B Y T TRNG I HC DC H NI NGUYN PHNG NAM NGHIN CU TNG HP TIU PHN NANO CHA BC KHA LUN TT NGHIP DC S H NI 2015 B Y T TRNG I HC DC H NI NGUYN PHNG NAM NGHIN CU TNG HP TIU PHN NANO CHA BC KHA LUN TT NGHIP DC S Ngi hng dn: 1. TS. Nguyn Th Thanh Bnh 2. ThS. Th Thanh Thy Ni thc hin: 1. Khoa Y-Dc, i hc Quc gia H Ni 2. B mn Ha Dc, i hc Dc H Ni H NI 2015 LI CM N Ti xin by t lng bit n su sc n : -TS. Nguyn Th Thanh Bnh -ThS. Th Thanh Thy Nhng ngi thy nhit tnh hng dn v ht lng gip ti trong thi gian thc hin kha lun tt nghip. Ti cng xin gi li cm n chn thnh n TS. V c Li, DS. Trnh Ngc Dng cng ton th cc thy c gio, k thut vin khoa Y-Dc, i hc Quc gia H Ni, cc thy c gio, k thut vin b mn Bo ch, b mn Hadc,ihcDcHNinhittnhgiptitrongqutrnh thc hin kha lun. Ti xin trn trng cm n: -Vin v sinh dch t trung ng -KhoaHa,trngihckhoahctnhin,ihcQucgiaH Ni gip ti thc hin mt s phn tch trong qu trnh lm kha lun. Ti xin chn thnh cm n Phng o to trng i hc Dc H Ni toiukinthunli,cnggianh,bnbngithnlunbn gip , ng vin v to mi iu kin thun li ti hon thnh kha lun tt nghip. Kha lun c thc hin t ngun kinh ph ca i hc Quc gia H Ni ( ti QG.14.58). H Ni, ngy 13 thng 5 nm 2015 Nguyn Phng Nam MC LC T VN ................................................................................................................. 1 CHNG 1. TNG QUAN .......................................................................................... 2 1.1.Gii thiu chung v bc ..........................................................................................................2 1.2.Gii thiu h gii phng ion bc ko di ............................................................................3 1.3. Tc dng ca tiu phn nano cha bc..............................................................................5 1.3.1.Tc dng khng khun ....................................................................................................5 1.3.2.Tc dng chng nm ........................................................................................................6 1.3.3.Tc dng chng virus .......................................................................................................6 1.3.4. Tc dng chng vim .......................................................................................................6 1.3.5. Tc dng cha bng v lm lnh vt thng..............................................................7 1.4 .Mt s ng dng ca tiu phn nano cha bc trong y dc v m phm ............7 1.4.1.Trang thit b y t ...............................................................................................................7 1.4.2. Thuc v m phm .............................................................................................................8 1.5.c tnh ca nano bc cloridv cc phng php xc nh ......................................9 1.5.1.c tnh ca nano bc clorid ..........................................................................................9 1.5.2. Cc phng php xc nh .......................................................................................... 10 1.5.2.1.S dng knh hin vi in t truyn qua .......................................................... 10 1.5.2.2Phn tch ph UV-VIS ....................................................................................... 10 1.5.2.3.Ph nhiu x tia X ............................................................................................ 11 1.5.2.4.Ph hng ngoi IR ........................................................................................... 11 1.6.Mt s phng php tng hp tiu phn nano mui bc .......................................... 11 1.6.1.Phng php tng hp ha l kt hp ....................................................................... 11 1.6.1.1.Mt s quy trnh tng hp ha l kt hp ........................................................ 12 1.6.1.2.c im ca cc phng php tng hp ha l kt hp ............................... 13 1.6.2.Phng php sinh tng hp tiu phn nano cha bc ........................................... 14 1.6.2.2. c im ca cc phng php sinh tng hp .............................................. 14 1.6.3.Phng php tng hp ha thc vt .......................................................................... 15 CHNG2.NGUYN LIU, THIT B V PHNG PHP NGHIN CU ........ 17 2.1.Nguyn liu nghin cu ....................................................................................................... 17 2.2.Thit b nghin cu ............................................................................................................... 17 2.3. Phng php nghin cu ................................................................................................... 18 2.3.1.Tng hp tiu phn bc clorid..................................................................................... 18 2.3.2. Khao st anh hng ca cht bn ho ti s hnh thnh tiu phn nano ......... 19 2.3.3. Xc nh tnh mt s c tnh ca tiu phn nano tng hp c ................. 19 2.3.3.1.Kch thc v phn b kch thc tiu phn .................................................. 19 2.3.3.2.Th Zeta ca tiu phn .................................................................................... 20 2.3.3.3.Hnh dng tiu phn ........................................................................................ 20 2.3.3.4. bn vi nh sng........................................................................................ 20 2.3.3.5.Bn cht ha hc ca tiu phn ...................................................................... 21 2.3.3.6.Tng tc gia tiu phn v cht bn ha ...................................................... 21 CHNG 3.KT QU THC NGHIM ......................................................................... 22 3.1 nh hng ca cht bn ho n s hinh thnh tiu phn bc clorid ..................... 22 3.1.1.Anh hng ca loi cht bn ha................................................................................ 22 3.1.2. Anh hng ca lng cht bn ha Polyvinyl alcohol .......................................... 23 3.1.3. Anh hng ca lng cht bn ha Polyvinyl pyrrollidon K30........................... 26 3.1.4. La chn cht bn ho v cng thc bo ch .......................................................... 28 3.2.Xc nh mt s c tnh ca tiu phn nano bc clorid va tng hp c ..... 30 3.2.1.Kch thc, PDI v th Zeta ......................................................................................... 30 3.2.2.Hnh dng tiu phn ...................................................................................................... 31 3.2.3. bn vi nh sng....................................................................................................... 32 3.2.4. Ban cht ha hc ca tiu phn ................................................................................. 34 3.2.5.Tng tc gia tiu phn v cht bn ha ................................................................. 35 BN LUN .............................................................................................................................. 38 KT LUN V XUT .................................................................................................... 40 DANH MC CC K HIU, CH VIT TT ADNAxit Deoxyribo Nucleic BPBritish Pharmacopoeia (Dc in Anh) CTSChitosan DVNDc in Vit Nam KTTP Kch thc tiu phn M(1..46) K hiu cc mu thc nghim PDI Poly dispersity index ( Ch s a phn tn hay phn bKTTP) PBKT Phn b kch thc PVA Poly vinyl alcol PVPPolyvinyl pyrolidon K.30 SEM Scanning electron microscope (Knh hin vi in t qut) TEM Transmission electronic microscope ( Knh hin vi in t truyn qua) TBTrung bnh USPThe United States Pharmacopoeia (Dc in M). DANH MC CC HNH V, TH Hnh 1.1. Thuc khng khun dng cho vt bng cha tiu phn nano bc clorid .... 3 Hnh 1.2.ng dng ca bc (bn phi) v nano cha bc (bn tri) trong y hc ..... 4 Hnh 1.3.Tc ng ca ion bc ln vi khun. ............................................................ 5 Hnh 1.4.Ion bc v hiu ha enzym chuyn ha oxy ca vi khun ......................... 5 Hnh 1.5. Ion bc lin kt vi cc base ca ADN ....................................................... 6 Hnh 1.6.Ming dn vt thng Acticoat. ................................................................. 8 Hnh 1.7. Bng cha tiu phn nano bc cha lnh vt thng ................................. 8 Hnh 1.8. Sn phm kh mi c th c thnh phn nano cha bc ............................ 9 Hnh 1.9.Dung dch ung tng cng min dch ASAP ........................................... 9 Hnh 2.1.Quy trnh tng hp tiu phn bc clorid .................................................. 18 Hnh 3.1. th mi lin quan gia lng PVA v kch thc tiu phn AgCl ..... 24 Hnh 3.2. th mi lin quan gia lng PVA v PDI ca tiu phn AgCl ......... 25 Hnh 3.3. th mi lin quan gia lng PVA v th Zeta ca tiu phn AgCl ... 25 Hnh 3.4. th mi lin quan gia lng PVP v kch thc tiu phn AgCl ..... 27 Hnh 3.5. th mi lin quan gia ng PVP v PDI ca tiu phn AgCl............ 27 Hnh 3.6. th mi lin quan gia lng PVP v th Zeta ca tiu phn AgCl ... 28 Bng 3.5.Cng thc bo ch mu M18 v M19 (n=2) ........................................... 29 Hnh 3.7.Anh chp SEM ca hn dch nano bc clorid .......................................... 31 Hnh 3.8.S thay i mu sc sau khi tip xuc vi nh sng mt tri ca hn dch nano bc clorid .......................................................................................................... 32 Hnh 3.9.Ph hp th UV-VIS ca mu hn dich M24 ngay khi iu ch ............. 33 Hnh 3.10.Ph UV-VIS ca mu hn dch M24 sau khi cho tip xuc nh sng ..... 33 Hnh 3.11. Ph X - ray ca nano tiu phn bc clorid .............................................. 35 Hnh 3.12. Ph IR mu hn dch bc clorid .............................................................. 36 Hnh 3.13. Ph IR mu hn dch trng ..................................................................... 37 DANH MC CC BNG, BIU Bng 1.1.c im ca mt s phng php tng hp ha, l kt hp ................. 13 Bng 1.2.cim ca mt s phng php sinh tng hp ................................. 15 Bng 2.1. Nguyn liu s dng nghin cu .............................................................. 17 Bng 3.1.Cng thc iu ch hn dch bc clorid vi mt s cht bn ha ........... 22 Bng 3.2.ng knh, th Zeta v PDI ca tiu phn AgCl vi mt s cht n nh (n=2) .......................................................................................................................... 23 Bng 3.3.ng knh, th Zeta v PDI ca tiu phn AgCl bo ch vi cc lng PVA khc nhau (n=2)................................................................................................ 24 Bng3.4:ngknh,thZetavPDIcatiuphnAgClkhiiuchvicc lng PVP khc nhau (n=2) ...................................................................................... 26 Bng 3.6.ng knh, th Zeta v PDI ca cc mu M18 v M19 (n=2) .............. 29 Bng 3.7: ng knh, th Zeta v PDI ca tiu phn nano bc clorid ................... 30 1 T VN Bclmttrongnhngnguyntctnhkhtrngmnhnhttrongt nhin,tnhkhngkhuncancchutrongbicnhtnhkhngkhngsinh ca vi khun gia tng. Do tc dng khng khun theo nhiu c ch khc nhaum t lkhngbccavikhunlrtthp.Tronglchs,bccsdnglmthuc dinhiudngkhcnhaunh:bcnitrat,bcprotacgon,bcsulfadiazin....Mi loiucunhcimring.Dngbcnitratchonngionbccaonhng thng gy c v tc dng ngn; bc protacgon c mu khng c a chung, n nh thp; bc sulfadiazin s dng di dng kem kh v sinh vt thng, thi gian tc dng ngn. Khi nim v cc th nghim u tin v cng ngh nano c hnh thnh t nhng nm 1970, t m ra xu th tng hp cc cht mc phn t v nguyn t. Ngy nay, vic to ra cc vt th vi kch thc nano (10-9 nm) tr nn ph bin,kchthcnycchtvtchtthhinnhiutnhchtlhartkhcso vi vt cht kch thc th. Nh ng dng cng ngh ny, nano tiu phn bc c tng hp v hin ang c ng dng trong nhiu lnh vc nhy t,mi trng,int,...Cchtnanobcvinnglngbmtlnckhnnggii phngttccionbcvotrongdungdch,nhvynanobcchiulckh khun ko di hn so vi dng keo bc. Tuy nhin dng tiu phn bc nguyn t c nhc im l kh nng gii phng ion bc thp. Gn y cc nh khoa hc dc pht trin thnh cng thuc s dng tiu phn nano ca mui t tan ca bc cho mc ch chng nhim khun. Cc thucny cu im l kim sotc tc gii phngionbcmctiu,trongthigiandi.Ccnghincutheohngny hin cha c trin khai trong nc. T thc t nu trn, chung ti tin hnh ti:Nghin cu tng hp tiu phn nano cha bc nhm cc mc tiu sau: 1. Xy dng quy trnh tng hp tiu phn nano cha bc clorid. 2. Xc nh mt s c tnh ca h nano cha bc clorid tng hp c. 2 CHNG 1. TNG QUAN 1.1.Gii thiu chung v bcBclmtnguynthahctrongbngtunhonccnguynthahc. BcckhiuAgvshiunguyntbng47,khilngnguyntbng 107,8682.Cchykhong200nmccnhkhoahcxemhuytthanhca ngi nh l mt dng dung dch keo, v vy h s dng keo bc lm cht khng khun ngay trong c th con ngi. K t keo bc c s dng rng ri cha cc bnh nm trn da, iu tr cc vt thng, vt bng, cc bnh rng ming, lm thuc nh mt. Tuy nhin, sau khi thuc khng sinh c pht minh (gia th k XX) vi hiu lc kh trng mnh hn, keo bc b thay th dn. Nhng ch 30 nm sau , ngi ta nhn ra rng c rt nhiu loi vi khun c kh nng chng li tc dng ca thuc khng sinh v vn nyngycng trnn ng lo ngi. Luc nytnhnngkhngkhuncabclicchudocphtcdngrngv khng b hn ch bi hiu ng khng thuc [13]. Nhiu dng bc c s dng lm thuc, in hnh l:-Dung dch keo bcyldngcsdngphbinnhttrcnm1960,cctiuphnion bc tinh khit, tch in, c phn tn trong mi trng lng. Cc ion tch in y nhau,vthchungcphntnngnhttrongmitrngngayckhibi thuc ln vt thng. -Phc hp bc protein Phchpbcviccproteinphntnhlmtngtnhnnhcaionbc trongdungdch.Tuynhinkhnngkhngkhunkmionbcvdocmts nhc im nht nh nn vo nhng nm 1960 chung nhanh chng c thay th bi cc mui bc. -Mui bc Bcnitrat0,5%tnglmtdungdchinhnhvphbinnhttr cc bt bng ngoi da. Dung dch mui bc th hin tnh khng khun cao, t b vi 3 sinh vt khng li v n cn c kh nng gim vim b mt vt thng. nng ln hn 1% dung dch bc nitrat c kh nng gy c vi t bo v cc m. -Bc sulfadiazin Bcsulfadiazincsdngnhiutrongnhngnm1970.Bcnitratv natrisulfadiazincphihptothnhbcsulfadiazinsdnglmthuc. Phc hp ny tc dng ln thnh t bo vi khun. Tc dng khng khun l tc dng hip ng ca c ion bc v ca sulfadiazin. Cc dng thuc phi hp cc sulfamit khcnhauvibccnghincuthnghiminvitro,ktquchothybc sulfadiazin cho tc dng tt nht [6]. Cc dng mui bc trn y, khi s dng lm thuc u c nhng nhc im nht nh. Chnh v th trn th gii c nhiu nghin cu pht trin theo hng bo ch thuc t cc tiu phn nano bc clorid trn tinh thn kim sot c s gii phng ion bc t h mt cch ti u nhm cho tc dng khng khun an ton v ko di [7],[9]. 1.2.Gii thiu h gii phng ion bc ko di Cc h kim sot gii phng ion bc ko di c pht trin trong nhng nm gnyvccoilmtcuccchmngtrongvicphttrinccsnphm chngnhimkhunvtthng.Nhiudngcabccsdngnghincu pht trin cc thuc dng ny: tiu phn nano bc; tiu phn nano mui bc; h cc ion bc v tiu phn nano bc phi hp vi cc polymer khc nhau Trong cc hnanotiuphncaccmuittancabccchunhiuvcpht trin thnh cng thnh thuc c nhiu gi tr. Hnh 1.1. Thuc khng khun dng cho vt bng cha tiu phn nano bc clorid 4 Ngy nay cng vi s pht trin ca cng ngh nano ni chung v ng dng ca cng ngh nano trong y dc ni ring to ra cc ht vt cht c kch thc nh(10-9nm).Tuynhinccsnphmtbctrnthtrnghinnayasit nguyn liu l cc tiu phn nano bc di dng bc nguyn t, do hn ch vic gii phng ra cc ion bc to tc dng khng khun, nu s dng cc dng mui dtancabcthskhkimsotvicgiiphngraionbc,dnnkhkim sot c c tnh gy ra. Cc ht tiu phn nano cc mui t tan ca bc, nh nng lng b mt ln, c kh nng gii phng cc ion bc vo trong dung dch cao, v nh vy cho hiu lc kh khun ng k ng thi c th kim sot c vic gii phng ion bc thng qua vic la chn mui bc c tan thch hp. Thi gian gii phng ion bc t cc nano tiu phn bc cn ko di hn nhiu so vi dng keo bc. Cc ng dng ca bc v tiu phn nano cha bc trong y dc c th hin trong hnh 1.2 [18]. Hnh 1.2.ng dng ca bc (bn phai) v nano cha bc (bn tri) trong y hc Bc nitrat cha chy Chng nhim khun: thuc nh mt 1% bc nitrate chng truyn bnh lu cu mt t m sang con. Chng vim: Bc nitrat dng cha tc nghn khoang mng phi (pleurodesis) Ti to da: Kem bc sunfadiazine dng cho cc vt bng v lot, ci thin kh nng ti to da Bc nitrat ngn cn s pht trin ca cc khi u ht sau chn thng Cc tiu phn nano bc c thm vo cc sn chng khun, cht st trng m bo mi trng v trng cho bnh nhn. Phng nhim khun: Nano bc c thm vo lp ph b mt cc ng dn trong gii phu thn kinh v ng thng tnh mch. Phng nhim khun: nano bc c thm vo xng gi v cc m cy khc. Chng nhim khun: ming dn vt thng cha nano bc chng nhim khun v tng cng lm lnh vt thng 5 1.3 . Tc dng ca tiu phn nano cha bc1.3.1.Tc dng khng khun Ion bc c tnh khng khun theo c ch trong hnh 1.3: Hnh 1.3.Tc ng ca ion bc ln vi khun. Cc c tnh khng khun ca bc bt ngun t tnh cht ha hc ca cc ion Ag+. Ion ny c kh nng lin kt mnh vi peptidoglican, thnh phn cu to nnthnh t bo ca vi khun v c ch kh nng vn chuyn oxy vo bn trong t bo dnnlmtlitvikhun.Dongvtkhngcthnhtbo,vvychungta khng b tn thng khi tip xuc vi cc ion ny. C mt c ch tc ng ca cc ion bc ln vi khun ng chu c m t nh sau: Sau khi Ag+ tc ng ln lp mng bo v ca t bo vi khun gy bnh n sivobntrongtbovphnngvinhmsunfuahydrinSHcaphnt enzym chuyn ha oxy v v hiu ha men ny dn n c ch qu trnh h hp ca t bo vi khun [29]. Hnh 1.4.Ion bc v hiu ha enzym chuyn ha oxy ca vi khun Ngoi ra cc ion bc cn c kh nng lin kt vi cc base ca ADN v trung ha in tch ca gc phosphate do ngn chn qu trnh sao chp ADN [32][33]. 6 Hnh 1.5. Ion bc lin kt vi cc base ca ADN 1.3.2.Tc dng chng nm Cc tiu phn cha bc c tc dng chng nm nhanh v hiu qu trn nhiu loi nm ph bin nh Aspergillus,Candida v Saccharomyces [33]. Hn na, tiu phn cha bc kch thc 13.5 2.6 nm cn hiu qu trong dit nm men phn lp t vu b b vim [23]. 1.3.3.Tc dng chng virus Cc tiu phn cha bc ng knh trung bnh 10 nm (5-20 nm) c ch s ti to ca virus HIV-1 [19]. Ngoi ra, Elechiguerrav cng s, 2005, ch ra rng tc dng dit virus ca tiu phn nano bc ph thuc vo kch thc tiu phn trong khong c tc dng l 1-10 nm [17]. Humberto H Lara v cng s, 2010, cho rng tiu phn cha bc pht huy tc dng khng virus HIV giai on u ca qu trnh nhn bn v giai on sau xm nhp ca virus [24].1.3.4.Tc dng chng vim Nhiunghincukhoahccthchinlmsngtcchchng vimcatiuphnnanochabc.ctnhchngvimcatiuphnnanocha bcctipcntrongiutrtich.Tcdngchngvimcatiuphnnano cha bc c th c lin qua ti kh nng gim gii phng cytokin [12] [16], gim s thmnhpcatbolymphovtbomast[22]vgythycctbovim [27][34]. 7 1.3.5. Tc dng cha bng v lm lnh vt thng Nhngthnghimlmsngngunhinnhgikhnnglmlnhvt thngbngnhanhcamingdnchananobc[3].Tiuphnnanobcgim ng k thi gian lnh vt thng vi thi gian trung bnh l 3,35 ngy v y li nhim khun cc vt thng nhim trng. Khng c tc dng khng mong mun no c quan st thy [20]. iu tr bng bng nano bc t hiu qu cao do c tiu phn nano bc c ch vi khun pht trin v phc hi tn thng nhanh. C ch c th l do cc ht nano bc c kh nng iu tit gii phng t t cc ion bc vo dch vt thng kch thchcccytokinhtriutrhoccchcccytokin,chophprutngnthi gian phc hi tn thng v khng li so [5]. 1.4. Mt s ng dng ca tiu phn nano cha bc trong y dc v m phm 1.4.1.Trang thit b y t Nh c tnh khng khun trong 1 thi gian di, nano cha bc thng c ph b mt cc thit b y t ghp vo c th nh ng thng trong tim mch, tit niu.Ngoira,nanobccngcdngtrongchtongrutdchnotytha trong phu thut thn kinh hayc thm vo cc cht hn xng. Ming dn vt thng cha nano bc c thng mi ha trong khong 2 thp k qua v c s dng trong lm sng cha cc vt thng trong c bng, cht t bo biu b, hi chng Steven-Johnson, ung th da mn tnh v bnh pemphigus. Tn thng minitingchodngsnphmnylActicoat,AquacelAg,ContreetFoam, PolyMem Silver, Urgotul SSD [5]. Mt s sn phm ng dng lm bng cha lnh vt thng c thnh phn nano cha bc hnh 1.6 v 1.7. 8 Hnh 1.6.Ming dn vt thng Acticoat. Hnh 1.7. Bng cha tiu phn nano bc cha lnh vt thng 1.4.2. Thuc v m phm Trong thuc v m phm, tiu phn nano cha bc c s dng rng ri trongccchphmlmp,stkhun,khminhsnphmcahngNano Cyclic(M),Nanopoly(HnQuc),NanogistCo.Ltd.(HnQuc),Nanobc cncsnxutdngdungdchbsungvilngnhdungdchASAP10 ppm.Mts sn phmmphm v thuc c cha thnh phn tiu phn nano cha bc th hin hnh 1.8 v 1.9. 9 Hnh 1.8. San phm kh mi c th c thnh phn nano cha bc Hnh 1.9.Dung dch ung tng cng min dch ASAP Nhiu loi thuc cha bc c pht trin t cc nguyn liu: bc nitrat, bcprotacgon,bcsulfadiazin.Gny,muibccloridcsdng pht trin thnh cng thuc chng nhim khun ngoi da c tc dng ko di (ti 72 gi). y l ch hng ti ca ti. 1.5.c tnh ca nano bc cloridv cc phng php xc nh 1.5.1.c tnh ca nano bc clorid Kch thc v hnh dng tiu phn nano thng c xc nh qua hnh nh TEM. Tiu phn nano cha bc sinh tng hp t vi toScenedesmus c dng hnh cu, kch thc 15-20nm khi quan st bng k thut TEM [21]. Nano bc c tng hp ha hc bng cht kh l acid ascorbic trong hn hp CTAB/NH3 cng c dng gn nh hnh cuvi nh hp ph UV-VIS cc i 422 nm [22]. ng Vn Phu 10 v cng s nghin cu ch to keo nano bc bng phng php chiu x Coban 60 s dng polyvinyl pyrolidon (PVP)/chitosan (CTS) lm cht n nh. Kch thc httrungbnhvtnsphnbkchthchtcxcnhbngphngphp m ht t nh TEM vi tng s ht t 500-1000 ht. Nhm tc gi nhn thy cc ht nano bc do h to ra ch yu c hnh cu, kch thc trung bnh l 15,96 0,51 nm (PVP), 5,55 0,25 nm (CTS), 2,92 0,05 nm (1%PVP/5% CTS), 11,442,07 nm (1%PVP/ Ethanol 1M) [2]. Tythucvohnnh,kchthc,phnbhtnanotothnhm nano bc hp th cc i cc bc sng khc nhau. Bc sng cc i ca nano bc trong cht n nh PVA v PVP ln lt l 410 v 418 nm [1]. 1.5.2.Cc phng php xc nh Ccctnhl,hacatiuphnnanochabccxcnhbimts phng php nh: -Hnh nh chp trn knh hin vi in t truyn qua . -o ph hp th t ngoi, kh kin . -Ph nhiu x tia X. -Ph hng ngoi IR [21]. 1.5.2.1.S dng knh hin vi in t truyn qua Knh hin vi in t truyn qua (TEM) hot ng trn nguyn tc ging thu knh quang hc, ch khc l s dng sng in t thay cho bc sng nh sng nn cbcsngrtngn)vsdngccthuknhint-magneticlensthaycho thu knh quang hc [14]. Anh ca knh hin vi in t truyn qua cho php ta quan st c hnh dng v xc nh c kch thc ca cc ht nano. 1.5.2.2.Phn tch ph UV-VIS UV-VISlphngphpphntchsdngphhpthhoctnxtrong phm vi vng cc tm cho ti vng nh sng nhn thy c. 11 Do cc thuc tnh quang hc ca dch lng cha ht nano ph thuc vo hnh dng, kch thc v nng ca ht, nn ta c th s dng UV-VIS xc nh cc thuc tnh trn. Vdvihtnaobc,ckchthcnhhn20nm,chcmtbmt plasmon duy nht nn trong ph UV-VIS ca chung ch xut hin 1 nh duy nht. Ngi ta x dng tnh cht ny xc nh hnh dng ca ht nano bc [21]. 1.5.2.3.Ph nhiu x tia X Nhiu x tia X l hin tng cc chm tia X nhiu x trn cc mt tinh th ca cht rn do tnh tun hon ca cu truc tinh th to nn cc cc i v cc tiu nhiu x.KthutnhiuxtiaXcsdngphntchcutrucchtrn,vt liu...[21]. 1.5.2.4.Ph hng ngoi IR Mt trong nhng u im quan trng nht ca phng php ph hng ngoi vt hn nhng phng php phn tch cu truc khc (nhiu x tia X, cng hng t,int)lphngphpnycungcpthngtinvcutrucphntnhanh, khng i hi cc phng php tnh ton phc tp [28]. 1.6 . Mt s phng php tng hp tiu phn nano mui bc C 3 phng php chnh tng hp tiu phn nano cha bc bao gm: - Phng php tng hp ha l kt hp - Phng php sinh tng hp - Phng php tng hp ha thc vt. 1.6.1.Phng php tng hp ha l kt hp Phngphpphbinnhtcsdngiuchnanotiuphnbcl kh bc nitrat bng cc tc nhn kh ha nh sodium borohydrid, tia UV, tia co-60(photoreduction),khH2,hydrazin,ethanol,ethylenglycol,ascorbicacidv aliphaticaminCctcnhnkhcmnhyukhcnhauchoccnanotiu phnbcckchthckhcnhau.Sodiumborohydridcsdngrtphbin do kh nng phn ng cao so vi cc cht kh khc. Kch thc nano tiu phn thu c tng i nh 1-15 nm [1],[4]. 12 Hn hp phn ng c thm cht n nh v cht nh v hn ch s kt t ca cc nano tiu phn. Cc cc cht n nh c s dng gm: tinh bt ha tan,dchkeosilica,polyvinylalcohol,polyvinylpyrrolidon,-cyclodextrin, chitosan, etylen glycol, sodium dodecyl sulfat , peptid, silica kch thc nano... Mc d c th s dng nhiu loi dung mi hu c khc nhau lm mi trng phn tn trong qu trnh tng hp nano tiu phn bc nhng nc vn l dung mi c s dng ph bin nht [3], [13]. 1.6.1.1.Mt s phng php tng hp ha l kt hp -Phngphp1:Dngtinhbthatanvalmchtnnhvalmcht kh kh AgNO3 thnh nano bc [30]. -Phngphp2:AgNO3cnhgitvochitosanhatantrongacid acetic. Ag+ trong dung dch c kh thnh tiu phn nano cha bc bi tia x v n nh bi chitosan [14]. -Phngphp 3: Kh nh sng dung dch [Ag(NH3)2]+ dng PVP (poly(N-vinyl-2-pyrrolidon) di tia UV [11],[15]. -Phngphp4:Dnginctruynnhanhqua2dybctrongnc kh ion lm bay hi cc tiu phn bc trn b mt v ngng t li trong dung dch [32]. - Phng php 5: AgNO3 c khuy vi dung dch peptid trong m cc pH khc nhau trong 12 gi trong bng ti, sodium arcorbat c thm vo kh Ag+ thnh tiu phn nano [19]. -Phngphp6:Phngphpdungdcht:nhgitttdungdch NaBH4vohnhpdungdchAgNO3vichitosanlmchtnnhti nhit phng, pH 5-6 v khuy mnh lit cho ti khi dung dch chuyn n sang mu vng nu [5]. -Phngphp7:HatanPVAvPVPvoncctnng0.1%(1g/l), thm isopropanol vodung dch vi nng 1 M. Thm 0.17g AgNO3 ha vo 1 l dung dch trn , chia vo ng nghim thy tinh c nut (10 ml/ng) v sc kh nit. Chiu x Co-60 [1]. 13 1.6.1.2.c im ca cc phng php tng hp ha l kt hp Mt s c im ca cc phng php tng hp ha l kt hp c th hin trong bng 1.1: Bang 1.1 c im ca mt s phng php tng hp ha, l kt hp PPKTTP (nm) u imNhc im 110 -34ngin.Tiuphnnanocha bc bn trong tinh bt > 90 ngy. Khngkimsotcphnbkch thc tiu phn. 24-5 Kchthcnh,PBKThp, chitosantrnhskttcatiu phnnanochabc,Tiam bo tng hp nano bc v khun. Chitosancthnhhngnc tnhcananobc.KTTPnhcth gy c. 34-6 KTTPrtnh,hnhcu,PBKT hp Khtngkchthc,PVPcth nh hng n c tnh sinh hc ca nano bc. KTTP nh c th gy c. 45-35n gin, hiu qu, PBKT nhTiuphnnanocthkttdo khng cha cht n nh. 519 Gimctnhcananobcdo sdngpeptidlmtcnhnn nh. Tiuphnnanocthtamts pH gy nguy him .Chui peptide c thgypngmindchkhng mong mun. 620 - 25 Ch to bng iu tr vt bng c tcdngtt,thigianphchi nhanh hn v khng li so li Cha c nghin cu, n nh, ng dng,hiuquditkhunnanobc to thnh 7< 20 Mi trng PVP th hin tnh n nh hn PVA ChiuxtrnngunCo-60,gy nguyhim.iukin,thitbphc tp 14 Nhnxt:Sdngccphngphphalkthptrongtnghpcctiu phn nano cha bc c nghin cu nhiu do cc phng php ny mang li kch thc tiu phn ti u, kim sot c kch thc tiu phn, hiu sut qu trnh tt, tiuphntnghpcnnhcaodocthsdngthmchtbnhatrong qu trnh tng hp. 1.6.2.Phng php sinh tng hp tiu phn nano cha bc Phng php sinh tng hp nano tiu phn bc bng nhiu chng vi khun v vi nm c cng b. Dung dch bc nitrat c thm vo sinh khi vi sinh vt, tcnhnkhha(nhhydroquinones)cmttrongsinhkhiskhAg+thnh nano tiu phn bc di cc iu kin xc nh. Nhc im ca phng php ny l kh khn trong vic tinh ch mu v cc sn phm chuyn ha ca vi sinh vt c thlivttrongsnphmcuicnggykhkhnkhingdngtrongngnh dc [4]. 1.6.2.1.Mt s phng php sinh tng hp-Phngphp1:ChtnicavikhunEnterobacteriagmccenzymv thnh phn kh AgNO3 thnh tiu phn nano bc. Phng php ny thu c ng knh kch thc tiu phn l 28 122 nm.-Phngphp2SinhtnghpdngPemicilliumbrevicompactumkh AgNO3.Phng php ny thu c ng knh kch thc tiu phn: 23-105 nm [30], [31]. 1.6.2.2. c im ca cc phng php sinh tng hpcimcaccphngphpsinhtnghptiuphnnanochabcc th hin bng 1.2 [32], [33]. 15 Bang 1.2.cim ca mt s phng php sinh tng hp Phng php KTTP (nm) Uu imNhc im 128 - 122 Thn thin vi mi trng Khng kim sot c KTTP C kh nng nhim mm bnh 223 105 Thn thin vi mi trng Tiu phn nano bc khng b kt t Khng kim sot c KTTP. Qu trnh kh ha ko di, mt nhiu thi gian. C kh nng nhim mm bnh

Nhnxt:Phngphpsinhtnghptnghptiuphnnanochabc hincngcnghincunhiudotnhthnthinvimitrngcaphng php. Nhng nhc im ca phng php l khng kim sot c kch thc tiu phn, n nh kch thc tiu phn km, hiu sut qu trnh thp, c th ln tp cht. 1.6.3.Phng php tng hp ha thc vt Gn y, vic s dng cc dch chit t thc vt lm tc nhn kh ha tng hp nano tiu phn bc cng c chu v pht trin do tnh thn thin mi trng, an ton cao v gi thnh r. Hn hp nhiu thnh phn trong dch chit va l tc nhn kh ha va l tc nhn n nh cho tiu phn nano tng hp c. Nhiu tc gi tng hp thnh cng tiu phn nano bc bng cch s dng cc dch chit t: cnghCurcumalonga,lchxanhCamelliasp.,lhungchanh(tnldy) Plectranthusamboinicus,lctkhtaTephrosiapurpurea,lbchttl Tribulusterrestris, v qu cam ngt Citrus unshiu, l l hi Aloe vera, r cy nhu Morinda citrifolia, v qu xoi Mangifera sp., l cy ngt ngho Gloriosa superba, l bch n Eucalyptus chapmaniana, v qu chui Musa sp., l mn kinh t Vitex negundo [13],[14]. Theo Ajitha B v cng s s dng dch chit l cy Tephrosia purpurea ng vaitrvalchtkhvaltcnhnbtgibc.Phngphpnythuc 16 ngknhkchthctiuphnl20nm.uimcaphngphplxanh, thn thin vi mi trng, d thc hin, sn phm nano cha bc c tc dng c ch chng Pseudomonas spp. v Penicillium spp. Bn cnh nhc im ca phng php l qua trnh chit tng hp tiu phn nano cha bc mang li hiu sut khng cao, c ln tp nhiu [6]. .Nhn xt: Phng php tng hp ha thc vt vi u im l thn thin vi mi trng, an ton cao, gi thnh r. Tuy nhin qu trnh tng hp mang hiu sut khng cao, c ln nhiu tp cht. Tiu phn nano bc hin ang l ti c chu nhiu khng ch trn th gii m c Vit Nam. Vi ph khng khun rng v t b khng, nano tiu phn bc m ratrin vng ln trong vic pht trin cc sn phm trong ngnhdc, m phm, trang thit b y t v sn phm dn dng. Thch thc ln nht trong iu chnanotiuphnmuibclvickimsotccthngsnhkchthctiu phn, phn b kch thc, hnh dng, hnh thi hc, n nh, thnh phn ha hc, cu truc tinh th, hiu sut phn ng, tp cht. ti t vn nghin cu iu ch tiu phn nano ca bc clorid l nguyn liu pht trin thuc chng nhim khun. V th chung ti mnh dn nghin cu tng hp tiu phn nano cha bc clorid da trnphngphphalkthpvimongmuntocquytrnhthchind thc hin, ph hp vi iu kin, trang thit b nc ta. 17 CHNG2NGUYN LIU, THIT B V PHNG PHP NGHIN CU 2.1 .Nguyn liu nghin cu Cc nguyn liu s dng nghin cu c trinh by trong bng 2.1: Bang 2.1. Nguyn liu s dng nghin cu STTNguyn liuNgun gcTiu chun 1Bc nitratTrung QucNh sn xut 2Natri cloridVit NamDc in Vit Nam 4 3Beta-cyclodextrinMUSP 34 4Polyvinyl alcoholHn QucNh sn xut 5Methocel E6MUSP 34 6Polyvinyl pyrrolidon K30Trung QucNh sn xut 7Carbomer 934MUSP 34 8Nc ctVit NamDc in Vit Nam 4 2.2 . Thit b nghin cu - My khuy t IKA-WERKE Khoa Y, Dc, i hc Quc gia H Ni. -MyoquangphUV-VISCaryUV-60-KhoaY,Dc,HQucgiaH Ni. - My ly tm EBA 21 - Khoa Y, Dc, H Quc gia H Ni. - Knh hin vi in t qut FESEM Hitachi S-4800 - Vin v sinh dch t trung ng. - My o ph hng ngoi FTIR-600 Khoa Ha, HKHTN, H Quc gia H Ni -MyophnhiuxtiaXD8ADVANCE-Bruker-KhoaHa,HKHTN, H 18 Quc gia H Ni. -MyokchthctiuphnZetasizerNanoZS90MalverinBmnBo ch, H Dc H Ni. -CnphntchSHIMADZUCORPORATION-KhoaY,Dc,HQGH Ni. - T sy chn khng HITACHI Khoa Y,Dc, HQG H Ni. - My lc nn SOTRAS- Khoa Y, Dc, HQG H Ni. - Buret, t lnh, nhit k. 2.3 . Phng php nghin cu 2.3.1.Tng hp tiu phn bc clorid Nguyn tc: S dng phng php ha hc iu ch cc tiu phn c kch thc nano cha bc clorid trong dung dch nc cha cht bn ho: Tin hnh tng hp tiu phn bc clorid t bc nitrat theo phng trnh: AgNO3 +NaCl AgCl + NaNO3 .Quy trnh nghin cu: Ha tan bc nitrat vo dung dch nc ca cht bn hackhost.Nhttdngdchnatriclorid0,1Mvo,tcnh 0,5ml/pht, va nh va khuy trn tc 500 vng/pht. Tip tc khuy duy tr trong 1 gi. Phn ng c thc hin trong iu kin trnh nh sng. Hnh 2.1.Quy trnh tng hp tiu phn bc clorid Cht bn ha/ Nc 19 2.3.2.Khao st anh hng ca cht bn ho ti s hnh thnh tiu phn nano Sdngccchtbnhokhcnhauiuchccmunanobckhc nhautheophngphpmc2.3.1.Mimuctinhnh2ln.Ngaysaukhi iu ch, nh gi h nano thu c da trn kch thc tiu phn, th Zeta v phn b kch thc PDI, t la chn cht bn ho ph hp. Sau khi kho st la chn chtbnhatiptckhostnhhngcalngchtbnhanhnhthnh tiu phn nano bc clorid. 2.3.3.Xc nh tnh mt s c tnh ca tiu phn nano tng hp c 2.3.3.1. Kch thc v phn b kch thc tiu phn Nguyn tc: Khi chiu chm tia laser vo cc tiu phn c kch thc khc nhausthucmctnxkhcnhau.Davomctnxcachmtia sau khi va chm vo tiu phn ta c th tnh c kch thc tiu phn theo thuyt Mie. Quytrnh:Ccmuhndchnanobcsaukhiiuchcoxcnh kchthctiuphn.Trckhio,mucphalong10lnbngncct lc qua mng c kch thc l xp l 0,2 m. Thit b o: My o kch thc tiu phn Zetasizer Nano ZS90 Malverin, s dng cuvet thy tinh, ch s khuc x RI l 1,34 v hp th l 0,001. Phpochoktqukchthctrungbnhcacctiuphn,mcng u KTTPth hin ch s a phn tn (PDI). KTTP c phn b trong khong hp nu PDI 0,3. Cng thc tnhPDI nh sau [27]: PDI = (

)2 Trong :PDI l ch s a phn tn hay ch s phn b KTTP l lch chun KTTPgia cc ln o d l KTTPtrung bnh 20 2.3.3.2.Th Zeta ca tiu phn Nguyntc:ThZetacxcnhgintipthngqualinhindica tiu phn nano khi t trong in trng. Tc tiu phn c xc nh thng qua vic so snh s sai khc v pha gia nh sng tn x (tia th ) v nh sng t ngun lade (tia chun ) nh ng dng nh lut Doppler [27]. Quy trnh: Cc mu o kch thc tiu phn xong c s dng tip o th Zeta ca mu. o th Zeta ca mu nano bc clorid trn my Zetasizer Nano ZS90 Malverin, s dng cuvet nha. 2.3.3.3.Hnh dng tiu phn Nguyntc:Xcnhhnhdngtiuphnbngknhhinviintqut (SEM). Chm in t qut trn b mt mu c thu li bi cc u d bin i thnh nhngtn hiu phn nh bmt,thnhphn camu a ramn hnh quan st. Cc hnh nh chp SEM c cc c im ca nh 3 chiu [21]. Quytrnh:Sdngknhhinviintqut(SEM)FESEMHitachiS-4800, in th gia tc l 5,0 kV. 2.3.3.4. bn vi nh sng Nguyn tc: So snh ph hp th UV-VIS ca hn dch trc v sau c khi chiu sng. Ph t ngoi v kh kin ca cc cht hu c gn lin vi bc chuyn electron gia mc nng lng electron trong phn t khi cc electron chuyn t cc obitan lin kt hoc khng lin kt ln cc obitan phn lin kt c mc nng lng cao hn, i hi phi hp th nng lng t bn ngoi [33]. Quy trnh: Qut ph hp th trong vng bc sng t 200 n 800 nm ca hn dch mi bo ch v sau khi c tip xuc vi nh sng mt tri trong 15 gi, s dng my o quang ph Cary UV-60 vi cuvet thch anh 1cm. 21 2.3.3.5.Bn cht ha hc ca tiu phn Nguyn tc: Xc nh bn cht ha hc ca tiu phn bng ph nhiu x tia X. Khi mt chm tia X c bc sng chiu ti mt tinh th cht rn di gc ti. Do tinh th c tnh cht tun hon, cc mt tinh th s cch nhau nhng khong u nd,ngvaitrgingnhcccchtnhiuxvtorahintngnhiux ca cc tia X [25]. Quy trnh: o ph nhiu x tia X ca mu trn my X D8 ADVANCE- bruker.. 2.3.3.6.Tng tc gia tiu phn v cht bn ha Nguyntc:Sbxcnhbnchttngtcgiatiuphnbccloridv cht bn ha bng cch so snh ph hng ngoi ca sn phm v cht bn ha. K thut ny da trn hiu ng n gin; cc hp cht ha hc c kh nng hp th bc x hng ngoi. Sau khi hp th cc bc x hng ngoi, cc phn t ca hp cht ha hcdaongvinhiuvntcdaongvxuthindiphhpthgilph hp th bc x hng ngoi [15]. Quy trnh: o ph IR ca mu trn my FTIR-600. 22 CHNG 3:KT QU THC NGHIM3.1. nh hng ca cht bn ho n s hinh thnh tiu phn bc clorid 3.1.1.Anh hng ca loi cht bn ha Cc mu hn dch c tng hp theo phng php m t trong mc 2.3.1. Mi mu c thc hin 2 ln trong nhng iu kin tng t nhau. Cng thc iu ch chung trong bng 3.1 nh sau: Bang 3.1.Cng thc iu ch hn dch bc clorid vi mt s cht bn ha STTHo chtLng 1Cht bn ho0,5448 g 2Bc nitrat0,0204 g 3Natri clorid2,4ml dd 0,1M 4Nc ct35 ml CcchtbnhocsdnglPVA,PVPK30,Beta-cyclodextrin, MethocelE6, Carbomer 934. y l cc polymer c s dng rng ri trong cng nghipdc,ctcdngtngnnhhndchbngcchlmtngnht dung dch, gim kt t tiu phn. V beta-cyclodextrin c khi lng mol xc nh nnsdngtlmolbeta-cyclodextrin:bcnitratl4:1.Ccchtbnhakhc khng c khi lng mol xc nh nn c ly bng khi lng beta-cyclodextrin. Cc cht bn ha nyto dung dch c nng khong 1,4% (kl/kl). Lngnc ct s dng l 35ml m bo ha tan hon ton cht c tan km nht l beta-cyclodextrin. S mol natri clorid (0,24 mmol) c s dng gp i bc nitrat (0,12 mmol) m bo chuyn ton b bc nitrat thnh bc clorid. Ngay sau khi bo ch, cc mu c o kch thc, th Zeta v ch s phn bkchthcPDItheophngphpmtmc2.3.3.1v2.3.3.2,lyktqu trung bnh. Kt qu c th hin trong bng 3.2. 23 Bang 3.2.Kch thc, th Zeta v PDI ca tiu phn AgCl vi mt s cht n nh (n=2) MuCht bn ho Kch thc TB (nm) Peak 1 (nm) Peak 2 (nm) % Peak 1 % Peak 2 PDI TB Th Zeta TB(mV) M1PVA85,783,11520896,93,10,238-13,2 M2PVP K3082,394,28010000,131-17,1 M3Beta cyclodextrin 324,4322,550629550,229-20,8 M4Methocel E6 274,2321,8010000,209-9,44 M5Carbomer 934 201,4237,8010000,148-14.7 Kt qu cho thy cc tiu phn AgCl c bo ch vi PVA v PVP K30 c ng knh trung bnh nh hn ng k so vi cc cht bn ho cn li. Vi PVA thu c kch thc tiu phn l 85,7 nm v PVP K30 thu c kch thc l 82,3 nm. Gi tr PDI ca M1 (0,238) cao hn hn so vi M2 (0,131) do xut hin mt pic ph5280nm(chim3,1%).TtcccmuucthZetam.MucthZeta mang gi tr tuyt i cao nht lM3 (-20,8 mV). M1 v M2 c gi tr th ln lt l -13,2 v -17,1 mV. Gi tr th m c th l do lp ion m Cl- hp ph ln b mt tiu phn. iu ny gi khi bo ch cc dng thuc khc nhau, c th kho st cc h m nh citrat, phosphat lm tng n nh cho hn dch. T cc kt qu ny, 2 cht n nh l PVP K30 v PVA tip tc c kho st nh hng ca lng cht bn ha n c tnh tiu phn. 3.1.2.Anh hng ca lng cht bn ha Polyvinyl alcohol CcmutM6nM11ciuchtheophngphpmttimc 2.3.1vilngPVAtngdnt0,0340gn1,0897g,ccthnhphnkhcgi 24 nguyn nh trong bng 3.1. Mi mu c iu ch 2 ln. o kch thc tiu phn, phnbkchthc,thZetatngmutheophngphpmtmc2.3.3.1v 2.3.3.2,lyktqutrungbnh.Ktqucthhintrongbng3.3vtrnhnh 3.1, 3.2 v 3.3. Bang 3.3.Kc thc, th Zeta v PDI ca tiu phn AgCl bo ch vi cc lng PVA khc nhau (n=2) MuLng PVA (g) Kch thc TB (nm) Peak 1 (nm) Peak 2 (nm) % Peak 1 % Peak 2 PDI TB Th Zeta TB(mV) M60,0340116,5142,5010000,19-17,9 M70,068198,9110,8010000,149-17,1 M80,136284,394,56491098,81,20,166-19,4 M90,272480,5177,66010000,251-21,7 M100,544885,783,11520896,93,10,238-13,2 M111,089692,85112,5010000,207-20,1 Hnh 3.1. th mi lin quan gia lng PVA v kch thc tiu phn AgCl 0204060801001201400 0,2 0,4 0,6 0,8 1 1,2Lng PVA(g)Kch thc (nm )25 Hnh 3.2. th mi lin quan gia lng PVA v PDI ca tiu phn AgCl Hnh 3.3. th mi lin quan gia lng PVA v th Zeta ca tiu phn AgCl Kt qu thu c cho thy, khi lng PVA tng t 0,0340g n 0,2724g, kch thc tiu phn gim dn. iu ny c th l do nng PVA thp, nht ca mi trng cng nh kh nng bao ph, to lp o ngn cn cc tiu phn kt t ca PVAthp,lmchokchthctiuphntng.TuynhinkhilngPVAtngt 0,2724g n 1,0896g,kch thc tiu phn tng dn. C th l dolng PVA qu 00,050,10,150,20,250,30 0,2 0,4 0,6 0,8 1 1,2Lng PVA (g)PDI-25-20-15-10-500 0,2 0,4 0,6 0,8 1 1,2Lng PVA (g)Zeta (mV)26 nhiu lm tng nht ca mi trng, gim kh nng phn tn v do lm tng kchthctiuphn.Trongccmuckhost,M9ckchthctiuphn trung bnh thp nht (80,51 nm). Gi tr tuyt i th Zeta ca M9 cng l ln nht (+21,7mV).GitrtuytithZetatrn20ccholtngibn,ckh nng ngn cn cc tiu phn kt t. PDI ca cc mu dao ng t 0,149 n 0,251 vPDIcamuM9l0,251chothysphnbkchthctiuphntngi ng u (di 0,3). Qua qu trnh thc nghim ny tm ra lng PVA bng 0,2724 g cng thc M9 l ti u nht. 3.1.3.Anh hng ca lng cht bn ha Polyvinyl pyrrollidon K30 CcmutM12nM17ciuchtheophngphpmttimc 2.3.1vilngPVPtngdnt0,0340gn1,0897g,ccthnhphnkhcgi nguyn nh trong bng 3.1. Mi mu c iu ch 2 ln. o kch thc tiu phn, phnbkchthc,thZetatngmutheophngphpmtmc2.3.3.1v 2.3.3.2,lyktqutrungbnh.Ktqucthhintrongbng3.4vtrnhnh 3.4, 3.5 v 3.6. Bang 3.4: Kch thc, th Zeta v PDI ca tiu phn AgCl khi iu ch vi cc lng PVP khc nhau (n=2) MuLng PVP (g) Kch thc TB (nm) Peak 1 (nm) Peak 2 (nm) % Peak 1 % Peak 2 PDI TB Th Zeta TB (mV) M120,0340106,7119,7010000,152-26 M130,068180,6891,51010000,162-13,6 M140,136275,6885,22010000,114-26,6 M150,272481,1192,87010000,145-7,19 M160,544882,394,28010000,131-17,1 M171,089687,26104,6010000,175-18,9 27

Hnh 3.4. th mi lin quan gia lng PVP v kch thc tiu phn AgCl Hnh 3.5. th mi lin quan gia ng PVP v PDI ca tiu phn AgCl 00,020,040,060,080,10,120,140,160,180,20 0,2 0,4 0,6 0,8 1 1,2Lng PVP (g)PDI0204060801001200 0,2 0,4 0,6 0,8 1 1,2Lng PVP (g)Kch thc (nm)28 Hnh 3.6. th mi lin quan gia lng PVP v th Zeta ca tiu phn AgCl KhisdngPVPlmchtbnha,banukchthctiuphngimdn khi tng lng cht bn ha.Lng PVP thp cng lm tng kch thc tiu phn l do nht ca mi trng cng nh kh nng bao ph, to lp o ngn cn cc tiuphnkttcaPVPthp.TuynhinkhinngPVPtngcaolilmtng kch thc tiu phn do lm tng nht ca mi trng, gim kh nng phn tn tiuphn.Trongccmukhost,M14lmuttnhtvikchthctiuphn trungbnhnhnht(75,68nm),PDInhnht(0,114)vgitrtuyticath Zetalnnht(-26,6mV).GitrPDInhhn0,3chothykchthctiuphn phn b tng i ng u. Gi tr tuyt i ca th Zeta M14 (+26,6 mV) tng i bn, kh nng kt t tiu phn thp. T kt qu thc nghim trn ta thy lng PVP bng 0,1362 g l ti u. Tccktquthuc,cngthccho2muM9vM14tiptcc kho st.3.1.4.La chn cht bn ho v cng thc bo ch c mu dng cho cc th nghim tip theo v tng lng mu iu ch-30-25-20-15-10-500 0,2 0,4 0,6 0,8 1 1,2Lng PVP (g)Zeta (mV)29 trong 1 th nghim, cng thc iu ch M9 v M14 c nhn ln 6 ln c cc mu tng ng l M18 v M19 vi thnh phn th hin trong bng 3.5.Bang 3.5.Cng thc bo ch mu M18 v M19 (n=2) Mu Ha cht M18M19 PVA1,6344 g0 PVP K3000,8172 g Bc nitrat0,1223 g0,1223 g Dd NaCl 0,1 M14,4 ml14,4 ml Nc ct210 ml210 ml Cc mu M18 v M19 c iu ch theo phng php m t mc 2.3.1. Mimuciuch2lntrongiukintngtnhau.okchthctiu phn,phnbkchthc,thZetatngmutheophngphpmttrongmc 2.3.3.1 v 2.3.3.2, ly kt qu trung bnh. Kt qu c th hin trong bng 3.6. Bang 3.6.Kch thc, th Zeta v PDI ca cc mu M18 v M19 (n=2) MuKch thc TB (nm) PDI TBTh Zeta TB (mV) M1888,240,117-11,3 M19284,80,337-38,1 Kt qu cho thy trong khi M18 c kch thc tiu phn v PDI( 88,24 nm v0,117)thayikhngngksoviM9(80,51nmv0,251)thM19lic kch thc tiu phn v PDI (284,8 nm v 0,337 ) tng cao ro rt so vi M14 (85,22 nm v 0,114 ). Gi tr tuyt i th Zeta ca M18 (+11,3 mV) gim i so vi mu M9 (+21,7 mV). Gi tr tuyt i th Zeta ca M19 (+31,8 mV) tng ln so vi mu M14 (+26,6 mV). Thc t trong qu trnh iu ch, hn dch M19 c dn nh vy 30 do c kt t to tiu phn kch thc ln. T kt qu trn, cng thc M18 c la chn bo ch cc mu nano bc clorid dng cho cc th nghim tip theo. Tm li, quaqu trnh th nghim chn lc cht bn ha, kho sts thay ilngchtbnhachungtithuccngthcM18ltiuvquytrnh tng hp tiu phn nano bc clorid nh sau: -Hatan1,6344gPVAvotrong210mlncntanhonton,lcqua mng lc 0,2 m c dung dch cht bn ha. -Thm 0,1233 g AgNO3 vo dung dch cht bn ha, s dngmy khuy t, khuy n tan hon ton -Nh t t 14,4 ml dng dch natri clorid 0,1M vo hn hp trn, tc nh 0,5ml/pht,vanhvakhuytrntc500vng/pht.Lngdung dch natri clorid nh xung l 14,4 ml. -Tip tc khuy duy tr hn dch trong 1 gi. Lu :Phn ng c thc hin trong iu kin trnh nh sng. 3.2. Xc nh mt s c tnh ca tiu phn nano bc clorid va tng hp c S dng cng thcM18 iu chccmu nano bc t M20 nM46theo phng php trnh by ti mc 3.1.4,sau tin hnh xc nh mt s c tnh ca tiu phn AgCl iu ch c. 3.2.1. Kch thc, PDI v th Zeta Tin hnh iu ch 3 mu nano bc clorid M20, M21 v M22. Tin hnh o kch thc, phn b kch thc, th Zeta ca tiu phn nano bc. Kt qu c trnh by trong bng 3.7. Bang 3.7: Kch thc, th Zeta v PDI ca tiu phn nano bc clorid MuKch thc (nm)PDITh Zeta (mV) M2089,570,101-8,21 M2187,220,190-11,6 M2289.660.118-9.74 Trung binh88,820,136-9,85 31 Kt qu cho thy, kch thc tiu phn bcTB l 88,82 nm, vi gi tr PDI TBl0,136(thpdi0,2)chothykchthctiuphnphnbtrongkhong hp. Gi tr tuyt i ca th Zetal9,85 mV. Gi tr th tuyt i th Zeta tng i ph hp v khi tng lng cc cht tham gia phn ng ln 6 ln th kh nng kt t ca tiu phn tng ln. 3.2.2.Hnh dng tiu phn TinhnhiuchmuhndchnanobccloridM23.Muhndchiu ch xong c tin hnh chp SEM trn knh hin vi in t qut FESEM Hitachi S-4800 ti Vin v sinh dch t trung ng.Kt qu hnh nh chp SEM mu hn dch nano bc clorid c th hin trong hnh 3.7. Hnh 3.7.Anh chp SEM ca hn dch nano bc clorid HnhnhchpSEMchothycctiuphncnhiuhnhdngnhngch yu l hnh lp phng. iu ny ph hp vi hnh dng ca tiu phn bc halid ni 32 chung(StephanGlaus,2003)[31].Bmtcctiuphnkhngcsccnhc th l do chung c bao bc bi PVA.3.2.3. bn vi nh sng Tin hnh iu ch mu hn dch bc clorid M24. Tin hnh o ph hp th UV-VIS ca M24 ngay sau khi iu ch v sau khi cho M24 tip xuc vi nh sng mt tri trong vng 15 gi. Qut ph hp th trong vng bc sng t 200 n 800 nm. Kt qu th hin hnh 3.8; 3.9 v 3.10. Hnh 3.8.S thay i mu sc sau khi tip xuc vi nh sng mt tri ca hn dch nano bc cloridTrongqutrnhtipxucvinhsng,hndchM24chuyndntmu trng nh xanh trong sang mu tm.Trc Sau 33 Hnh 3.9.Ph hp th UV-VIS ca mu hn dich M24 ngay khi iu ch

Hnh 3.10.Ph UV-VIS ca mu hn dch M24 sau khi cho tip xuc nh sng PhhpthUV-VIShnh3.9vhnh3.10chothy:Hndchnanobc cloridmiiuchchhpthmnhtrongvngbcsngnhhn300nm.Sau khi cho tip xucvi nh sngmt tri, khnng hp th cahndch trongvng 34 400-600nm tng ln ng k. AgCl khng c kh nng hp th bc sng trn 400 nm v c nng lng vng cmtrc tip v gin tip ln lt l 5,15 eV ( 240 nm) v 3,25 eV ( 380 nm). Pick hp th rng bc sng trn 400 nm xut hin l do cng hng plasmon b mt, mt hin tng hay gp cc tiu phn nano kim loi ckchthct2n100nm.AgClcthchuynthnhAgkhitipxucvi nh sng, bm t trn b mt tiu phn nano gy ra hin tng ny. iu ny chng t tiu phn AgCl khng bn khi tip xuc vi nh sng mt tri, i hi qu trnh th nghim v bo qun hn dch trong iu kin trnh nh sng mt tri. 3.2.4.Ban cht ha hc ca tiu phn Bn cht ha hc ca tiu phn c xc nh da trn ph nhiu x tia X. Quytrnh:Tinhnhiuch20muhndchchananobccloridM25 nM45.Tinhnhlytmlyta(trnmylytmlnhEBA21,tc5000 vng/phut,thigian15phut),gpta,rata,syvemophnhiuxtiaX trnmyD8ADVANCE-Bruker(ngphttiaXbngngvibcsngK= 1,540, in p 40 kV, cng dng ng pht 30 mA, nhit 25, gc qut 2 thay i 20-80o, tc qut 0,2 /phut, ti phng th nghim vt liu- Khoa Ho - HKHTNHQGHN). Kt qu c so snh vi vch chun AgCl trnmyv th hin trn hnh 3.11. 35 Hnh 3.11. Ph X - ray ca nano tiu phn bc clorid Kt qu o ph nhiu x tia X cho thy c 8 peaks nhn tng ng trng vi 8 vch chun gc 2 ca AgCl chun trn my xc nhn s c mt ca AgCl trong hndchv8peaksnhnphnnhmtphngcakhilpphngtmcatiu phn AgCl. 3.2.5. Tng tc gia tiu phn v cht bn ha Xcnhtngtcgiatiuphnvchtbnhaquaophhngngoi trn myFTIR-600. Tinhnh iu chmu hndch nano bc cloridM46. Song song tin hnh mu trng l dung dch PVA v NaCl trong nc ct. Tin hnh o ph hng ngoi IR ca mu hn dch v mu trng. Ph IR ca 2 mu ny c th hin trong hnh 3.12 v 3.13. Faculty of Chemistry, HUS, VNU, D8 ADVANCE-Bruker - Sample AgCl say01-085-1355 (C) - Chlorargyrite, syn - AgCl - Y: 82.94 % - d x by: 1. - WL: 1.5406 - Cubic - a 5.54900 - b 5.54900 - c 5.54900 - alpha 90.000 - beta 90.000 - gamma 90.000 - Face-centered - Fm-3m (225) - 4 File: Duong BK mau AgCl say.raw - Type: 2Th/Th locked - Start: 20.000 - End: 80.000 - Step: 0.030 - Step time: 1. s - Temp.: 25 C (Room) - Time Started: 9 s - 2-Theta: 20.000 - Theta: 10.000 - Chi: Lin (Cps)01020304050607080901001101201301401501601701801902002102202302402502-Theta - Scale20 30 40 50 60 70 80d=3.197d=2.770d=1.387d=1.961d=1.672d=1.601d=1.239d=1.27136 Hnh 3.12. Ph IR mu hn dch bc clorid Ccnhhpthphhngngoimuhndchlnltti( max/cm-1 ): 3278,99; 2104,34; 1637,56; 680,87; 466,77; 422,41; 405,05. 37 Hnh 3.13. Ph IR mu hn dch trng Cc nh hp th ca ph IR mu hn dchtrng ln lt ti ( max/ cm-1 ) 3741,90;3273,20;2382,09;2308,79;1923,03;1641,42;1514,12;1454,33; 1340,53; 1087,85 ; 1045,42; 707,88; 447,49; 430,13; 405,05. GiaphIRcahndchnanobccloridvphIRcamutrngkhng thycskhcbit.NhvycthktluntngtcgiatiuphnAgClv PVA l tng tc vt l, khng c tng tc ha hc xy ra.38 BN LUN Quaqutrnhthcnghimchungtinhnthyqutrnhbochnanobc clorid ph thuc vo nhiuyu t: quytrnhbo ch,mi trng th nghim, cht bn ha. C th nh sau: -Quytrnhtnghpnanobccloriddavophnnghahcgiabc nitratvnatriclorid.toctiuphnnanophitinhnhnhchmnatri clorid vi tc chm 0,5 ml/phut, ng thi tin hnh trong iu kin khuy trn lintcbngmykhuyt.Tckhuycduytr500vng/phut,nugim tckhuysgimkhnngphntncctiuphnbccloriddnnktt thnhtiuphnkchthclnhn,nutngtckhuyqunhanhslmtiu phn chuyn ng nhanh hn qua c thcc tiuphn s kt tto kch thc lnhn.Doiukinthnghimnnchungtichsdngkhuyvilng khong250mldungdch,nutnglngthtchhnthskhngmbovic khuy trn ng u v lm gim kh nng phn tn tiu phn bc clorid. -Mi trng th nghim tin hnh trong iu kin hn ch ti a nh sng mt tri, do kh nng b phn hy ca mui bc chuyn mu en di nh sng mt tri. Bc nitrat c ha tan trong bnh cha dung dch PVA c bc kn en m bo hn ch nh sng mt tri. Qu trnh cn bc nitrat v bo qun cng phi chu hn ch ti a nh sng. Qu trnh thc hin th nghim s dng nc ct, dng c sch, hn ch bi hay tiu phn l trong thc nghim.-Qutrnhtothnhnanobccloridckchthcphthucvochtbn ha v lng cht bn ha s dng. Qu trnh thc nghim tm ra v la chn PVA l cht bn ha mang li kch thc ti u nht trong s cc cht bn ha kho st. KhilngPVAthpthnhtcamitrngcngnhkhnngbaoph,to lp o ngn cn cc tiu phn kt t thp, lm cho kch thc tiu phn tng. Ngc li, khi lng PVA qu nhiu s lm tng nht v gim kh nng khuy phn tn ca my khuy t, do cng lm tng kch thc tiu phn. Bn cnh cng 39 thcM14cthsdngPVPlmchtbnhatnghptiuphnnanobc clorid cng mang li kch thc ti u, v c th thc hin cc nghin cu tip theo vnnhtiuphnnanoAgCltrongchtbnhaPVP.Tuynhinkhitng cngthcbochln6lnthkchthctiuphnAgCltrongPVPlitngln kh nhiu.Khi tng lng cc cht ln 6 ln th th Zeta tuyt i cng thc M18 gim hn (+10 mV) lm tng kh nng kt t tiu phn, iu ny gi nghin cu c th thm cht chng kt t, tng n nh cho tiu phn tng hp c nh h mcitrat,hmphosphat.Saukhitnghptiuphnnanobccloridgp nhngthchthckhngnhtrongvicboqunsnphm(snphmdbphn hyngoinhsng),trongnhginnhcatiuphn(tiuphnckh nng kt t) v nghin cu ng dng tiu phn bc clorid tng hp c trong bo ch cc ch phm dc hc. Cn c nhng nghin cu tip theo v vic nghin cu thm cc cht chng kt t tiu phn, hay nghin cu tng hp tiu phn bc clorid dng ng kh tng kh nng n nh ca tiu phn. 40 KT LUN V XUT KT LUN Chung ti thu c cc kt qu theo ung mc tiu ra nh sau: 1.Xydngcquytrnhtnghptiuphnnanochabccloridckch thc tiu phnthu c nm trong khong 90 nm. +Cht bn ha la chn: PVA +Cng thc tng hp thu c:Ha chtLng PVA1,6344 g Bc nitrat0,1223 g Natri chlorid14,4 ml dd 0,1M Nc ct210 ml + iu kin th nghim: Hn ch nh sng mt tri. +Quytrnhtnghptiuphnbccloridthucnhsau::Hatan 0,1223 g bc nitrat vo dung dch nc ca cht bn ha PVA c kho st. Nh ttdngdchnatriclorid0,1Mvo,tcnh0,5ml/phut,vanhvakhuy trn tc 500 vng/phut. Tip tc khuy duy tr trong 1 gi.Sn phm thu c bo qun trong iu kin hn ch nh sng mt tri. 2. xc nh c mt s c tnh ca tiu phn nano bc clorid thu c: + Kch thc tiu phn bc TB thu c l 88,82 nm,vi gi tr PDI TB l 0,136. + Th Zeta TB hn dch nano bc clorid thu c c gi tr l -9,85 mV. + Hnh nh chp SEM cho thy cc tiu phn c nhiu hnh dng nhng ch yu l hnh lp phng. 41 + o ph hp th UV VIS nhn thy s thay i bc sng cc i hp th khi hn dch di nh sng mt tri. T a ra iu kin hn dch bc clorid phi bo qun trong iu kin trnh nh sng mt tri. + o ph nhiu x tia X thu c: Xc nhn s c mt ca AgCl trong hn dch. + o ph IR cho thy: Khng c tng tc v mt ha hc xy ra gia AgCl trong mi trng c PVA, NaCl, NaNO3. XUT Do iu kin hn ch v thi gian nn cc kt qu ca kha lun ch l bc uxydngquytrnhtnghp,xcnhvnhgimtsctnhcatiu phn nano bc clorid. Chung ti xin a ra mt s xut nh sau: 1.Tip tc theo doi n nh ca tiu phnnano bc dng hn dch iu ch c 2. nh gi kh nng khng khun ca tiu phn nano bc tng hp c 3. 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WhiteJML.,PowellAM.,BradyK.,etal.(2003).Severegeneralized argyriasecondarytoingestionofcolloidalsilverprotein.ClinExpDermatol,pp. 254256. PH LC Phlc1:ngknh,PDItiuphnAgCliuchviPVA(M9)vPVP (M14). Ph lc 2:ngknh, PDI tiu phn AgCl iu ch vi PVA(M18) v PVP (M19. Ph lc 3: ng knh, PDI ca tiu phn AgCl iu ch vi PVA (M20, M21, M22). Ph lc 4: Th Zeta tiu phn AgCl iu ch vi PVA (M20, M21, M22). Ph lc 1: ng knh, PDI tiu phn AgCl iu ch vi PVA (M9) v PVP (M14) M9 M14 Ph lc 2: ng knh, PDI tiu phn AgCl iu ch vi PVA (M18) v PVP (M19) M18 M19 Ph lc 3: ng knh, PDI ca tiu phn AgCl iu ch vi PVA (M20, M21, M22)

M21 M20 M22 Ph lc 4 Th Zeta tiu phn AgCl iu ch vi PVA (M20, M21, M22). M20 M21 M22