Thus, no only olic acid p o ec ion du ing passage h ough he s omach can be achie ed, bu also
p og essi e elease sys ems.
4. Conclusion
Adminis a ion o encapsula ed olic acid can be achie ed by using mesopo ous ma e ials MCM-41.
This sys em p o ec s he i amin om he ex e nal en i onmen du ing passage h ough he s omach,
being able o elease in he in es ine. This e ec can only be achie ed h ough encapsula ion due o he
low solubili y o olic acid in acid pH, o by using molecula ga es.
When con olled elease o e ime is needed, i is necessa y o use molecula ga es, being able o
secu e a p og essi e elease o olic acid along he diges ion pe iod, using a pH ga e-like. Once his in
i o beha io has been desc ibed, "in i o" s udies should be done in o de o ela e he con olled elease
wi h an imp o ed bioa ailabili y o he i amin.
Re e ences
[1] Lucock, M. & Ya es, Z. Folic acid o i ica ion: a double-edged swo d. Cu Opin Clin Nu Me ab Ca e 2009; 12:555-564.
[2] Casasús, R.; Ma cos, M.D.; Ma ínez-Mañez, R.; Ros-Lis, J.V.; So o, J.; Villaescusa, L.A.; Amo ós, P.; Bel án, D.;
Guillem, C. & La o e, J.. Towa d he de elopmen o ionically con olled nanoscopic molecula ga es. J. Am. Chem. Soc 2004;126,
8612–8613.
[3] Be na dos, A.; Azna , E.; Coll, C.; Ma ínez-Mañez, R.; Ba a , JM.; Ma cos, MD.; Sancenón, F.; Beni o, A. & So o, J..
Con olled elease o Vi amin B2 using mesopo ous ma e ials unc ionalized wi h amine-bea ing ga e-like sca oldings. J. Con ol
Release 2008; 131:181-189.
P esen ed a ICEF11 (May 22-26, 2011 – A hens, G eece) as pape FMS1286.