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Nghin cu chto ht nano cu trc li vnhm ng dng trong ysinh

Nguyn ThThy

Trng i hc Cng nghLun vnThS. ngnh:Vt liu v linh kin Nano

(Chuyn ngnh o to th im)Ngi hng dn: PGS.TS. Trn Hng Nhung, TS. Nghim ThH Lin

Nm bo v: 2012

Abstract. Tng quan vcc ht a chc nng; cc phng php chto silica SiO2xp bng phng php sol-gel; ht nanoshell Fe3O4@SiO2@Au; c trng ca htnano v cc ng dng ca ht nanoshell. Nghin cu cc phng php thc nghim:Chto lp bc SiO2 cho ht nano tto ht nano cu trc li vv cc phng

php kho st (Phng php knh hin vi in ttruyn qua (TEM), phng php tk mu rung (VSM), phng php o tn x nh sng ng hc (Dynamic LightScattering - DLS), phng php nhiu xtia X, Hp thhng ngoi, phhp thv

phhunh quang). a ra kt quv tho lun: Ht nano Fe3O4@SiO2; ht nanoa chc nng v ht nano SiO2 pht quang.

Keywords. Linh kin nano; Ht nano; Cu trc li v; Y sinh hc

Content

MUCc ht nano Fe3O4vi nhiu c im l th, c rt nhiu ng dng trong i sng

nh: cht lng t, cht bi trn, vt liu hp th sng in t (trong cng nghip), x lmi trng: lc nc, thu hi cht thiv c bit trong lnh vc Y-Sinh hc. Ht silica nnh v cu trc, khng c, c kh nng tng thch sinh hc cao v nht lb mt cachng d dng c chc nng ha bng nhm NH2do c th to c lp in mi Au

bng lin kt cng ha tr. Vi lp Au bao ph, ht nanoshell dng hot ha gn kt vicc phn t sinh hc nh amino acid, protein, enzyme, DNA v cc phn t thuc thng quacc cht c cha nhm SH.Nh vy ht nanoshell c ththc hin nhiu chc nngc sdng trong cchn on v ciu tr. Va dn ng, va pht hin v mang thuc, va lmgiu, phn tch v chn lc tbo, va sdng phng php t nhit v quang nhit.

V vy, mc tiu chnh ca lun vn ny l nghin cu v ch to ht nanoshellFe3O4@SiO2@Au nh hngng dng trong lnh vc y sinh hc. Trong lun vn ny chngti trnh byccphng phpch toht nanoshell Fe3O4@SiO2gm phng php Micellthun, micell o v phng php Stober. Sau chc nng ha b mt bngcch s dngAPTES, ri ph lp vng ln.

Lun vn c b cc nh sau:

Chng 1: Tng quanChng 2: Cc phng php thc nghim

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Chng 3: Ktqu v tho lun

Chng 1. TNG QUAN1.1 Tng quanv cc ht nanocu trc li v1.1.1 Cc ht nano

Ht nano l vt liu c kch thc t vi n vi trm nano mt, bao gm hng trmn hng nghn nguyn t ging nhau. Do kch thc nh nn cc tnh cht ha l ca chngb ph thuc nhiu vo trng thi b mt hn l th tch khi. So vi vt liu khi th vt liuc kch thc nanomet c nhiu tnh cht mi v l.

Tuy nhin cc vt liu ng dng trong sinh hc phi c p ng cc yu cu vcctnh cht dc l, ha hc, vt l, ng nht ca cc ht khnng ha tan c ca cchtv phi c tnh c thp. Do cc ht nano phi c kch thc nhc thddng khuch tn qua m, thi gian lng ng di, din tch bmt hiu dng cao; cc ht t

phi dbc bo vchng khi thoi ha do tc ng ca mi trng v c khnng tngthchsinh hc vi mi trng1.1.2 Cc ht nano Fe3O4

Ht xit tFe3O4 c cu trc tinh thferit lp phng cu trc spinel o, thuc nhmi xng Fd3m, hng smng a = b = c = 0.8396 nm. Sphn ttrong mt c sZ = 8, gm56 nguyn ttrong c 8ion Fe2+, 16 ion Fe3+v 32 ion O2-.

Nu kch thc ca ht st tgim n mt gi trno , tnh st tv ferri tbinmt, chuyn ng nhit sthng thv lm cho vt liu trthnh vt liu siu thun t. ivi vt liu siu thun t, td v lc khng tbng khng. trng thi siu thun tvtliu hng ng mnh vi ttrng ngoi nhng khi khng c ttrng ht nano trng thimt ttnh hon ton.1.1.3 Cc ht nano vng

Ht nano kim loi nh ht nano vng, nano bc c sdng thng ngn nm nay.Ni ting nht c thl chic cc Lycurgus c ngi La M chto vo khong thktht trc Cng nguyn v hin nay c trng by Bo tng Anh. Chic cc i mu tythuc vo cch ngi ta nhn n. N c mu xanh lc khi nhn nh sng phn xtrn cc vc mu khi nhn nh sng i ttrong cc v xuyn qua thnh cc[11].

Hnh 1.3.Mu sc ca cc c thl xanh lc hoc khi nhn nh sng phn xtrncc hoc nhn nh sng i tbn trong xuyn qua thnh cc

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Hnh 1.4. Cc php phn tch ngy nay cho thy cc mu sc cc ht nano vng phthuc vo kch thc ca chng [9].

1.1.4 Ht nano cu trc li v(nanoshell)

Ht nanoshell c thc cu trc a dng, nhng thng thng gm c hai thnhphnchnh l li v v. Hnh dng v cc tnh cht ca li v v, theo l thuyt cho thy c thc iu chnh bng cch khng chcc thnh phn v cc thng schto. Lp vc vaitr bo vv nhm khc phc mt snhc im ca phn li. Do chng thng cchto tnhng vt liu tr ha hc, c n nh cao, bmt c khnng tng thch sinhhc nh cc polimer, cc cht v c nh SiO2.

a, b, c, d,

Hnh1.6. Cc dng nanoshell: a)ht nano cha nhiu nhn ging nhaub) Ht nano cha nhiu nhn khc nhauc ) Ht nano cha mt li ng nht

d) Ht nano c bao quanh bi nhiu lp v.Trong cc dng ny, vt liu chn lm li/v, tlv kiu kt li/vl ccyu tc

bn to ra cc cu trc khc nhau ca ht nanoshell.1.2. Ht nanoshell Au@SiO2@Fe3O41.2.1. Phng php ch to phn li Phng php ng kt ta

Phng php ng kt ta iu ch cc ht oxit bng cch kt ta t dung dch muica cc cation kim loi di dng hydroxit, cacbonat, oxalatVic trn hn hp ferit v ferohydroxit trong mi trng dung dch di tc dng my khuy t, hn hp c kt ta vi

baz (NH4, NaOH, KOH) to ra cc ht hnh cu c kch thc ng nht [14].

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1.2.2 Phng php to ht Fe3O4@SiO21.2.2.1 Qu trnh sol-gela) Phn ng thu phn.

Phn ng thy phn thay thnhm alkoxide (OR) trong lin kt kim loi alkoxidebng nhm hydroxyl (OH) to thnh lin kt kim loi hydroxyl.

Theo phng trnh phn ng sau rt gn sauR-Si(OR)3-x(OH)x+ H2O R-Si(OR)2-x(OH)x+1+ ROH (1.2)Trong x l 0, 1 v 2

Cc thng snh hng chyu n qu trnh thy phn l pH, bn cht vnng ca cht xc tc, nhit , dung mi, tsH2O/ precusorb) Phn ng ngng t.

Phn ng ngng tcng xy ra theo c chthnucleophine lng phn tSN2 xy rahoc do sloi ra mt phn tnc do phn ng gia hai nhom silanol ho c do loi ra mt

phn tru do phn ng gia nhom silanol v nhom alk oxit. Giai on ngng tny nhmto ra cu lin kt siloxane ( Si-O-Si ), o l mt n vc bn ca polime v c.1.2.1.2.Phng phap Stober

Phng php Stober l qu trnh sol-gel tc l phn ng thy phn v ngng tca ccsilicon alkoxyde c pha long nng thp trong dung mi nc v cht ng dung minh acetone, ethanol, propanol v n-butanol hoc trong hn hp cc ru cng nh trong ccte. Da trn c sca qu trnh thy phn v ngng t, chng ta c thchto ht nanoshellFe3O4@SiO2bng phng php Stober. Sdng nguyn liu: ethanol, nc, NH4OH, TEOS.1.2.1.3.Phng phap micelle

Micelle l h gm 3 thnh phn: cht hot ng b mt, nc v dung mi. Trong ,cht hot ng b mt l cht m phn t c hai ugm mt u k nc v mt u anc. Do , ty thuc vo pha ca h l nhiu nc hay nhiu dung mi m s hnh thnhcc h micelle thun hay o.

Hnh 1.7. Cc hmicelle a. Hmicelle thun. b. Hmicelle o[8]1.2.3 Phng php ph lp Au

Qu trnh pht trin lp vi vi ht nanoshell l qu trnhph lp mng ln trnmt bmt ht nano v kt hp nhiu qu trnh ha hc trong mt dung dch keo: tlp rpha hc tip theo l mkhng in ly.

Hnh 1.8. Cc trng thi trongqu trnh pht trin lp vvng ln ht silicaLi nano silica vi ng knh khc nhau t khong 50 nm n 1 m c chto

hoc tng hp hoc tcc ngun thng mi. Cc li ht nano c bn cht tr. trthnhchc nng, cc b mt phi c thay i v mt ha hc bng cch s dng mt chtsiloxane c nhm amin (NH2) hoc thiol (-SH). Khi qu trnh to lp chc nng ny kt thc,cc bmt ht nano c mt cc nhm chc nng c khnng lin kt vi ht kim loi. Htnano vng kch thc nh1-2 nm c a vo mt dung dch silica l dung dch mkim

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loi khng in ly, kim loi c thbkhtrn bmt ca ht silica. Qu trnh ny to ramt vkim loi ng nht trn li in mi, vi dy khc nhau, ttrong khong t5 n20 nm.

1.3 c trng ca ht nano1.3.1 Tnh cht t

Khi gim kch thc ca ht xung di mt gii hn nht nh, td khng cngitheo cc hng xc nh bi dhng hnh dng hoc dhng ttinh thca ht na.Cc ht ttnh trthnh siu thun tkhi bn knh gim xung di 20nm. ng cong tha ca ht siu thun tl mt ng thun nghch c td Mr=0 v lc khng tHC=0.Lc khng tphthuc rt nhiu vo kch thc ca ht. Khi kch thc ca ht gim th lckhng ttng dn n gitrcc i ri tin gim rt nhanh v0 nh hnh v(Hnh 1.11).

Hnh 1.11. Hcphthuc vo ng knh ht1.3.1 Tnh cht quang

Khi kch thc ht nano vng dng cu tng th tit din tn xtng, do tng titdin tn xngi ta chto ht vng bc silica.

Hnh 1.12. Phhp thca cc ht SiO2/Au vi cc tlli/ vthay i[15]

Trn hnh 1.12ta thy khi ng knh li cnh, thay i dy lp vvng c thiu chnh vtr vng cng hng plasmon bmt (SPR) ca ht trong di bc sng mongmun. iu ny c l gii nh sau:1.4 Cc ng dng ca ht nanoshell1.4.1 Dn truyn thuc

Ht nanoshell c tnh tng hp sinh hc c gn kt vi thuc iu tr. Lc ny htnanoshell c tc dng nh mt ht mang. Khi cc ht i vo mch mu, ngi ta dng ttrng ngoi tp trung cc ht vo mt v tr no trn c th. Mt khi h thuc/ht ctp trung ti v tr cn thit th qu trnh nh thuc c th din ra thng qua c ch hot ngca cc enzym hoc cc tnh cht sinh l hc do cc t bo ung th gy ra nh pH, qutrnh khuych tn hoc s thay i ca nhit .

Hc

Dp Ds D

a menn men

Siu thu n t

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1.4.2 Lm giu v phn tch chn lc t boTrong y sinh hc phi thng xuyn tch mt loi thc th sinh hc no ra khi

mi trng ca chng lm tng nng khi phn tch hoc cho cc mc ch khc. Phntch t bo s dng cc ht nanoshell c t tnh l mt trong nhng phng php thngc s dng. Qu trnh phn tch c chia lm hai giai on: nh du thc th sinh hc

cn nghin cu v tch cc thc th c nh du ra khi mi trng bng t trng.1.4.3 Phi hp t nhit, quang nhit

Trong phng php t nhit,cc ht nan t tnh c phn tn trong cc m mongmun, sau tc dng mt t trng xoay chiu bn ngoi ln (cng v tn s) tcdng ln ht nano chng to ra nhit nung nng nhng vng xung quanh. Nhit khong42 C trong khong 30 pht c th git cht cc t bo ung th. Cc yu t nh hngn hiu ng t nhit l lu lng mu vphn b ca cc m.

Hnh 1.16. Qu trnh t nhit bng ttrng xoay chiuHoc s dng liu php quang nhit in vivo, tc l dng ngun laze chiu vo.

Trong trng hp ny ch s dngngun sng hng ngoi gn (NIR) v ch loi nh sng nymi c kh nng i su vo c th qua ca s ca da (800 -900 nm) do y l vng cc tiuca hp th ca hng cu v ca nc.

Chng 2. CC PHNG PHP THC NGHIM

2.1 Ch to ht nano Fe3O4@SiO2@Au core-shell-shell2.1.1 Ch to ht nano st t Fe3O4

Trong lun vn ny, chng ti s dng ba ht nano st t nh sau: Fe3O4bc trongoleic, Fe3O4bc axit Citric, Fe3O4 trn.

Hnh 2.1. Cc ht nano st t2.1.2 Ch to ht nano Fe3O4@SiO2core-shell2.1.2.1 Ch to ht nano Fe3O4@SiO2core-shell bng phng php micelle thuna) Cc nguyn vt liu ha cht:- Precusor: methyltriethoxysilane (MTEOS): CH3-Si-(OC2H5)3, > 97%, Merck

- Hot ng b mt: Aerosol-OT (AOT): C20H37O7NaS v ng hot ng b mt Butanone-2: C4H8O (Trung Quc).

- Amine: aminopropyltriethoxysilane (APTEOS): (C2H5O)3- Si- C3H6-NH2 , > 98% Merck,ng thi y cng l ng precuosor.

Ht nanoshellFe3O4@SiO2@Au Nam chm

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b) Quy trnh ch toHn hp hot ng b mt AOT v Butanone-2 c pha vi nhau theo t l 0.11g

AOT: 200 l Butanone-2 (c 0.31ml hn hp hot ng b mt) hay v t l mol l 1:9,sau c rung siu m cho ti khi dung dch trong sut v mt quang hc v ng nht vmt ha hc. Sau , cho Fe3O4trn vo v rung siu m 20, tip n l cho 200l MTEOS

vo hn hp trn, rung siu m 10. Tip theo, 10ml nc ct hai ln c thm vo v khuyt 1h thu c h micelle: (Fe3O4 + MTEOS)/AOT (butanone -2)/ nc. Mi h micelleny s l mt trung tm phn ng, phn ng sol-gel s xy ra trong mi micelle khi lng xctc APTEOS c thm vo trong h. Sau khi cho xc tc (APTEOS), pH ca hn hp tngln khong 9. Cc ht nanoshell Fe3O4@SiO2 hnh thnh vi nhm lin kt amine (-NH2).Sau ht nanoshell c ra 3 ln bng ly tm.2.1.2.2 Ch to ht nano silica bng phng php micelle oa) Cc nguyn vt liu ha cht

- Fe3O4bc oleic phn tn trong toluene (kch thc 10nm)- Hot ng b mt, Triton X-100 v ng hotng b mt: Hexanol-1- Dung mi: Cyclohexan. Tetraethyl orthosilicate (TEOS- Xc tc NH4OH- Amine: Propylamin-triethoxysilan (APTEOS)

b) Quy trnh ch toTrn hn hp TritonX-100 1.77ml, hexanol-1 1.6ml, Fe3O4, cho vo rung siu m 30

pht, sau thm 7.5 ml Cyclohexan v cho hn hp vo khuy t trong 45 pht. 430l H2O,100l TEOS c a vo hn hp trn v khuy t trong 1 gi. Sau cho dung dch

NH4OH, ri khuy t trong vng 10 gi. Cho tip 20 l amine APTEOS, chc nng hab mt cc ht nano silica vi cc nhm chc amine. Qu trnh chc nng ha b mt cc htnanosilica c thc hin trong khong 20 gi. Sau loi ht dung mi cyclohexan, hotng b mt bng ra ly tm 3 ln.2.1.2.3 Phng php Stobera) Cc nguynliu vha cht

- Fe3O4bc axit Citric- Dung mi Ethanol (C2H5OH)- Precursor: Tetraethyl orthosilicate (TEOS) cng thc phn t SiC8H20O4- Xc tc NH4OH- Amine: Propylamin-triethoxysilan (APTEOS)

b) Quy trnh ch tob) Quy trnh ch to

15ml Ethanol trn vi Fe3O4bc axit Citric, rung siu m 20, tip n cho TEOS,NH4OH khuy t 30, cui cng a APTEOS khuy t 20h.

Phn ng thy phn v ngng t ca TEOS c trnh by cng thc 2.5 v 2.6.Cht xc tc NH4OH cung cp nc cho phn ng thy phn (cha khong 70% lngnc), v nng pH ca hn hp phn ng thc y qu trnh ngng t, cn APTEOSng vai tr l ng precursor nhm to nhm chc NH2trn b mt ht nano silica2.1.3 Tng hp ht nanoshell Fe3O4@SiO2@Au2.1.3.1 Ch to ht vng kch thc nh lm v ht

Ht nano vng kch thc t 1-2nm l cc ht nano vng c ch to bng phngphp Duff-Baiker trong mi trng citrate, cc ht nano vng c hp phtrn bmt ht lithng qua tng tc tnh in.2.1.3.2 Qu trnhgn mm Au ln ht nanoFe3O4@SiO2Qu trnh pht trin mm hnh thnh lp v nano vng

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2.2 Cc phng php kho st2.2.1. Phng php nhiu x tia X

Khi chiu chm tia X ti, tia X b tn x trn cc nt mng theo mi phng, hai tiatn x song songc th giao thoa vi nhau v to thnh cc cc i v cc tiu nhiu x. Vvy, theo cc phng ta u c hin tng nhiu x nh sng, tuy nhin ch theo phng

phn x gng (phng c gc phn x bng gc ti) mi quan st c hin tng nhiu xv theo phng cng nhiu x ln. Phng php phn tch cu trc tinh th v phabng nhiu x tia X da trn hin tng nhiu x tia X bi mng tinh th khi tha mn iukin phn x Bragg:

2dsin= n.(2.7)

Trong l gc phn x, l bc sngtia X, n l bc nhiu x.

Hnh 2.7. Hin tng cc tia X nhiu xtrn cc mt tinh thcht rn2.2.2. Phng php knh hin vi in t truyn qua (TEM)

Knh hin vi in t truyn qua(TEM) l cng c rt hu hiu cho cc nghin cu vtliu v y hc.Khi o TEM ta c th xc nh c hnh dng, kch thc trung bnh ca htv s phn tn ca ht trong mi trng cht. Khi chm in t chiu ti mu vi tc caov trong phm vi rt hp, cc in t b tn x bi th tnh in gia ht nhn nguyn t vlp my in t ca vt liu gy ra nhiu x in t. Chm in t nhiu x t vt liu ph

thuc vo bc sng ca chm vt liu ti v khong cch mt phng mng trong tinh th,tun theo nh lut Bragg nh i vi nhiu x tia X.2.2.3. Phng php t k mu rung (VSM)

Thit b o VSM cho php ta kho st s ph thuc ca t vo cng t trng,dng ng cong t tr t ta c th bit c lc khng t Hc. S dch chuyn ca ngcong t tr cho ta bit c Hint, Hexdo vy c th kho st c lc tng tc trao i giacc lp st t vi st t, st t vi phn st t. M hnh b tr h o c th hin trn hnh2.9.

Hnh 2.9. S cu to ca my o VSM2.2.4. Ph hp th

Ph hp th l ng biu din s ph thuc ca h s hpth ca mi trng vtvo bc sng ca nh sng ti. Khi chiu mt chm tia sng n sc c cng I 0song

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song vo mt mi trng vt cht c b dy l (cm) v nng C (mol/l), chm tia ny s bmi trng hp th, tn x hoctruyn qua. Cng I ca chm tia truyn qua mi trngny b gim theo quy lut theo biu thc ton hc ca nh lut Beer-Lamber sau:

ln (I0/I) = Kn (2.13)

hay: ln (I0/I)= lC (2.14)

Trong : K- l h s hp thn- s mol cht nghin cu ttrn ng i ca bc xi lng ln(I0/I) gi l mt quang (D) hay hp th (A), l h s hp th mol.

2.2.5. Hp th hng ngoiPh hp th hng ngoi chnh l ph dao ng quay v khi hp th bc x hng ngoi

th c chuyn ng dao ng v chuyn ng quay u b kch thch. Tt c cc phn t ccu to t cc nguyn t ni vi nhau bng cc lin kt ha hc. Tn s dao ng khngnhng ph thuc vo bn cht ca tng lin kt ring bit nh: C -H hay C-O, m cn phthuc vo c phn t v mi trng xung quanh n. Bin dao ng ca mt h s tng lndi tc ng ca s va p. Tng t nh vy, di tc ng ca trng in t (sng hngngoi) th bin dao ng ca cc lin kt ha hc v cng vi chng l dao ng ca ccin tchcng thay i. Do cc phn t ch hp th cc sng hng ngoi c nng lng tngng vi khong cch gia hai mc nng lng dao ng ca nguyn t nn bin dao ngtng khng lin tc m nhy bc.

Chng 3. KT QU V THO LUNTrong chng ny, chng ti strnh by mt skt quthu c tthc nghim.

Cc tho lun c tp trung chyu vo phn tch cu trc, thnh phn ho hc, tnh chtt, quang v kch thc ht.

Cc ht nanoFe3O4bc axit Citric, bc oleic v Fe3O4 trn c k hiu l F1, F2,F3

3.1. Hnh thi b mt v kch thc ca htnanoshell qua nh TEM3.1.1 Hnh thi b mt v kch thc ca ht st t qua nh TEMCc mu o c phn tnvo trong dung minc, toluene chp nh TEM di

dng cht lng. Phn b kch thcv hnh dng ca ht c kho st thng qua php ony.

Hnh 3.1 a, b, c l nh TEM cacc mu F1, F2 v F3 cng thang o 100 nm. Tnh TEM ta thy cc ht trong mu c dng hnh cu, kch thc kh ng u v phn tnkh tt trong dung mi.

Hnh3.1. Kt qunh Tem ca ht nano st tmu F1, F2, F33.1.2 Hnh thi b mt v kch thc ca nanoshell Fe3O4@SiO2qua nh TEM

a) nh Tem ca ht nanoshell Fe3O4@SiO2 ch to bng phng php Micell thun

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Hnh 3.2. nh Tem ca ht nanoshell Fe3O4@SiO2 ch to bng phng php Micell

thunb) nh Tem ca ht nanoshell Fe3O4@SiO2 ch to bng phng php Micell o

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Hnh 3.3. nh Tem ca ht nanoshell Fe3O4@SiO2 ch to bng phng php Micell

oc) nh Tem ca ht nanoshell Fe3O4@SiO2 ch to bng phng php Stober

Hnh 3.4. nh Tem ca ht nanoshell Fe3O4@SiO2 ch to bng phng php Stober

KT LUNLun vn trnh by cc kt quca vic chto v nghin cu cc tnh cht ca cc

ht nano Fe3O4@SiO2@Au cu trc li vc cha tm mu RB v FITC. Lun vn thc

hin c cc cng vic sau:- Sdng nhiu loi ht nano st tcho cc phng php khc nhau nh: ht tbc

axit oleic, ht tbc citric v ht ttrn-

Ch to cc ht nano Fe3O4@SiO2 cu trc li v cha tm mu RB bng phngphp micell thun

-

Chto cc ht nano Fe3O4@SiO2cu trc li vcha tm mu FITC bng phngphp micell o.

-

Chto cc ht nano Fe3O4@SiO2cu trc li vphng php Stober.- Chc nng ha bmt ht Fe3O4@SiO2bng nhm NH2- Tng hp thnh cng lp vnano Au quanh li in mi t tnh Fe3O4@SiO2. Ht

nanoshell c to ra va c tnh cht siu thun tca ht nano Fe3O4, va c nhcng hng plasmon trong vng nh sng nhn thy rt ph hp cho cc ng dng ysinh

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- Kho st hnh thi v kch thc ht.

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