7. RESULTADOS
7.3 Efecto del consumo de los suplementos sobre
at~ • • qainat a blo.nk ot wator on Unioam Sp. 500 apeotrophoto~~~etar.
11.
(a) Speotrtl ShU't
Clitt'ord &D4 ieee (1967b), J<i.Qs enl Ri.oholaon (1969) uud the
lhltt in the utra'f'iolet apeotral peak ot aflatoxin :a,_ aa criterion tr:.o oonolwUIII that binding between 41"l.atoxin ~ ani the tollodna
•terlala had taken plaoea DRA, purinea, pyr1.addi.nea , 11.1oleoa1doa and pol.JDJoleotidea.
'l'!w oritarion wu alao used in this imeatisation ot the
1nter-Mt10D 'between DIU and atlatoxina ~ ' c 1 and a o• ot their oo-motabolitoa.
lilA wu di.uol'f84 1n 0 . 0 l U trU-}\Ydroohloride butter (pH . 7 .o ).
1he apeotrua ot DNA aolution waa tirat determined uaina a
Perkin-lya'-UltraTiolet"'Yiaible apeotrophoto lllltter 1} 7 uv . The DNA solutio n
wu then .U.4 ritb ODII ot the abo.,. ~nt1uned oompowda. At'tor tho ll1n1IZ'e ba4 'b•n allowed to atam tor about )0 minutea, ita ultraYiolot
lpeotz,aa wu plotted on the aaca oiBrt u that ot the pure DNA aolution.
4 lb1.tt iD the wanlen,th ot the peaka wu talcan u indioatin ot bond f01 a\ioo betw"n DnA au1 the fluoro :toent compound involnd .
('b) Rielxtit I'
'!be dial,raia uthod or Blaok Al1l Jirsenaoo (1967) utilised to
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-88-ill n•'artl \a. 1..-_...tiOD 'behllll atlatoen ~ 1 hiaton . . &ll4 D NA
- ?'npte&. lelaUou ot t • ..,.n oom entrwtiona ot llliA, mtA, prot ein,
... ·a. eD4 ... peuio .. ot oell-..brane, o oll ulo ae, ..teylroar bo.a;y-cellu-1111, pboapM aall•loae, oellulon a cetate and eo t eol&-4 ell alo . . w ax ..
Jll1al"e&• Tbeae ... eMe4 to aq~ aolUtlon ot t.be tluoruoent
11 1)'10n4t u4er iareati&&tion (i.e. Qf'lata d..na I:L c 1 aD4 5 otMI'
oo-MaMlltea 4edpw.te4 S, o 3 , Jls• c
5 m1 o 6 ). Allquota ot t he llixturu ware ti"AAIIaterred to 4'•17ait tubin& {2 ot: . diai'Ot er ) . The tlabea n N
eupenllecl in an eqlol&l Yolu.e o t 4btlll o4 watezo. n. llixtiU'ea oontaini.Jl6
0 0
.rA am W were plAoed in a re~ator at ~ -8 C while thoee imolYi.D&
\be oth er ~ 80leou.lar weiaht o OilJlOU.Dla were kept at roo• tODperAture (25-28°c). Pol- eaob intenotion betwoon a .aorozcol eoule and a tluoro:.oent OCMIJIJOI'D21 1 It 41al.rtia UIIUlioa woro i~"pcLI'ed o Dail,r wet1mt1oa at tbe
oonoentration ot tbe oocpoum. 1o t he inn14e and outa1cle ooapa.rtrerdl ot dhl,rab Tue ola w ore carried out. 'lhau •• ti•tlons were ooaUmc4 aut:U tb. yaluaa or the oonoentratio .. ot the tluor eaoent ooliQC)ull!a in tbe 1na14e and outa1cle oo~~part•nt• 1nd 1oated t he a ttai nt or en
eqaUib.naia (Donnon eq, l1librii.I.Cl)o .For eao h tluoresoe.nt o o11p0 \l.Dd tor wld.ob equilibra.i!l biLl been 1.ndioat.o4, oird Jar de to.rl:d.DAtic=. o t U•
M••auaUou 1D t.be inaUe aD! outa1cle OO=P&riaenh n~ oarr1e4 out.
A at.cnttioantl,y ht s bor oonoantrat 1oa within tbe tub~ t haD oa\&14• it. na hkeo aa ilxlioatbe ~ a b inli"<J interaotion be tnen
\be pvUeular t'lllonaoeDt oOIIpOW'ld ani the ~Uro ooleo ulo imoln4 la \be "'•'7•11.
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- 1°C tor ..., ~ '!!
·~•U• . , 20 , 000 •·
111 tria-lluftG' (pH. 7~). w..
~ll'etioa of tbe Yarbur1 SUif'FI , . . .
1' iat-oln4 pud.llf tile ot • tilter "per tol414 ia
••tet-11 ill ~ tnll
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...
:ra .._ \o rn "• ol4 II'•••• tro• ·amMtera aD! the arbuzoa
t1M1J wwe wube4 with paoline md t he aoaked in o broaio o .o14
e. • ,..,... '!'he obroeo ao14 n.a J-l'ep .reel u 01.1tH ned b7 Oabzoeit,
l&aU .... Stau.tfG' (1961.) .
'!be "'',_.are wu tbeD aeneroual,y rinaed w ith tap--.rater follow ed
t 111 ~ 41atUle4 w a tezo , The mz M~~etera in odcUt1on WIZ'I tluabod with
0 0
..noae Won be' q clriecl at 90 - 100 c .
(o)
... ioall,y the mthod ot Sobale a (19/.4) u 110dit1e4 by S&ntia.{ o
llftaola &D4tA&tllne4 b7 U llbreit , Bun-ia IUid St..u1'fer (1961.) wu uaed tor
Mwltll'l ._.. inolined at about .30 0 to the boriaonhl with
till etopoook ooouprinl the lower ~Dition . The atanoe wu a uob that
- - 6101'~ " . . . Joint tor the t' arburs t l uk ... direotl,r abou the arm ot m ?tezo b~ the 150 • • reference ~~ark . A l'\l'bb er teat i 1'Ule4 wl\h paritied -1'0111'7 . . . aentl,y pu.~ hacl into t be inbt tube ot the
z ruz1ter replated b7 the tbr ... n,y et . ook. .&a t be teat wu p \l .ahed
Sa f'uoU.r 1ato tbe inlet tube , the c.erour y ro ae tow&rlb the two
l'etawoe a r b u.atil the lnela juat tell aho rt o t t he r4l'ka . A t tb1l jr •"'*• a MrW-Ollp w u ti t tecl to the hat an4 llo.l,y a4Juated to
I dJ aQe EINU1'7 1Dto the n Mllter , Jb' &d juat~~eut o t th:11 aor11 ollp '- Ji Ill ~ . . aa the te&t e01.1pled with alla bt a4Juatraent o t the iDoll·
.. ., •• ot \be • muter, tbe lrrela ot Ml'OUJ7 were brouah t to the two
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-91-tufww??R -.rb. At tbia po1Dt the otoJioock wu tur.oe4 in ordco to
~ \be MI"'UI"J iD the -.no.etor t'rotz that in tbe rubber teat.
(tile teat wu N~WHe4 &Dil tbe 8IZ'Oury uaooi&ted ritb it lib ri.ae . )
!Ill 7 I"'I.li'J in the 7 no=ptcr WU let out 1Dto a tared 50 al, beaker
netptaole. '.L'hb wu ni&Pe4 aa1 ita tetrpel'Rture recorded. 'l"he aeoond thp 1Droln4 tillinc the n .. k which went ritb the J.&rli.oular 7 mmt C!r
to .soh a U.el tbat wben t1tte4 CD the .ano~cr, the arou:r1 roae Juct
to the reterenoe ..rk &bon the quiok- tit Joillt tor the f'leek , n.
....uzo;r in tbe tluk wu thS'I woisl*l and ita tqerature reoordod , These
zdd;nl.Uon were repeated to p:re weiahh in duplloate . t~ •ol~a tar
* two auploa ot •roury wore ocaloulated tram tboi.r reapeotin te~~pera•
. . . . at tba t ! . Of Wei&biJll • 'cba IWII Of t.haae two YOli.I.CMII su n the
total TOl!!RI ot a sinn fi aak to&otbtr with ita MnOIMtar up to tba )50 m.
FM'k . 'lbR T&lun obtained ware aubatitutod. 1n the tlaak oonatan\
eqnation (A) attar aubtraotion ot' the Yolun 1 ot' liquid pre11nt 1n tbe tl•ak dur1zlt reapintion aperiaenta
I •
•
'• • ' •
1:13
X • b Yl • 04-V t
'1' • bk - - ( A )
Toluae ot ,... taken up
tba •oln r o&lou.lated abo•• - t~ total •olu. ot
Uqu14 1D nut
tt '*'tur• ot the both to whiob the maMTMten
are tixed ,
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-92-• • Mnhr4 pruaure wbiob b 10,000 ca. keb a or IC • lol• .. llit1 ot p.a imolud in r uotion . . natrawa •
(apaoea . . cl in a1. su/al l iqu14 wbc:n gu i.e at a
Jll"••ur• ot 760 • Rs at th. teapc.nture '1') K • !he tluk oonatant requirocl.
aua penaion:
!be t e at llioro orp.nia11, were obtainnd troa aaar alanta in RDuaa
~ W t IODte4Di"' ' " 110re q a.r thap l.lDual , 'fh powth obtdJMCl attar l8 hou.ra o t ino1.1bAt1on at 30°C vu wuho4 ott
till aluta w1 tb aterile Jtzoeb phoaphate buffer. The r 'f&l or the
11uten.1 ooloDita ott the ala.a t aurtaoao vu tuUi t&hd by tho uoa
ot a sterile J,.1bapri s laea-1'04. The a uapena i oa obta1DD4 wu
MDtr1t'uae4 at 10, 000 S• i n ot orilo oaatrit'usa tubea t or 20 lliriJt •
a. IU,PVnAtUit wu 4iloiU"de4 an ! tho aodi&:ont re4iaporee4 in J'reoh
n.ru. lre~pbo1pbata butter. f bia proo .,a w u o&n'iad out thri oa.
'lbe fSrwl lluterial auapeaai? n wu utiliae4 ill cn:;rpn uptw atudha.
·l·'t·~
I t ' Uog ot 2JV!9 uptt ft , bz acmoao tct! teobniqut
'1o tbe z So oo ... rt~ ot aterUe , oloan , dey a.nd 8J'OU e4 arburs
t'leea~ 1 111 . ot aterila &q\I.IOlll aol utiom o t the oollllOIUld.e boin&
...,, .. , baTil)s the aaleota4 oooo.utraticma were aMed u eptUall,y.
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