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Ten de tal: NGHIEN cmr UNG DUNG PHUONG PHAp sAc KY LONG HI~U NANG CAO (HPLC) VA DO QUANG PHO UV-VIS DE DINH TINH VA DINH LUONG CAC HOAT CHAT TRONG . MQT s6 THUOC CO'Ttr2 DEN 5 THANH PHAN
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T.S. THAI nUY THIN

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Co quan chu trl: TRUONG DAI HOC nuoc
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Co quan ph6i hop chinh: VIEN KlEM NGHIEM THUDe BO Y Tt . . .

BAO CAO TONG KET DE TAl NGHIEN COO KHOA HOC CAP BO . .
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NGHIEN ccu UNG DUNG PHUONG PHAp sAc KY LONG N HI~U NANG CAO (HPLC) VA DO QUANG PHO UV-VIS DE DINH TiNH VA DINH LUONG cAc HOAT CHAT TRONG . MOT s6 THU6c CO·Ttr2 DEN 5 THANH PHAN
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Ten d~ tal:

ChiI nhiem de tal: Thai Duy Thin Danh sach nhUng nguOi thl}'c hi~n chinh:

1. PGS-TS Thai Phan Quynh Nhu 2. NCS-DS Trdn vt« Hung 3. ThS Nguyin TUUngVy 4. ThS vs sn n« 5. ThS Nguyen Tudn Anh, 6. DS CKI Hoang Vein DUe.

ThOi gian thuc hien de mi: Tit thang 1-2003 den thang 12 -2005. Tdng kinh phi dll(_1c duyet: 190.000,000d. T6ng kinh phi dll(_1c Ap: 190.000.000d. c

MUCLUC . NQidung Phin A. TOM TAT cAc KET

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Trang 1
1 2 2 3

QuA NOI B~T CUA DE TAL 1. K€t qua n6i b?t cua d~ ali.

2. Ap dung vao thirc ti~n xa h9i. 3. Danh gia tlnrc hien de tai d6i chi€u voi de cuong nghien ciru da dtroc phe duyet. 4. Cac

y kien

de xuat.

Phin B. N(n DUNG BAo cAo CHI TIET. 1. D~t vAn d~. 2. Tang quan d~ tai,
2.1. Tinh hinh nghien ciru trong va ngoai mroc. 2.2. T6ng quan

4 4 6
6 6 18 18 18 19 22 23

v8 cac plnrong

phap phan tich.

3. DAi ttrC}'llg phirong phap nghien cuu. va 3.1. Thi€t k€ nghien ciru, 3.2. Chon mftu, ca mdu va d6i nrong nghien ciru. 3.3. Phuong phap nghien ciru. 3.4. Phuong phap xu

Iy s6li~u.

4. KSt qua nghien ciru. 4.1. Dinh tinh va dinh hrong paracetamol, bromhexin hydroc1orid, pseudoephedrin hydroclorid, clorpheniramin maleat va cafein bang phtrong phap HPLC. cs« tri: PGS-TS Thai Phan Quynh Nhu. 4.2. Dinh hrong d6ng thai paracetamol (PA) va acid mefenamic (AM) trong vien nen bao phim Pamesic bang phuong phap HPLC.

23

en: tri: TS. Thai Duy

Thin.

27

4.3. Nghien ciru dinh hrong ddng thai paracetamol (PA) va cafein (CA) trong thu6c da thanh phfuJ bang HPLC. Chil tri: TS. Thai Duy Thin. 4.4. Nghien ciru dinh hrong d6ng thai paracetamol (PA) va ibuprofen (IB) trong thuoc da thanh phan. Chi: tri: TS. Thai Duy Thin. 4.5. Dinh hrong d6ng thai paracetamol CPA) va quinin sulfat (QS) trong vien bao Antigrip F bang phuong phap HPLC. ci« tri: TS. Thai Duy Thin 35 31

38

58 va acid 69 79 85 4.6. Tai li~u tham khao. 46 52 cs« tri: NCS-DS Trim n« Hung. Dinh hrong d6ng thai paracetamol mefenamic trong vien nen Pamesic bing do quang ph6 MeA. o« tri: NCS-DS r-s« vi« Hung. Dinh hrong dong thoi theophylin.4. Dinh IUQ11g Ang thai paracetamol va quinin sulfat trong vien nen d Antigrip F bing phuong phap phan tich da cAutir (MCA). ephedrin hydrochlorid va 41 phenobarbital trong thuoc h6n h<JPtri hen suy€n bang phirong phap s~c Icy long hieu nang cao. 7. 87 . Chu tri: PGS. clorpheniramin maleat va phenylpropanolamin hydroc1orid b~ng phuong phap HPLC.8. Thai Duy Thin. Thai Duy Thin. Ti€u de tai 10.9. Dinh lUQ11gd6ng thai paracetamol. 4. 4. 8. 4. Phl} lue.7.TS Thai Phan Quynh Nhu. cs« tri: TS. ChiLtri: TS. Dinh IUQ11g d6ng thai paracetamol va ibuprofen trong vien nen bang phirong phap toan ph6.10.

vi nghien CMU khoa hfJc 12 cong trinh khoa hoc co lien quan da dircc cong b6..7-. --_ .Ch i Minh. GUO . --. >- TOM TAT CAC KET QUA NOI B~T CUA BE TAl ? X _ ? ~. d6 la phirong phap HPLC va phirong phap Quang ph6 tir ngoai phan tich da cAu tir MCA. •:. Tom tiit cae . nhb trao d6i thong tin. sre dung thu~t toan th6ng ke trong viec danh gia cac k~t qua phan tich .__ .. tap chi diroc hoc .. •:. Chung toi da khai thac. .. HQi nghi direc Dong dirong IV 1112005 0 Thanh ph6 He... . Dang ti~p tuc nghien ctru. b. (rng dung thanh ceng phirong phap. . da diroc cong b6 tren cac tap chi chuyen nganh nhu tap chi kiem nghiem thuoc.._____________________ Phdn A. ... thfun dinh la co gia tri khoa hoc va y nghia tlurc ti€n.... . Kit qua C(l thi ... Trong d6 c6 7 qui trinh sir dung plnrong phap HPLC va 3 qui trinh sir dung phirong phap quang ph6 tir ngoai pharr tich da cAu ttl MCA .__ cua _ .. K~t qui n6i b.--_ -.-- PHAN A. c. va tham gia bao cao trong cac hQi nghi v€ diroc n6i rieng va hoa hoc n6i chung (xem phfut phu luc). •:. . ---.1- .... •:. nhieu hon mQt cong thirc so voi du ki~n trong d€ cuong nghien ciru.2 cong tlnrc thuoc co ba thanh phfut ph6i hQ'Pva 1 cong tlnrc thuoc c6 nam thanh phfut ph6i hQ'P .:.__ _. Da khao sat tren 4 cong tlnrc thu6c c6 hai thanh phfut ph6i hQ'P.. 1. Da xay dung diroc 10 qui trinh phan tich dinh tinh va dinh hrong cac hoat chAt trong 7 cong thirc thu6c ph6i hQ'Pph6 bi~n tren thi tnrong... kit .. Xay dung duoc phuong phap. qui trinh phan tich da xay dung vao viec kiem nghiem mQt s6 cong thirc thu6c c6 til 2 toi 5 thanh phfut hoat chAt.l anh.__ oJ tai . Cac nghien CUutren d€u da duQ'Ccac chuyen gia v€ ki~m nghiem kiem tra... irng dung hai phuong phap phan tich cong C1.:. mo hinh qui trinh phan tich dinh tinh va dinh luong n6i chung d6i voi cac hoat chAt trong mQt cong tlnrc thuoc ph6i hop sir dung hai phirong phap tren . HQi nghi hoa hoc A Au 0 Ha NQi 10/2003.... danh gia tren mQt s6 cong thirc thuoc ph6i hep khac.. Dong gop mUi cila iI2 ta.:. ki~m clnrng va ban lu~ v€ phirong phap lien quan toi cac nQi dung c6 trong de tii cua chung toi. n6i b(it .t ella d@tii a. d~c bi~t la cac thuoc c6 nhieu hon 5 thanh phfut boat chAt.. Tham gia 3 hQi nghi khoa hoc qu6c t~: HQi nghi diroc Dong dirong III 5/2003 0 Thai Lan... hien m dai vito viec kiem nghiem cac thu6c c6 nhieu han mQt thanh phfut hoat chAt.

_?~4'!_:1~ t!. diroc vi~n kiem nghiem va m{>ts6 trung tam kiem nghiem img dung de kiem tra chdt hrong cac thuAc da thanh phfut dang hru hanh tren thi tnrong. d. Htrong a noi cong tac (vien kiem nghiem thuoc. MQt s5 chirong trinh dang diroc cac co so tham khao d~ xay dung tieu chuan cho san phAro cua minh. •:. ThVChifn cdc mflc tiiu nghien cu-u Da hoan thanh ddy dU theo muc tieu va cac nQi dung nghien ctru dllQ'ChQi dang khoa hoc cua be>phe duyet. hieu qua.:. tu« d{j hodn thdnh DS tai hoan thanh dung ti~n dQvS thoi gian dang kY. C6 12 sinh vien lam cong trinh t5t nghiep phuc vu cho nQi dung d~ tai. D~ tai da t~p hop va huy dQng duQ'c mQt sA hrong 1611 can bQ tre cua tnrong D~ hoc diroc Ha nQi~Vien kiem nghiem thu6c va mQt s6 Trung tam kiem nghiem dia phirong tham gia. .2- ..'!!_~4~ _! _~~~_M~ _1)_4!_k(iL~~q_~~_!!!~ _q_~!! d: Dong gop dao t(lo can b{j cho nganh . con thuc hi~n them diroc 1 cong tlnrc so voi dS cuong. trong d6 co 3 nh6m sinh vien tham gia bao cao tai hQi nghi khoa hoc tu6i tre cua tnrong. Cac chi tieu khoa hoc dB ra trong dS cuong duoc ton trong va thirc hien ddy duo c. MQt s6 chirong trinh diroc nghien cuu la de ap dung lam tieu chuan cho cac co so. Kit qua l(lo ra so v6i dfl"kiln cua ban td cumrg Cac k~t qua thu diroc hoan toan phil hop voi du kien. chi tiBn dS tai theo dung qui dinh. Banh gia v. 2. trung tam ki~m nghiem cac tinh. Ap dung vao thjre tiin sin xuAt va dM sang xi h{)i K~t qua nghien CUu cua chung toi Iii co tinh kha thi va c6 y nghia thuc ti6n. dan nhieu sinh vien tham gia thuc nghiem khoa hoc phuc vu cho dS tAi co k~t qua. phong kiem nghiem xi nghiep. Dauh gia thVC hifn di tai dai chi@u v6i di CVO'Dgughien ctiu di dtrqc phe duyft a. •:. gop phfut nang cao trlnh dQ va kinh nghiem chuyen mon cho cac can bQ. Gop phfut dao tao diroc 2 thac _ sy duoc hoc va mQt diroc sy chuyen khoa cdp 1. MQt s6 sinh vien sau khi t6t nghiep da phat huy t6t chuyen mon thanh) . 3.?c sir dpng kinh phi Da sir dung kinh phi dung muc dich. •:. b. 1 sinh vien tham gia bao cao hQi nghi diroc dong duong.

jrng dung hai phuemg phap dAxay dung cho nhi€u cong thirc Ian thuoc da thanh phAn khac. •:. irng d1J11g nhieu hem trong thuc ti~n d~ co co sa d~ nghi dua rnQt s6 chuyen luan thu6c da thanh phftn vao duoc diSn Viet Nam trong xudt ban too. Chung t6i se tiep tuc nghien ci'ru d~ hoan chinh va rna rQng hem trong nhtrng d€ tai khac. tiSp we danh gia cac phirong phap. qui trinh va k~t qua thu duoc. 4. Ti~p tuc rna rQng kSt qua nghien CUu cho mot s6 thuoc co nhieu thanh phful hoat chdt hem._q_. TiSp tuc nghien ciru. the -3- . RAt rnong ducc S1.:. d~c biet la plnrong phap quang ph6 ill ngoai phan tich da cAu ill MCA.!!_ ~4~ ~UIl!~_1}_4!_!!_ti_~ ~q_~~_!?!~ _ £~~_ £ .l' quan tam cua BQ Y t~ va cac co quan de d€ tai co duoc ap dung rQng rai hem va rna rQng cho nhi€u san phAro khac. Cae y kiin d~ xuit ._r~1J_4~r. •:.

dira tren phirong phap HPLC va phirong phap quang phd tlr ngoai phan tich da cAu tlr MeA . vi~c nghien ctru d~ dira m{>tsf.V. " PHAN B. Xay dung qui trinh phan tich m{>t s6 thu6c da thanh phfut dB: chon (chua co tieu chufut hoac co tieu chudn nhimg chua dAy du hoac co nhuoc di6m). Trong s6 do. gop phdn nang cao chAt hrong thucc hru hanh tren thi wang. Theo th6ng ke cua C\lC quan Iy Duoc Vi~t Nam. M\lC tieu nghien Ctlu . tieu chuan cho cac thu6c loai nay vao duoc di6n trong cac Ian xuftt ban toi lit rAt cfm thiet.. tren the gioi.. •:. cac thu6c ha nhiet giam dau.fi_~_~4~!!~ ..2. thuoc s6t ret. Nhu v@y viec nghien ciru phirong phap kiem nghiem cac thuoc da thanh phfm 1ft m{>tviec rAt cfm thiet d6i voi cac nha san xuAt.. v . cac vitamin.D. Vi viY. thu6c chong lao V. hien da co khoang 6 nghin m~t hang thu6c trong nuoc va khoang 4 nghin thu6c ngoai nh~p (iroc tinh co tren 800 hoat chAt) dang hru hanh tren thi tnrong Viet nam. thu6c da thanh phfm thirong duqc ph6i hop cac hoat chAt trong cung m{>t nhom thuec nhir: cac khang sinh.. cac thu6c co nhieu thanh phfut dang diroc san xuAt va sir dung ngay cang nhieu. hau nhu chua co cac chuyen Iu~ ve thuec nhieu thanh phAn.N. Cling co th~ k€t hQP cac hoat chAt cua 2. cac nha nghien ciru v€ kiem nghiem va cac co quan quan Iy chAt hrong thuoc. Tinh cAp thi~t cin nghien Ctlu cua d~ Ud san xudt va kinh Trong nheng nam gAn day. Cac thudc ngoai duqc cAp sd dang Icy hru hanh ngay cang nhieu.. -4- . Trong diroc di~n Viet Nam moi nhAt (D.:.v . thi wang doanh. chung toi dii tiSn hanh nghien ciru d~ tai: "Nghi2n CNU . ? A CUuDE 1.Vis ttl lIinh tinh va djnh lurmg cac hopt chat trong m{jt sa thuac co til' 2 din 5 thanh ph8n" 1.1. 3 nhom thu6c nhir vitamin voi thu6c ha nhiet. H~T TAIcApBQ vAN DE thuoc dang phat trien nhanh ca v~ 1. . thu6c nho mAt h6n hQP. D~ ph6i hop tac dung diroc Iy trong m{>tch€ pham thu6c va ti~n sir dung cho benh nhan._!_·__1?. Noi chung.mg dpng phrmng phap sdc kj long hi?u niing cao (HPLC) va do quang phD UV. cac thu6c da thanh phfm dB: va dang chiem m{>t tY Ie cao. Ap dung cac qui trinh tren d~ ki~m nghiem cac thuoc da thanh phAn co cong thtrc ttrong 1\1'. x X. NQI DUNG BAO CAO cm TIET KET QUA NGmEN 2. thu6c dung ngoai hem hop v. III). XuAt phat tir yeu cau thuc tS va cAp thiSt do.. i:J mroc ta. khang sinh voi thu6c chong viem.

clorphenirrarnin maleat va phenylpropanolarnin hydroclorid.. + Paracetarnol va ibuprofen.l·L-t (L '£~ t\A~ t-~~: [L~ ~ f'{_ c. d. Thuec c6 5 thanh phan +Paracetarnol. ephedrine hydroclorid va Phenobarbital..__l?.A ) tr ) ... + Theophyllin. + Paracetamol va quinin sulfat.p bo chung toi chon mot s6 cong thirc thu6c c6 ill 2 den 5 thanh phlin d~ nghien ciru (da diroc bOphe duyet). brornhexin hydroc1orid va clorphenirrarnin maleat. t p4:_ 1Af MLC~ML C <UCW-[ (-'\l1L{ _ ) + Paracetarnol. d6la cac thuoc han hQ'Psau: Thu6c c6 2 thanh phfuI + Paracetarnol va cafein. cafein._!. Muc tieu nghien CUll cu th~ Trong pham vi 1 d~ tai - .. -5- . Thuoc c6 3 thanh phdn {Ul.~~_'!41J_!!~ •:.. pseudoephedrin hydroclorid..

Tinh hinh nghien eu-u trong va ngoai nwe sa va cAp bQ v~ ki~m Nghien Trong nhtrng nam qua. 27. Diroc dien Anh (BP1998 va BP2000) co ghi mQt s6 thuoc da thanh phk chuyen lu~ thudc multivitamin. quang ph6 dao ham de dinh hrong cac thuec da thanh phfin [7. nhirng phan 160 chi dirng a mire tieu chucln co sa. Qua trinh site kY x~y ra co th~ thea cac co ch€: hftp phu. 28] . dang ti~u phan hoac mQt chAt long phu len mQt chAt mang rAn hay mQt chdt mang dd duoc biBn dbi bfutg lien kBt hoa hoc voi cac nhom htru ca.. chu yeu ghi cac dang thudc dan thanh phan. USP 24. -6- . 8]. nhir cac My (USP 23.. Tuy nhien. trao dbi ion hay ray phan ttl.. Trong cac duoc di~n 160 va thong dung nhir BP.. ciru dinh lUQ11gd6ng thai cac vitamin trong dAu trong thuec h6n hop multivitamin - Nghien cuu irng dung quang ph6 UV-VIS. cac ch€ phfun thuoc da thanh phfin duqc nghien ciru rAt nhieu. Cac mroc trong khu V\lC Dong Nam thuoc tiem lidocain va adrenalin ..] .!_!~~_~~~~ 2._~·__!_4~g_q~l!_. T6 clnrc Y te the gioi cling dd co nhtrng chirong trinh d~ t6 chirc hoc t~p va trao d6i gitra cac nuoc v~ vdn d~ nay. USP 27 .. mQt pha dQng va met pha tInh. USP.1.2. Tren the gioi. Pha tinh chua trong cQt 1(\mQt chAt rAn dd direc phan chia duo. so voi tdc dQ phat tri~n cua thi tnrong thudc hien nay thi chua du dap img. JP. phan bd.. Khai ni?m sac kY long hi?u nang cao La mQt phtrong phap tach hoa ly dua vao ai hrc khac nhau cua cac chAt khac nhau voi hai pha luon tiep xuc va khong dang tan voi nhau. Qudc t€ [19. Pha dQng 1(\chAt long chay qua cQt voi mQt t6c dQ nhdt dinh. con rdt nhieu nQi dung cfin phai duqc nghien ciru d~ img dung trong thirc ti€n... TONGQUANTAILIBU _ 2. 2. ) co sd thudc da thanh phfin nhieu nhom thudc ha hon va t~p trung vao mQt s6 nhom thuoc ph6 bien nhir: cac vitamin. 25. a Vi~t Nam ta dd co nhi~u cong trinh nghien crru a cAp co nghiem cac thuoc da thanh phfin nhir: tan trong mroc va vitamin tan bang phuong phap HPLC [10]. A dd co nhtmg hQi nghi v~ cac phuong phap kiem nghiem ngoai diroc di~n.2.. 24. EP. Phu01Ig ph tip siic kj long hifU nang cao (HPLC) a. Nghien ciru dinh hrong dang thoi cac hoat chAt trong mQt s6 thucc ha nhiet giam dau bang phirong phap HPLC [6. 23. Rieng duoc di~n nhiet giam dau .1. Phap. TAng quan vi eac phuong phap phan tieh 2.

. ___________~ -------..:..phan b6 nay dtroc d~c trung boi can bang phan b6 voi h~ s6 phan b6 duoc tinh theo cong thirc sau: K= Cs CM Trong d6: Cs... Khi pha dQng chay qua cQt vm mQt t6c de) khong d6i thi thai gian hru co th~ thay th~ bang th~ tich hru.W.... "-------·-.. Th~ tich luu la th~ tlch pha dQng thu duoc sau cQt trong khoang thai gian nrong irng voi thoi gian hru.K ••• LIe so p h an b._._?_. ~ .l _---_ ..thoi gian hru thuc (thoi gian luu hieu chinh).._---. Thbi ....to Trong do: t'R hi. -7- ..~--~... Vi v~y thai gian hru la d~i hreng dinh tinh cac chat.~~----------1· lIOl...-------.. Trong cung mQt di~u kien sAc kY eta chon.. eM la nAng de)cua chAtphan tfch trong pha dQng va pha tinh 6 thai di~m can bang.... .~ .~--..1 I ._-_..:t .I ...--....._x'_4~gq~q_'!_~~~X~~1}_ b._-_ ..... '''.. ._ ......6..-.ian lltu ... ----._..~~--------------. S\!.. Cdc ilQiltamg ilgc trung . 10la thai gian ch~t (thai gian khong hru gift). cAu tao va tinh chAt cua chAt do. siic ky db cua hai chAtva cac thong s6 d@ctnrng ..~.. va thi coo qua trinh sAc 10/ tich lltu Thai gian hru cua mQt chAt la thai gian tinh til Iuc tiem mdu vao cQt d~n khi chAt do ra khoi cQt dat gia tri nAng dQcue dai va cho pic tren sAc kY dA_ N~u goi tR la thoi gian hru gift cua me)tchAt thi : t' R = tR . thai gian hru cua m6i chAt la hang dinh va cac chAt khac nhau thi thoi gian hru khac nhau tuy thuoc vao ban chat. .--.__·•••• _ .f 0 • rIA A Trong qua trinh sAc Icy luon co su phan b5 cua chAt tan gitra pha dQng va pha tinh. :i Hinh 1.

Tlnrong phan tich trong dieu kien a thi trong khoang 1. M6i tAng dUQ'Cgill lap pha tinh co chieu cao H. Chien rQng pic do (] mra chieu cao pic. nhu v~y a luon 16'11 han 1.5 d€n 2 .:. 86 dia ly thuyet N diroc tinh theo cac cong thirc: N ==16 X (!!L)2 W hoac 5.. Sa ilia IVthuv2t va ehi2u eao ilia IV thuv2t Hieu hrc cQt tlnrong diroc bieu thi qua hai thong s6: se rna Iy thuyet (N) hoac chieu tAng su phan cao dia ly thuyet (H). a==-==-= KB sb chon IQc cho r.V M lit the tich pha tinh va pha dQng. Chieu cao cua dia Iy thuyet dugc tinh thea cong tlnrc: H=!::.. QM lit hrong chAt tan co trong pha tinh va pha dQng. -8- . . •:. Thira _ sa dung /U'(!IIg Thira s3 dung IUQ'Ilg lit d~i IUQ'Ilgbi~u thi khll nang phan bb cua chdt tan trong hai pha cQng vai strc chua cua cQt. Trong do L la chieu cao cua CQtsAc ky. nrc lit tY sA gitra IUQ'IlgchAt tan trong pha tinh va IUQ'Ilg chAt tan trong pha dQng a thoi diem can bang k'== Qs == Kx Vs QM VM Trong do Qs.~~!1. Dfa ly thuyet dircc dinh nghia nhu lit mQt khu V\lC cua h~ thong phan tach mit trong do thi€t l~p mQt can bfutg nhi~t dQng hoc gina n6ng dQ trung binh cua chAt tan trong pha tinh va trong pha dQng. CQt sAc ky diroc coi nhu co N tAng Iy thuyet. C6 the tlnh k' thea mQt cong tlnrc khac: . dinh nlnr mQt a m6i b6 chAt tan vito hai pha lai d~t den mQt trang thai can bang moi. k'A I RB -10 IRA -10 KA I'RB ==I'RA o day ta quy uoc chAt B bi hru gift manh han chAt A. Hi sA ehvn lee Hai chdt chi diroc tach ra khi chung c6 cac gia tri k' khac nhau. a cang 16'11 kha nang tach cua hai chAt CMg ro. V s. chieu cao dla 19 thuyet (H) va s6 dia 19 thuyet (N) lit hang dinh d6i voi m6i chAt phan tfch._Lr4~g_p'!'f!_.:. N V6'i mQt dit~u kien sAc kY nhAt dinh.!_~~~!.54 X (_!L_)2 ~/2 Trong do W ~/2 : Chieu r{>ngpic : a day pic. h~ bi~t hieu qua tach cua h~ thbng sAc kY.

__. PhU'dng trinh Van-Deemter Phuong trinh Van-Deemter mo til anh huang ella tac dQ dong pha dQng va cac thong s6 dQng hoc khac d~n hieu hrc cua CQtsAc kY H=A+-+Cxu u B H u : Chi8u cao rna ly thuyet. Be Dhlin cidi (R~ DQ phan giai la d~i hrong bieu thi dQtach ella cac ehAt ra khoi nhau trong mQt di8u ki~n sitc Icy da cho. Trong d6: B/u: M6 ta anh huang cua su khuech tan doc cua cac phan tiT chAt tan theo phirong dong S1)' chay cua pha dQng (longitadinal diffussion).•0'. B. C : Cac h~ s6 thay d6i phu thuoc vao tirng CQtsAc kY.licua pic d€n chan dirong cong phia tnroc. May ghi tin hi~u hoac may vi tinh. 2.18 x {tRB . chuyen khai (mass transfer).._ __ __~ _. = 2xVRB -tRA) WA+WB 1. B(>phan tiem mdu: tiem vao cQt mQt the tich mdu nhAt dinh.'2(A) + WII2(B) ho~c --x a-I k' a l+k' B B x4 -IN . A: M6 Ul anh huang cua su khuech tan xoay (eddy diffusion). T6nq quan tai lieu . •:. A. : Tac dQ dong pha dQng.:. Bam cao ap: dAypha dQng qua CQtsAc ley. H~ th6ng HPLC: ~ Binh chua dung moi (pha dQng). Detector. H~ thang may HPLC c6 cac bQph~ chinh sau: -9- .__ _ __ __ __ __ _ L ... DQ phan giai ella hai pic canh nhau diroc tinh theo I trong 3 cong thirc sau: s. C: M6 ta anh huang cua c. n6 duoc tinh theo cong thirc: T=Wz 2A Trong d6 Wx: la dQrQng day pic do a 1120chieu cao ella pic. CQt tach (pha tinh).:. He sa dJi xfmg (V Cho bi€t mire dQ can d6i cua pic sAcley.. A : la khoang each til dirong vuong g6e ha til cue df. 6 tai 1120chien cao pic ..tRA } W.

5. n~u pha tinh la chdt cua pha tinh va pha dQng. 2. Detector.10- .:.10 um. Khdi ni€m Pha tinh trong HPLC lei chAt nh6i cQt d€ lam nhiem vu tach mQt h6n hop chAt phan tich. dien tich b~ m~t hat thuong ill 50 ~ 500 m2jg . ngtroi ta chia n6 thanh hai loai. Module di~u khien (Control module). Can cu theo trang thai rAn hay long cua pha tinh. B{} tiem mdu 1\1' d{}ng(Autosampler). phan b6. --Hlnh 2. Bam cao ap (Pump). sAc ky pha dcio (pha tinh it phan cue con pha dQng thi phan cue). Pha tinh . 4.!_!?!~_~~~~ -------------. 7. d. Can cir theo c~u tnic x3p cua pha tinh la cac hat rAn. . Phiin loai Can cu theo ban chAt chinh cua qua trinh sAc kY trong cQt tach. kich tlnroc hat rAt nho. Cac thanh phfut co ban cua mQt may s&ckY long hien dai -_ - -- 1_Khay chua binh dung moi (Solvent cabinet). nguoi ta chia n6 thanh hai C\lC kY long-ran (LSC). x3p. dirong kinh ca hat ill 3 ._?__!4~g_!J!I_l}_. N61A nhirng ch~t rAn. Can cu theo de) phan thuan (pha tinh phan d6i ion. c6 cac loai: sAc Icy pha C\lC con pha dQng it phan eire). 3. 6. BQ di~u nhiet cho cQt (Columm compartment). •:. BQ du6i khi chan khong (Vacuum degasser). n~u pha tinh hi chAt rfuI ta c6 sdc long ta c6 sdc Icy long-long (LLC). h~ th6ng HPLC hoan chinh Hlnh 3. sdc kY pha dao tao c~p ion va sAc Icy trao loai Ia xop toan phan hat va xop b8 m~t hat (xop chi lap v6 ngoai). trao d6i ion va ray phan tiro Tirong irng voi loai chAt nh6i nhu th~ nguoi ta co mQt loai sAc kY rieng trong ky thu~t HPLC. ngiroi ta chia n6 thanh nhieu loai nhu hAp phu.

Pha dQng la mQt trong nhtrng yeu tA quyet dinh hieu suAt tach sAc ky cua mQt han hQ'Pm§:u. N6 cling c6 th~ Ia dung dich cac mu5i c6 chua cac chAtd~m. . T6c de)dong pha dQng. Pha dc)ng la dung mol dung de rna giai chAt tan (chdt cAn phan tich) ra khoi cQt tach d~ thirc hien mQt qua trinh sAc kY_ Day la mot y€u t6 rAt linh dQng va d€ dang thay d6i. Rira giai gradient: pha dQng lien tuc thay d6i (do tY I~ cua cac thanh phAn tao nen pha dQng thay d6i) trong su5t qua trinh rna giiti. Y iJ sAc kY trao d6i ion va sitc kY cap ion) .. lJiiu kien d6i vu. chAttao phirc . . Cae ylu ta ehinh can chu V Irong lila chen pha ilOng - Ban chAt cua dung moi lira chon lam pha dQng. kY nhdt dinh. N6 c6 th~ la mQt dung moi hoac h6n hop nhieu dung moi trQn Ifill voi nhau thea nhtrng tY I~ nh~t dinh. Thanh phAn cac chAt tao ra pha dQng. Pha dong . la ~c trung x5p cua no). . Tinh chAtb8 mat phai dn dinh (d~c biet wong d6i dAng nhAt. Hoa tan diroc chAt cAnphan tich. Qua Irinh rira giiii Rihl giii ding dong: pha de)ng khong thay dbi trong su6t qua trinh nra giai. •:.:. lJi€u kien iliii vUi mot pha dong Phai tro dAi voi pha tinh. Phil hQ'Pvoi Ioai detetor dung d~ phat hien cac chAtphan tfch. Phai nhanh dat cac can bang trong qua trinh sAcky. C6 tinh kinh t€ va khong qua hiem . Luc nay.pH cua pha de)ng (d~c bi~t chu •:. Khai niem .N6 quyet dinh thai gian hru gift cac ch~t mau va hi en qua su tach sic Pha dQng c6 th~ anh huang d€n: DQ chon IQc cua he pha. de) phan cue cua pha dc)ng cling se hi bien d6i va thuong la tang hi~u quit tach . •:. mOt pha tlnh Phai tro va h6n vtrng voi cac di6u kien cua moi tnrong sitc kY.co. C6 de)tinh khiet cao.. N6i chung m6i loai sitc kY se c6 cac he dung moi rita giai rieng dS c6 duoc hi~u quit phan tach t5t nhAt. .:.11 - kYo .Can hang dQng hoc cua su tach phai xay ra nhanh va I@plai t6t. B~n vtmg thea thei gian. Thai gian hru gift ella chdt tan. C6 kha nang tach chon IQc mQt h6n hop chdt tan nhdt dinh trong di~u kien s~c ._?__!4!!g!l¥_q_'!_!?!~_~~f¥_ •:.hat phai e.

. •:. Trong HPLC. K~t qua cua chAt chua bi~t dircc tinh toan so vci miu chufin da bi~t tnroc nAng dQhoac suy ra tlr direng chuan . g.. Phuong phap nay co th~ dung cho cac pic khong bi troi duong n~n va ca pic co dirong nen b] troi.Do r(lng va S\I can d6i cua pic sAc kY . phirong phap nay chi dung khi co S\1' dap trng cua detector tren cac chAt la nhu nhau. f each ilanh gia pic do. Phlrung phap ngo(li chunn: La phirong phap dua tren co sa so sanh m§u chufin va miu thir trong cung di~u kien. hien nay ngtroi ta thuong dung may tich phan di~n ill gAn vol may vi tinh (sai s6 khoang 0.them vao) va dQ tang cua dien tich hoac chieu cao pic .n diem dau va diem cu6i cua pic dtroc . Danh gia chiiu cao pic: Khi pic c6 dang khong dAi thi chien cao pic (khoang each gitra duong n~n va dinh pic) lit mQt dai hrong the dung d€ danh gia sdc tY l~ voi dien tich pic va n6 ding co kY db._?_·__!4!!g__q~q_. •:.!1. . •:.!_!?!~_~~f. Phuong phap nay chi ap dung khi cac chi s6 k' la hang dinh. Voi pic co duong n~n bi nhiBu hoac pic hep thi vi~c xac dinh chien cao pic se dB dang va chinh xac hem vi~c xac dinh dien tich pic. •:. •:. N6 direc tach hoan toan va co nang dQ gfut bang nllng dQ cua chdt phan tich va co cAutnic hoa hoc nrong tu. each tinh kit qua .:.:. n~u khong nhu nhau khi do voi m6i chAt can c6 h~ s6 hi~u chinh. hilt chan khong. D~ tinh dien tich pic. Hic sieu fun . Phuung phtip them chunn: Chu yeu duoc sir dung trong kY thu~t HPLC khi c6 vftn de anh huang cua cac chdt phu (vi du : ta diroc).5%) hoac may tich phan co hoc (sai s6 khoang 1. rna trong cung di~u kien sAc kY n6 co thai gian hru gdn c voi thoi gian hru cua chAt cAnphan tich trong miu thir. DQ phan giai cac chAttrong mQt pha tinh. - Hieu 19c cua CQttach.3%). Vi khong khi hoa tan trong dung rndi se 410 bot khi trong detector va gay nhiBu tin hieu. . Dung dich miu thir diroc them mQt lUQ11g ac dinh chit chuan. K~t qua ducc tinh toan dua vao str chenh l~ch nbng dQ (luong chAt chufuJ. Phlrung phdp tinh theo phlin tram dien tich pic: Nang dQ cua mftu thir diroc tinh toan dua tren dien tich pic tinh thea tY 1~phftn tram dien tich pic chAt thir tren tAng dien tich toan bQ pic co trong sAc kY db.12- . Tdt ca cac dung moi dung trong HPLC (k~ ca pha d(lng) d~u phai duoc dU6i khi bang each dun nong. . Phuong phap nay chi d. vira va cao . Cac pic thu diroc cua cll hai dung dich miu thir va x mfiu thir them chAt chufut phai duoc do trong cung mQt dieu kien slic ky. Dank gia dien tick pic: Dien tich pic cua mQt chAt nrong (mg voi tlmg IUQ11ghAt c nhan ra chinh xac va cho k~t qua tat d6i voi nang dQtrung binh. Phuung phap n6i chunn: La plnrong phap cho them vao miu chuan va miu thir mot IUQ11ghAt khong d6i.

[K] lit rna tr~ tY Ie. _ 2. voi n A2 = Kl. Gia sir m > n. ta se thu duoc h~ g6rn m phirong An sf. D~ co th~ giai duoc phirong trlnh (4) ta can phai chuyen rna tr~ thanh rna trin vuong bang each nhan voi rna tr~ chuyen vi cua K.n c.d duqc goi la h~ sf.n Am. C: n6ng dQ dung dich.:.C (voi K = E. . tY 1~( Km. v~ n6ng dQ ( Cn) va n. Neu ta gia sir h6n hop cAn do co n thanh phan thi theo nguyen ly tren ta co: Tai buoc song thit nhcft: Al trinh.2 he sf... .13- .I K2.1 KL2 •. (5) chuyen vi cua K lit [K']m.n ta diroc Nhan ca hai ve cua phirong trinh (5) voi rna tr~ .. Nguyen tdc chung Co so Iy thuyet cua phuong phap dua tren nguyen Iy "D(j hap th¥ quang A coo mot h6n h(1JJ trong dung dich tai m6t buue song m6i thanh phim tai buac song do".nCn (3) AI A2 = KI. ta co th~ viet gon lai nhu sau: A = K. m An sf. v~ h~ sf.n en [A] lit rna tr~n dO hAp thu quang.d (1) E: dQ hAp thu rieng tai biroc song A.n •••• ~ •••••••• a 111 ••••••••••• CI x C2 (4) Am Trong do: Km.nCn .2C2 + + Kl. diroc xac dinh tnroc bdng each do ph6 cua cac dung dich chuan h6n hop d§ hiet tnroc n6ng dQ. d: b~ dAy cf.c do. K2....nCn + K2. khi do ta co th~ viet phirong trinh (4) dum dang sau: ehu nh~t [Klm. A = E._?__.lCI + K1. Co . KI.Beer: lien he voi nhau A: d{>hAp thu quang tai btroc song A. diroc do tren may. a bat 10/ bfmg tang do hap thu quang m6i cua Trong dung dich.2. ) (2) Khi de>hAp thu quang duqc do tai m buoc song.2C2 + _ + K1.l Km. 1 =[Klm.C.2. • .. Km.. tY 1~). [C] la rna tr~ n6ng dQ cua cac thanh phAn trong h6n hop. dQ hAp thu quang va n6ng dO dung dich co theo dinh lu~t Lambert .1CI + K1. Phuong phdp quang phD tii"ngo{1iph8n tich iIa c8u tii" (MeA) so lY thuyit .n K2.2C2 + = K2_1C1 + K2.2 ".4!!g_!l!l_l}_'!_~~~L~~'!: ! a.n Al = Kl.

tuyen tinh.l = [T]m.14- .n ···a n. _ .t = [T] m. d~ xac dinh nbng de>chua hiet cua rndu tlnr.t • •:.l a2. khi d6 ta se c6 h~ n phuong trinh n he nay ta dung phirong al.nxn= b2 he sf.l -1 (6) Nhir v~y.n. Tone quan fai li~u _ . day chinh An (nghia lit ta phai c6 m ~ n). ---- ----_. Trong wang voi m = n.2x2 + + al. _-or - ..1xI + a1._ .n (7) Phuong phap Gauss la phuong phap giai bang each dung cac bi€n d6i sa cAp d~ co rna tr~ h~ sf.n A= o a2. Phlin dch chuan. [Z]n.11 ••• a2. au au .1x] la he Cremer An va rna tr~ [K] lit rnQt rna tr~ vuong quen thuoc trong dai sf.__ 2.2x2 + a2..2 ••• dua he da cho vS he tam giac co dang: a1. Xn..lx] + a2. + a2..t Trong thirc te cua bai toan..m .nXn= bI + a2. a1. v~y ta c6 th~ viet phirong trlnh tren duoi dang nhir sau Cn. - - - - - - - - - - - - - - - -- - - - _. diSu quan trong la ta cdn phai giai ducc phirong trinh (6) dua tren viec xac dinh rna tr~ qua do de>hAp thu h~ s6 hAp thu [K]m.nthong Am. va Xl a plnrong thull trinh dAu tien . Gitii he Cremer bdng ohufTIIg ohao Gauss Me>t h~ phuong trinh dai s6 tuyen tinh chi tim diroc nghiem.2x2 + + al..2 a2.n . th€ len cac phirong trinh ti€p tren tim ducc Xn-l.nXn= b2 a1.i thieu (least square method) se lam cho dQ dung trong viec phan tich chuan cao hon vi n6 lam giam thieu S\l' sai khac gitra n6ng dQ tlnrc (entered concentration) va nAng de>tinh toan duqc (calculated concentration).nXn= b.Tir phuong trinh cu6i ta se tim duoc xn. co rna tr~ Xet h~ phuong trlnh Cremer sau: + aI. .2··· a2. khi do viec giai he tam giac tren rAt don gian.f 0. au ~. ---+ [Z]n.. •:.1I (8) oo Voi gia thiet ~._______ 0..2x2 + A= a2. Cn. danh gia chat lup?y! cda phlin tich chuan bdng cach ap dung binh phufTllg tAi thiiu va cdc thOng k2 tUfTllg quan Thuat binh phtrong tf... khi s6 plurong trinh hop t6i thieu cua h~ 100 han hoac bAng s6 (kich cO'n x n).2 . D~ giai loai phap Gauss. C lit nang dQ chua hiet cdn tim cua rndu tlnr.

co nghia la ta dd phai giai 10 he co 4 phuong trinh d~ xac dinh 4 An s6 la cac h~ s6 tY Ie K cua 2 chAt tai hai buoc song.n. ta co ly binh phirong tbi thieu = K2.[A] .15 - (10) . tinh duQ'c rna tr~ trung binh [K]tb va se xac dinh duoc nong dQ cua m6i thrum ph§n trong dung dich chuan h6n hop theo cong thirc: [C] = [K]tb-1. Neu gia thiet chi do tai hai btroc song thi se co phuong trinh.cu the nhtr sau: Tai biroc song I dung djch han hop chuan 1: Al Tai buoc song 2 dung dich h6n hop chuan 1: A3 = K2. ta phai sir dung tbi thieu 2 dung dich h6n hop chudn va do rn~t d9 quang tai t6i thieu 2 buoc song khi do ta se co 2 x 2 = 4 phuong trinh xac dinh cac gia tri cua h~ s6 tY Ie Km. d~ phan tich chuan chung ta se dung 5 ChUM hon hop._?__!.2C4 s6 cdu tU (p>n}".!_!~~_~~~l}_ Gilt thh~t s6 hreng cac h6n hop chuan duoc sir dung Ia p. khi do m6i I§n chung ta chi can dung gia tri phan tich cua n dung dich chuan trong s6 p dung dich sir dung (p en = P p > n).2C2 dung dich h6n hop chudn 2: ~ Tren tlnrc t€. nghiem. de giai loai h~ nay ta dua v~ viec giai h~ Cremer.2C2 dung dich han hop chuan 2: A2 = Kl. .overdetermined system".2C4 = Kl.lC1 the giam sai s6 trong phan tich chuftn bang each ap dung nguyen "su dung sa /U(Y11g dung cac dich h6n hap chudn nhiJu han K2. Til cac gift tri K nay. v~y chung ta g~p bai toan t6 hop ch?p n cua p ph§n tiro Theo cong thirc tinh t6 hop ta co: p! nL(p-n)! (9) p Ket qua la ta se co thu duoc . V~ nguyen lAc.IC1 + K2.1C3 + KI. en tnrong hop._4~g_q~f!_. m6i nghiem gAm n2 gia tri nrong img voi n2 phcln tir cua he sf> T ~ . nghia la ta se co nrong ung en h~ Cremer va e. Vi d\1: vfut voi tnrong hop han hop co 2 chAt 6' tren. Chung ta d§ bi€t muon xac dinh duoc rna tr~ he sA [K] cua h6n hop gAm n chAtthi ta phai sir dung t6i thieu n dung djch han hop chuan va do tai tbi thieu la n biroc song . Theo dang rna tr~n cua h~ (4) 6' tren. mai he c: 5= 10 tnrong hop va se co 10 gift tri rna tr~ chuan hoa K. Vi du: muon xac dinh cac h~ s6 K cua h6n hQP gAm 2 chAt. khi do he tren se tro thanh ~~he tren xac dinh. (rn = n = p) de co n2 phuong trinh xac dinh rn9t nghiem duy nhAt cua h~ gAm n2 phcln tir cua K.lC3 + + K1. tY Ie K.

·_ _!. rna tr~ [Kltb duoc sir dung d~ tinh true ti~p nAng de.:l!__. m6i IAnta chi cAn sir dung gia tri di) hAp thu quang do tai n buoc song trong sa m biroc s6ng pharr tich. sau do tien hanh phan tich cac miu chufut va miu tlnr. tinh diroc d~ danh gia sai sa cling nhu chAt hrong dtrong chuan . detector chu6i diot dUQ'cnbi voi bQ truyen va bien dc3idft li~u tac di) eao. Ph6n mJm cho phdn tich da ctiu tu MeA 15 gia tri cho n6ng dQ cua m6i thanh phAn va k~t qua se la gia tri trung binh cua IS gia tri rna ta Th\I'Cte. cilng nhu han ch~ v~ thi~t bi xir Iy sa li~u d§n d~n ket qua phan tich khong chinh xac hoac co k~t qua. Sau do.._. •:. VS phan cirng. ta co th~ dung cac thong ke nrong quan ta. Ih. co sa Iy thuyet cua phuong phap pharr tich da cAu til (MCA) cling nhu cac thuat toan d~ giai quyet no da: eo tic lau. D~ phan tich MCA tren the h~ may nay. cac may diroc trang bi bi) vi xir Iy tac di) cao.16- the do duqc chinh xac nhung lai phai kha cong phu trong viec tinh toan ._4!!g_!l!. Theo nguyen Iy binh phuong tai thieu. do han ch~ v~ di) ehinh xac cua cac thi~t bi phan tich. d~ giam sai sa efta phep do. ta lai g~p bai toan t6 hop ch~p n cua m phfut tiro Vfut a vi du tren. DU 7000 cua hang Beekman hay HP8452._ _. Phlin tich mflu Ihir va ap dung Ihu{il loan hinh phu01Ig mduthir a tren va nAng dQ thirc cua cac dung dich chuAn h6n hop. cAu hinh quang hoc cai ti~n. cho phep phan tich ph6 nhanh hon. tnroc h~t chung ta phai lua chon mi)t sa buoc song pharr tich thich hop dai voi m6i thanh phArr. HP8453 cua hang HP e6 su cai tien ro ret ca v~ phAn cirng va phArr mem. va sai sa se cang nho khi sa bircc song pharr tich cang tang. b. Nhu v~y. trong tnrong hop dung 6 bmrc 2 C6 = -" song pharr tich cho hon hop gem c6 hai chat. chung ta se giai duoc tinh duoc._7. _ So sanh cac gia tri C nay voi gia tri nAng de. chua bi~t cua miu thir. cuai cung la xir Iy k~t qua dua vao phArrmem tren may tinh di~n nr. . Nam 1989-1990: Hang Beckman gioi thieu phAn mem pharr tich quang ph6 MCA tren cac may quang ph6 Beckman DU thong thuong. Tuy nhien.!_!~~_~~~1}_ . va da: c6 thS thuc hien diroc ngay tit khi co may quang ph& UV -Vis. chung ta se dung sa hrong cac buoc song phan tich Ian hon sa cAu tir trong h6n hop (m > n) bang each sir dung them nhieu thong tin hon cua ph&.2u Irong phlin tich Sau khi phan tich chuan d~ xac dinh rna tr~ h~ sa tY l~ [Kltb va chAt hrong cua no dira vao cac thong ke da: neu. khi tang sa buoc song pharr tich se giam duqc sai sa cua phep do (giam gia tri t&ng binh phtrong cua su sai khac gitra n6ng dQ trung binh va n6ng di) cua nrng tnrong hop t& hop biroc song). _. d~ tinh n6ng dQ chua bi~t cua miu thir. tang di) nhay va di) l~p lai cua biroc song.. rna ta pha ban dAu cho m6i thanh phAn trong h6n hop ta se tinh duqc S\l' sai khac gitra nAng de. Cac th~ he may quang ph6 hien d~i sau nay nhu: DU 650i. Do do.

Hinh 5. Hinh 4. dit li~u ph6 trong khoang biroc song da chon dung cho phan tich._~·__4!!g_q~q_. Su cai ti~n ve giao di~n phfut mem.!_!~~_~~f.'!':. t6 hQP de tren xac djnh-overdetermined) xu ly k~t qua cho phep ap dung ky thuat MCA de phan tich duoc mQt s6 chc§pham c6 ill 2 d~n 3 thanh phfuJ.17- . xu ly du li~u ph6~ cling nhir Ia ap dung cac thong viec tinh tom k~t qua tra nen dan sa kiem ke toan hoc da lam cho gian han voi sai s6 t6i thieu. vi v~y chung toi da thi~t l~p mQt phdn mem hApthu quang tren may quang ph6 thong thuong. a nhieu co nghiem chua dugc trang b] cac th~ h~ may quang phA hien dai nhu tren. Thic§tl~p cac thong s6 chung . r d6ng thoi thu diroc toan be. Chuang trinh h~ plurong trinh Cramer. dung thuat toan chuyen h6a chu6i Fourier trong Hien nay . xu Iy nay rna thuat giai h~ Cramer (h~ cua n6 dua tren viec giai quyet hai bai toan chu y~u~do la: plnrong phap Gauss de khir xu ly cac kc§tqua thu duQ'c til nhieu cung voi viec loai be cac kc§tqua qua sai l~ch theo nguyen ly cua thuat binh phuong t6i thieu (least square method). Cira s6 chuang trinh phan tich da cAu til MCA.

NQi dung nghien ciru: . M6i tieu d~ tai chon t6i thi€u la 2 mdu thuoc d~ nghien ciru voi lam cac khao sat thuc nghiem. •:. Thu6c co 3 thanh phan: + Paracetamol. Thir nghiem tim kiem ky thuat hoac giai phap d~ loai trir anh huang cua cac thanh phfut khac (k~ cft ta duoc). clorphenirramin maleat va phenylpropanolamin hydroclorid.. khai thac cac irng dung cua phuong phap s~c ky long hieu nang cao (HPLC) va phirong phap quang phi. bromhexin hydroclorid va clorphenirramin maleat. cafein. Thi~t k~ nghien ctru f)~ tili (hrgc thi~t k~ theo mo hinh nghien cnu thvc nghi~m t~li pheng thi nghiem. Mfii chuong trinh cho mQt cong tlnre thn8c se Ii mQt ti~n d~ tili. 3.:.. DOl TUONG vA PHUONG PHA!> NGHIEN CUU 3.2._. . •:. s6 hrong dli d€ lJtJi tlf(1llg nghien cau Cac mau thuoc ap dung tuy theo thi tnrong cung c~p. 3.!l9l cho cac ti€u d~ tai. •:. ephedrin hydroclorid va phenobarbital. Chon min. •:. + Theophyllin. crr min Chpn mau va cO mau va dBi ttrQ'Dgnghien ctru Chon cac m~u thudc di€n hinh.. tinh chinh xac.1._ P_~U_l[_(!!l_g_ _v_~ l!!!'!l!!!g p_~_qp__. pseudoephedrin hydroclorid. cu th~ nhu sau: Thuoc co 2 thanh phan: + + Paracetamol va cafein. D\1 kien xay dung phuong phap dinh tinh va dinh luong mQt s6 thuoc co chua tir 2 d~n 5 thanh phdn. UV -Vis hien dai van viec phan tich cac thuoc da chon . Nghien ciru phirong phap xu Iy mdu. U ng dung cac phirong phap da xiiy dung vao viec kiem nghiem mQt s6 thuoc dang hru hanh tren thi tnrong. . Danh gia ich lei va tinh kha thi cua phirong phap .}_. Paracetamol va quinin sulfat. + Paracetamol va acid mefenamic..!g!!~~!! _ c:_0!_ .18 - . + Paracetamol va ibuprofen.. Nghien ciru xay dung cac phirong phap dinh tinh va dinh hrong d6ng thai nhieu hoat chAttren cac thi~t bi HPLC va quang ph6 UV -Vis hien co . chirng minh bing thuc nghiem tinh dung. khao sat tinh tuyen tinh. Thu6c co 5 thanh phdn: + Paracetamol..__. co tir hai thanh phan tra.

Phuong phap nghien a. pH . d~c biet lit co kha nang quet ph6 va do nhieu buoc song cung mt)t luc. <IQ xfip hat .Vis (l90nm (DAD: diode array detector). Phuong phdp xac dinh cac chi lieu nghien cuu . detector). cac thong s6 v€ Ioai pha tinh. Ph6 bien nhAt lit detector UV. detector). . detector) .. detector huynh quang (FLD: fluorescence co th€ g~p mQt s6 loai detector detector chiet suat vi sai. PhU'01lgphap siic kj long hi?u nang cao (HPLC) b. Loai ct)t dung phan tich (pha tinh): cQt cua hang nao. 3. do do d6i voi mot h6n hQP gbm nhieu thanh phfut ta co th€ chon cho m6i thanh phan mt)t bu6c song ph at hien thich hQP nhftt. sau: detector hdp thu tir ngoai . detector hftp thu tir ngoai (UV). detector detector do do wavelength wavelength a a . Chi lieu nghien cuu Cac chi tieu nghien ciru duoc mo ta 6 sa db sau." n~u hi rna giai ding + Thanh phan va chuang trinh dung moi n~u lei rna giai gradient..1. Pha dt)ng: + Thanh phan va dong (isocratic). detector ion hoa ngon hra.3. kich thtroc cQt ra sao. ..kha kien (UV.19 - .:. cO"hat. em.900nm) nhir: detector mang diode nhieu song bUGC song thay dbi (MWD: multi (VWD: variable buoc Trong do detector mang diode hi loai detector hi~n d~i co dt) nhay rAt cao..3. detector hong ngoai (IR) .Vis). tY l~ cac thanh phfut cua pha dt)ng._~·__!_~U'!_f{!!g__':~l!!?'!_'!!}gp_~_4P__1Jg!?!~!!E~ f 3. LU'a chon di2u kien sac kV Thit~t bi HPLC V6i may HPLC ta quan tarn nhat toi detector...

C + b hoac h = a. daub gia dQ tuyen tinh thong qua h~ sf>nrong quan r . sau do pha loang dung dich g6c nay d~ thu duoc m(>t day dung dich chuan bi~t truce n6ng dt). Khao sat do chinh xtic CUD phU01lg phap Be> chfnh xac tlnrong duoc xac dinh thong qua d(> l~p lai cua phirong phap (each thong thirong nhdt Hi xac dinh d(> lech chuan nrong dbi cua k~t qua phan tich RSD .C + b (a. Tien hanh khao sat d(> l~p lai bfing each dinh hrong rieng biet tren nhi~u mau gi6ng nhau ella chat can phan tich r6i xu Iy ket qua bang phirong phap thong ke._ p~~ L _~l!_f!!!g__~~ p!!l!'!!}g_p~_qp_ _. b la h~ s6). Ti~n hanh pha mt)t dung dich g6c. nong de>cua m6i chftt phan tich deu co m6i quan h~ tuyen tinh voi dien tich hay chieu cao pic sitc kY (trong mQt khoang n6ng dQ nhdt dinh nao do).!g!!~?!!E~ Luu pH .. 50Jll. neu cac pic tach khoi nhau hoan toan. l?p phirong trinh h6i qui.. thay d6i •:. Th~ tich tiem: thtrong la 20Jll. ta dich chuan va cac dung dich thir cho qua trinh sitc ky . Bmrc song phil! hien: phai thich hop voi m6i thanh phftn. S = a. Chu8n hi cae dung dich phlin tich Trong phtrong phap HPLC. Khao do turin tinh CUD cae thimh phOn can phlin tich Ta bi~t rang. . •:. thong thtrong ta tinh k~t qua bang each so sanh mau chuan va mftu thir trong cung dieu kien. Do do. Bern cac dung dich chuan nay chay s~c kY d~ thu duoc gia tri dien tich hay chien cao pic. Nhi~t dQ c(>tphan tich . •:.20- . Chung ta can phai xac dinh khoang tuyen tinh nay bang thirc nghiem tir do lam co sa d~ pha cac dung djch phan tich co n6ng dt) nam trong khoang tim diroc. y: Pha d(>ng la m(>t y~u t6 rAt linh d(>ng. Khao sat tinh thich h{!JJclio he thAng Yeu cftu a buoc nay co thich hop khong? la phai kiem tra xem cac di~u kien site ky ta da lua chon Ta thtr chay me>t mftu phan tich vai dieu kien sitc ky da chon. gon la duoc . din chuAn hi cac dung •:. pic nho. T6c de>dong pha de>ng: khao sat d~ chon t6c d(>phil hop nhat.theo cac duoc di~n hoac sai s6 nrong d6i). ta co th€ d€ dang thay d6i dS thu dtrcc k~t qua tach t6t nhAt bang each thay d6i tY l~ cac thanh phan. Slit . Nghia la trong khoang n6ng d(> do thi dien tich (S) hay chieu cao (h) pic phu thuoc vao nbng d(>(C) theo mt)t ham b?c nhdt. .

Them vao mdu thir met hrong ehinh xac ehdt chuan da bi~t tnroc ham hrong sao eho dung dich eu6i cling co nbng d{>nk sat.3.:. Khao sat Dhb hap thu UV tlch a sa db sau. chon cac huuc song. Do dO hap thu quang cua cac dung dich chuan han hem . danh gia. hoac gil'. Danh gia do dung va do chinh xac cua phuO'ng phap Phdn nay tuong 1\1 nhu d6i voi phirong phap HPLC. Phon Ilch mau thu . KhaO sat phA hAp t11\1 UV va IvachQn elk btr6c song.:.}.2. 0 ] J CcO ] ) . Phuong phap quang phD phon tick da cdu tIC(MCA) a.:.:. [hoac giai song phan tich. __}_~Ul!'_f!!1:g_ _~~ p!!'!_(!!!Z p_~!P__. tai uu duirng chunn hdng phOn m2m tTen may tlnh . Xac dinh khoang nang do phon tlch . trong khoang tuyen tinh da khao 3. song phon b._~ .!g~!?~_ ! t.:.:.:. Chuan hi day dung dich chunn hDn h(!fJ . Xoy dU'ng. Chi lieu nghien ctru Cac chi tieu nghien ciru duoc mo til . Phuong phdp xac dinh cac chi lieu nghien cUu va Iua .:. Khao sal do dung cua phuO'ng phap _ __ - 0 De> dung duoc xac dinh thong qua kha nang thu hbi cua k~t qua phan tich khi dung phirong phap them chuan.21 - . xac dinh phan tram tim lai.

:.4. str dung cong cu chuan va thir.!?. Str dung phuong phap h6i qui tuyen tinh.22- . thuat binh phirong t6i thieu d~ danh gia Cac 49c trung th8ng k2 re xu Ij kit qua phiin tich: x:.!gp~_qp:_ng~~~!!_. Phuong phap xir - ._ _f!~U'!_(!:!g_l!~l!!!~. Danh gia phirong phap phan tich va cac k~t qua phan tich: sir dung phuong phap thong ke.!_~ L 3.:."'x n 1 + Gia tri Trung binh: n .~ i i-I + DQ lech chuan (standard deviation): + Sai s6 chuan: + DQ lech chuan tuong d6i (relative standard devitation): n -1 RSD (%) = ~ x 100 x . _ Iy s6 Ii~u h6 tro lit chuong trinh Microsoft Excel.

5 urn).5 ml acid nitric (69 . Dieu kien sac leY .28%) va 21. chung toi dl1 nghien CUu phirong phap dinh hrong 5 hoat chdt tren trong vien h6n hop bang phuong phap HPLC voi di~u kien sitc ky long pha thuan. clorpheniramin phap HPLC maleat va cafein bing phuong Vien nang ha nhiet. _ va djnh hr4]llg paracetamol. . KET QUA NGHI"EN CUu 4.45 urn. .~0. E¢!. . hydroclorid. .5 (884 ml mroc cho them 94 rnl Amoniac d~m d~e (25 . Ce>tLichrosorb Si 60 (250 x 4 mm. A. dung miJi.VIS.5 bang acid nitric hoac Amoniac. giam dau. b. .'.c nghi~m A. Hod chdt dung moi Methanol tinh khiet site kyo Dung dich d~m Amoni nitrat pH 9..May sac ky long hieu nang cao HP 1100 . chua thdy san pham nao c6 cong thirc nhu tren.. - Dung moi A: h6n hop Methanol: Nuoc (1 : 1). f)i~u chinh pH d€n 9.:t. q'!_q. !!&~!?_'}. hoa chat a. 4.2.__ .1. . Th. .' . LQc qua mang loc 0. chua ho "Cotussin" do cong ty diroc pham Tien Giang nghien diu san xudt c6 cong thirc nhu sau: Paracetamol (PA) Cafein (CA) Pseudoephedrin hydroc1orid (PE) Bromhexin hydroc1orid (BH) Clorpheniramin maleat (CM) 280 mg 30 10 8 4 mg mg mg mg Trong cac Uti lieu tham khao.1 mg.23 - . May lite sieu am. Phu01Jg phap a.. .1. Djnh tinh .. khuay d~u. May moe. A. . d~ nguoi. detector UV va chuang trinh sAc ky d~ng dong.1:.Hewlett Packard.71%). Vi v~y de giup cac co s6 xay dung tieu chuan cho san pham cua minh. May moe dung C{l May site ky long hieu nang cao HP 1100 voi detector UV . bromhexin pseudoephedrin hydroclorid. Can phan tich voi de>chinh xac 0.

_ . Dung dich thir - sdc kY Dung dich thu dS dinh luong PA: Can chinh xac mQt hrong bQt vien tucmg irng voi 30mg PAvao binh 50ml. pH 9.. .. . l~c khoang 10 phut.VIS: 257 nm do PE va CM. 4. .. Pha dong: MethanolT6c dQ dong: 1. 1. them 30ml pha dQng.______ .. Cdc dung dich Hem ..24- . PE. Them Cloroform Cloroform. 10ml NaOH IN. them chinh xac 5mI dung dich CM. tir doan " dung 30mi mroc d€ chuyen h~t h6n hop nay vao binh gan ...45Jlm..:. _ T CQt Lichrosorb 4mm.5 (45 : 6) Detector UV .. Dung dich dem Amoninitrat - Th€ tich tiem: 20 ul.... PA. .5 ml/phut do PE va CM.... . them 30 ml pha dQng. 1. 5 urn). Dung dich chuan d€ dinh hrong CA.. __ .. hoa tan trong pha dQng vita du 25ml + Can chinh xac khoang 40mg BH. l~c ky rbi chiet bang 3 IAn x 30ml._... them pha dQng toi vach.. Dung dich thir dS dinh hrong CA. BH va CM: Tron d~u va nghien min hrong bQt vien lfiy tir 20 nang. IN rbi them methanol vira du 200mI.45 urn ....._. uie khoang 10 phut. _.. 298 nm do CA va BH. Dung 30 mi mroc d€ chuyen h6n hop nay vao binh gan. khuay cho tan h~t rbi xir Iy ti~p nhu mau thir. PE. sau do IQc qua mang loc 0. 15mI pha dQng. hoa tan trong pha dQng vira du 25ml (dung dich BH) + Can chinh xac khoang 560mg PA.. b. ~::::l . Si 60 (250 X . sau do IQc qua mang IQc 0... _0': _. l~c d~u. l~c d~u. l~c d~u. •:.. GQP dich chiet.0 ml/phut do CA va BH. them pha dQng d~n vach.. b6c hoi tren each thuy d~n h~t Hoa tan c~n trong 20mI dung dich HCI 0.. 20mg PE. . LQc qua gifiy IQc thuong. . ~ aUG .lglml. Dung dich chufut Dung dich chuan d€ dinh hrong PA: Pha chfit chuan PA trong dung moi pha dong d€ thu ducc dung dich PA co nong dQ chinh xac khoang 30J.0 ml/phut do PA. Hut 2ml dung dich tren cho vao binh dinh mire 20ml. IQc qua gifiy IQc thuong. 60mg CA. 10mI dung dich BH. CM va BH: + Can chinh xac khoang (dung dich CM) 40mg CM. " Tinh k~t qua: Dtra vao dien tich hoac chieu cao cua pic chuan va tlnr tirong (mg. cho vao mot c6c co mo. Kit - nehien cUu ... Can chinh xac mQt hrong bQt vien nrong irng voi 2 vien.

. waveJengffi=25Tnm\THANH\UU9.2. 40 ..9998 0..1'0 . s~c ky d6 da:tach cac chat CA va BH.7.1. .. ~ ~ U) 80· ~ <ri 1 .1 va hinh 1..._ _ 4...3 phut B. Bang 1.5 phut.-.9985 Khoang tuyen tinh dff khao sat (ug/ml) 80 .--_~.n V6i chirong trinh s~c ky da:neu.6x + 118.9985 . Ket qua diroc gioi thieu a bang I .1. Ket qua khao sat d9 tuyen tinh Thanhphk CA CM Phuong trinh h6i quy y y y y y H~ s6tuong quan 0.24x .300 25 .400 20 .125 20 .n do va loai be Paracetamol dll ducc khao sat kha nang chiet h~t bing each: sau khi chiet 3x40ml Ian 40ml Cloroform .9999 0. trong khoang n6ng dQ da: khao sat.. L B. . 5 .---~--~'---.. -" .__rnf[) {I .'--vwm'A.•. S~C ky d6 cua CA va BH Hinh1.--.. h6n hQ'PCA va BH. L6p Cloroform chi~t l~n 4 nay .1.99x + 20.1. Khii nang chiit hit hO{ltchat Trong phirong phap xu Cloroform thi chi~t them 1 ly m§u.• ~-~r-'--~~~~--.9987 0.2. Cac ket qua tren cho thay.82 B.30x . co su nrong quan tuyen tinh gitra gia tri dien tich pic va nong dQ (r = 0..~~.. S~c Icy d6 cua PE va CM PE : 4.9999 0.9999). Kh q_ua nghien cuu _ _ _ _ _ _ _ _ _ __ .2. . _ .24 = 0. .8 phut BH: 4....120 60 ..120 = 2. .9.. PE va CM voi cac nong dQ khac nhau.88 = 17..57x . CA: 6..0..27. Khiio sat khoiing tuyJn tinh Chung t6i da:khao sat su phu thuoc tuyen tinh gitra n6ng dQ cac hoat chat va dien tich pic tren 5 dung dich chuan cua PA. 201 10·t i "' .6 phut. K~t qua va bifn lu.._ . . PE va CM nhu hinh 1. C --. . 40··: . N6ng d9 dung dich chuan va dung dich thir cua chung toi nam trong khoang tuyen tinh dll khao sat..25 - nfra. CM: 5._-- ) 5 10 rniFl Hinh 1. J .11 = 15. giai doan chiet cac hoat chAt d._ .. mAU 120 . __ _ • .88 = PE BH PA 11...

.38 9.31 99. B.2.lN them Methanol vim du 20ml.2.1 Thanh phan CM PE 4.~~·. hoat chdt dll dircc chi~t h~t 6 3 IAn dAu.21 0.63 0. hrong chuan them vito 20% m6i chat) CA 29.~ ' .4 K~t qua khao sat cho th§y dQ l~ch chuan nrong d6i tuy thea nrng tnrong hop tir 0. Nhir v~y.05 0.0 BH 7. SKS: 071199.0 GiA tri trung binh X ( mg I vien ) DC) l~ch chuan S DQ l~ch chuan nrong d6i Ty l~ tim l{lidiroc ( %) ( n = 3. K~t qua khao sat dQ l~p lai va dQ dung tren mftu COTUSSIN sA li~u thAng ke PA 278. ducc b6c hoi tren each thuy d~n kho.17 1. sai sa va dp dung Cac thir nghiem nay ducc thirc hien tren mfiu vien Cotussin cua cong ty Diroc phAm Ti~n Giang. hoa tan cin trong 5ml dung dich HCI O. ~~ ~g_~!~'}~ ~~~~ ~ " .26 0. K~t qua diroc gioi thieu trong bang 1.q'!_q~ ~~ .4 0.07 98.~~4!. rbi do HPLC theo chU011g trinh tren.2.85 98.38 1. Kha nang chi~t duoc dam bao.5 99.08 0.23 99.~ ~ "~" ~..3. Dp I~p l(1i.4 • 99.23 .07% va tY l~ tim lai diroc hoat chdt tir 98.1%.15 2.83 1. K~t qua thir tren 3 binh d~u khong xUdt hien pic tren s~c ky db. Bang 1.

_. 250mg. Chftt chuan paracetamol va acid mefenamic do Vi~n Ki€m nghiem .ng dung moi pha dong d€ diroc dung dich chuan co n6ng dQ cua PA khoang 80 ug/ml va cua AM khoang 40 ug/rnl. nUG ::t _. T6c d9 dong 1 ml/phut..2. . tinh kh6i IUQ11g trung binh vien.. _ ... Trong cac diroc dien hien hanh [19.. tetrahydrofuran loai dung cho HPLC. .. Th€ tich tiem 20111. 4.. Can chinh xac met luong bOt vien nrong (rug voi kh6i IUQ'l1g .. sa xay dung tieu A. 27. fJiJu kien sac IcY - CQt Lichrosorb RPI8 (250x4 mm.. . _ .... . Chu{m bi dung dich - Dung dich thir: Can 20 vien. Dung dich chuan: Can chinh xac khoang 0. 10Ilm).. Pha dong: Acetonitril . - - .2 bang acid phosphoric. _0_ ... Acetonitril. 4. N- ......." _.2. _ .] chua co chuyen luan v~ hai thanh phan nay.I.. ... Vi v~y chung toi da:nghien CUu phuong phap dinh tinh va dinh IUQ11g 6ng d thai eel2 thanh phan tren trong h6n hop thuoc voi muc dich cling co chuan cho san pham maio A. C9t Lichrosorb RP18 (250x4 mm. ..._ .tetrahydrofuran (3 : 8 :9) dieu chinh d€n pH 2. nehien cuu ... Binh hrong dAng thm Paracetamol (PA) va Acid mefenamic (AM) trong vien nen bao phim Pamesic bing phmmg phap HPLC Vien bao Pamesic do cong ty v~t tir Yt~ Ti~n Giang nghien ciru san xuit co thanh phan nhir sau: Paracetamol (PA) Acid mefenamic (AM) 500mg. 23.27- .. Thi~thi. Acid phosphoric tinh khi€t phan tich.1 g chdt chuan PA va khoang 0...mroc . Kit ~.. hod chtit May sAcky long hieu nang cao HP 1100 voi detector UV -Vis..... . hoa tan bang 5 ml THF.. 1011m)... Hai chit tren co de) tan nhir nhau (d~u d6 tan trong kiem) nen rftt kho chiet tach.. sau do pha loang ba. . . Th\l'c nghifm A.. - _.. . Detector UV d~t a buoc song 279 nm. Phuong ph tip nghien cau a. 29. nghien thanh be)t min va tron deu. Be) IQcdung moi va IQCmftu voi duong kinh 16mang IQc 0.. _ .. h..05 g chit chuan AM.45 11m.... .BQ Y t€ cung cap..

.26 R =0.. LQc qua mang IQc 0..07 .l....35 x + 20... Or __ .. hoa tan bang 5 ml THF..... K~t qua B. pha loang bang dung moi pha d¢ng d~ duoc dung dich thir co nong de>cua PA khoang 80 ug/ml va cua AM khoang 40 ug/ml lac d€u....1 cho thay mAU N 160 140 120 100 80 60 40 20 o~----~~--------~--~--~ Hinh 2. cua khoang 50 mg AM. k~t qua khao sat de) a bang sau: Bang 2....1 938..9 3 4 5 Khoang nong d<) tuyen tinh d1lkhao sat Phuong trinh hoi quy H~ so nrong quan = 19. ---..451J.ffi.28 - = 46.... - -- - . - _ . Kit :::l nehien cuu aua 0 ~ . -_ ..__________ 4..6 1568. _ __ .. _ ..4 AM Nang d<) (ug/ml) 10 20 40 80 100 16 -7 80 Y Dien tfch pic 456.. - _ . - - -_ ...6 1883.4..9996 ..8 3098.1 3650.9999 R=0.n dinh hrong voi dien tich pic tuong irng tren cac chftt chuan PA va AM. Ket qua khao sat de>tuyen tinh STT 1 2 PA Nang d<) (ug/ml) 20 40 80 160 200 20 -7 200 Y Dien tich pic 401. - -- - - - _ . s~c ky db thu diroc PA va AM diroc phan tach hoan toano a hinh 2..1.3 4711. Chuan bi me)t day dung dich h6n hop chuan trong pha de)ng tuyen tinh diroc trinh bay co nong de) cua PA bi~n Thien trong khoang 20 - 200 ug/rnl va cua AM bien thien trong khoang 10 .1.... Khiio sat khoang tuy2n tfnh Chung t6i ti~n hanh khao sat de>tuyen tinh gitra neng dQ cac chat d. B. S~c kY db thu duoc cua dung dich hem hop PA va AM B.65 x ....6 803. Khao sat tinh thich h{1JJ cua h? thang sAc kj V6i di€u kien s~c ky du chon nhtr tren.100 ug/ml.1..5 3901.. - - - - _ ....

49% - S =4.n xet: Trong khoang nbng de>da khao sat co SI! nrong quan tuyen tinh gifra dien tich pic va n6ng dQ voi h~ s6 nrong quan r~t gful 1 (r = 0.2a.46 506. Db thi bieu di~n S\l phu thuoc tuyen Hinh 2.50 250.78 247.75 248.15 499. - - --- --.~ .____ 4. Khao sat 49 /(ip /{li cua phuang phdp DQ l~p lai cua phuong phap diroc ti~n hanh tren hai miu thir vien nen bao phim Pamesie.2. Paracetamol Acid mephenamic 4500 4000 3500 3000 2500 2000 1500 1000 500 0 0 50 100 150 200 250 2000 1000 0 0 50 100 150 I 5000 4000 3000 Hinh 2. B.21 RSD%=0.3.55 S = 1.3.22 RSD%=O..97 So lieu thong ke X . K~t qua khao sat dQ l~p lai tren miu vien nen bao phim Pamesie MJu 1 SKS: 011102 SIT 1 2 3 4 5 6 PA AM So lieu thong ke Ham hrong (mg/vien) 511.79 508.. Db thi bi€u di~n su phu thuQC tuyen tinh nAng dQ va dien tich pie wang irng cua PA tinh nAng dQ va dien tich pie nrong irng cua AM Nh.. __ _ ~ T ~ _ r L L-+~ -+_ - - --. - - • - - • - - ---- - -+ ~+- - - -- - --- .9999).95 504.9996 .29- .54 247.32 505.83 Ham hrong (mg/vien) 247.08 X = 248..= 506.75 248.0. Kit qua ng_hien cuu . Chung t6i chon n6ng dQ dung dich thir va chuan d~ dinh IUQ'Ilg nim trong khoang tuyen tinh da khao sat (PA khoang 80 ug/ml va AM khoang 40 ug/ml). K~t qua khao sat de>l~p hili diroc trinh bay (] bang sau: Bang 2.83% . ----..

n xtt: Ket qua tren cho thAy phirong phap co dQ l~p lai cao v6i dQ l~ch chuan tuong d6i khoang tir 0. .25 488.32 20.8 98.02 252.021202 PA SIT 1 2 3 4 5 6 AM (mg/vien) So lieu thong ke Ham luong 498.21 495.58% Nb.77 9. Khdo sat 49 dung Khao sat dQ dung bang phuong phap them.7 98.31 250.07 503.68 253.08 (mg/vien) So li~u thong ke X = 495.99 9..9% va cua AM la 98.32 20. ----------------l __ '!.-_.7 97.3.9 98.09 19.~~_i_~'!.3 99.. Bang 2.:t__i~~~~q _<:~ --------------------------_.25 % tim lai 98.-----------------------.30 501.88 S = 6.97 20.2%. them vao miu thir da: hiet wac thir. Ket qua khao sat dQ dung PA AM SIT Luong them vito (mg) Luong tim thay (mg) 20.49 .30- .71 Ham hrong 255.36%_ B.2% 1 2 3 20.99 TB Nhin xet: PhU01lg phap co de>dung cao vei kha nang tim lai cua PA la 98.88 %timl~ 97.------------------------.55 253.36% S = 1.13 487. Ket qua khao sat de>dung ducc trinh bay iJ bang sau : ham hrong m9t luong chinh xac chAt chudn tuong img v6i khoang 20% hrong co trong miu Bang 2.46 RSD%=0.9 98.75 RSD%= 1.63 252.9% Luqngthem vito (mg) Luong tim thay (mg) 9.1.32 9.76 9..2h.99 9. Ket qua khao sat dQl~p lai tren miu vien nen bao phim Pamesic Mau 2 SKS..61 X = 252.4.

Chudn hi dung dich Dung dich chuan: Chuan bi mQt dung dich h6n hop chuan trong dung m6i A co nong dQ cua PA khoang 600 ug/rnl va cua CA khoang 20 ug/ml. lOf-Lm). Pha dong: Dung dich acid phosphoric 0. t6n nhieu thai gian. acid phosphoric (PA). Chung t6i dff nghien ciru mQt chuang trinh HPLC dan gian va tach t6t han chuang trinh dff de c~p trong cac tai lieu tham khao [29] de dinh tinh va dinh hrong d6ng thai 2 thanh phdn tren.11. Phuong phap nay co nhiroc di~m la phai qua biroc chiet tach kha phirc tap.45f. A. IQc qua giAy IQcthirongfbo khoang 20 ml dich IQc d§u). Be>IQc dung moi va IQcmftu voi dtrong kinh Methanol loai dung cho HPLC. dung bi May s~c kY long hieu nang cao HP 1100 voi detector UV. ChAt chuan paracetamol va cafein do Vi~n Kiem nghiem cung cftp. vi du vien Pasepan cua CTDF Ha Tay hay vien Sedapa ella XNDFTW 2.3. hoa tan va them dung moi A d~n vira du 100 ml. De dinh tinh va dinh hrong cac hoat chat trong h6n hop thuoc tren thirong ap dung theo tieu chan cc sa (chiet tach r6i do quang) [2. Kit qua nehien . Dung moi A: Ethanol. Hoti chiit.. Dung dich thir: Can 20 vien. A.cUu 0 . CQt Lichrosorb RPI8 (250x4 mm.mroc (35:65).16].I.mroc (1:1). _____ 4. nghien thanh bQt min va trQn deu.. 16 mang IQc 0. 4. thiit - de m~c sai s6. __ __ __ .1m.. .\t6 buoc song 273 nm. Detector UV di.75% trong h6n hop dung moi methanol . tinh kh6i hrong trung binh vien. Ethanol tuyet d6i (PA)._ _ _ _ _ ./ . T6c dQ dong 1 ml/phut. Thuc nghi~m A. Binb hrqng dAng tbm paracetamol phAn bing phmrng phap HPLC (PA) va cafein (CA) trong thu6c da thanh Thu6c da thrum phdn bao g6m paracetamol va cafein la mot dang ph6i hop thong dung. The tich tiem 20f. Sau do IQc qua mang IQc 0. __ __ __ _ .2. 10f-Lm). Can chinh xac mot hrong bQt vien urong img voi kh6i hrong cua khoang 60 mg PA. h.45 urn.31 - -- ? . Phuong phtip nghien cuu a.. DiJu kien sac Icy CQt Lichrosorb RP18 (250x4 mm. moi va hoa chat.

..... .. K~t qua khao sat d9 tuyen tinh STT 1 PA Nang dQ (pg/ml) Nang dQ (~ml) Dien tich pic 2 3 4 5 Khoang 150 300 450 600 750 2370 4701 7094 9412 11423 10 20 30 40 50 10 ~ 50 Y = 46.-...-..__... - - - _. chung toi thu dircc sAcky a6 cho thay PA va CA diroc phan tach hoan toan nhir hinh 3...9996 470 978 1432 1901 2330 nong de> 150 ~ 750 tuyen tinh dff khao sat Phuong trinh hoi quy H~ so tuong quan Y = 15.9 r = 0.._.... - .. _0 . nehien .qua .3 r= 0..._.......750 ug/ml va cua CA bi~n thien trong khoang 10 .. Sllc ky d6 thu dircc cua dung dich h6n hop PA va CA B.J .-. B.. Chuan bi mot day dung dich h6n hop chuan trong dung moi A co n6ng d9 cua PA bien thien trong khoang 150 ...2x + 154....50 ug/ml.... - - - _ .... Kit .-....... _ - -<'.9995 -32- .... K@tqua V6'i di~u kien s~c ky ad chon nhir tren.. 4. Kit qua khdo sat d{J tuyin tinh Chung toi tien hanh khao sat dt) tuyen tinh gitra nong dt) cac ch~t cdn dinh hrong voi dien tich pic nrong ung tren cac ch~t chuan PA va CA.......1...l..-_ ... ...... mob 3.... k~t qua khao sat dt) tuyen tinh duoc trinh bay a bang sau: CA Dien tich pic Bang 3...1.... .4x + 29.._. cUu ....

82 9.63 9.1 288. DQ l~p lai diroc khao sat tren hai miu vien nen SEDAP A do XNDPTW II san xudt c6 thanh phfut: Paracetamol: Cafein: 300 mg.51% (mg/vien) - 9.81 301.86 hrong (mg/vien) So li~u thong ke 1 2 3 4 5 X = 302.83 9.66 9.36 303.93% Bing 3.64 = 0.53% (mg/vien) S = 0.. MJu 1 SKS: 350900 SIT PA CA (mg/vien) So li~u thong ke Ham hrong 291.52 RSD%=0.44 .75 . vira du 1 vien Ta duoc: Mltu 1 SKS : 350900 Mltu 2 SKS : 361000 Bing 3.97 RSD% 6 301.! phu thuQc tuyen Hinh 3.54 RSD%= 0.6 287. K~t qua khao sat de>l~p lai tren miu SEDAPA.9 287.09 RSD% - 9. Db thj bi~u di~n S.+ Paracetamol T __ - -- - T _- - - -~ _ --- - _ .40 302.3. 10 mg.52 9.09 = 0.71 9. Db thj bi~u di~n su phu thuoc tuyen tinh nbng dQ va dien tich pie tirong ling cua PA tinh nbng dQ va di~n tlch pie tuong ling cua CA B. Kit qua khao sat i/{j 19P19.37 Ham 9.4.-- - -- .3 288. ____ •Kit nUG ::t nehien cuu O~ .92% 9.33 - 9.3.71 luong (mg/vien) So li~u thong ke 1 2 3 4 5 6 X = 288.4 X (mg/vien) S = 1.30 305.57 9.2.-- _.83 = 0.76 9.7 286. --- - ---- Cafein 2500 2000 1500 1000 15000 10000 5000 0 500 0 0 200 400 600 800 0 20 40 60 Hinh 3. Mdu 2 SKS: 361000 SIT PA CA So li~u thong ke Ham hrong (mg/vien) 301.62 X S (mg/vien) S = 1.2. - - --- - ---- - -- - - --- - - --- - - .-- - - - -- - - --- - --- - .2.9 Ham 9. K~t qua khao sat dQ l~p lai tren m§u SEDAPA.

9% STT 1 2 3 4 Luong them VaG 20 20 30 30 40 40 5 6 TB Bang 3.898 % tim lai 100.90 0.896 0. K~t qua khao sat dQ dung tren mdu SEDAP A..7 39..8 100.7 97.90 them Luong tim thay (mg) 0.1 20.884 % tim lai 101.4 101.30 0.2 % Lugng vao (mg) 0.293 0.2 97.60 0.1 98.60 0.0 97.9 d~n 99.298 0. Mau J SKS : 350900 PA CA Luong tim thdy (mg) 20.2 98.. K~t qua khao sat dQ l~p lai a tren eho thdy sai s6 tirong d6i til 0.07%._.305 0.582 0.... _..1 29..30 0.5 98..2 % TB K€t qua tren cho thdY phftn tram tim lai cua PA va CA tren hai mfiu dat tir 98.. phirong phap co dQ dung t6t.4 va 3._...600 0.4 98..3 97. ~__.__________ 4..60 0.595 0.1 99.: .9 100..300 0.8 99. ..5 % tim lai 100.. Kit qua nehien..592 0. - --- . CUll _ 0.u SEDAPA k€t qua diroc trinh bay a bang 3.0 98. . - - _.30 0.1 39..8 99. _ -< .1 (mg) %timl~ 101.. B.6 30.90 Luong tim thdy (mg) 0.3 98. Bang 3.3._.3 98.30 0. MJu 2 SKS : 361000 PA CA Luong tim VaG SIT I 2 3 4 5 6 Lugng them (mg) 20 20 30 30 40 40 thdy (mg) 20..7 99.886 0.3 29.4. ...4 40..34- . voi sai s6 nay eho phep ap dung duoc phirong phap tren....5 99..._..1 100..90 0. Ket qua khao sat dQ dung tren ma.0 99..5.5 99.0 99." .__•• . a tren bing phuong phap them chuan..54 den 1...7 99.6 29.u SEDAPA.2 19.7 39..5 %. ..5. Kit qua khao sat dp dung Khao sat dQ dung tren hai ma.5 % Luong them vao (mg) 0.60 0.

.35- .acid phosphoric 0.. - Ch~t chuan paracetamol va ibuprofen do vi~n kiem nghiem cung c~p. Detector UV d~t a biroc song 224 nm.1. Diroc nr Y t~ Tra Vinh. I~c d~u. Th\lc nghi~m A. D6ng thai mong duoc gop ph~ xay dung cac tieu chuan chdt IUQl1g ella nganh cling nhu cua QU6c gia cho cac A. 4. IQc (bo khoang 20 ml dich 19C dfiu). hoa tan va them pha dQng d~n vira du 50 ml.A. hoa chat 655A-12 voi detector UV- May sAc kY long hieu nang cao MERCK-HITACHI Vis 655A-22. Kit ..5 mg PA. Th~ tich tiem 20Jll..0 mI. Muc tieu cua chung toi la nghien ciru mQt s6 phirong phap dinh hrong danh gia cac plnrong phap eho cac dang thuoc co thanh phan nrong nr.1 % (60:40). Dung dich chuan : Chuan bi mQt dung dich h6n hop ehuAn trong pha dQng co nong dQ ella PA khoang 65 ug/ml va cua IB khoang 40 ug/ml.). vien nen diSn Vi~t Nam cling paracetamol va nhu cac ducc diSn khac hien nay dSu chua co chuyen lu~n cho dang bao ch€ nay.0 ml dich IQc tren cho vao binh dinh mire 20.. 10Jlm).IN va O. _ __ __ __ __ __ __ -__ -- -- -- - . Pha dong: Acetonitril. dung dich NaOH O. them pha dong d~n vita du. 4. tinh kh6i IUQl1g trung binh vien. natri sulfat khan.45Jlm.___ . A.OlN.. nghien thanh bQt min va trQn d~u.45 urn. C<Y ibuprofen co trong thanh phk mot dang thuoc voi muc dich lam phong phu them va sa hra chon ap dung. LQc qua mang loc 0. I urn). Thi2t - hi. CQt Lichrosorb RP 18 (250x4 mm. ° BQ IQcdung moi va IQcm~u voi dirong kinh 16 mang IQC0. Chudn bi dung dich cua khoang 32.. Can chinh xac mot hrong bQt vien nrong irng voi kh6i hrong b. ethanol vc''. Phllung phap nghiin cuu a. Nghien Ctlu dinh hrQ'llg dAng thM paracetamol thu6c da thanh phin bAng phmrng phap HPLC (PA) va ibuprofen (IB) trong MQt trong nhtrng dang thong dung tren thi tnrong hien nay la h6n hop paracetamol va ibuprofen (vi du: vien nen Alaxan cua xi nghiep United Pharma Vi~t nam [II].. __ o~ . Hut chfnh xac 2..2. vi en nen Pi-Antalvic [5] cua cong ty Diroc v~t MK protamol cua cong ty cb phan Duoc pham Me Kong . T6c dQ dong 1 ml/phut. Dung dich thir: Can 20 vien. DiJu kien - sdc kY CQt Lichrosorb RP 18 (250x4 mm._ . Diethyl ether.__ aua nehien elm __ _.4.

"~>"..l.:.. K~t qua khao sat dQ tuyen tinh STT PA Nbng dQ (ug/ml) Dien tich pic IB N6ng dQ (ug/ml) Dien tich pic 1 2 3 4 5 Khoang nong dQ tuyen tinh dll khao sat Phuong trinh hoi quy H~so tuong quan -.. k~t qua khao sat dQ tuyen tinh duoc trinh bay a bang sau: Bang 4. S~c kY db thu diroc cua dung dich h6n hop PA va IB B.9999 --._.--_._(l!i!_ q~4_lJ.g~j~'!_ _c:~ B. 20 40 60 80 100 Hinh 4.4x + 343.3.80 ug/ml.1. -------.0 R = 0.~--- 1970 3628 5475 7168 8922 16 32 48 64 80 16 ~ 80 Y = 37.2.".130 ug/ml va cua IB bien thien trong khoang 16 .C)":::i[:'"·'.lx + 199. cluing t6i thu diroc s~c ky db cho thely PA va IB duoc phan tach hoan toano r . D6 thi bieu di€n sir phu thuoc tuyen Db thi bieu dien SlJ phu thuoc tuyen tinh n6ng dQ va dien tich pic nrong img ella PA tinh n6ng dQ va di~n tich pic nrong irng cua IS -36- ._-- 26 52 78 104 130 26 ~ 130 Y = 67.4 R = 0.9997 958 1521 2148 2711 3356 Paracetamol 10000 l+J.L·>-"'~~~'_."FiC':"%· Ibuprofen 4000 3000 2000 1000 8000 6000 4000 2000 o o 50 100 150 o o Hinh 4. K~t qua ------------------------------------ V oi di~u kien s~c ky da chon nhu tren. Hinh 4.1. Khao sat khoang tuyin tinh Chung t6i ti~n hanh khao sat dQ tuyen tinh gitra nAng dQ cac chelt cAn dinh hrong voi dien tich pic nrong irng tren cac chelt chuan PA va IB_ Chuan bi mot day dung dich hem hQ'P chuan trong pha d¢ng co nong dQ cua PA bi~n thien trong khoang 26 .

50 6.87 4.2 % SIT Luc.50 6.95 3. B.6 % 4.2 98.50 6.43 6.9 96. SKS : 003501.6%.0.3 S = 2. K€t qua khao sat de)l~p lai STT PA Ham hrong (rng/vien) 328.8 100.65% B. Chung toi chon nang de) dung dich thu va chuan d~ dinh hrong n~m trong khoang tuyen tinh da khao sat (PA khoang 65 ug/ml va IB khoang 40 ug/ml).00 4.0 331. Khdo sat ap lpp l{li ella phuang phap D<}l~p lai cua phirong phap duoc ti~n hanh tren miu thir la vien nen Alaxan.9 336. tim (mg) 3.9 99.0. K~t qua khao sat dQdung PA IB Luc.37- .2.mg tim (mg) 6.00 4. KSt qua khao sat dQchinh xac duqc trinh bay iJ bang sau : Bang 4. - Nb.76 RSD%=0.--------.1 206.7 331. .9997 .3.00 blnh Nbtn xet: Phuong phap co de>dung cao voi kha nang tim lai cua PA 18.9999).5 96.2% va 9 ella IB Iii.2.mg them Lugng Luc.34 RSD%=0.8 97.83% X =204. Bang 4.6 98.90 3. ay % tim lai 98.0 203.n xet: Trong khoang nAng de)da khao sat co S1.2.4 333.7 205.9 S = 1.65 .3.28 6.83.3 206._~·__l!:4!_qlf_4_ ~g~!~.1' nrong quan tuy~n tinh gitra di~n tich pic va nAng de) voi h~ s6 nrong quan rAtgAn 1 (r = 0.3 So lieu thong ke 1 2 3 4 5 X =332.!~~.50 6. K~t qua tren cho thAy phuong phap da xay dung co dQ l~p lai cao voi dQ l~ch chuan tuong d6i khoang 0.98. them va mdu thti' da bi~t tnroc ham hrong mQt hrong chinh xac chAt chuan nrong trng voi khoang 20% hreng co trong miu thtr.50 thAy % tim lai them vao (rng) .00 4.6 99.98 3.43 6.00 4. K~t qua khao sat dQ I~p lai duoc trinh bay iJ bang 4. 8.mg vao (mg) 1 2 3 4 5 Trung 6.---- ----------------------------------------------------------------'------------.5 98.4 98. Khao sat t1P aung Khao sat dQ dung bang phuong phap them.32 6.3 IB So lieu thong ke Ham hrong (rng/vien) 203.46 97.02 thJ.

. ChAt chuan paracetamol va quinin sulfat do Vien Kiem nghiem cung cAp. Acid acetic 4%.!1.- -------_.. Dung moi: Ethanol . -. _. Vi v~y chung t6i ti€n hanh nghien ciru dinh tinh va dinh hrong dAng thai paracetamol va quinin sulfat trong vien nen Antigrip F bang phuong phap HPLC voi di~u kien sAc ky pha dao tao cap ion.. Nhiet dQ phong. Cot Lichrosorb RP 18 (250x4 mm. Cac phirong phap nay thtrong phai chiet tach phirc tap. Pha dQng: Ll g natriheptansulfonat 4%-diethylamin ti trong 1 lit h6n hop methanol-acid acetic Ie (650: 350: 1).2...!c nghifm A... diethylamin.. 29J co phirong phap d6i voi ch~ pham don thanh phfut.. Vien nen Antigrip F co cong thirc nhu sau: Quinin suifat Paracetamol Ta diroc vd 30 mg 150 mg 1 vien chi De dinh hrong cac thanh phan nay. . nhirc dau. A..----_. h§.. .. hod chat May sAc ky long hieu nang cao HP 1100.. 4..---------_--... --- --. dau minh may.5.. 19.. A.VTYT Ti~n Giang (Tiphaco). Sum)....lm). 15. . T6c dQ dong: Irnl/phut... Detector UV: 235nm._o~ . DiJu kien - sdc kY CQt Lichrosorb RP 18 (250 x 4 mm. ThS tich tiem: 20f..----------_...I... ____K€t qua .. natriheptansulfonat..38 - .45 urn.mrcc (1: 1) 16 mang lQC 0. Th.--. 27.. PhU'lIng ph tip nghien c«u a.VTYT Tra Vinh ph6i hop paracetamol va quinin sulfat dS di~u tri cac clnrng dun s6t. Thi€t - hi.___ ._ _ nehien cUu _. . Be>loc dung moi va loc mdu voi dtrong kinh Methanol loai tinh khiet s~c ky.4... phirong phap chuan de>moi tnrong khan. . vien Anogin quinin cua CTD... phuong phap acidbase. con voi cac ch~ phAm da thanh phan thi tieu chuan cua cac nha san xuAt dira ra rdt khac nhau: Phuong phap quang ph6 hApthu tir ngoai-kha kien... t6n nhieu thai gian [3].. Djnh hrqng dang thei paracetamol (pA) va quinin sulfat (QS) trong vien bao phim Antigrip F bAng phuong phap HPLC Tren thi tnrong hien co mQt s6 che pham nhu vien AntigripF va Tiphagrip cua CTD...u h~t cac diroc dien [1. 10J..

22 2214.9999 .. _ .90 1131. ..4 x + 22. ...9999 Dien tich pic 107... Dung dich thir: Can 20 vien.......... Or .u 350 300 250 200 150 100 '0 j n B .. .mroc tY Ie 1:1).. .-.39 - . .._....1. hop hai thanh phan B... . SAc kY db cua dung " dich h6n .50 Nong dQ (ug/ml) 1.. .2 ....5 r= 0.cUu ...mroc (1: 1) co n6ng d<) cua paraeetamol 30Ilg/ml va cua quinin sulfat khoang 6 ug/ml.30 2794. Kit qua khao sat dp tuyin tinh Chung toi ti€n hanh khao sat de) tuyen tinh gitra n6ng d<) cac chAt cAn dinh hrong voi dien tich pic tuong irng tren cac chAt chuAn PA va QS......_.a.... -_ T b.. Chudn bi dung dich Dung dich chuan: Chuan bi mot dung dich chuan cua h6n hop hai thanh phan tren trong dung moi ethanol . hoa tan trong 50 ml h6n hQ'P dung moi (ethanol ..4 6 12 30 1.. so 0 Hinh 5..3 r= 0.l. . B..§ !il .50 13882.. . .16 x + 19.26 897.... .... IQc qua mang loc 0...-_ .... _ .. Kit A . nehien ~__... aua Q~.2 .. k~t qua khao sat d<) tuyen tinh duoc trinh bay 6 bang sau: Bang 5._.::.1.. .09 194..30 ug/ml y = 73. cac pic can xirng .150 ug/ml SIT 1 2 3 4 QS Dien tich pic 576.. xac dinh -kh6i IUQ11gtrung binh cua vien. Chuan bi met day dung dich h6n hop chuan trong pha d<)ng co neng dQ cua PA bien thien trong khoang 6 150 ug/ml va cua QS bi~n thien trong khoang 1.. Pha loang 20 l§n dich IQc tren bang dung moi pha de)ng... nghien thanh b<)t min rbi tre)n d~u_ Can chinh xac mQt IUQ'Ilg bQt vien tirong irng voi khoang khoi hrong cua khoang 30 mg paracetamol.. . lQC. tren sAc ky db thu diroc hai pic tach khoi nhau hoan toan... K~t qua V oi chuang trinh sAc ky nhu tren.______ 4._...2 2.10 5 Khoang nong dQ tuyen tinh da khao sat Phuong trinh hoi quy H~ tirong quan so y = 92. .30 ug/ml......88 458. K~t qua khao sat de) tuyen tinh PA Nong dQ (ug/ml) 6 12 30 60 150 6 .45 11mtruce khi tiem sAc ky..0 . . _ .50 5566.

~~ T.48 1. Kit qua khao salll(j dung Khao sat dQ dung bAng phuong phap them. ..n Hlnh 5. Kit qua khao Stit dp /(ip /(li DQ l~p lai cua phuong phap diroc tien hanh tren mdu thtr la vien nen bao phim Antigrip F.52 % tim 1~ 98.5 Luong tim tM_y(mg) 1.5 1.66 S = 0.n PA Ham luqng (mglvien) 153.60 150.2. Nb.n phap 1. Paracetamol 15000 10000 5000 0 Quinin sulfat 2500 2000 1500 1000 500 0 0 50 100 150 200 0 10 20 30 40 Hinh 5. Db thi bieu di~n su phu thUQC tuyen tinh nbng dQ va dien tich pic nrong img cua PA Nh.0 QS Luqng vao (mg) them IB xet: Phuong ella QS Icl99.2. Nong dQ dung dich chuan va dung dich thu cua chung toi nk trong khoang tuyen tinh dii khao sat. Db thi bieu di~n S\l phu thuQC tuy~n tinh nbng dQ va dien tich pic nrong img ella QS xet: Cac k~t qua tren cho thdy.0 (mg) 7.79 149.0 98.15 xet: Phuong phap So lieu thon_g_e k X = 150. c6 su nrong quan tuyen tinh gifra gia tri dien tich pic va ndng dQ (r = 0.6 101. k~t qua diroc trinh bay 6 bang sau: Bang 5.43 7.89 152.81.88 29. ~ - - _ .36 28. k~t qua khao sat dQ l~p lai duoc trinh bay a bang sau: Bang 5.5 7.1. - + _ . trong khoang ndng dQ khao sat. Kit aUG ::t nehien cUu O~ .2.4 99.0% va ...17 28.5 1. - - ~ .3 99.10% - QS Him lugng (Il!glvien) 28..01 29.. 4.3. K~t qua khao sat dQ l~p lai SIT 1 2 3 4 5 6 Nb. B.45 So li~u thong - ke X -28.3%.47 1.06 148.52 RSD% = 1.3.5 Luong tim tMy (mg) 7.66 RSD% = 1.5 7..40 - .48 % tim 1~ 99.10 .0 98.3. K~t qua khao sat dQ dung SIT 1 2 3 PA Luqng them V3.3 co de>dung cao voi kha nang tim lai cua PA la 99.12 149.. .7 99.60 S = 1..38 7.81% co dQ l~p lai eao voi dQ lech chuan nrong d6i 1.~.10 28. B.9999)..

l. Thtre ngbi~m A. (C6ng ty DuQ'C va v~t tu y te Ti~n Giang). Be) IQc dung m6i va IQCmdu voi dirong kinh 16 rnang IQc 0. Asmin (Xi nghiep Duoc pham trung irong 25). IOum). 10llm). Tiphasmyl phan theophylin.45 urn.I.6. Methanol loai dung cho HPLC. chung t6i da ti€n hanh nghien ciru phuong phap Ephedrin hydroclorid. hoa chat May sic kY long hieu nang cao MERCK-HITACHI 655A-12 voi detector UV- Vis 655A-22. Phenobarbital trong thuoc dinh hrong d6ng thai Theophylin.base.acid .Phap).5: hoa tan 12 g NaH2P04 trong 1000 ml rnroc Natri heptansulfonat Dung dich d~m phosphat cfU. Sl. A. ou« kien sac kf Ce)t Lichrosorb RP18 (250x4 mm. Binh hrQ'llg dang thot theophylin. dinh IUQ1lg trong m6i tnrong chuan dO.5 bang acid phosphoric. Detector UV dij. pH 3. .C chenh lech v~ hrong gitra cac thanh ph§n rat Ian trong do hrong Phenobarbital qua kho chinh xac phu thuoc nhieu vao nguoi thtrc hien. plurong phap (C6ng ty DuQ'C va v~t ill y t~ Ben Tre).41 - . di~u chinh v~ pH 3. Tedralas (Labomed a Viet Nam va met s6 tai nghiem cac dang bao ch~ don thanh phdn cua cac hoat chat tren va phirong phap dinh hrong co th~ la: do m~t dQ quang. £)~ rat nho him cho k€t khAc phuc nhiroc di~m nay. CQt Lichrosorb RPl8 (250x4 mm. phtrong phap so mAu.2._~·__f~!_q'!_q_!.45 urn. Phuong phap nghien cu-u a. Cac chat chuan theophylin. ephedrin hydrochlorid va phenobarbital do Vien Kiem nghiem cung cAp. epbedrin bydroclorid -----------------. co nhi~u loai thuoc chua hen co 3 thanh phenobarbital nhir Etadasmyl. A. Asmalin di~n hien co (Xi nghiep Diroc pham trung irong 5). tinh khi€t sAc ky. Trong cac Diroc li~u tham khao khac chi co chuyen luan kiem khan. Antasma . va phenobarbital trong thuAc han hQ'P tr] hen suy~n bing phuong phap HPLC Tren thi tnrong ephedrin hydroclorid. Nhir v~y muon dinh hrong cac thanh phan trong h6n hop thi phai thirc hien vi~c chiet tach plnrc tap. Thiit hi. h6n hop bang phuong phap sic ky long hieu nang cao.t a biroc s6ng 240 nm.g~j~r:_£~ I 4. Hon nita. Dasmin (Xi nghiep Diroc pham trung trong 2). LQc qua mang IQc 0.

3. T6e dQ dong 1 ml/phut.1 mg/ml.2. phenobarbital ephedrin.!1.0 g natri heptansulfonat d~m phosphat pH 3._ i·_ »« ql}_l!_ ~g_~!~l}_~0f Pha dQng: Hoa tan 1.6 mg/ml. tren sic ky db thu diroc cac pic tach khoi nhau hoantoan Hinh 6.4 .HCI 0.6. va b.05 phut) B.3 . them vien. lAc ky. Dung dich thir: can 20 vien (d6i voi vien bao dirong phai rica h~t lap vo bao cua vien. 12.mroc (70 : 30) d~ duoc dung dich h6n hop chuan co nbng dQ cua theophylin khoang 0. Khao Slit tIP tuy€n tinh Chuan bi m<)t day dung dich chuan cua h6n hop 3 thanh phan tren voi nbng dQ cac chAt trong h6n hop bien thien nhtr sau: 0. nghien min rbi trQn d~u.1.2.6. Can chinh xac mQt hrong bQt vien nrong irng voi kh6i hrong cua khoang 80 mg theophylin. Dung dich chuan: Hoa tan cac chAt chuan theophylin.36 phut.5 mg/ml d6i voi ephedrin. B. Chudn bi cac dung dich tiem trong h6n hop dung moi ethanol .HCI ephedrin hydrochlorid khoang 1.!m.5 (38 : 62). 1 d~n 0. 6.HCl 7. 15 ml mroc cAt. rbi IQc qua mang lee 0. trong 1 lit h6n hop cua methanol - .1. tinh kh6i hrong trung binh cua cho van binh dinh mire 50 ml. - Th~ tfeh tiem 20J.45J.44 phut va cua phenobarbital 4.42- va lJ cac hinh 6. IQc qua gidy IQc thuong.2 mg/ml va cua phenobarbital - cua khoang 0. SAc ky db cua h6n hop 3 thanh phan (tr (theophylin) ephedrin. Kit qua V 6i chuong trinh sdc ky nhu tren.4 d~n 2.25 mg/ml d6i voi phenobarbital K~t qua khao sat S1)' nong do cac chat duoc phu thuoc tuyen tinh dien tich pic nrong irng thu duoc va trinh bay a cac bang 6. them ethanol tuyet d6i d~n vira du th~ tich. sAy a 50°C trong 1 gio rbi moi dem can). lAc d~u.0 mg/ml d6i voi theophylin 0.05 d~n 0.1. 6.

43- .05 52.25 258. D6 thi bi~u di8n su phu thuoc tuyen tinh n6ng dQ va dien tich pic cua theophylin Dien tich pic N&ng dQ (mg/ml) 1 0.1 10.4. Bang 6.__ o .0.. .5 Hinh 6. Ket qua khao sat dQ tuyen tinh dbi voi ephedrin STT Ephedrin.._~~~4 H1nh 6.2. Kit .0 5 2.lijp lai tren hai mdu vien nen va vien bao dirong: Mfru 1: Vien bao duong Etadasrnyl.1 .3 2 0.5 5 0._ .5(mg/ml) Phuong trinh h6i quy : y = 111..3 Hinh 6.0 4685.1 3 4 1.2 0. Khiio sat iI(j J(ip J{li Chung toi ti~n hanh khao sat de.0.5 2 2.5 5 Khoang nfmg dQ tuyen tinh : 0.20 208.5 H~ s6 nrong quan : r = 0.3 32.4 42.2 2851.05 .1 0..2 20. .1.6 3736.5 4 0.0 0.10 3 0.9998 .3.8 x -1.9993 Bang 6..2 x + 2.4.Nong dQ (mg/ml) Dien tich pic 1 0._ . SKS : 011099 Mfru 2: Vien nen Typhasmyl.2.2 I 1840..9997 Bang 6._.0 (mg/ml) Phuong trinh h&i quy : y = 2337.54 H~ s6 tirong quan : r = 0.2. Nong dQ (mg/ml) Dien tich pic 0. D6 thi bi~u di~n su phu thuoc tuyen tinh n6ng dQ va dien tich pic cua ephedrin r-----------.2. SKS : 010600 .8 2 1.1 3 4 0.1 0.43 H~ sA nrong quan : r = 0._ . Ket qua khao sat dQ tuyen tinh doi voi theophylin STT Theophylin 5000 4000 3000 2000 1000 o o 0.15 160.9 x + 8.5 1. 0:~ ~0~2_. D6 thi bi~u di8n su phu thuec tuyen tfnh n6ng dQ va dien tich pic cua phenobarbital B.5 2 105.. K€t qua khao sat dQ tuyen tinh d6i voi pheno barbital STT \ 300 Phenobarbital 250 200 150 100 50 o o 0.5 55..25(mg/ml) Phuong trinh h6i quy : y = 1030.2 Khoang nong dQ tuyen tinh: 0.3 0.- -- .3.0 0.4 .3 Khoang nong dQ tuyen tinh: 0.-- __ -- -- -- -- .HCI 60 50 40 30 20 10 L_.4 957.x aua _________ nehien cuu 0": .

55 2.04 0.88 4.4.90 7..12 1.53 10.82 41..HCI 7.95 4.40 10.47 2.2 mg/ml Phenobarbital: 0.12 7. Ket qua khao sat dQ l~p lai tren miiu Typhasmyl STT 1 2 3 4 5 6 Ham hrong heat chat trong mQt vien (mg) Theophylin 80.00 4.02 80.59 10.HCI: 0.n xet: Phuong phap co dQ I~p lai t6t.4 va 6. Chuan bi cac dung dich h6n hop chuan va thir co n6ng dQtuong ung nhu sau: Theophylin: 1.16 1.67 10.83 8.10 80. .78 Ephedrin. ----.6 0..71 10.90 0.52 Phenobarbital 4.20 0.51 0.18 41.41 41.90 4.5 diroi day Bang 6.31 Ephedrin.51 4.04 80.47 2.49 41.1 mg/ml Kch quit khao sat dQ l~p lai duoc trinh bay (] bang 6.HCl 10.57 2.43 0.59 Tinh thong ke X S RSD(%) Bang 6.50 2.79 7.91 7.46 81.91 5.-----.6 mg/ml Ephedrin._~-_l_ !i!_ql}_q_!'_g_~!~f}_~0! --- ----.6 0. K~t qua khao sat dQl~p lai tren mftu Etadasmyl SIT 1 2 3 4 5 6 Ham hrong hoat chat trong mot vien (mg) Theophylin 41.81 Nh.32 0.90 Tinh thong ke X S RSD (%) 80.04 1.92 0.78 81.52 Phenobarbital 2.68 41..48 2.85 7.5.02 40.72 10.

K~t qua khao sat dl) dung tren miu vien bao dirong Etadasmyl Theophylin Tim Them vao thAy (mg) (m_g) 20.0 1.0 40. Khiio sat ilP dung Khao sat dQ dung bfutg phirong phap them.__f:~!_q_1!_q_ !l_g_~j~1}_ 1 £~ .0 98.5 99.98 2.0 2.3.0 38._.99 2.7 97.0 4.0 39.li 98.99 3.95 3.0 1.5 Nh.6 40.8 Trung binh % I~ 99.6 va 6.7 dum day: Bang 6.99 thay % tim lai 98.6 Phenobarbital Them TIm vao (mg) (mg) 0.0 5.0 0._~.1 20.0 19.1 Ephedrin. B.97 1.0 97. K~t qua diroc trinh bay 6 bang 6.7.0 Bang 6.0 1. them vito miu thir mQt luong chinh xac chdt chuan bang khoang 20 .30 % hrong co trong miu thir.0 98.10 5.HCI Tim Them vao thAy (mg) (mg) 5.5 99.6.0 2.li 99.3 99.3 99.0 99.0 Phenobarbital Them Tim vao (mg) (mg) 1. K~t qua khao sat dl) dung tren mlm vien bao dirong Typhasmyl Theophylin Tim Them thfty vao (mg) (rng) 39.7 98.99 % tim Il.8 20.HCl Them Tim vao thfty (mg) (mg) 2.0 thay % tim l..45- .0 101.0 4.0 19.85 5.9 40.93 % tim Il.0 99.98 1.0 97.n xet: Phuong phap co dQdung cao voi ca ba thanh phan.. ti~n hanh nrong nr phep thir dl) l~p lai.1i 99. .0 98.0 3.0 97.0 0. ---.li 102.0 98.5 20.5 98.6 tlm Ephedrin.94 2.0 97.0 Trung blnh % tim ll.

pipet thuong. A. 27... maleat bing phuong phap HPLC ----. Dung cu thuy tinh: binh dinh mire 1000. May do PH Meter 3305-Jenway. Can phan tich Mettler AB 204 dQ chinh xac 0.!m). 6ng dong. 29] chu y€u chi co dang thuoc dan thanh phan..46- . kY khac nhau. lO. Djnh hrqng dAng thM paracetamol. c6c co mo .._~. loai tinh khiet phan tich. 12]. Th\l'c nghi~m A.. . Nham tim met phuong phap chung.Oml. Acid phosphoric loai dung cho HPLC. Trong diroc di~n My [29] va tieu chuan co sa [4.45 urn... UV diode array. . BQ loc dung moi va bQ IQc mau voi dirong kinh 16 mang IQc 0.HCL Clorpheniramin Voi nhiing hoat chAt tren.. clorpheniramin .Oml. . pipet chinh xac. trong cac duoc di~n thong dung hien hanh [I. nguyen li?u Vien nen Decolgen co cong thirc nhu tren. 99.. 20. Mdu 1 co s6 kiern soat: 402221 Mau 2 co s6 kiem soat: 404971 ChAt chuan cua Vi~n Ki€m nghiem: chuan paracetamol (ham hrong 100%). Hod chat. 100.I. 25.-_ .66%). co th~ dinh tinh va dinh hrong nhanh 3 thanh trinh sitc ky co A.7.... 50.. co sft dung phirong phap HPLC d~ dinh hrong cac hoat chiit trong h6n hQ'Ptren.. nhirng m6i chAt phan tich duoc su dung mQt chuang trinh sitc phan trong h6n hQ'P thuoc.. - May lAc sieu am-SONOREX. trong do vien nen Decolgen Paracetamol (PA) (PH) : maleat (CM) do cong ty United Pharma san xuat co cong thirc: 500 mg 25 mg 2 mg Phenylpropanolamin.Oml._ ..Oml. Thi2t hi.-_ hydroclorid phenylpropanolamin va Tren thi tnrong hien nay co nhieu dang thuoc co k~t hQ'P cac thanh phAn nhu tren. loai dung cho HPLC. chung toi nghien ciru xay dung mQt chuang th~ phan tich d6ng thai cac hoat chat tren..Oml. d(lng C(I May sitc ky long hieu nang cao: DIONEX-Detector CQt Nuc1eosil C18 (250mm x 4mm.86%). ~¢!_ t!. Trimethylamin Acetonitril Niroc cAt.1 mg.2. chuan - phenylpropanolamin hydroclorid (ham hrong chuan clorphenirarnin maleat (ham hrong 99. 19.g_~j~t}_~0! __ q'!_q_ 4.Oml.. 5J..

5 0:' LD N 0 Hinh 7. 0.c · • c 1. " " <3. PhA hApthu UV -VIS cua CM trong dung moi pha dQng Sau qua trinh thtrc nghiem. PhA hApthu UV-VIS cua PH trong dung moi pha dQng 1. Khao stit Iva chon ili2u kirn sdc kf Khao sat ph6 til ngoai cua cac ehftt phan tich: O._~.4 '" 0.•.47- . CQtNucleosil C 18 (250 mm x 4 mm.0 .3.1. . ~ ~ 0. B. K~t qua --- ----------------------------------------------------------------------------_.2 0 200 300 Hinh 7.. tnrong Dai hoc diroc Ha noi.B '! g c .5 025 Hinh 7.5 2 ::. .e 5 0.75 1. 5 um). 50 . chung toi da Iva chon ducc cac di€u kien s~e dinh hrong 3 thanh phfut PA. 0.1.5 5 0 1 125 . Ph6 hftp thu UV-VIS cua PA trong dung moi pha dQng 2._!f4!_q_l_I_q_~g~!~'Y_-:~ ! B.2.0 ~ .5 :.~ .75 0. ' ~ . PH kY d~ va CM nhu sau: May sie ky long hieu nang eao DIONEX-Detector UV diode array cua bQ mon Hoa diroc.

5 g kali dihydrophosphat trong 1 lit h6n hop cua nirrrc acetonitril phosphoric. I &3 c.9991 20 40 Nong d9 (ug/ml) Dien tich pic 3. -- - - - - - - _. K~t qua khao sat dQtuyen tinh gitra nbng dQva dien tich pic cac chat 3 41 40. __ '__'_W_''__'I . Cac pic da: tach kh6i nhau hoan toan.0 ID 11-e.4.jl II ~ 1- '~!i8Il Ii II i I Ii I II ~"_'_" ~(l ! \.2 ml/phut.4.lg.082 H~ so wang quan: r = 0.56 100 17.4 N 6f!g_ d9 (~g.09 Phuong trinh hoi qui: y = 0. r~t I Hinh 7.. ~~e'-"""'I~. nehien 0' Clfu .(_m!) Dien tich pic 8.-..62 N6ng d9 (f.-.-.24 1.~y~~-..2 10.2..1 5. Tien hanh phan tich voi dieu kien sac ky nhu tren.48- a bang 7.(_m!) Dien tich pic 0...-.-.. 10.-.7 b~g acid - T6c dQ dong 1. .. ta thAy di€u kien s~c ky la phil hop.25 5 82 76. Pha dQng: Hoa tan 2.65x .16 H~ so tuong quan: r = 0.74 PH eM 8. gon. sic ky d6 cua dung dich chuan PHva eM a buoc song 216nm PA.-..53 H~ so wang quan: r = 0.-..35 16.72 160 27..4.-.9999 3. SAc ky db cua dung dich chuan PA song 244nm a buoc Dira vao cac k€t qua thu duoc.2 16.._ -op + - _ ...-.17x + 0.6-+16 ug/ml.96 8...-.. . ta thu ducc cac stlc db nhir sau: - ~::.5..'"~ .4.6 61.93 2. dieu chinh den pH 2.. - Detector UV (] 216nm d~ do PH va CM. B. Bang 7. Nhiet d<)phong.triethylamin (968:30:2).-- __ . (] 244nm d~ do PA....89x + 3.45 4 65.4 Phuong trinh hoi qui: y = 0.. K€t qua khao sat duoc trinh bay Hoat chat PA Dung dich 1 2 8.14 12. Khtio sat 49 tuyln tfnh cua phseong phap Pha day dung dich chuan cua h6n hop ba thanh phftn co nbng dQ cua PA bien thien trong khoang 8-+80 ug/ml.79 200 34. Kit nUG __________ :::t.9999 . Minh 7.44 .9 19. Th~ tich tiem mdu: 20 ul.--.0..69 7. dang pic d5i xirng.63 Phuong trinh hoi qui: y = 0. cua PH trong khoang 20-+200 ug/ml va cua eM trong khoang 1..

76 8.13 17._.Lm.5038 0. Bang 7.2012 0._.40 17.10mg Luong can bQt vien (g) 0. K~t qua khao sat dQchinh xac diroc trinh bay 6 bang 7.2657 Dien tich pic PA 44.03 lieu th6ng ke X = 2.64% CM Ham hrong (rng/vien) 520.50 sA Ham hrong (rng/vien) 2.05 517.-.57 519.64%.45%.0 K~t qua trinh bay 6 bang 7. LQe qua mang Ioc 0.49 25.02 4.2632 0. Khao sat dp chinh xdc ciia phuang ph6p .-.08 S = 0. 7. ella eM IA 1.---_.22 17.__-_..3._.03 522..45 25.0448 Nong dQ (ug/ml)..51 S = 0..24 100. 7.'---. B._.45% - - - 1.l~/!_ !!g_JJ!~.64 4..76 44.-_-_.56 44.'.2636 0.023 RSD%= sA X=519.09 2.41 25.96 Bang 7.6.._.96 Di~n tich ~e 43.110.54 Bang 7.62 CM 4.0 ml. Duoc dung dich thfr d~c.41 25..62 4._.5. K~t qua khao sat dQchinh xac ella phirong phap PA IT 1 2 3 4 5 6 PH sA li~u th8ng ke Ham hrong (mg/vien) 25. q.!C!_ .2636 0. them mroc d~n vach. Can chinh xac kh6i luong bQt vien tuong irng voi kh6i hrong cua khoang 2/5 vien cho vao binh dinh mire 100...07 2.6..60 4. Dung dich thir loang (dung d8 do PA): Hut chinh xac 1.85 PH 17.-_-'.70 4.09 2.47 25..0 ml. xac dinh kh6i luong trung binh vien. Luong can bQt vien va di~n tieh pie cua dung dich thir SIT 1 2 3 4 5 6 M =658. them mroc.45 J.164 RSD%= 0._.0 ml dung dich thtr d~c eho vao binh dinh mire 50.08 2.34 RSD%"'" 0.69 44..-"_.._. 40._. N6ng dQva dien tich pie cua dung dich chuin CMtchufut LUQ'Ilg can (g) PA PH eM 0..-_.11 % .43 S"" 2..17 17. ella PH lit 0... them mroc d~n vach. lAcsieu am d~ hoa tan khoang 10 phut.09 2.34 516.08 521.:.5.----_.23 17. Dung dich thu d~c (dung d~ do PH va CM): Can 20 vien.!_ ~0f.19 17.63 4.-.. . nghien thanh bQt min . •:.7.fac d~u.91 44.89 44. lAc d~u.7 eho thdy dQ l~eh chuan nrong d6i cua PA la 0.84 lieu themgke X =25.2668 0.._'!_·.----..___.--_.7.2628 0.

61 PA PH eM 1 0..73 100.91 2.30 2.30 Luqng Ty l~ thu chuan tim h6i (%) th~ (m_&) 101.01 101.-- -..0129 0...61% d~n 100.36 99.._ -- - ..8: Bang 7.50 0..40 23...18 9.__nehien. K~t qua dinh hrong duoc trinh bay cac hinh 7. Kit ::l. Khiio sat tIP dung ciia phuong phtip DQ dung cua phuong phap duoc xac dinh bang phirong phap them chuan.65%..13 98.30 2. B.0137 3 Trung binh 1 0.52 2... .041 0..84 10.-.7 va 7.8 eho thAy kha nang tim lai til 99.81 9. PH va Clvl tren mdu 2 vien nen Decolgen.0129 0.26 2.34 23. B.53 0.043 23. . Cac s~c ky db thu diroc cua dung dich mdu tlnr va mdu chuan diroc trinh bay & -~~==~~--------~--------------~~ .__ . .9.-- .0131 3 0.84 10._. ap dung d~ dinh IUQ'l1g eho k~t qua t6t.09 9.0129 2 0.8.70 2. Hinh 7.65 99.01327 Trung binh K~t qua & bang 7._ aua o_ CUll __ __ __.52 23. a bang 7.52 23.0131 2 0.. K~t qua khao sat dQ dung Ho~t chAt STT Luong bot vien (g) Luong co san Luong trong mdu thir chudn them (mg) vito (m_£) 10.275 98.040 0. 7.6.__ _..0137 Trung binh 1 0. K~t qua khao sat dQ dung diroc trinh bay a bang 7._.S.0131 2 3 0. Sdc ky db cua dung . PH va vien nen Decolgen eM trong Chung toi ti~n hanh dinh tinh va dinh hrong dbng thai PA.. Ung d(lng di dinh tinh va dinh /U'p7Igd6ng thm hnn h(YpPA.6.95 99..51 0..-.260 98.-.___.12 9.38 99.34 9.84 101.84 0.338 101.65 23. .4.50- dich mdu chuan ._ '" 4. co nghia la plnrong phap co dQ dung cao..8.75 10...

...09 514."' '·13..0 104..-.. "WVL.5 104.09 2.50 25..9...03 2.-..08 2.50 102. .__________ ::.97 103.. K€t qua a bang 7..30 25.0 103...11 513._..0 103. PH va CM.19 512.-..36 25.42 101.-.20 101.06 2. SAc kY db cua dung dich mdu thu do PH va CM (y burro s6ng 216nm I·RDeO Hinh 7.62 102..17 25.41 (%) nhan so 103.88 512..S.06 2.0 Tir cac hinh 7.Q 11. SAc leY db cua dung dich mdu thir do PA Bang 7..82 102.0 100.9 cho thAy ch~ pham dem dinh hrong dat tieu chuan v~ ham IUQ11g cua PA.0 11.7 va 7. Kit aua nehien Czlu 0'..-..64 (%) nhan so Ham hrong vien (mg/vien) 2. n'_o 14.44 101.36 25.44 102.. .35 Ham hrong v6i 1 2 3 4 5 6 TB (%) 102.:21t -t c .-.. ~.68 101..2 PH Ham hrong " Vat Ham hrong so vien nhan (mg/vien) 25..68 102.... 7.51 - ...-.83 518.48 101. t• . -- W". K€t qua dinh hrong Ho~t chat STT a btroc song eM 244nm PA Ham hrong vien (mg/vien) 512..:{ 4.04 2...:) Hinh 7.6.58 102.42 25.-.06 Ham hrong voi 101..8 cho thAy gia tri thai gian hru cua cac chAt a mdu thir so voi mdu chuan la nrong duong nhau.10 509.--.7.-..-..0 101.-..

phirong phap HPLC). ThVC ngbi~m A. B. trQn d~u. chung tbi da nghien ciru thanh cong phuong phap phan tich toan phd d~ dinh hrong d6ng thai paracetamol va ibuprofen.1. Khio sat va xiy dl}'llg phuong phap B.2.1 eho th~y ibuprofen (1) h~p thu cue dai tai 222nm.1mg. dflng cfl - May quang phd UV-VIS Beckman DU 650i. . paraeetamol (2) hdp thu eire dai tai 243nm va dQ hdp thu cua dung dich h6n hQ'P(3) i. Dung dich chuan g6e ibuprofen (IG): Can chinh xac khoang 50 mg ibuprofen. Cac dung cu thuy tinh chfnh xac: Binh dinh rmrc. A. Hod chat. Mftu tlnr: Vien nen Dibulaxan. tron d~u._(_~~!_q_~q_~g_~!!_'!_t:~ 4. pipet. Tren hinh 8. B. Pha cdc dung dich chu8n g6c paracetamol va ibuprofen . nguyen vpt /i~u Chdt chuan: paracetamol.2. SKS: 020601. hoa tan trong binh dinh mire bang d~m pH 7 vira dil 100ml. hoa tan trong binh dinh mire bang dem pH 7 vira dli 100ml.:. ket qua ph6 ghi diroc nhu & hinh 8. Binb hrqng dang tbOi paracetamol va ibuprofen trong vien nen bing _ phmmg phap toan pba Ngoai cac phirong phap da noi a phfm tren (phirong phap quang ph6 UV-VIS thong thuong sau khi chiet tach. Ldy 10 ml dung dich pha loang bing dem pH 7 vira du 100ml. 1rQnd~u. .:1 biroc song diroi . 1rQn d~u. Thiit bi.52- . tich co dQ ehinh xac 0. Xac dinh giai song phdn tich Ghi phA h~p thu quang cua m6i dung dich thanh phfut rieng re ill 200nm d€n 300nm.1.8. diroc dung dich chuan g6c paraeetamol co n6ng dQ 80 meg/ml. diroc dung dich chuan g6c ibuprofen co nbng dQ 50 mcg/m!. L~y 10 ml dung dich nay pha loang bang d~m pH 7 vira dli 100ml.:. Xtiy dvng phuang phap a. ibuprofen dat tieu chuan BP 1993.1.can phan A. Dung dich d~m phosphat pH 7 pha thea DDVN III. Dung dich chu§n g6e paracetamol (PG): Can chinh xac khoang 80 mg paracetamol.

.an VavelBngtb (n..37445 ..3348 ~_._ _.300 ~__ ._.686 11.-l._.. pha mQt day..._.._L~_. K~t qua duoc trinh bay (y bang 8. . Ibuprofen I ~~_. Paracetamol uz/ml .5066 0... Bang 8. Lmm~~--------------------~--~ SIIo:Iothin9._ ".._. D222 ....n. PhA UV-VIS ella ibuprofen (1) n6ng dQ 7.n .752 .. 5.0186 ·_·.".__ Duong hoi quy D = 0._. t:5' Ie phbi hQ'P gitra hai eh~t trong cong thirc ella vien nen Dibulaxan._.C .n.__ ~ __ .._·_ .._. .._.._. c._. Kit ::::t nUG __________ nehien cuu 0 .. __I 1.. ! 0._0.~. sir dung d~m pH 7 lam dung mei pha loang." __ .168 i 10...._._._._.29743 __ ".._.457 ."._. ~_~4.. .._."..._._._"..'. .__ ._.u_~ __ _..n...n...15._.) Hinh 8._." D243 i 0.u __J 0. chung toi chon khoang nong d~ phan tich cho paracetamol tir 8 ug/ml d~n 12 ug/ml va ibuprofen til 5 ug/ml d€n 8 ug/rnl..L Til k~t qua khao sat m6i tirong quan gitra dQ hAp thu quang va n6ng de) cua m6i thanh phan._.. .. ~.._. .._.._.. ! .u_.... __ ._ ---_ _--..l! ____.i 0.~__ .__ i 6.. .._..9994 uz/ml ..'._+I__.•. ."~ . __ Duong hoi quy D = O..__ _~_ .. .576 ug/ml paraeetamol (2) n6ng dQ 10. ~..843 .7." +_ ._.._.543 ug/ml va h6n hop ella cluing (3) trong d~m pH 7 b.._~nf.C ._. .".n.. .~"~ I.n. " " . ..._.22317.~. _ -----_ -_.~-+-n_. .52569 0..' . til 8 ug/ml d~n 15 ug/ml cho paraeetamol..047374..J:"._.l __ . 215nm la luang d6i Ian (> 1.0._._ 8.._._ _ .'.. __ ~ __. None Functions. u_··..__~cn._...__ I __. __ ~ __ .1._._.U_L~_. ..._. Do do chung toi chon khoang buoc song phan tich eho hai ch~t til 215nm d~n 300nm._.L r i _'_"""'_n.u..504 __. __. Pha day dung dich chudn h6n h(IJJ Dimg pipet chinh xac phbi hQ'Ptil dung dich chuan g6c PO va 10. ._.._ "~ .__ .._..u.~ .__ ."..9999 1..45714 i 0.'.. Ti~n hanh do m?t dQquang cac dung dich ibuprofen tai 222nm va paracetamol tai 243 nm.... ~__. .53- . Trong khoang n6ng dQ d§ khao sat cho th~y co su luang quan tuyen tinh gitra n6ng dQ va d{)hAp thu quang ella m6i ch~t tai biroc song khao sat.__ .··U.n._ "._"'cn._..._."__ .J._.n_..u.~_.6763 . ...1.8498 _...n.06621....·'·_~n. Xae dinh khoang n6ng d9 co luang quan tuyen tinh VGid9 hJp thu cua m6i chdt tai cac buac song qrc dai Chung toi da pha day gbm 5 dung dich chuan co nang dQ bien thien til 5 ug/ml d€n 11 ug/ml cho ibuprofen.. Sc::...071 __ n._ ".._" "----!_ .. __ _~_~_.g5m 7 dung dich h6n hQ'Pehufin voi cac ._. ._.....----_ -----_ ---_ --.n..._~_. i 9. __ ..0019 V01 r = 0._n_n.0.7.. ! 0..00679 voi r = 0.n._._._....._.920 i __ ".._.n._. l. _ _._. _.n.0).4._. -._._..._..1. "__..."..... ._ !. . Mbi nrong quan gilra n6ng dQ va m~t dQ quang..336 I 12._._.U.

.82!>'& 6 1324 10.iUKle.0_ 300 ...canQa«a.6t'#S 8.) ua ..OOa7 -0.~13 li. Kit nehien .0234 0_0040 'l'....."l:. . khong phai loai bo dung dich chuan nao.1 6_1140 1. ~. -=III:tIJiu '1M! DO 6&OJ. Phdn tich chudn Dung d~m pH 7 lam m~u trang.01. Nxl.• ..ut 4 0.:l4!:. K~t qua phan tich chuan nhu hinh 8. D."00 6_6434 1I.. _ . \ PAlttlR1 1IrIIS1:19 S8Ipl~ ul..r..B.7 Hinh 8. - -- - - - - - - - - - - - - --- - - - -- - - - - - _ .4816 6 U75 10_ 6749 6. 'dill Wi! '>'Sa ~ .1ng ~ .0. chusn nay dircc khang dinh chdt lUQ11gbang mQt nhom 3 h6n hop chuan cho sai s6 chuan tien hrong SEP tuong dirong Nhu v~y dirong chuan xay dung ITtren diroc danh gia la co chdt ltrong nit t6t va duoc sir dung d~ phan tich cac m~u tlnr. d.au . them 75ml d~m pH 7 lic .2.... _ .034 0... 4&00 8. _41 _4 . __ czm .~~=..0049 M:. l§n luot dira cac dung dich chuan theo thir ttr vao phan tich.c=.54- ky khoang 15 . Can voi khoang 50 mg chinh xac m9t hrong dirong Paracetamol.~4U 8..S 9.s ..2...Q30 0034 O.2"000 CXH:BJIIPItI\oUON Cal..2 ta thAy chenh I~ch n6ng dQ cac thanh ph§n cua tAt ca cac h6n hop chuan rna may tinh tOM duoc (Calculated) so voi n6ng de) thuc (Actual) la rAt nho.0307 0.01411 tVI £VI Iy) o 0266 _<15 _d _d't 4> ~ ~ 'IZC DIU __ l!!o\JUIIClI.0011 0_0(»1 -0_0044 -000&6 0.OOSG nnJ~ ~DIJJiIIICII'1II 7..&'aL' r ~J4lI. e. K~t qua phan tich cua 7 dung dich h6n hop chuan Tren hinh 8._000_~ "'.0w.OlS . 0144 -O. _ .. .aUG __________ A.D21I .ND IIiCIf T~ ~OlOPt 11..o. ~" 7 0 4 3lJ COiiop.ual 8. ~JIX.81lGO It.2040 ..noe 0..... n6ng de) cua paracetamol thay d6i ill 8 ug/ml d~n 12 ug/ml..o... 1.022 0. a.. Or .)OOO &. Chudn bi mdu thit Can 20 vien..201. cho vao binh dinh rmrc IOOmI.CIt.1t 1 a. nghien bi}t vien da: nghien min tuong ky thanh bQt min....al lJaiti v... Duong voi sai s6 chuan cua uoc IUQ11g SEE..d o:.. ~ MKiliCB'fIlN l:IIUPJlQJJIH ~JIX' DttJiftalo"I!Ilf ~AH IZrIJF!ICI'lDl 5:U.1"4 9..Cn U. "1Ib. lolL 211>. __ II. 4 ~ __4 l 0.012. O~ 02 _h:>d iItoan<ing _. ~ 0_018 O... - - - - - - --- n6ng de) cua ibuprofen thay dAi ill 5 ug/ml d~n 8 ug/ml.....zg~ rn fVl (VI S_ISU>1 11.0200 7. dlwJ.. tinh kh6i IUQ11gtrung binh vien. 4._ - __ - - - - -- - - - - -- - - _ .OaJ E!td.6700 0.. A. IUIIM liII:lIH:tiIx nat.·1I00Q IVI 8cl7of# B.Z.~ ot e•..p::melE o\.5 ·0.0C'9 .. 1..r 01:~...73 1.cu4ead l JCG/ Dite.0 ..'PAX ~ • O.1eg2 0. I:.. 0.

54 207.0234 9.q_~8.2.2142 6.6091 7. Ket qua khao sat dQ l~p lai cua ParacetamoI BQt vi en IT 1 2 3 4 5 6 7 8 9 (g) va Ibuprofen.1027 0._~·. Ham hrong ibuprofen {[It] c. tri)n d~u.1112 0..59 211.5 mg S = 1. duoc tinh bang each thay [Pt] bang [It].73 332.08 207.38 330.62 207.6429 10. Bang 8.98 205.~0!.1313 = 2. Iii.nAng dQ cua ibuprofen doc duoc tren may (mg/ml)}.25 330.1147 0.899 RSD%=0. Dung dich thit duoc chuAn bi nhu muc tren vii.6795 6. VT Iii.055 RSD% = 0.1179 0. tien hanh phan tich theo phtrong phap dd xay dung. Ham lUQT1g trong mot vien: Xtrng/vien) Trong do: [Pt] Iii. .\ kh6i hrong bot vien da din d~ dinh hrong (gam).3768 6.91% Ket qua tren cho thdy phirong phap co dQ l~p lai cao d6i voi ca hai thanh phAn.6907 gam) voi 9 phep thir.0825 6.7072 12.2. Xdc iljnh ilP /9P /{li cua phuung phdp SKS: Tien hanh thir dQ l~p lai cua phirong phap tren mdu vien nen Dibulaxan 020601 (Mtbv = 0. them d~m pH 7 den vach. S6 li~u thong ke Paracetamol £)0 Ibuprofen £)0 duoc Hv hnmg (mg/vien) 329.01 328.2927 6._ _ -.lugng (mg/vien) 209._.3472 10. la kh6i IUQTlgtrung binh vien (gam). __ .60 Paracetamol Xtb == 329.1046 0. 1.1731 7.32 209.1010 0. LAy chinh xac 2ml djch IQc pha loang bang d~m pH 7 vira du 100ml. dtroc dung dich thir.55- .62% Ibuprofen Xtb = 208.1113 0.37 207.03 diroc H.9063 9. _ _ _.1038 0._.24 328.~4!_ql!.9619 6. tron d~u. _ _ __ >- __ •••• _._.. phut. Kit qui thl}'C nghi~m Cl. B6 khoang 30ml dich 19C dAu.5693 10.59 326.45 210.94 329. n6ng dQ cua paracetamol M m = [Pt]x M xVT m doc duoc tren may (mg/ml). IQc.9604 10. the tich pha loang cua dung dich thir (VT =100xIO0/2 = 5000 ml). Cac ket qua duoc trinh bay trong bang 8.01 332.69 mg S (ug/ml ) 9.8929 0.6947 11.4714 (ug/ml ) 6._ .~j~f}.

B CO!IIpCInIl: p.1862 9. ..073 '. UUlIJIIIOIIBN '.3768 ------ 10. Hlnh 8.. 6.- _ .._- Xtb = 330.CB'fAH DtlPR:)1IlDf PAJaCB7AH PAMCIlftK UUlIC>PDf ~'!NI DQllJIDPJDf ~'IAX 2: KMn-_ 3 3 1 2: )Cun: w. phan tieh trong cung di~u kien ella cung mQt miu K~t qua a bang 3 cho thay dung dich co dQ dn dinh cao.nh 4p In dinh cua dung dich Chuan bi mQt miu thu nhu tren.1g(!lll) . .---- 3 4 r--c-----~ _.01'6 0. q_l_I_q. .065 2: 2: 1 Ht. .81 -209.·····_·' __ L· ___ 330."'.3374 6.12% ."83 6.II .013 0._ 6..01'6 0.".-------- 330. 2 -- -~--'. - 6.3..:.0177 6 _ --. K~t qua duoc ghi a bang 8.lon ~ %3tJJI1OI'IIN PAaCZ-. c:~ ~4!_ _' .- 210._ . $TANDARDDItOZ OF : : '-" .2940 ---_.1'731 11.20 330.-_ -~---_."2 7. K~t qua khao sat dQ 3n dinh cua dung dich.2P38 9..--E$nw..0268 --- _ .. 0.3609 --6.f .078 100.- 10.01'1 0. C2. Bang 8.B.!l_g_~j~f} . .3473 - -- --.. 330.. 029 14 IQI' ~bm 1t0lp • DUur..CIS'MK .57 209.0234 ...A~ o.w.05 _ _-- ---. dQ l~eh chuan nrong d6i ella phep thir la kha nh6.924 RSOOIo == 0.016 0.6Sl08 10.2927 9..20 --_.2361 11.014 0.G" 6. Xdc d. .33 - --_ .910) t._-- .CP48 1 1 1 PAMCIS'lNf 0.- Xtb == 209. .:2:142 10..V. ---. _ .42 S == 0. Cll' 10 phut doc miu 1 IAn trong cung di~u kien..8a: 7.9~0 6.OGS 0.acaWoX ~ 4 4 1 2 Hun: DOPICPJIM PuP. Paracetamol n 1 f)QC deoc Ibuprofen rng/vien Tinh thong ke DQc dugc (~_gIml ) mg/vien 210.15" 0..c:II'fAaI %BOJIR:)PIIN »MCIl'I'NC muJlJIOPDf ~ca'rIIX :tJIUPR:)1'IIN PADCII'f'AIC DlJDR)1IJDI 9. -..- 331. Ket qua phan tich ciia 5 phep thir. - 10. PAaCIIBK DUJiROnN PADCIl'INII IJlUPlIIDPIIN PAMca"."-.31 _.076 0.44% -- --- 330..0042 5 --_ ---_--10.SlQJ4 10.~1P 10. -.---._.082 0.II S. ----.967' 0..3.3C.75 6. - ..3402 Tren cung mQt duong chuan.076 0.- .56 ~ . c:cacc~uon 9_~33 6._. -.385 RSD% =0.5"1'00 '.066 a Kean: ~ ~ O.SI~8 10. Ti~n hanh phan tich mAu tren duong chuan da xay dung trong vong 60 phut._.94 --.-.02 207.G4? .al : -. .3472 1.0009 _'.IIft.24 209.54 --- Tinh thong ke _{J..013 0-018 l25Ol«1OPIIN 0.2023 11o.0011 10.-._- 10.9063 '.3._291C 9.n t .3..SQJ3 '.--.- 'lI::Ital S..58 S = 0. . __ .02:3 A.c!u. ._~ ____ .~----..sa6 6.012 0.-..562 S S 1 2: )C_..

.g_~j~1J_~ ~ C..---.._...-."3 ..4. mQt hrong xac dinh ch~t chuan Ibuprofen.6440 4.0. K~t qua dugc trinh bay trong bang 8.--~ -_ _.-.-.----.4.... Bang 8.3......047 100..··· ··················· .-. Xae t1..._ -----_ 4....-. Ty 1~tim Paracetamol bi~n thien tir ."5()"f··-···-·········-····· 101.. dQ dung cua phuong phap ch~p nh~n diroc d6i vci ca 2 ch~t. _._ 10.-f ···-······· ·--·.-~~-~.-. 110% rbi tien hanh nhu muc 2..2% va 100.. . K~t qua khao sat dQdung ciia plnrong phap TT BQt vien (g) Chuan them vao (ug/ml ) Tong do diroc (ug/ml ) Luong tim thay (ug/ml ) Ty l~ tim thdy % .2% d~n 99..2%.-. ·················----100.623 ··················--5-:45-6"·············------T6"~4·37-8-----·-······-···-·-·-····--·-··s...-...1 -.----...··-····· .1% d~n 101.-.. --····-o~i6·i4---··· -·-·~r~···· """"'0}027--' TB: 98.nh ------------------------------------------------ella phulYng phap dp dung Khao sat dQ dung bang each them chuan..8 '-~-~"~5Jj'O~·--~·' ----·--iO~096T--~--· ·---···5j94--~ ···-· ._--_.1045 2 0.---.8% Nlnr vay..--....1052 ._---- 1 . 100%.-.0346 .-.. 5..~--._.4% PARACET AMOL 5....-.2.-.-...-..-. Paracetamol bing 90%. 0.---.-T6T..-.·····.040 ._..h ro ham hrong._~. th~y cua Ibuprofen bi~n thien til 97.!q_!l.575 9.-----~. Them vao hrong bQt vien da bi.-------..-.-._Fl!_9.. TB: 100.--. .57- .3.5 sao cho nbng dQ cua m6i hoat ch~t mung (eng khong nim ngoai khoang nang dQ trong day dung dich chuan va ti~n hanh phan tich bang plnrong phap da chon..

Phuong phap nay tAn nhieu thai gian va cong sire. HITACHI 3210 (Nhat Ban). ChAt dbi chien hoa hoc: Quinin Sulfat va Paracetamol chuan (do Vien kiem nghiem . l~c ky khoang 15 phut. 500ml.t phan tich da ciu tir (MCA) B. dung dich Hel O.IN. .9. Vi v~y chung toi da: nghien ciru xay dung mQt qui trinh dinh hrong dAng thai paracetamol va quinin sulfat trong vien nen Antigrip F bfuIg phuong phap phan tich da cAu tir (MCA) tren may quang phb thong thueng..I.IN vira dil t6i vach. 30mg. . lOmI. Binb hrQ11gdAng tbOi Paracetamol va Quinin sulfat trong vien nen Antigrip F bAng phuong phap phan ticb t1a cAu tir (MCA) Vien nen Antigrip F cua Tiphaco san xuAt duoc sir dung lam thu6c ha s6t co chua 2 thanh phdn voi cong thirc nhu sau: Paracetamol Quinin sulfat 150mg._'.!_·__4.IN (theo tieu chuan DDVN III - ky thu. hut chinh xac lOmI dich Icc ti~p theo cho vao binh dinh . Sml. Bb sung HCI O. Khao sat pha hlip thy UV va llfa chon cdc btrue song phlin ttch can chinh xac khoang 75mg Paracetamol cho vao binh dinh mire lOOmI.BQ Y t~ cung d. IOOmI. Dung cu: pipet ImI.!_!l. so sanh k~t qua thu duoc voi phuong phap do quang phb thong thirong (bang chi~t tach) thea tieu chuan co sa. 4. mircopipet: 10m!. bo 20mi dung dich dfiu. B. .!I_q_!!g_~j!_. . Phuong phap HPLC cho k~t qua nhanh va kha chinh xac song thi~t bi chua th~t phb bien 6 tAt ca cac co sa ki~m nghiem trong rnroc va chi phi cung kha tfm kern. . TbuAc thir. LQc. Xay dl}'ng phirong phap t1inb hrqng dAng thOi Paracetamol trong h6n hQ'p bAng va Quinin sulfat vien nen Antigrip F (Tipharco) Thuoc tlnr: ethanol tuyet dAi. phai sir dung nhieu dung moi hfru co dQc hai (cloroform). phuong phap quang ph6 thong thirong [3] va s~c ky long hieu nang cao (HPLC) da: diroc sir dung. .p). Dinh hrong bang quang phb uv-VIS thong thuong cAn thiet phai chiet tach rbi do rieng tirng chAt tai cac cue dai hAp thu d~c tnmg. D~ dinh hrong hai thanh phan hoat chAt tren. xir Iy k~t qua bAng phftn mem tv xay dung. . hoa chit.58- . 2mI. Thiet bi: May quang ph6 ttr ngoai kha kien GBC Cintra 40 (Australia). binh dinh mire cac loai: 50ml. dyng cy Nguyen v~t lieu: "Thu6c thir"). can phan tich. . them 70mI HCI O. l~c deu. A..!_£0!.

_!_·__If4!_q_'!_4_!'..g_~!!_l}_E.0!

.

---.

mire 100ml rAi bd sung dung dich He] 0, IN vira du, litc deu. LAy chfnh xac Sml dung dich vira pha cho vao binh dinh mire 50ml, them dung dich H'Cl 0, IN cho toi vach, litc deu, diroc dung dich Paraeetamol 7,5f-Jglmltrong moi trutmg HCI 0,IN Can chinh xac khoang 40mg Quinin sulfat cho vao binh dinh mire 100ml, them 70ml dung dich ethanol 96%, litc ky khoang 15 phut, Bd sung ethanol vira du toi vach, litc d€u. LQc, bo 20ml dich IQc dau, hut chinh xac 20ml dich IQc ti~p theo cho vao binh dinh mire 100ml rAi bd sung dung dich Hel O,IN vira du, lAc deu. Ldy chinh xac Sml dung dich vira pha cho vao binh dinh mire 50ml, them dung dich Hel O,IN cho toi

vach, litc d€u, duoc dung dich Quinin sulfat 8 f-Jglmltrong moi truong HCI 0,IN.
Ti~n hanh ghi phd hAp thu UV rieng biet cua m6i dung dich tren trong khoang biroc song nr 200nm toi 400nm voi miu trang la dung dich HCI O,IN. K~t hop ca hai phd ghi diroc tren cimg mQt dA thi ta thu dtroc k~t qua nhu 6 hinh 9.1 .

\:/j;

. -1\~~1.~..--.-..",
A/
22~O

--......
3392 ,

_J

Hinh 9.1. Phd hdp thu quang cua Pracetamol (1), Quinin sulfat (2)
trong moi tnrong Hel O,IN.

200 0

'
2~OO ZlS.O 300.0

. .c ... '--sas.o
3500 :l"1S.o

~oo.o
"'"

J

.r

Nh{in xet: Trong moi tnrong Hel O,IN, Quinin sulfat co btroc song 208nm, 250nm, 314nm, 341nm con Paracetamol
bmrc song nh6 han 215nm Paracetamol buoc song

C1,lC

dai hAp thu tai 4

co met eire dai hAp thu tai

buoc song 241nm. Noi chung, hai thanh phan co su phan bi~t nrong d6i

ve phd,

a

a vung

co de>hAp thu quang Ian nen ta khong chon

nay. Dua tren du li~u ph6, chung toi chon 6 btroc song phan tich

cho hai chAt la: 215nm, 231nm, 241nm, 250nm, 314nm va 340nm.

B.2. Xac tlinh khoiing nang
va S dung dich Quinin hydrochloric bay trong bang 9.1.

tip phOn tfeh
(khoang nAng de>5-25 ug/ml) trong dung dich acid nAng de> 1-10 ug/ml)

Do dQ hdp thu quang cua 6 dung dich Paracetamol sulfat (khoang

,

0,1 N

a ca 6 buoc

song dd chon, k~t qua khao sat dQ tuyen tinh diroc trinh

- 59-

_____•____

4. Kit qua nehien elm _.... O~_.

....

....

......

..

......

.......

-

- - - - -- -

... ... _. _-

Bang 9.1. Tfmg hQ'P k~t qua khao sat m6i wang quan gitra dQ http thu quang va nfmg dQ cua Paracetamol Paracetamol va Quinin sulfat tai 6 biroc s6ng da chon,

Bmrc
song
_frun)

H~

tuong

so

quan (r)
0,99940 0,99990 0,99997 0,99997

215 231 241 250 314 340

Phuong trinh h6i qui y=ax+b _y == 0,036x + 0,037 y = O,066x + 0,032 y = 0,082x + 0,030 y = 0,075x + 0,027 Khong hAp thu

Quinin sulfat Btroc H~
song
(run)

so

tirong

_guan(l"}
0,99997 0,99997 0,99997 0,99999 0,99982 0,99955

215 231 241 250 314 340

Phuong trinh h6i qui _y=ax+b y = 0,080x + 0,002 y = 0,039x + 0,001 _y = 0,064x + 0,003 Y = 0,099x + 0,001 _)'_ 0,015x - 0,001 = Y = 0,018x + 0,001

Nh(in xit: nhin bang ta thAy c6
s6ng k~ tren (h~ s6 nrong Paracetamol quan r

S1,l'

tuong quan tuyen tinh rAt ra rang gitra d<}hAp va Quinin sulfat tai 6 biroc Tai hai biroc song 314nm va 340nm

thu quang va khoang nAng de>khao sat cua Paracetamol

=

0,9999).

hAu nhir khong hAp thu (hinh 1), tuy nhien tren thuc t~ dieu nay khong (5-25 ug/ml) va Quinin sulfat (1-10 ug/ml).

anh huang toi phirong phap phan ttch, do do chung toi chon khoang nAng de>phan tich cho Paracetamol

B.3. Chuan hi day dung dich chulin hOn hpp a. Dung dich chudn g6c Paracetamol
Can chinh xac khoang 25mg ParacetamoI them 35ml HCI O,IN, lAc kg khoang chuan cho van binh dinh mire SOml, 15 phut, Them dung dich HCI O,IN vira du toi

vach, lAc deu. LQc, bo 15ml dich loc dAu, hut chinh xac Sml dich IQc ti€p theo cho vao binh dinh rmrc 50ml khac, them dung dich HCI O,IN cho tci vach, lAc deu, duoc dung dich chuan g6c Paracetamol (PG 50 ug/ml).

h. Dung dich chudn g&c Quinin sulfat
Can chinh xac khoang 2Smg Quinin sulfat cho vao binh dinh rmrc 50ml, them 35ml ethanol 96%, lAc kY khoang 15 phut, Them ethanol vira toi vach, lAc d€u. LQc, bo 15ml dich JQc dfiu, hut chinh xac 5ml dich loc ti€p thea cho vao mot binh dinh rmrc SOml, them dung dich Hel O,IN cho toi vach, l~c dSu. Lai lAy chinh xac lOml dung dich nay cho vao binh dinh mire SOml khac, them dung dich HCl O,IN vira du, lAc deu,

ducc dung djch chuan g3c Quinin sulfat (QG 10 ug/ml).
c. Pha day dung dich chudn h6n h(/JJ D~ tang d<}chinh xac cua chuan, loai tnr sai s6 mac phai khi do ta se ap dung thuat binh phuong t6i thieu bing each sir dung sf> hrong dung dich chuan him hop nhieu han s6 cAu tir, Boi v~y chung Wi da pha m<}t day gAm 5 dung dich chuan h6n hQ'Pdung cho phan tich chuAn.

- 60-

_~.__44!_9.'!_q_~g~!~1}_ £~

-

---- --------------------------_.

Dung pipet chinh xac phoi hop til hai dung dich chuan g3c (] tren trong binh dinh rmrc 50m] va sir dung dung dich HC] O,IN da pha loang ta thu duoc me}t day dung dich chuan h6n hop theo bang sau:

Hang 9.2. Pha day dung dich h6n hop chuan.
Dung dich chuan h6n hop s6 ml dung dich chudn g6c cdn Paracetamol (PG 50 ug/ml ) Quinin sulfat (QG 10 ug/ml )

idy

Cl C2 C3 vao binh dinh mtrc 50ml 6 10 7 9 8 8

C4 9 7

C5 10 6 10 1,2

N6ng tJ(J cua m6i thanb' phdn trong dung chudn h6n h(1JJ tuong tsng ( pg/ml ) Paracetamol 6 7 8 9 1,4 Quinin sulfat 2 1,8 1,6 B.4. /)0 d{j hlip th(l quang cua cae dung dich chufIn han hpp

Ti~n hanh do d{>hdp thu quang cua 5 dung dich chuan h6n hop vira pha tai 6 biroc song da chon. Sir dung mfru trfutg la dung dich HC] 0, IN ta thu diroc cac gia tri de}hdp thu quang nhir 6 bang 9.3.

Hang 9.3. Gia tri de>hAp thu quang do diroc cua cac dung dich chuan.
Buoc song (nm) / d2 chuan Cl C2 C3 C4 C5 215 0,2896 0.3098 0,3299 0,3390 0,3491 231 0,3219 0,3676 0,3987 0,4196 0,4522 241 0,4308 0,4841 0,5120 0,5318 0,5732 250 0,4641 0,5085 0,5281 0,5400 0,5753 314 0.0224 0,0206 0.0190 0,0169 0,0154 340 0,0251 0,0234 0,0216 0,0185 0,0165

Nh(in xet: nhin vao hang de}hAp thu quang ta thfty

a cac

biroc song tir 215 - 250 (n6ng d{>ph3i hop tang
-

nm thi de>hAp thu quang cua cac dung dich chuan til C] - Cs tang ddn, di~u chirng to tai

cac btroc song nay thanh phdn hdp thu chinh la Paracetamol
vay ta thdy tai hai buoc song nay Paracetamol

dAn). Con tai biroc song 314 va 340 nm de>hAp thu quang til C1 Quinin sulfat hdp thu la chu y~u (n6ng de>ph3i hop giarn ddn). B.S. Xay drplg duO'ng chufin bdng phan m2m tTen may tinh

C, lai giam ddn, nhu

hAu nhu khong hdp thu

rna

chi co

a. Thiit ltjp cac thong s6 phon tich
DAu tien ta vao chuang trinh bang each nhay kep bieu nrong cua chuang trinh tren man hinh desktop, sau do ta chon

rna ij.p

tin maio Cira s6 chuang

trinh nhir (] hinh

9.2, chon "Dfmg y" da

rna t~p tin maio

Ta nhap cac thong s6 dil' li~u chinh nhu sau:
- 61 -

N~u cAn ta e6 the sira d6i cac dlr lieu da thi~t oJt' hoac "X6a".. _ __ _ __ _ 4.5. sau m3i IAn nhap chon "Them" chon "D6ng - s" de hoan thanh cira s6 nhap. (hinh 9._ -- . Thi6t l~pcac thong s6 chung.5) d6 chuyen sang nhap buoc song moi. mo tao (hinh 9.. Hlnh 9. __ .. Cac thong s6 chung: nguoi thuc hien. __ . Kit __ __ __ . (hinh 9. (hinh 9. __ . . S8 li~u v~ cac buoc song pharr tich. Nh~p s8 li~u v~ cac chdt thrum phk.. eu6i cling N6ng de>cua cac dung dich chuan h6n hop: lam nrong W nhu phan nh~p bmrc s6ng.3. . c~u tir MCA..2.4.. ten miiu.7) Khi da nh?tP xong cac du li~u cAn thiet eho qua trinh phan tich. Hinh 9. __ .4) Cac btroc song dung trong phan tieh: nh~p IAn hrot theo thir t\1" 6 biroc song.62- . Cira s6 chuang trinh phan tich da Hinh 9. ta hru lai bang each nhAn nut "Nhdn" hoac "D6ng l~p bang each chon "Thay y".6) De>hAp thu quang cua dung dich chuan va dung dich tlnr. Hinh 9..3) Ten cua m3i chAtthanh phan: Para + Quini .aua nghien Clfu __________ ::t. __ __ __ __ . (hinh 9.

.~~~.S.~. • - _ . .4.. Phdn tieh chudn Sau khi dA nh~p du dfl li~u phan tich ctra sf.63- .. b. tl6 ella dung K~t qua cua m6i lfin phan tich duoc bieu di€n trong khung "Nfmg phai dich chudn". rbi lAy gia tri trung binh ella cac n6ng de) tinh toan duoc. K~t qua phan tich eho 5 dung dich chuan h6n hQ'P tai sau biroc song dA chon diroc tinh toan va t6ng hQ'P trong bang 9. Dtr li~u n6ng dQ m6i thanh ph§n trong dung dich chuan h6n hop.. Nhap s6li~u v~ dQ hftp thu quang do duoc tai cac buoc song. Nong de) thuc Cr cQt ben trai va nbng de) tinh to an dircc nam ngay (] cQt ben canh.. Phsn mem chi cho phep phan tich n6ng de) ella tAt ca cac dung dich chuan tai hai biroc song cung luc.':. ••••• ~.' __I!:4!_ 9. ta bitt dftu ti~n hanh phan tich chuan bang each nhAp vao nut "Phdn tieh" tren thanh menu hoac tren thanh cong cu._1. ..' ._:.~_~~~:L: ~' ... ella sa phan tich dung dich chuan va tlnr.:~:_:. . do do tfch ci de do diroc tai 6 biroc song ta eftn phai phan = 15 lfin. #w·i. "'~'. Hinh 9.8.6.7._c:~ ---------------------------------_.~. Hinh 9...--------_. phan tich xuAt hien nhir dn thiet.'-': - '.l!t!_ t!g~j!''!..·_.__ :.----------------------------_. . •• ~~~·'::.. ~ .. Hlnh 9.••• < ••••• . . a hinh 9. •• . .

015 0.000 7.785 8._ :t_-__ 44!_ 9_l_I_Cf_ !l_g_~!~'!_!:~ Dung dich chuan heln hop Cl C2 . N~u dirong chuan chua thirc t6t thi ta co the t6i dung dich chuan. _.4. K~t qua phan tich 5 dung djch chuan. N~u dirong chuan da co chat Iirong.010 -0.10. S\l sai khac gitra hai gia tri n6ng dQ thuc va nAng dQtinh toan diroc th1JCt~ co the chAp nhan. c. Thimh phdn Para Quini Para Quini Para _Quini Para Quini Para Quini Nongdo th\l'~ml}_ 6.800 8.981 1. Nlnr 0 hinh 9. Loai bo mQt dung dich chuAn. Bang 9. T6i uu hoa duong chulm Ta co the kiem tra va t6i Uti hoa dirong chuan bing each co the thay d6i d€ sai s6 gitra n6ng de>th1JCva nbng dQ tinh toan diroc cang nho thi dirong chuan se cang chat hrong. Vi du: ta Uti hoa dira tren S\l thay d6i s6 co the bo b61 rnQt each ngdu nhien mot hoac hai dung dich chuan nao do.000 1.012 -0.014 -0. .020 1.1g/mlJ -0.200 Nong dO tfnh duqc~ml) 5._ .390 9.014 -0.209 SI! sai khac(J. Tinh toan lai dtrong chu~n.600 9.024 -0.978 2.986 1.000 1. no se bi loai bo khoi cQt "Dung dich chudn chon".000 1. .022 0.' _ .9 ta chon dung dich chuan 5 sau do nhap vito nut "«".976 1.009 C3 C4 C5 NhQn xet: Dua vito bang ta thay su chenh l~ch nong d9 cac thanh phan cua tat ca cac h6n hop chuan rna ph:1n mem tinh toan diroc so vai nbng de>thuc Iii khong dang k~ nen ta khong phai loai bo dung dich chuan nao. cit 5 dung dich chuan d~u co the dung cho phan tich MeA.020 0.000 2. thi khong cdn phai thay d6i lai.400 10.9.614 8.019 0. Hinh 9. ------__.64- Hinh 9.012 6.000 1.

5. tinh kh6i hrong trung binh vien.bo 20ml dich IQc d§.pthu quang tai sau buoc song da chon. Ket qua kiem tra. lAc ky.tnho.5mg Paracetamol cho vao binh dinh rmrc 100mI.unhu la r§. B.800 8. cu6i cung dem so sanh chung voi gia tri nAng dQthuc.l sai khac. can 20 vien._ -------------_. ddnh gia chdt luang dutmg chudn Dung pipet chinh xac d~ ph6i hop ill hai dung dich chuan g6c PG va QG. ta se do d<)h§. Dua vao su thay d6i ta se phat hien ra dung dich chuan gay ra sai s6 va loai b6 no.619 8.409 SlJ sai khactug/ml) -0. Dem IQC.u.022 -0. £)0 il9 hdp thu quang ella mdu thit Dem dung dich thir nay di do d<)h§.IN vira du.10. Kit aua __________ ::t. lAcdeu.:.010 0. lAcd6u.017 0.IN lam dung moi pha loang. d~ sieu am 10 phut. Kidm tra.000 1.400 Nong dQ tinh dUQ'c(~g/ml) 6. sir dung dung dich HCI O. Phan t{ch mbu thir Antigrip F .lAy chinh xac IOml dich IQcti~p then cho van binh dinh mire 100ml.4.978 1.IN.65- . them Hel 0. LAy chinh xac mot hrong bQt vien da: nghien min luang irng voi khoang 7. them HCI O.000 1. e4• Coi day la cac dung dich thir.y str sai khac h§. Nhpn xet: K~t qua cho th§.009 Thanh ph&n Para Quini Para Quini Para Quini Bang 9. nhir v~y duong chucin da xay dung diroc daub gia 1(\co chAt hrong t6t.p vao nut "Phdn tich". nghien ky thanh bQt min trong c6i sir. Chudn hi mJu thit Cao bo lap vo boc duong ben ngoai.6.983 1.019 -0. dung duong chuan da xay dung d~ tinh toan k~t qua .p thu quang cua chung. pha 3 dung dich h6n hop kiem tra co n6ng d<)cac thanh phan gdn gi6ng cac dung dich chuan C2.600 9.000 1. •:.5.010 0. NongdQ thlJc()J. --- Sau do ta nh§. •:.790 8. K~t qua duoc cho boi bang 9. nehien cuu 0 ----------------_ _ .. e3. danh gia dua tren Dung dich han hop kiem tra KTI KT2 KT3 S1. Cdch tinh kit qua Ham hrong cua Paraeetamol va Quinin sulfat trong mQt vien duoc tinh then c6ng thirc sau: .glml) 7.010 1. sau do sir dung dirong chuan vira xay dung dS tim lai cac gia tri v€ nAng dQ. d. danh gia chat hrong dtrong chuan. cac gia tri v~ nAng d<)se diroc tinh toan lai nlur 0 hinh 9. ducc dung dich tlnr . them 70ml dung dich Hel O. IN vira du. ghi lai cac gia tri dQ hAp thu quang d€ chuan bi nhap van phan mem.

__l!/!_ q~4_~g_~!~'!_ ~ ~ HL (mg/vien) Trang do: C 10 M : hi n6ng dQ cua Paracetamol : la h~ s6 pha loang. Vao phdn mem nay bing each chon bieu nrong cua no tren desktop. Tinh toan thong ke cac k€t qua thu diroc co the th1JC hien nhanh chong bing each sir dung cac ham thong Sir dung cong thirc (*) a phfut tren de tim ra ham hrong ke (statistical) Trong co s~n trong phfrn mem Microsoft h9P thoai "Insert Function" Excel. Kit quiz phdn tich tren mdu Antigrip F Sau khi phan tich tren may tinh ta se thu diroc gia tri v~ n6ng de) m6i thanh phan trong dung dich thir (ug/ml). chon Insert\Function. (*) = m m : la luong bqt vien da: can trong dinh hrong (gam) .11._j_. hoac Quinin sulfat doc duoc tren may (ug/rnl). Xu ly thong . hoat chelt trong mqt vien. M C x 10 x - . . •:.11) MQt s6 ham thong dung: Gia tri trung binh DQ I~ch chuan Phuong sai So sanh hai phirong sai So sanh hai gia tri trung binh ham Average ham Stdev ham Var ham Ftest ham Ttest chon mQt 0 belt ky chon "Statistical". (hinh 9.66- ke tren Excel. : Ia kh6i hrong trung binh cua mQt vien (gam). Hinh 9. (cell).

trong thuc nghiem nay chung toi dtl chon plnrong phap do quang ph6 ill ngoai dinh hrong rieng hai thanh phdn Paracetamol va Quinin sulfat lam plurong phap so sanh.7.98 0.03mg a 29. . Nhu v~y phuong phap phan tich MeA co th~ su dung d~ phan tich mdu Antigrip F. tren mdu Antigrip F.51 30.41 29. sai s6 tuong d6i my thee tirng tnrong hQ'Pla 0..71% d6i voi Quinin sulfat. Con dQ dung duoc xac dinh thong qua kha nang thu h6i cua k~t qua phan tich khi dung phuong phap them chuan.49 0.68 :to.34 30.60 0. .68 0.13 147.90 147.40 0.51mg_ Dc) l~ch chuan Sai so chuan Sai so nrong doi Khoang tin c~y n=6'p=095-1'J .03mg con Quinin sulfat la 29.67- . Ti~n hanh so sanh d9 chinh xac va dQ dung cua hai plnrong phap thong qua test F (so sanh hai phuong sai) va test T (so sanh hai gi:i tri trung binh).69 28. phirong phap B). =257 .6.70% d6i voi Paracetamol va 1. D~ danh gia phuong phap MeA. tIP chinh xdc elia phuang pluip B.60:t 1. .:.53 29.43 147. Bdnh gid dp dung.theo cac duoc di~n hoac sai s6 nrong d6i). NhQ.93 148. Chung ta cling co th~ danh gia mQt phirong phap phan tich (Phuong phap A) b~ng each so sanh k~t qua phan tich voi met phirong phap phan tfch khac (duoc tien hanh cung phong thi nghiem va cung khoang tho. ciing nhir cac ch~ pham chua hai thanh phfm nay co tY l~ gdn giong nhir tren._~·__Jf4!_q1!_4_!!g_~!!_'!_!:~ Bang 9. SKS: 050801.71 29. Nguyen tdc M{>tphuong phap phan tich thirong diroc danh gia bang each khao sat d{>dung (accuracy) va dQ chinh xac (precison).mg_ hoat chat fronK m(>tvien (m_g)_ Quinin sulfat Paracetamol 29. _ qua dinh hrong S6 thutt.60 ± 1.68 ± 0.70 147.n xet: K~t qua cho thAy de) l~p lai vci cit hai thanh ph§n la nrong d6i t6t. gian da:dtroc danh gin va chap nhan.51mg voi rmrc tin c~y 95%.85 146.r 1 2 3 4 5 6 Tinh thong Ire Gia tri trung binh X Ham /llt. Ham hrong cua Paracetamol trong vien la 147.69 148. K8t .10 146. D{>chinh xac thirong dtroc xac dinh thong qua d{>l~p lai cua phirong phap (each thong thirong nhAt la xac dinh d{>lech chuan nrong d6i cua k~t qua phan tich RSD .20 1.

33 < 1'(0.9). Phuong y nghia thong ke 6 mire a phap x n S2 Paracetamol UV(B) 149.1O = Test t Nhfjnxit: 1..__ __.-...--- -- -- -- -- -_.79 29.9.05 (Bang 9.93 29..05.23 2.05 FTN 0. Bang 9.71 < 1.68 6 0.5.00 % 99. So sanh dQdung.77 29. y nghia So sanh 2 gia tri trung binh (test T) cho thdy k€t qua dinh hrong gitra hai phuong phap khac nhau khong co y nghia thong ke d6i voi cit hai thanh phftn Paracetamol va Quinin sulfat (P=O.. Kit _:t _ _. STT 1 2 3 4 5 6 Paracetamol (mg/vien) 149.79 150. _ _.4 _.23 - So sanh hai phuong sai bfing test F cho thdy chung khac nhau khong co thong ke (P=O._~ ..10=2.05 Test F SI. .98 148.38 ITN= = 5. tu« hanh Hai thanh phan Paracetamol va Quinin sulfat da: ducc cac tac gia dinh hrong bang phirong phap tach rieng hai thanh phfut sau do do quang ph6 nr ngoai..84 149.65 28. .. Ket qua khao sat dQdung theo phtrong phap B.05 Quinin sulfat (mg/vien) 28..S..60 6 0.95).05..23 6 0.5) Quinin sulfat UV (B_) 29.06 151. Bang 9.__.35 MCA(A) 147.. Ket qua khao sat dQ l~p lai cua 6 miu thir theo phuong phap B.67 6 1.32 28. de>chinh xac cua hai phuong phap.2 FTN =0.49 ITN = 1. __ cuu aua nghien __ __.96 1'(0.68- .95.5. Nhir vfty de>dung va dQ ehinh xac cua phuong phap MCA khong khac so voi phtrong phap do quang ph& hap thu tir ngoai dinh hrong rieng tirng thanh phdn.95). Paracetamol (n = 5) Quinin sulfat (n = 5) 99.7. __. .46 148.__ .5) = = 1.03 % Bang 9.24 5. cac tac gia da: khao sat dQ dung va dQ chinh xac.17<10.S...95.18 MCA(A) 29.89 Ket qua so sanh gitra phuong phap MCA va phuong phap quang ph6 thong thuong (da diroc danh gia) cho thdy su khac nhau khong co = 0.:.7 va S. Ket qua phan tich dircc trinh bay trong bang 9.94 < to.

Khiio sat phD hap thfl UV va i1!'achpn cdc bUtTe song phlin tich Cit PA va ME d~u co kha nang hAp thu quang trong moi tnrong kiem va moi tnrong acid! ethanol. may quang ph6 ill ngoai kha kien GBC Cintra 40 (Australia).. Thi~t bi: can phan tich METTLER AB 204.69 - .8 (theo tieu chuan DDVN III .. 100ml. ME chuan (ham hrong: 99.. I~c d~u dircc dung dich co nong oQ khoang 20J. may l~c sieu am BRANSON 5210. Do do chung toi khao sat dQ hAp thu cua dung dich PA va ME trong moi tnrong d~m phosphat 5.. gifty IQc. may quang ph6 Hitachi U-321 0 (Nhat Ban). A. . mircopipet: 10ml._ -. Dung pipet hut chinh xac 5ml dung dich vita pha cho vao binh dinh rmrc 50ml.IN. dung C\l Vien bao phim Pamesic cua Tiphaco sf>10 san xuat: 02 02 03 Cong thirc: Paracetamol Acid Mefenamic Ta diroc 500mg 250mg vira du vien Thu6c thir: ethanol 96°."Thubc thir").1. c6c co mo. Them 120ml ethanol 96°.8 vira du. d~m phosphat 5. may thu dQhoa tan PHARMA TETS PT III. Them dung dich d~m phosphat 5. 4. Hic sieu am d~n tan hoan toano B6 sung ethanol 96° vira dti den vach. B.__ O~__ . lOml._ . MOi trutmg d?m phosphat 5. dAng tbiri Paracetamol 1 va acid Mefenamic trong vien nen Pamesic bing do quang pho MCA Ngoai phirong phap HPLC da trinh bay i:J tren.__ -- _ - -_ .10. Kit__qua nehien_ cUu ___. - Dung cu: pipet lmI. Cac ta diroc cua vien (Tiphaco cung cAp).8 va mei tnrong NaOl-l O.. ChAt df>i chieu hoa hC)C:PA chulm (ham hrong: 99.8 - Dung dich PA: Can khoang 40mg PA cho vao binh djnh mire 200mI.._ " " _ __ __ _ __ __ .BQ Y t~ cung cap). trong d~ tai nay chung toi gioi thieu mQt k€t qua nghien ciru trng dung plnrong phap MCA diroc sir dung tren phfm mem sAn c6 cua may quang ph6 GBC Cintra 40 d~ dinh luong d6ng thai paracetamol va acid mefenamic trong san pham thuoc Pamesic. •:. " _.. 5ml.. Hic d~u duqc dung dich P' 0 c6 n6ng d<) khoang 200jJg/mI. dung dich NaOH O. Thu&c thif. 21. binh non.Djnb hrqng . Xay dung phuong phap djnh hrQ'DgdAng thiri PA va ME bing phuong phap quang pb6 MCA B.52%). 2ml.68% do Vien Ki~m nghiem . hoa chit._ 4.20~g/ml). binh dinh mire cac loai: 20ml.1g1ml(n6ng dO thuc h123.IN .. ph~u. 50ml..

. Them 120ml dung dich NaOH O. ~ .Lglml.~ .Lglml nong dQtlnrc lit 15.IN. Them 120ml dung dich ethanol 96°.84J. l~c deu duoc dung dich co n6ng de>khoang l Oug/ml (nong de>thirc la 9.1 va 10.lN vira du d~n vach.1. l~c deu diroc dung dich P'l co n6ng dQ khoang 200J. Them dung dich NaOH O.IN vira dii.. Dung micro pipet hut chinh xac 4ml dung dich vira pha cho vao binh dinh rmrc 50mI.... ( Dung dich ME: Can khoang 20mg ME cho vito binh djnh mire 200mI.. l~c sieu fun d~n tan hoan toano .. ~ •• ~".Dung pipet hut chinh xac Sml dung dich vira pha cho vao binh dinh rmrc SOmI.Lglml).8 vira du. Them dung dich NaOH O..2.IN.2.~.65~glml) ..~J""".~ :- Hinh 10. Hic deu dtroc dung dich M' 0 co n6ng de> khoang 100J.~"..8 (8. l~c deu dUQ'cdung dich co n6ng de>khoang 16J. Phi>cua PA (l5. Them 120ml dung dich NaOH O. PA ME ~~.IN vira du d~n vach. ~ .IN ..:z- . Quet ph6 cac dung dich tren (] buoc song til 220nm d~n 400nm va chong ph6 cua hai ph6 P A va ME 6 cung moi tnrong ta duoc cac ph6 db nhir hinh 10. moi wang d~m phosphat 5. Phi>cua PA (23.84). l~c deu duoc dung dich M'J' Dung micro pipet hut chinh xac 4ml dung dich vira pha cho vao binh dinh mire 50mI. •:.Lg/ml. l~c sieu am d~n tan hoan toano B6 sung dung dich NaOH O.70 - . Them dung dich dem phosphat 5.18Jlg/ml).65J.Lg/ml)..1 8Jlglml) trong moi truong NaOH O._(!f4!_q_l}_4_1}_g_~!~1J_ 5:~ Dung dich ME: Can khoang 20mg ME cho vao binh dinh mire 200mI.lN Dung dich PA: Can khoang 40mg PA cho vao binh dinh mire 200mI. l~c deu duoc dung dich co n6ng dQ khoang Sug/ml (nong de>thuc la 8. B6 sung dung dich ethanol 96° vira du d€n vach.20 ug/ml) va ME Hinh 10.. l~c sieu fun d~n tan hoan toano B6 sung dung dich NaOH O.lN vira dii..lg/ml) va ME (9. MOi trutmg NaOH O.

Dc) hAp thu quang cua dung dich (] moi tnrong base t6t hon moi tnrong acid duoc chirng minh ca trong ly thuyet va thuc nghiem.8 279.0006 Bu6c song (nm) 257 H~ so 285 0.3 Abs) 205.IN se thao tac pha ch€ don gian hem so voi dung dung dich d~m phosphat.0060 y = 0.2986 0.43 360. a -_.1..8nm voi dQhAp thu la 1. __.5 - Trong moi truang NaOH O..1 0.IN.IN vita du den vach H22 2 2 H33 I I Do de> hAp thu quang cua cac dung dich thu diroc (] 2 biroc song lei 257nm..0213x .2.999985 ME Phuong trinh h6i qui y=ax+h y = 0. .3479 1. Mat khac khi tien hanh voi dung moi la NaOH O..1269Abs (All 334.-------__.2.H33 Bang 10.lN PA c6 dinh pic ME a 257. 4 ql!. .0._.0715x + 0. Vi v~y.4 220.999998 Phuong trinh h6i qui y=ax+h y = 0.1682 0.0008 y = 0.q_ _c::~ - . M2._f_.-----_.1.71 - nrong quan (r) 0.. P3.2953 = 309.0366x .5 285..2. voi cac dung dich chuan PA la Ph P2.0. Til ph6 d6 ta chon dai song phan tich til 220nm d~n 300nm.52 134.0004 . cac dung dich chuan ME lei Mj.__ ~!_ ~g_~!~'!.23 714. ---. 285nm.8 PA c6 dinh pic ME 246.8 0. Khiio sat khoiing tuyJn tinh va tinh chAt cpng phJ cua PA va ME Pha day dung dich chuan va dung dich chufut h6n hQ'Ptheo bang 10.---------_.0.---------------------- Trong moi trutmg ilcrn phosphat 5.0346x . K€t qua khao sat khoang tuyen tinh PA Biroc song (nm) 257 H~ so nrong quan (r) 0.5713Abs (AI 1= 677.3407 Nhin xet: . Pha day dung dich chuan va dung dich chuan h6n hQ'P Dung dich chuan I PI I P2 I P3 I M1 1M2 1M3 HI1 So ml can lay vito binh dinh mire 50mI -I P'l(ml) 5 12 15 I1 l l M'l(ml) 5 12 15 II I I I Them dung dich NaOH O.999996 0. B. chon moi tnrong thuc nghiem lei dung dich NaOHO. K€t qua thu diroc a bang 10.999999 285 . M3 va cac dung dich chuan h6n hQ'Plei HI h H22..6 416.Inm voi dQhApthu la 1. Bang 10.3 Abs) 353.

. __ .. Kc§tqua khao sat tinh chAt cQng phd BUGe Nong dl) (ug/ml) PA-ME 20 ..IN vao hai binh dinh rmrc tren dc§nvach.4355 0.._.. .. ..5818 0..3652 0. ...wml) p 11(pIg/ml) 8 10 12 14 16 Mll(!Jig/mI) 8 7 6 5 4 . I~e d~u diroc cac dung dich luang (rug la Pll va Mll..l Oug/ml._-_ - Bang 10.IN vao m6i binh...0845 0...__ .1 N d€ pha loang thu diroc day dung dich nhir sau: Bang 10.6922 0.331 1..88 100..2117 0..-.666 0.2894 0..02 99.72 - CI . bd sung dung dich NaOH O.648 0.. c Trong dung dich h6n hop PA va acid ME dQhdp thu quang co tinh eQng tinh. . sir dung dung dich NaOH 0.. Diiy dung dich h6n hap chudn Can 50mg PA ehuin vao binh dinh mire 50ml (ky hieu la PI)' Can 25mg ME chuan vao binh dinh mire 50ml (ky hieu la MJ)' Them khoang 30ml dung dich NaOH O.0725 TongdQ Mp thu PA+ME 1. l~e d~u tao hai dung dich luang dirong la PI va MI....057 0..1461 0.058 0.20Ilg/ml va cua ME Ia 2 . Kit qua nehien cuu _ 0':" ......0415 0. binh MI vao binh dinh mire 50mI...647 0.lPA 1...21 DQ hap thu h6nhqp PA. Vi v~y chon nong dQ dinh IUQ'I1gua PA la 4 ..28 Thdng ke XTB = 99.422 0.__ ..__________ 4..211 HH xl00 T 100. B...IN vira du.10 8-4 4-2 20 -10 8-4 4-2 song (nm) 257 De>hap thl... Uie d~u dc§ntan het....:.05% 0=3 XTB= 100.332 1.ME 1. Ldy chinh xac 5ml Ldy chinh xac Sml (y (y binh PI vao binh dinh mire 50mI. Them dung dich NaOH O. Dung micopipet ph6i hop 2 dung dich Pl1 va MI J vao binh dinh mire 50ml.665 0.. Xay dtplg phuang phap i/inh 1U'(TIIg dang Ihm PA va ME b&ng plucang phap MeA tren may quang phd GBC Cintra 40 a...._.98% RSD=O. ...4.07% RSD = 0..05 99.03 100.. .94 100....9999) gitra n6ng dQ va dQ hAp thu cua PA va ME trong khoang nbng dQ khao sat.285% 0=3 285 Nh{in xet: Kc§tqua cho thAy co Sl1 nrong quan tuyen tinh rAt chat che (r = 0.2767 0..-. Chudn bi mdu .3. Pha day dung dich chuan d€ xay dung dirong chuan Dung dich hon hop chuan C2 C3 C4 Cs So ml dung dich chuan goe can lay vao blnh dinh mire 50ml Pl1(ml) 4 5 6 7 8 Mll(ml) 7 8 6 4 5 Nong dQ cua mlii thrum phdn trong_ dun_gdieh chuan han h<JP nrong imgf... __ -...1379 De>hap thu ME 0.423 0.3.

3.ld~n vach diroc dung dich thir. tren man hinh hien thi cira sA chuang trinh nhir hinh 10.:.:. •:.IN.IN vita du. Hlnh 10.:. Dung dich thir Cao lap phim boc ngoai cua vien. chon irng dung Multicomponent.IN vira du. . Thi~t l~p cac thong s6 cho phirong phap trong muc "setup" -<} Method .3. L~y chinh xac lOml dich IQcvito blnh dinh mire lOOml. Vito Menu APplication. them dung dich NaOH O. xac dinh kh6i hrong vien trung binh.. din 10 vien.IN vita d1... L~y chinh xac mQt hrong bQl vien tirong trng voi mot vien chua khoang 500mg PA va 250mg ME cho vao binh dinh mire lOamI. L~y chinh xac l ml dung dich thir 2 nay cho vao binh dinh rmrc SOmI. be. trong 6 "Description" cua cira s6 nhap ten phirong phap phan tich mdu. Nghien ky thanh bQt min trong cBi sir.them NaOH O. I~c sieu am 10 phut.73 - . b.__!:~!_ql}_<!_ !:!g_~j!_'!_ _c_. Trong muc "General": + S6lAn I~p lai (Number of repeats): + sa IAndo lai mdu chuan (Number 1 of standard replicates): 1 ug/rnl 1 + S61An do lai mdu tlur (Number of sample replicates): + Don vi nong dQ (Concentration units): . them khoang 70ml dung dich NaOH O. Ldp phuong phdp phdn tich tren may B~t may Quang phA. Hic d~u dem loc. 20ml dich loc dAu. lAc d~u duoc dung dich thir 2.!!! 1 . Dung dich kiim tra -. them dung dich NaOH O.-__. nhay dup vito bieu tuong chuong trinh do Quang cua may Quang ph6 GBC Cintra 40 tren desktop cua may tinh di~u khien ._~._ Dung dich kiem tra I(ky hieu lit KTI): 7ml dung dich Pl1 + 6ml dung dich Ml1 Dung dich kiem tra I(ky hieu lit KT2): 6ml dung dich Pl1 + 7ml dung dich Ml1 . Cira sA "Application Control" .

q_ ~g_~!~~. Lap duong chudn tren may. Hlnh 10. do mdu thit . ... . ... Cua s6 "Sample Setup" . . Trong muc "Component": Nh~p ten cac thanh phfut tham gia phan tich: PAvAME. .74 - ....:._. V~ tfic quet (Scan speed): lOOO.6. -. mdu cAn do va nbng dQ thuc cua chuan.._ f._·__If~!_ q1!. . .5.. _.4. ... Nhap cac dir lieu v~ ten cac chuan. Hop thoai "Method Setup" Hinh 10. Trong muc "Algorithm": -¢- Chon "K matrix" Instrument . Cua s6 "Instrument" c. Hlnh 10.Onmlphut.. Nhftp nut "Sample" tren khung "Setup". Dtt li~u nao la chuan thi tich dftu (X) vao c<)ttrong muc "Std" cua cira s6. kiim tra chdt luang duong chudn. £~ _. Phuong phap do (Operation Mode): chon quet ph6 (Wavelength scan) . Ki€u do quang (photometrict Mode): chon m~t d<} quang (Absorbance) . DAi song (Scan range): chon ph6 til 220 d€n 300nm.

7 .4.8. may 10._1_·_!f4!_q'!_q_!'_g_~!?_1}__C:~ •:.S.10 va dUQCxu Iy se thong bao k~t qua phan tich nhu hinh a bang 10.3 man hlnh se xudt hien hQP thoai nhir hinh 10.9.75 - . cho dung dich mdu tlnr vao thi~t hi quang hoc r6i nhdp nut "OK" trong h<)p thoai nhir hinh 10.9. Hinh 10. Hinh 10. Hop thoai xac nhan nhap mau thu Sau khi do xong cac dung dich.:.5. £)0 dung djch KTI va KT2 d~ kiem tra chdt hrong duong chuan cua may da xay dung bfutg each cho mdu vao thi~t bi quang hoc rbi nhap vao nut "OK" trong hop thoai nlnr hinh 10. £)0 m§.7.6 va bang 10. sa hinh 10. Sau m6i IAn do xong mt)t chuan lai c6 mot h<)p thoai tuong tir va l~p lai qua trinh nhu chuan thu 1 d6i voi cac chuan thu 2. •:. Cach tinh kSt qua: Ham hrong cua PA ME trong vien tinh theo cong thtrc: Mg/vien Trang do: C 5000 : m He s6 pha loang m N6ng de>cua PA hoac ME dtrec doc tren may Kh6i hrong b<)t vien dff diroc can =C * 5000 * M TB MTB: Kh6i hrong trung binh mt)t vien . •:. Nhay num "Run" tren cira ----------------- .3.u thir. HQP thoai xac nh~n nh~p m~u kiem tra Hinh 10.7_ Cho chuan tlnr 1 cAn do vao thiet bi quang hoc r6i chon "OK" tren ht)p thoai nay. Hop thoai xac nh~ nhap chuan .

24 14. N.ev-..aeJ~ Olea .19 6.__ __ __ __ .41 ~_13:T~ S11 5.5...42':0.n xit: K€t qua cho thdy s\!' sai khac gitra nbng dO thuc va nang dQ do dtroc rdt nho.D (ug/ml) 12. Vi v~y dirong chuan dugc danh gia lit kha t6t va dang tin cay.. III U1 N:l_ 6: 1Cl'1 NJ. "'wleiJIi : DiCO .~-: 1~17 14'B 1 1 Ml K1IfIIm 1 ! S&td!tdl: 111> U4 susn 4: 1117 7.24 N6ng de>do diroc .JlOIIMIi: ~_t -~~ SIlIIiIn!I: SIl!ltatd 1: .. .lO:B lOS5 U.N'«~ N.aIIpnU: NIIu· d. K€t qua kiem tra drum gia chdt Iuong dirong chuan Dm.14 7.__ .d: 31it~'dh: 6.__ 4.b!r o!iiaIml~ U!le~ Dim reriOal: Ntab!!fd aepJt npliCiIei: Slimlifa C~batU N.23 Sai sA cua phirong phap 0..Tlff*100% ME PA ME Nh.__.a!II.tg djch kiem tra KTI KT2 TP PA N 6ng de>thuc .71 dQC Hinh 10..llT4 ~ UT5 Iua 1139 11.!:m 11141:24 }419 6.49% 0.. 1()Q') tIIl.~M! .. K€t qua tren may GBC Cintra 40 Bang 10.28 14......T (ug/ml) 12.__ Q __ __ StiG~ _ __.afa4 5.kd!: N. khong dang k~.&iIl !Qs..k<t_Mk Nalw d t..28 416 5':» S.1~ lJ.nmew: p l.__.28% 0.55% 0.13 i!'9 ill ill i96 N:J ~: Tl 1137 l'\b..l~ Rl.10. Kit qua nzhien cuu . __ __ Or __ __ .. 100_ ~ ~~ B9IIlr.20 7.76 - .23 6.16% ID .

74 5.26 12.96 5.05 Gin tri tnmg binh DQ l~ch chudn Sai s6 chuan Khoang tin c~y Sai s6 tuong d6i N6ngdQ Xu ly thong ke do dtroc (ug/ml) 5.39 11.7.90 505.8935 1.90 98.71 Gin tri tnmg binh 5.1227 0.98 98. Pamesie ta co k~t qua 6 bang 10.5 99. .89 5.78 101.30 97.9853 0.51 98.0311 1. B.0120 1.S.6 ta thfty ham IUQ1lgcua PA va ME trong vien mau Pamesic hoan toan dat chi tieu chftt hrong (PA til 98.23 2 3 4 5 6 Til bang 10.88 247. Dung dung dich da dem xac dinh dt) l~p lai lam dung dich thu (T 1) co n6ng de> ehinh xac 13.62 5.17 12.81 5. Bang 10.7.3%). SIT 1 mCan (g) 0.55 ME 255.37 10.35 98.91 ME 102.77 - .82 5. B.55 12.85 DQ I~ch chuan Sai s5 chuan Khoang tin c~y Sai s6 tirong d6i 0.35 Xu ME ly th5ng ke 12.74 5.82 101.:.26Jlg/ml).54 99.72 .71 PA 98.05 253.73 5.78 d~n 102.62Jlg/ml).p l{li clia phuang ph tIp Ti~n hanh pha va phan tieh 6 mdu thu.38 12.9%.88 11.75 244.16 11.41 PA 493.PA (12. Dt) l~p lai cua phtrong phap PA STT 1 2 3 4 5 6 N6ngdQ do diroc (ug/ml) 12. II 5.1 102. ME til 97.4.n xet: K~t qua khao sat dt) dung cua plnrong phap eho thfty phirong phap cho k~t qua dt) lap lai kha t6t voi cit hai thanh phdn PA va ME._ (Fl!_ q_7!_q_~g_~!~.9798 0.!_~~ Bang 10.45 492.1288 1.93 248.98 d~n 102. K~t qua phan tich mau Pamesic Nang dQ do diroc (ug/ml) Him hrong tinh diroc (ug/ml) ME 5.55 494.09 12.0762 0.0320 0. ME (5.80 Nh.30 514.6.27 0.3947 0.78 5. Xir 19 nhu do mau Pha chinh xac mot dung dich h6n hop thir nrong irng voi 12(Jlg/ml) PA nhu muc phan tich mftu Pamesic.9876 % dat diroc (%) PA 11.0501 0.39 12. XtIc dinh tIP lq.95 245.70 497. Xtic dinh dp dung clia phuang ph tIp Ti~n hanh: .

Hoa tan va them vao vira du d~n vach bfutg NaOH O.2094 Xu 1j thong ke DQ l~ch chuan Sai s6 chudn Khoang tin c~y Sai s6 tuang d6i Nh{in xet: K€t qua khao sat dQ dung cua phuong phap cho thAy phuong phap co dQ dung cao.87%.2150 0.11 4._f~:4!_9.65 99.19 7. Them Iml dung dich chuan C vao binh TC 1 2 3 dirong chuan vira tao. Them NaOH O.54 99.8._i. ty l~ thu hdi df>ivoi PA la 99.03 3.9597 2. Til dung dich Tl hut chinh xac 50mI cho vito 3 binh dinh mire 100mI dtroc ky Can chinh xac 30 mg PA va 15 mg ME vao binh dinh mire 100mI.2230 0.61 99.53 6.78 - .------.IN.9609 2.2972 0.25 0.67 100.1288 0. TC3_ --- ------__---. DQ dung cua phirong phap Nong de) do dircc Mdu do TC1 TC2 TC3 (Ilg!_ml) PA Nong de) them vito _(a) (Il_gt'ml) PA Nang de) thu hoi (b) (ug/ml) PA Ty l~ thu hoi a/b*100% PA ME 9.34 12. .40 9.:.87 ME 100.02 ME 3.59 Gia tri trung binh ME 99.87 15. ME (150 ug/ml) .8.43 100. TC3_ Bern do va xu Iy tren a bang 10.04 3.IN vira du cac binh TCI. df>ivoi ME la 100. •:. TC2. Bang 10.05 100.1!/!_ !!g_~!~'!_~z!!!_ hieu lit TC 1.06 6. Hic deu diroc dung dich chuan C co PA (300 ug/ml). TC2.-----_. •:.-----.06 9.25%. ta thu duoc k~t qua Te2 TC3 .07 4.5148 0.67 1.01 1.83 3.33 4.53 5.

.79 - . .mroc . kY long pha dao voi he dung moi phan C(J CQt RPI8 la loai cQt thong dung trong cac s6' kiem nghiem da diroc sir dung d~ phan tich cac duoc pham Qua nghien ciru..-.2 bang acid phosphoric. Dinh tinh va dinh hrong d6ng thai paracetamol (PA) va ibuprofen (IBu) chung toi chon diroc di€u kien sAc kY sau: CQt Lichrosorb RP 18 (2~Ox4 mm. dQ dung t6t (zy 1~thu h6i 98. .. 5.16 - 99.. ?l!<J!!.TC hoac chdt co tinh acid).1.. = Phirong phap da duoc daub gia la co dQ chinh xac cao (sai s6 nrong d6i % 0.. lOJlm).g __ __. Da s6 cac chAt phan tich (cac thuoc) da khao sat la nhtrng chAt co tinh phan (la cac muoi cua base htru chirong trinh sllc khao sat. khao sat bang tlnrc nghiem. - -. Phuong phap da duoc irng dung lam tieu chufut cho vien nen bao phim Pamesic. Detector UV d(lt 6' biroc song 224 nm. Vi v~y chung toi thuong chon Cl. 10Jlffi)...c!!!. . nhfrng noi da duqc trang bi may sllc ky long hieu nang cao. Uu di~m chung cua phirong phap HPLC Ia co su 6n dinh va co de>chon lee cao.. Tinh tuyen tinh d&direc khao sat trong mQt khoang luang d6i rQng.TC.2. . /J.16%). . C:!T!:~. T6c de) dong 1 ml/phut.51-1.. Tinh thich hop cua h~ thong dat yeu cdu..· :5. va co th~ ap dung rQng rai cho cac trung tam kiem nghiem thuoc. dtroc sir dung rAtrQng rai trong phan tich va kiem nghiem diroc pham.. Phuong phap dinh hrong d6ng thoi paracetamol va acid mefenamic da duoc xay dung voi cac di€u kien sau: CQt Lichrosorb RP 18 (250x4 mm.1.acid phosphoric 0. C(J Cl.tetrahydrofuran (3 : 8 :9) di~u chinh d8n pH 2. chi cAn lua chon diroc dung moi hoa tan va pha loang thanh cac dung dich thir (hoac dung dich chuan). Th~ tich tiem 20Jll. Viec chuan bi mftu tlur cilng don gian. 5. T6c dQ dong 1 ml/phut. Detector UV d~t a buoc song 279 nm.. chung toi d& xay dung duoc chuong trinh phan tich bang HPLC cho cac h6n hop nhir sau: 5. Phuong phap sic ky long hifu nang cao (HPLC) Day la mQt phirong phap phan tich hien dai. Pha dQng: Acetonitril..1.1 % (60: 40). . BANLU!N CHUNG 5._.1.'!. Pha dong: Acetonitril . . ti~n hanh nhanh chong va co th~ ap dung d~ dinh tinh va dinh hrong cac hoat chdt trong h6n hop rna khong ccln phai chiet tach rieng biet.42).

. Phuong phap xu Iy m§. Detector UV: 235nm.. H~ s6 nrong quan tuyen tinh r=0.3. Han hop paracetamol (PA) va cafein (CA)..5 %.3%.__ .0. Pha dQng: 1. 10J.mroc (35: 65).-. di~m han va co th~ Slr dung cho cac cong thirc thu6c khac c6 thanh phfuI nrong nr._ -_ -_ . - -_ --_ -_ -_ --.--_ ..lg natriheptansulfonat 4%-diethylamin ti I~ (650: 350: 1).. dQ dung cao (ty l~ thu h6i 98% . -_ -_ -. cac pic c6 dQ phan giai ro rang. - Th~ tich tiern 20J..ll. Detector UV d~t 6 buoc s6ng 273 nm. trongl lit h6n hop methanol-acid acetic Tren sAc ky db.9999)..9 den 99.1.. Sum)..4%). 5.--.. -Elm . Tuy dien tich pic cua cac hoat chdt thu ducc co dQ chenh I~ch I6n do ham hrong cac hoat chAt co tY I~ khac nhau va co dQ hdp thu tu ngoai khac nhau... cua quinin sulfat la 1.-~---. phan tach t6t cac chdt trong h6n hop...98. _ ..81). phfuI tram tim lai cua PA va CA tren hai m§.-- ludn chung -..80 - . Phan tich dbng thai paracetamol (PA) va quinine sulfat voi dieu kien sAc kY la: CQt Lichrosorb RP 18 (250 x 4 mm. Th~ tich tiem 20111. Ti~n iru hanh so sanh ket qua voi phirong phap chiet tach (tieu chuan co sa) ta thAy phuong phap da sir dung d~t dQ chinh xac nrong dirong.16%. T6c dQ dong: lrnl/phut..0% den 99.lm).07 %).u til 98. DQ l{lp lai voi sai s6 nrong d6i cua paracetamol la 1. dQ l~p 5& tuong d6i 1. Th~ tich tiem: 20111. 5. -.54 den 1.5...88% va dQ dung tir 99. co nhieu che pham Alaxan hru hanh tren thi tnrong.u dan gian._ . Phuong phap c6 th~ ap dung d~ dinh hrong dbng thai PA va CA trong thuoc c6 chua hai thanh phan tren.. d€ thuc hien trong thai gian ngan.9999 voi met khoang khao sat kha rong cho phep ap dung voi nhieu ch€ pham nrong tu co tY I~ cac thanh ph§n khac nhau. vay phirong phap c6 dQ dung t6t. dieu kien sAc ky da ducc khao sat bao g6m: CQt Lichrosorb RP18 (250x4 mm. Nhiet dQphong. .4. T6c dQ dong 1 ml/phut.03 ..-_ -_ -_ -_ . Va irng dung d~ phan tich K~t qua khao sat danh gia cho thdy phirong phap co dQ l~p lai t6t (sai s6 urong d6i til 0. = Da khao sat tinh tuyen tinh trong khoang n6ng dQ phan tich (r lai t6t (sai 0.1.75% trong han hop dung moi methanol..-. Pha dQng: Dung dich acid phosphoric 0. Nhimg cac pic each xa nhau chirng to khong co str anh huang Ifillnhau khi dinh hrong.

Co thS dung chuang trinh nay de dinh tinh va dinh hrong d6ng thai paracetamol. Pha dt}ng: Hoa tan 1.1. H6n hop 3 thanh ph~n gdm co paracetamol (PA). 5. Pha dQng: Hoa tan 2. theophyllin va _ phenobarbital chung toi dung chuang trinh sAc ley nhu sau: Da:ti~n hanh khao sat tinh tuyen tinh. i. Da: img dung phan tich co k~t qua tren hai miu vien nen va vien bao duong la vien Etadasmyl. dQ dung.6. hydroc1orid.67%. Tuy nhien trong thirc t~.dem phosphat pH 3. Detector IN d~t 6 biroc song 240 nm.81 - . cua CM la 1. . 5.7 bfuIg acid phosphoric. T6c dQ dong 1.2 ml/phut.4 7%. SKS : 010600. sir dung chuang trinh sau: May sAc ley long hieu nang cao DIONEX-Detector IN diode array.~!!_q_~!:'. CQt Nuc1eosil C 18 (250 mm x 4 mm. . Kha nang tim lai duoc kha t6t (dao dQng til 99. T6c dt} dong 1 ml/phut. phenylpropanolamin hydroc1orid (PH) va clorphenylamin maleat (CM). cua PH lit 0.-Ull).1}g f Phuong phap nay co thS dung d€ dinh tinh va dinh luong cac thanh phan trong h6n hop thay cho phirong phap chiet tach trong tieu chuan co-se. SKS : 011099 va vien Typhasmyl.19%. di~u chinh d~n pH 2. 5 urn).triethylamin (968 :30 :2).Th€ tich - Nhiet dQphong. Vi v~y phirong phap nay co th€ irng dung thay cho phuong phap cua tieu chuan co s6.r 244nm dS do PA..5_ Han hop thuoc co 3 thanh phful ephedrine Ct}t Lichrosorb RP 18 (250x4 mm. Detector UV d~t a biroc song 216nm dS do PH va CM. Th€ tich tiem 20l-l1.65%). 10J. Sai s6 nrong d6i cua PA la 0.5 g kali dihydrophosphat trong 1 lit h6n hop cua mroc acetonitril . dQ l~p lai. do ham paracetamol la qua cao so voi hai chdt con lai nen phai do rieng chdt nay voi dung dich tlnr da: dircc pha loang 50 lfut. phenylpropanolamin hydroc1orid va clorpheniramin maleat trong vien nen.0 g natri heptansulfonat trong l Iit han hop cua methanol .61% d~n 100. Phuong phap dinh co dQ l~p lai cao.5 (38: 62).1._~·__J_q!!_?lf-. tiem: 20 I-ll.

- _ ... - - - - - - - -- - _";. _.- - - -- - - - - -- -

5. es« ludn chung

-- ......

-,._

- - - -- - - -- --- - -- -- - - -- -- - - - -- - -- - -- - - - -_ .... --_ .... - - - - - - _ ....

-- -- -

... .. - - -- - - -

-- - - - - - - -- -- -- -- - - -- --_ .. -- _. la: paracetamol,

5.1.7. Dinh hrong h6n hop 5 thanh ph§n trong nang thuoc colussin cafein. Dieu kien siic ky: May sAc leY long hieu nang cao HP 1100 - Hewlett Packard. CQt Lichrosorb Si 60 ( 250 x 4mm; 5 urn ). Detector UV - VIS: 257 nm do PE; CM va PA. 298 nm do CA va BH.

bromhexin hydroc1orid, pseudoephedrin hydroclorid, clorpheniramin maleat va

Pha dQng: Methanol- dung dich d~m amoninitrat pH 9,5 (45: 6). T6c d{>dong: 1,5 ml/phut do PE va CM. 1,0 ml/phut do CA va BH. 1,0 ml/phut do PA.

-

The tich tiem: 20 ul,

Da khao sat S\l' nrong quan giua nAng dQ dinh hrong voi dien tich pie, dQ l~p lai va dQ dung nitro trong gioi han eho phep, Do paracetamol la mQt thanh phfu1 co 1)' l~ IOnnen tnroc khi do HPLC cho 4 thanh phfu1can lai phai loai bo bat ra khoi h6n hop. Voi chuang trinh nay, ngoai vien nang tren, chung ta co th€ ap dung cho cac che pham khac co chua til 2 den 4 thanh phfu1 tren (CA, BR, CM va PE). Dan gian han nua, kh6ng co paracetamol thi kh6ng cfu1phai chi€t tach khi chuan bi dung dich thir va dung dich chuan, Tuy nhien, tnroc khi ap dung cho mot che pham cu the khac ta phai khao sat anh huang cua ta duoc. Phuong phap nay co the (rug dung eho ca dinh tinh va dinh hrong. Qui trinh da duoc sir dung lam tieu chuan co

sa.

5.2. Phuong phap ph in tieh da ciu til'MeA
Voi cac thuoc da thanh phan, dinh hrong bang HPLC v§n la mot phuong phap t6i
iru,

cho k€t qua nhanh chong va chinh xac, tuy nhien phuong phap nay doi hoi phai co

thiet bi HPLC d~t tien, chi phi cho dung m6i kha t6n kern. Phuong phap tach rieng cac thanh phan va dinh hrong rieng re t6n nhieu thai gian va c6ng sire, ngiroi thuc hien phai ti€p xuc voi cac dung m6i htru cc dQc hai, Phuong phap MCA co nhieu
Uti

diem

nhir phan tich nhanh chong, chinh xac, ti€t kiem duoc thai gian, c6ng sue cilng nhu dung m6i, hoa chAt. D~c bi~t phuong phap rAt thuan lei khi ta phai phan tich hang loat m§u thu gfu1giong nhau. Ngay nay, su phat trien manh me cua khoa hoc ky thuat trong linh V\l'C thi€t dien til co kha nang canh tranh va thay th€ ky thu~t HPLC trong met s6 twang hop the. RAt nhieu diroc pham co chua til 2 - 4 thanh phan hdp thu quang c6 - 82 -

hi
C1,l

phan tich, ky thu~t phan tich quang ph6 UV - Vis MCA voi SlJ tro giup cua may tinh

the ap dung

_~._l_ _?1}_{!!!: _c;_~!!.,!g !i!!!:
cling c6 mQt s6 han ch6 nhdt dinh. .. Trong cac thu6c da thrum phfm,

..

...

.

.. -

._- .. ---_._----_-----

phirong phap phan tich da cAu tir MeA. Tuy nhien, trong thuc t6 phirong phap nay

tY

1~ph6i hop gitra cac duoc chAt trong mQt

cong thirc thuoc doi khi rAt khac nhau roy thuoc vao nbng dQ cua duoc chdt hay nbng dQ gay tac dung duoc ly mong muon, Vi du: vien nen Trivitamin (BI + B6 + B12) duoc ph6i hop thea chAt la kh6 c6 th~ thuc hi~n diroc. ,. Khi phan tich mot cong tlnrc thuoc bdt ky, neu mQt chdt nao do co

tY

1~nrong

irng 250: 250: 1 (mg), V&i tY Ie chenh lech den v~y, phan tich MeA cho ca ba

tY l~ nho

va

diroc coi nhu "tap chat" thi doi khi lai co anh lnrong kha 1&0,anh huong nay tang thea s6 hoat chAt duoc ph6i hop trong ch6 pham. Trong mot sb tnrong hop thi S\I anh htrong do du la nho cung khong th~ bo qua. Vi du: h6n hop chua ba thrum ph§n la Paracetamol, Aspirin va Cafein, cac tac gia cling chi ap dung phan tich MeA thrum cong tren cac thuec con moi va

tY Ie acid

Salixylic nho diroi 1%, con n6u ty Ie nay tren 1% thi khong th~ ap

dung phuong phap MeA. .. MQt di~u dang chu

y

ntta la: trong cac diroc pham d~u co ta diroc va cac chAt

bao quan, neu cac chAt nay cling co kha nang hAp thu quang thi viec phan tich bang MeA co th~ cling khong thuc hien diroc. Vi du: cac che phsm thuoc tiem co 4 thanh phcln Vitamin B 1; B2, B6 va PP co ty l~ ph6i hop gitra cac thanh phfin kha

ly luang

cho viec phan tich d6ng thai

b~g MeA, nhirng vi trong h6n hQP cac thanh ph~ nay con co mQt ta diroc la a1col benzylic va mot s6 chAt bao quan khac d~u co kha nang hAp thu quang, do do viec phan tich hang MeA la khong th~ thuc hien duoc . ... Ngoai ra, voi S\I xmi! hien cua cac he phan tan rfut va he phan tan long nhfim ph6i hop cac hoat chAt co tinh hoa tan rAt khac nhau trong mQt dang bao che,
~~~~~~~~~~~~~~~~

thich hop, Cac hoat chdt nhieu khi lai co tinh chdt hdp thu tir ngoai kha ki€n khac nhau trong cac dung moi khac nhau (acid hay kiem) lam cho viec phan tich MeA trong nhieu tnrong hQP cu th€ khong co tinh kha thi. Vi vay chung toi da: nghien CUu xay dung met s6 quy trinh phan tich dbng thai (khong qua chi~t tach) m{)t s5 hoat chdt trong cac thu6c c6 2 thanh ph~ bang phuong phap phan tich toan ph6 tren cac may quang ph6. Tir do irng dung cac quy trinh da xay dung d€ phan tich mot s6 mdu thuoc chua hai thrum phcln nay dang luu hanh tren thi tnrong trong ca mroc.

- 83 -

_~._!!..q!!_?7!..~!!_£~¥'1Jg___., __., __.. __. __.. __.. __.

...

.

..

.

--. _--. ---. ----------------.

5.2.1. Dil xay dung duoc phirong phap dinh Iuong d6ng thai paracetamol va ibuprofen trong vien nen han hop bAng phuong phap phan tich toan ph6, cho k~t qua

nhanh, don gian do khong phai chiet tach. Phuong phap 6n dinh, co dQ chinh xac (ham
lirong paracetamol la 329,69 ± 1,58 mg, ibuprofen lit 208,5 ± 1,46 mg voi rmrc tin c~y 95%) va dQ dung chAp nhan duoc

(ry Ie tim

thAy cua ibuprofen bien thien tir 97,2%

d~n 99,2% va paracetamol bien thien tit 100,1% d~n 101,2%).

Phuong phap da dugc trung tarn ki~m nghiem duoc pham va
hanh tren th] tnrong tai dia phuong.

my pham Lang

san

tri~n khai co k~t qua dS phan tfch mot s6 mdu thuoc chua 2 hoat chAt tren dang hru

5.2.2. Dil xay dung mQt qui trinh dinh hrong d6ng thai paracetamol va quinine sulfat trong vien nen Antigrip F bang phirong phap da cAu tir (MCA) tren may quang ph6 thong thirong, xir ly k~t qua bang phfut mem tv' xay dung. Da ti~n hanh so sanh k~t qua thu duoc voi phuong phap do quang ph6 hAp thu ill ngoai thong thtrong (tieu chu§n co sa) sau khi chiet tach tirng thanh phan, Phuong phap co
iru

diSm lit nhanh, chinh xac, it tbn hoa ch~t, dung moi, V6'i h6n

hop chi chua 2 thanh phfut tren chi cAn do dQ hAp thu

a 4 buoc

song da khao sat la co

thS thtrc hien ducc phep dinh hrong co k~t qua t61. Han ntra, phuong phap nay chi yeu cdu co sa kiem nghiem co may quang ph6 thong tlnrong khong diroc trang bj sfuI phdn mem phan tich MCA. 5.2.3. Phuong phap MCA dtroc sir dung tren phftn mem sfut co cua may Quang phd GBC Cintra 40

aa diroc irng dung co k~t qua d~ dinh hrong

d6ng thai paracetamol

va acid mefenamic trong ch~ pham Pamesic. Phtrong phap dinh hrong diroc ti~n hanh trong moi tnrong kiem NaOH O,IN, voi dai biroc song phan tich tir 220nm d~n 300mn. Phuong phap cho k~t qua chinh xac (sai s6 nrong dbi la 1,05 va 0,8%) va dang tin c~y (zy 1~thu h6i Ia 99,87 d~n 100,25%). Chung toi hy vong co thS sir dung phuong phap phan tich quang ph6 MeA d~ b6 sung them hoac thay th~ cho phirong phap HPLC phdn nay khong xen phu nhau qua nhieu,

a co

sa khong co thiet bi HPLC. D@c

bi~t voi cac thuoc han hop co chua cac thanh phfut hApthu quang va ph6 cua cac thanh

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Cac phirong phap nay d~u co th~ ap dung dA dinh hrong d6ng thoi cac heat chdt thay cho phuong phap HPLC._. (PA) va acid mefenamic (AM).__- - .+ ... 6. Ung d\lng phuong phap HPLC Chung tai da xay dung dircc chuang trinh dinh tinh va dinh IUQ'11g hoat chAt cac cho cac h6n hop thu6c sau: Paracetamol Paracetamol Paracetamol Paracetamol Paracetamol maleat (eM). • 6..2...1. . dn ducc ung dung co ket qua d~ dinh hrong hai hoat ch~t tren trong che pham Pamesic. Ung d\lng phuong phap quang pb6 UV-Vis va ibuprofen phirong phap phan tich toan ph6.. Xay dung dircc phirong phap MCA dA dinh hrong d6ng thai paracetamol va acid sir dung phdn mem sin co cua may quang ph6 GBC Cintra 40._~~~. (PA) (PA) va va ibuprofen (IBu). dan va quinine sulfat Da xay dung duoc phuong phap dinh hrong d6ng thoi paracetamol trong vien nen h6n hop b~g gian do khong phai chiet tach. clorpheniramin dung. Da xay dung mQt qui trinh dinh IUQ'11g d6ng thoi paracetamol trong vien nen Antigrip quang ph3 thong thuong. Paracetamol. _r • " .. cafein. KET LuAN vA DE NOHl 6. quinine sulfat (QS).85 - .- - - - - -- . pseudoephedrin hydroclorid.__ . phenylpropanolamin va Cac chuang trinh tren d~u ducc danh gia v~ tinh tuyen tinh. ..__. ~_Q __. _ .. TAt ca d~u bromhexin maleat hydroclorid. hydroclorid (PH) va clorphenylamin (PA) va cafein (CA). DQ l~p lai: sai s6 nrong d6i (hoac dQ l~ch chuan nrong d6i) d~t Dc) dung: kha nang tim lai d~t 2: 98%. Kha nang » >- s 2%. theophyllin va phenobarbital.999. . dc) chinh xac va tinh cr rmrc chAp nhdn diroc (xem phfut ban lu~ cr tren)..~ 6.__ or - - - - . voi h~ s6 tuyen tinh d~u dat r > 0. Kit luan .._!_ - va ki~n nehi .. Ephedrine hydrocIorid. (PA). xu mefenamic F bing phirong phap phan tich da cdu tU (MCA) tren may Iy ket qua bing phdn mem nr xay dung. M6i phirong phap d~u bao dam cac yeu cAu sau: >- Dc) tuyen tinh: x§y dung duoc plurong trinh h6i quy. cho ket qua nhanh. » ap dung: xay dung phirong phap co thA ap dung diroc cho nhieu san phAm co cung thanh phdn hoat chat..

da: ducc vien kiem nghiem va mQt s6 trung tam ki€m nghiem img dung de kiem tra chdt hrong thuoc dang hru hanh tren thi tnrong. ve duoc va hoa MQt s6 chuang trinh dircc nghien crru Ia de lam tieu chuan cho cac co chuang trinh dang diroc cac co san pham sa tham sa. cac bao cao hQi nghi hoc trong mroc va qU6c t~.86 - . tap chi diroc hoc.3. ti€p tuc danh gia cac ket qua d~ d€ nghi dua mQt s6 chuyen lu~ vao diroc . Banh gia chung va d@nghj Cac nghien ciru tren d€u da diroc cac chuyen gia v€ kiem nghiern kiem tra. Tiep tuc nghien ciru plnrong phap quang ph6 MCA de co the ap dung cho nhtrng cong thirc co tren 2 thanh ph§n hoat chdt. MQt s6 khao irng dung de xay dung tieu chuan cho cua minh. K~t qua nghien ciru cua chung toi la co gia tri thuc ti~n. tharn dinh la co gia tri khoa hQCva da: dtroc phep gioi thieu tren cac tap chi chuyen nganh nhir tap chi kiem nghiem thuoc.6. Da hOM thanh theo muc tieu va dAy diI cac nQi dung nghien ctm da d€ ra rna h¢i d6ng bQ da phe duyet cho de tai. rna rc)ng trong nhtrng de tai khac. Chung toi mong diroc ti€p TI)C nghien CUud~ hoan chinh va di~n Vi~t nam trong Iftn xudt ban t6i.

Cong ty duoc pham Hit tay._?_·_T. Trfu:tVi~t Hung. 8. Tieu chuan co s6 vien nen Sedapa. (2001). Cong ty diroc TW Hu~. Xi nghiep duoc pham TW 2 (1999). lien hi~p duoc H?u giang. Nghien ciru dinh hrong cac vitamin tan trong mroc va tan trong dAu trong thubc Multivitamin bang sAc Icy long hieu nang cao. Tieu chuan co 18. 6.2001. Thbng ke irng dung trong cong tac diroc. sa vien nang acid sulfat. . Trinh Van Quy..2002. Duoc di~n Vi~t nam III. Be>y t&. Trftn Th~ Phuong. 5. sb 1-2002. Dinh hrong d6ng thai sulfadoxin va pyrimethamin trong vien nen Fansida bAng phtrong phap HPLC va phirong phap quang ph6 dao ham. Trinh Van Quy. sa vien nen Alaxan. Tieu chuan co sa vien bao dirong Antigrip sa vi en nen Desilogen. The United Pharma. Tap chi phan tich hoa. tnrong dai hoc duoc Ha nQi. (1999). Tr.Truong Dai hoc dtroc Hit nQi . sa vien nen Dibulaxan.i!!_!!f_l!_~Jy_q_'!'__I~!!{l_q 7. Tieu chufin co sa vien nen Lopenca. 14. Thong bao kiern nghiem. de tai c§p be>. The United Pharma. Tieu chuan co mefenamic. 13. Xi nghiep diroc pham TW 2. 9. 2002. Thai Phan Quynh Nhu.. T?p 7.Iy va sinh hoc. Tieu chuan co 12. 11. 2. 10.87- . Thai Duy Thin. Dang Trftn Phuang H6ng. TAl LIEU THAM . 15.CT Diroc VTYT Tra vinh. (2001)_ Nghien ctru dinh luong mQt sb thubc da thanh phAn co chua paracetamol bang phirong phap HPLC. Cong ty Duoc VTYT Ti~n giang. Tieu chuan co s6 vien nen Pasepan. KHAo Tiing Vi?t 1. 7.Phan tich duqc pham bang ky thuat quang ph6 UV-VIS MeA. D3 Thi Oanh. 4. 76-81. (1997). Thong tin khoa hoc cong nghe diroc. . (2002). sb 4. Thai Phan Quynh Nhu.Ky y~u cac cong trinh NCKH 1996-2000. (2002) Tieu chuin co s6 vien nen Pi-antalvic. Xi nghiep duoc pham TW 5. Til sach sau dai hoc. Nguyen Thanh Dat. BQ Y t~. Xi nghiep sa vien nen Quinin 16. Xi nghiep dtroc pham TW 2. Tieu chuan co 17. (1995). Tieu chuan co sa vien nen Decolgen. Nguyen Ti~n Khanh. Vien ki~m nghiem. 3. Tieu chuen co F.

Sena. p. 20. 25. Clarke's isolation and identification of Drug (2th Edition) 1996. 26. Hamide Ceniuva and Tunce1 Ozden Determination (2002). The United State Pharmacopeia XXVII-2004. 29.C! _ Ti2ng mruc ngoai 19. Tantishaiyakul. The International pharmacopeia III~1981.88- . 2001. February 2002. British pharmacopeia 1998. V. Nouvember of paracetamol. Poppi. Journal of Chromatografic Sience. Spectrophotometry. K. phenylephrin maleate in pharmaceutical dosage forms. ISSN 0021-9665 Volume 40. Pharmacopeia of the people's Republic of China 1997. 23.. M. pp. paracetamol and caffein. R.. Number 2. Simultaneous High Performance Liquid Chromatographic HCl and chlorpheniramine 97-100. 05G6-Analytical Abstracts. 30.. 880. M.2000. The United State Pharmacopeia XXIV . 21_European pharmacopeia III 1997. p. Poeaknapo. P. Sirisuppanonnon. . Pharmacopee Francaise X. 2000. 237-243. May 2004. N-way PLS applied to Simultaneous Spectrophotometric determination of acetylsalicylic acid. 11G 178-Analytical Abstracts. 24. Sribun. 27. J (2004). Simultaneous determination of dextromethophan HBr and Bromhexine HCl in tablets by first derivative 1998. C._?_·__'[_q!_!!fH_~!t_q~_~4. 28. (1998). The Japanese pharmacopeia XIII ~1996. XIV ~2001. 22.

Nguyen Tirong Vy. Hung. Nguyen Thi Thanh. Tran Vie. Thailand. Dinh tfnh.ll-l4. 7 Ian 8 Thai Duy Thin va cac CS. 4-7. Vo Nbi Ha. HQi nghi hoa hQCtoan quoc IAnthu IV.19-21. Dinh tinh va dinh hrong d6ng thai paracetamol va quinine sulfat trong vien nen antigrip F bang phirong phap HPLC. Hoang Van Dire (2003).Thai Duy Thm. pseudoephedrin hydroclorid. 2003. Nguyen Thi Huang. Tap chi diroc hoc so 7. Thai phan Quynh Nhu (2003). 6 Thai Duy Thin va cac CS (2003). clorpheniramin maleat. 19-21. Dinh tfnh va dinh luong d6ng thOi paracetamol va quinine sulfat trong vien nen antigrip F bang phtrong phap phan tfch da cau tir MCA. . Thai Phan Quynh Nhtr (2001). May 20-23. Tap chi ki~m nghiem thu6c s6 4. Thong tin khoa hoc cong nghe dirocTnrong D~ hoc diroc Ha noi. pp136-143. Tr. 2 3 4 5 U Quang Thao. Tr. Tr. 10/2003. 279-285. Tap chi kiern nghiem thu6c so 4. Dtnb hrong dong thoi paracetamol va ibuprofen trong vien nen bang phuong phap phan tich loan phd. bromhexin hydroclorid. Dinh hrong paracetamol. Tr.8. D6 Thi Oanh. 10-2003. Research on application of HPLC and UV spectrophotometry for quantitative and qualitative analysis of some multicomponent farmaceuticals of paracetamol. Nguyen Thanh Dat (2003). va cafein trong nang cotusin bang phtrong phap HPLC. Thai Duy Thin. Bangkok. TrAn The Phuong. The third Indochina Conference 9 on farmaceutical Sciences. Phung Ngoc Quang (2004). Tran Thi Thuy. dinh Iunng paracetamol va quinine sulfat trong che' pham h6n hop chua 2 thanh phan. Tr. Thai Phan quynh Nhu (2004). (2003). Thai Duy Thin (2003). Bili Thu Hue. Tr.Truong Dai hoc duoc Ha noi. 76-81. Thong tin khoa hoc cong nghe diroc. Nghien ctiu dinh luong mot s6 thu6c da thanh phan c6 chua paracetamol bang phirong phap HPLC. Dinh luong dong thci paracetamol va acid mefenamic trong thu6c vien bang phirong phap HPLC. Tieng nuoc ngoai: Thai Duy Thin. Thai Duy Thin. 99-105. Nghien CUu dinh hrong paracetamol va ibuprofen trong thuoc chua 2 thanh phan. Hoi nghi hoa hoc toan quoc tlnr IV. CONG B6 LItN QUAN DEN Dt TAl 1 Nguyen Thanh D~. Thai Duy Thin. Tr. Tap chi Diroc hoc so 5. Thai Phan Quynh Nhu. PHl) Ll)C cAc CONG TRINH NGmtN Tieng Vift: coo Dt\. Thai Duy Thin.

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