B Y T TRNG I HC DC H NI ---------- V TH TH NH LNG NG THI CEFOPERAZON V SULBACTAM TRONG CH PHMBNG QUANG PH O HM KHA LUN TT NGHIP DC S

H NI - 2013 B Y T TRNG I HC DC H NI ---------- V TH TH NH LNG NG THI CEFOPERAZON V SULBACTAM TRONG CH PHMBNG QUANG PH O HM KHA LUN TT NGHIP DC S

Ngi hng dn:1.TS. V ng Hong 2.DS. Nguyn Th Loan Ni thc hin:B mn Ha phn tch - c cht Trng i hc Dc H Ni

H NI - 2013 i LI CM N Trcht,emxingilicmnchnthnhvsuscnhttiTS.V ng Hongv DS.Nguyn ThLoan,nhng ngi trc tip hng dn, ht lng truyn t kin thc v tn tnh gip em trong sut thi gian thc hin ti nghincumangtn"nhlngngthicefoperazonvsulbactamtrongch phm bng quang ph o hm". Em xin chn thnh cm n s quan tm v gip nhit tnh ca cc thy c gio v cc anh ch k thut vin b mn Ha phn tch - c cht trng i hc Dc H Ni trong sut thi gian em lm thc nghim ti b mn. Em xin chn thnh cm n hi ng chm kha lun tt nghip Dc s i hc dnh thi gian xem xt, gp v sa cha kha lun tt nghip ca em c hon thin hn. EmxincmnBanGimhiuNhtrngvPhngototoiu kin tt nht em hon thnh tt kha hc i hc 5 nm ti trng. Cuicng,emxinchnthnhcmngianhvbnblungip, ng vin, khch l v h tr em c v vt cht ln tinh thn trong sut qu trnh lm ti cng nh trong hc tp v cuc sng. H Ni, thng 05 nm 2013 Sinh vin V TH TH ii MC LC LI CM N ....................................................................................................i MC LC .......................................................................................................... ii DANH MC CC CH V K HIU VIT TT ........................................iv DANH MC CC BNG BIU ...................................................................... v DANH MC CC HNH V V TH ......................................................vi T VN ................................................................................................... 1 CHNG 1: TNG QUAN ............................................................................ 2 1.1. TNG QUAN V I TNG NGHIN CU ..................................... 2 1.1.1. Tng quan v cefoperazon ....................................................................... 2 1.1.2. Tng quan v sulbactam........................................................................... 3 1.1.3. Mt s phng php nh lng cefoperazon v sulbactam ................... 3 1.1.3.1. Phng php sc k lng hiu nng cao (HPLC) ................................. 3 1.1.3.2. Phng php quang ph UV ................................................................. 6 1.1.3.3. Mt s phng php khc ..................................................................... 7 1.2.PHNGPHPQUANGPHHPTHTNGOI-KHKIN (UV-Vis)............................................................................................................. 10 1.2.1. nh lut Lambert-Beer ........................................................................... 10 1.2.2. nh lng dung dch nhiu thnh phn bng phng php quang ph o hm .............................................................................................................. 11 1.2.2.1. Phng php ph o hm .................................................................... 11 1.2.2.2. Phng php ph o hm t i.......................................................... 13 1.3. PHNG PHP SC K LNG HIU NNG CAO (HPLC) .............. 14 1.3.1. Nguyn tc chung ca HPLC ................................................................... 14 1.3.2. Cc thng s c trng cho qu trnh sc k ........................................... 14 CHNG 2: I TNG V PHNG PHP NGHIN CU ............ 19 2.1. I TNG NGHIN CU, NGUYN LIU V THIT B ............... 19 2.1.1. i tng nghin cu............................................................................... 19 iii 2.1.2. Nguyn liu v thit b ............................................................................. 20 2.2. PHNG PHP NGHIN CU ............................................................... 22 2.2.1.Xydngphngphpnhlngngthicefoperazonv sulbactam bng quang ph o hm ly HPLC lm phng php i chiu .... 22 2.2.2. ng dng cc phng php quang ph nu v HPLC nh lng ng thi cefoperazon v sulbactam trong ch phm ........................................ 23 2.2.3. X l kt qu thc nghim ....................................................................... 23 CHNG 3: THC NGHIM, KT QU V BN LUN ..................... 24 3.1. KT QU THC NGHIM V NHN XT .......................................... 24 3.1.1. Phng php quang ph o hm ............................................................ 24 3.1.1.1. Chun b dung mi ................................................................................ 24 3.1.1.2. Chun b mu chun v mu th ........................................................... 24 3.1.1.3. Xy dng phng php nh lng ...................................................... 26 A. Chn bc sng nh lng .......................................................................... 26 a. Xc nh khong cng tnh ............................................................................. 26 b. Phng php ph o hm bc 1 ................................................................... 27 c. Phng php ph o hm t i ................................................................... 28 B. Kho st khong tuyn tnh ........................................................................... 30 a. Ph o hm bc 1 .......................................................................................... 30 b. Ph o hm t i ......................................................................................... 32 3.1.1.4. lp v ng ca phng php..................................................... 35 3.1.2. Phng php HPLC ................................................................................. 36 3.1.2.1. Chun b mu nghin cu ..................................................................... 36 3.1.2.2. Xy dng phng php HPLC .............................................................. 36 3.1.3. Kt qu nh lng ................................................................................... 38 3.2. BN LUN ................................................................................................ 39 KT LUN V KIN NGH............................................................................ 41 TI LIU THAM KHO ............................................................................... I iv DANH MC CC CH V K HIU VIT TT ACN BQ CP HD HPLC MeOH NSX PH PHT Ptl RSD SB SD SK : : : : : : : : : : : : : : Acetonitril Bo qun Cefoperazon Hn dng Sc k lng hiu nng cao (High Performance Liquid Chromatography) Methanol Ngy sn xut Ph o hm Ph o hm t i Phn t lng lch chun tng i (Relative Standard Deviation) Sulbactam lch chun (Standard Deviation) S ng k SKS STT TBAH : : : S kim sot S th t Tetrabutyl ammonium hydroxid TEAA TLTK : : Tetraethyl ammonium acetat Ti liu tham kho TMAH:Tetramethyl ammonium hydroxid TNHH:Trch nhim hu hn UV:T ngoi (Ultra Violet) UV-Vis:T ngoi - Kh kin (Ultra Violet - Visible) [1] (1.1) : : Ti liu tham kho s 1 Cng thc 1, chng 1 (2.1):Cng thc 1, chng 2 v DANH MC CC BNG BIU Bng 1.1. Mt s c im l ha ca cefoperazon .......................................... 2 Bng 1.2. Mt s c im l ha ca sulbactam ............................................. 3 Bng 1.3. Phng php sc k lng hiu nng cao (HPLC) ............................. 3 Bng 1.4. Phng php quang ph UV ............................................................. 6 Bng 1.5. Mt s phng php khc ................................................................. 7 Bng 2.1. i tng nghin cu ........................................................................ 19 Bng 3.1. Kt qu kho st khong tuyn tnh ca CP vi gi tr PH bc 1 .. 30 Bng 3.2. Kt qu kho st khong tuyn tnh ca SB vi gi tr PH bc 1 .. 31 Bng 3.3. Kt qu kho st khong tuyn tnh ca CP vi gi tr PHT bc 1 .......................................................................................................................... 32 Bng 3.4. Kt qu kho st khong tuyn tnh ca SB vi gi tr PHT bc 1 .......................................................................................................................... 33 Bng 3.5. Kt qu kho st khong tuyn tnh ca CP v SB ........................... 34 Bng 3.6. lp v ng ca cc phng php quang ph o hm .......... 35Bng 3.7. Thng s qu trnh HPLC ca hn hp CP 20 mg/l + SB 20 mg/l ... 37Bng3.8.Ktqunhlngccchphmbngquangphohmv HPLC .................................................................................................................. 38 vi DANH MC CC HNH V V TH Hnh 1.1. Phhp th (a), pho hm bc2 (b) v bc 4 (c)(diph nt chmccngvtrvcngnhngrnggpi);phhpthca trans-stilbenetrongcyclohexane(d)vphohmbc2(e),bc4(f) tng ng ............................................................................................................12 Hnh 1.2. S tch 2 cht A v B trn 3 ct c phn gii khc nhau ........17Hnh 3.1. Ph hp th ca SB 20 mg/l, CP 20 mg/l, ph cng v ph hn hp SB 20 mg/l + CP 20 mg/l ...................................................................................26 Hnh 3.2. PH bc 1 ca dy dung dch SB nng thay i (10- 35 mg/l) v ca dy dung dch CP nng thay i (10 - 35 mg/l) .................................27 Hnh 3.3. PHT bc 1 ca dy dung dch hn hp CP (10 - 35 mg/l) + SB 20 mg/l vi s chia l SB 20 mg/l v ca dy dung dch CP (10 - 35 mg/l) vi s chia l SB 20 mg/l .......................................................................................... 28 Hnh 3.4. PHT bc 1 ca dy dung dch hn hp SB (10 - 35 mg/l) + CP 20 mg/l vi s chia l CP 10 mg/l v ca dy dung dch SB (10 - 35 mg/l) vi s chia l CP 10 mg/l .......................................................................................... 29 Hnh3.5.thbiudinngchuncaCP(10-35mg/l)vigitr PH bc 1 ........................................................................................................... 30 Hnh3.6.thbiudinngchuncaSB(10-35mg/l)vigitr PH bc 1 ........................................................................................................... 31 Hnh3.7.thbiudinngchuncaCP(10-35mg/l)vigitr PHT bc 1 ..................................................................................................... 32 Hnh3.8.thbiudinngchuncaSB(10-35mg/l)vigitr PHT bc 1 ..................................................................................................... 33 Hnh 3.9. Sc k ra gii ca hn hp chun CP 20 mg/l + SB 20 mg/l ..... 36 1 T VN Tnh khng khng sinh ca vi khun ngy cng gia tngvi nhiu c ch khcnhaunh:sinhmenkhngthuc,thayitnhthmmngtbo,bini ch tc dngTheo T chc Y t th gii th Vit Nam nm trong danh sch cc nc c t l khng khng sinh cao nht th gii, do , vn s dng khng sinh an ton v hiu qu hin ang l mt trong nhng vn mang tnh thi s. Cefoperazonlkhngsinhbntnghpthucnhmcephalosporinthh III,cdngtheongtimvctcdngditkhundocchstnghp thnh t bo vi khun. Cefoperazon c th c kt hp vi sulbactam trong cng mt ch phm thuc bt pha tim. Sulbactam l cht c cu trc tng t nh beta-lactam, c kh nng gn kt vo beta-lactamase do vi khun tit ra v lm bt hot enzym ny. Do vy, sulbactam bo v cc khng sinh beta-lactam khi b phn hy bienzymny.Skthpnygpphnduytrhotphvgimtnhtrng khngthuccacefoperazontrnlmsng.Hnhpcefoperazonvsulbactam thng c ch nh iu tr cc nhim khun nng do cc vi khun Gram (-), Gram (+) nhy cm v cc vi khun khng cc khng sinh beta-lactam khc. nh lng ng thi cc ch phm thuc a thnh phn, sc k lng hiu nngcao(HPLC)vquangphohmthngctrinkhainghincu.Vi mong mun xut phng php nh lng ng thi hn hp hai thnh phn c kh nng ng dng trong cng tckimnghim thuc, chng ti tin hnh ti: "nhlngngthicefoperazonvsulbactam trong chphm bng quang ph o hm" vi hai mc tiu: -Xydng php nh lng ng thi cefoperazon v sulbactam bng quang ph o hm, ly phng php HPLC lm phng php i chiu. -ngdngccphngphpnynhlngngthicefoperazonv sulbactam trong cc ch phm: Sulperazon, Jincetam, Bacamp v Cefactam. 2 CHNG 1 TNG QUAN 1.1. TNG QUAN V I TNG NGHIN CU 1.1.1. Tng quan v cefoperazonBng 1.1. Mt s c im l ha ca cefoperazon c imCefoperazon Ngun gcL khng sinh bn tng hp thuc nhm cephalosporin th h III. Cngthc cu to C25H27N9O 8S2 Ptl: 667,65 Tn khoa hc5 - thia - 1 -azabicyclo [4.2.0] octo - 2 - ene - 2 - cacboxylic, 7 - [[[[4 - ethyl- 2,3 dioxo- 1 - pyperarizyl) carbonyl] amino] - 4 -hydroxyphenyl)acetyl]amino]- 3 - [[(1 - methyl - 1H - tetrazol - 5 - yl) thiol] methyl] - 8 - oxo, monosodium salt (6R, (6,7 R*)) Tnh chtBt mu trng, khng mi v c v ng. Tantttrongnc,tantrongmethanol,tanttrongalcoholv hu nh khng tan trong cc dung mi hu c. 3 1.1.2. Tng quan v sulbactam Bng 1.2. Mt s c im l ha ca sulbactam c imSulbactam Ngun gcL khng sinh nhm beta-lactam. Cng thc cu to C8H10NNaO5SPtl: 255,22 Tn khoa hcsodium (2S,5R) - 3,3 - dimethyl - 7 - oxo - 4 - thia - 1 - azabicyclo[3.2.0]heptane-2-carboxylate-4,4- dioxide Tnh chtBt mu trng, tan tt trong nc 1.1.3. Mt s phng php nh lng cefoperazon v sulbactam 1.1.3.1. Phng php sc k lng hiu nng cao (HPLC) Bng 1.3. Phng php sc k lng hiu nng cao (HPLC) STTCht phn tchc im phng phpTLTK 1Cefoperazon- Pha ng: H2O : ACN : acid acetic 1 N : (14 ml triethylaminv5,7mlgracialacetictrong1000 ml) (876 : 120 : 2,8 : 1,2) - Ct: 4 mm x 300 mm- Tc dng: 2 ml/pht [29] 4 - Detector UV: 254 nm 2Sulbactam- Pha ng: 0,005 M TBAH :ACN (1650 : 350) - Ct: 4 mm x 300 mm - Tc dng: 2 ml/pht - Detector UV: 230 nm [29] 3Sulbactam Cefoperazon - Pha ng: m phosphate pH 3,5 : ACN (35 : 65) - Ct: Kromasil C8 5 m, 150 mm x 4,6 mm - Cht chun ni: ornidazol - Tc dng: 1ml/pht - Detector UV: = 215 nm [11] 4Sulbactam Cefoperazon -Phang:mphosphate20mMpH3,5: ACN (35 : 65) -Ct:Phenomenexphenylhexyl5m,250 mm x 4,6 mm - Cht chun ni: ornidazol - Tc dng: 1 ml/pht - Detertor UV: = 190 nm [30] 5Sulbactam Cefoperazon - Pha ng: nc pH 3,2 : ACN (40 : 60) -Ct:PhenomenexGeminiC185m,250 mm x 4,6 mm- Cht chun ni: Captopril - Tc dng: 0,5 ml/pht - Detector UV: = 210 nm [23] 6Sulbactam Sultamicillin Ampicillin -Phang:MeOH25%trongTMAH0,005 MciuchnhnpH3,4viH3PO4, siu m loi kh trong 10 pht [7] 5 Cefoperazon Cefaclor Tosylat - Ct: Spherisorb ODS 2,5 m, 120 mm 4,0 mm - Cht chun ni: salicylamid - Detector UV: = 230 nm - Tc : 1 ml/pht 7Cefoperazon -Phang:mamoniacetatepH5,5: MeOH (30 : 70) - Ct: Supelco C18 5 m, 250 mm x 4,6 mm - Tc dng: 0,5 ml/pht - Detector UV: = 250 nm [25] 8Cc penicillin Cefoperazon -Phang:MeOH:NaCl0,2M(35:65 65 : 35)- Ct: Alltech C18 10 m, 250 mm x 4,6 mm- Tc dng 1,5 - 2,2 ml/pht - Detector: in ha sau phn ng phn hy do ngun bc x Photronix Model 816 UV - Cht th c pha trong dung dch NaCl 0,9 M thnh dung dch c nng 20 mg/ml [26] 9Ampicillin Cefoperazon Sulbactam -Phatnh:phalinkt|-cyclodextrinCyclobond I, 5 m, 250 mm x 4,6 mm- Pha ng: MeOH : m TEAA 5mM pH 4,5 (35 : 65) - Detector UV: = 280 nm - Tc : 0,8 ml/pht [28] 10Cefalexin Cefoperazon Ceftriaxon Ceftazidim - Pha ng: KH2PO4 (50 mM, pH 4,6) : ACN (80 : 20) -Ct:WatersBondapakC1810m,250 [12] 6 Cefepim Cefoperazon Sulbactam mm x 4,6 mm - Tc dng: 1 ml/pht - Detector UV: = 230 nm 1.1.3.2. Phng php quang ph UV Bng 1.4. Phng php quang ph UV STTCht phn tchc im phng phpTLTK 1Cefoperazon-Dung mi: MeOH -o quang ti = 230 nm -odintchdingcongphhpth trong khong 225 - 235 nm [31] 2Cefoperazon Sulbactam -Dung mi: nc - Ph o hm (A = 4 nm) bc 1, bc 2: theo php o giao im khng 277,5; 267,5; 272,5; 290 nm v php o th 267,5 nm / 290 nm; 272,5 nm / 290 nm [8] 3Cefoperazon TophnngviFolin-Ciocalteautrongmi trngkim(Na2CO3)voquangtimax ca hp cht mu xanh 652 nm [20] 4Cefoperazon TophnngviFolin-Ciocalteauphenol trong mi trng Na2CO3 v o quang ti max ca hp cht mu xanh 668 nm [27] 5Cefoperazon o quang ti- Nitrathavtophcvitcnhninhn (I): 390 nm -Nitrathavtochalatekimloi(II):520 [19] 7 nm - To lin kt vi diazo (III): 435 nm - Phn ng vi mui ng v chit phc chelat to thnh vo cloroform (IV): 415 nm 6Cefoperazon - o quang trong 1,2 - dicloroethan ti 364 nm sn phm ca phn ng vi iod -oquangtrongmethanolti460nmsn phmcaphnngvi2,3-dicloro-5,6- dicyano - p - benzo - quinon -oquangtrongacetonitrilti843nmsn phmcaphnngvi7,7,8,8- tetracyanoquinodimethan [14] 7Cefoperazon oquangti=510nmsau30phtphn ngviquercetincoxyhabiN- bromosuccinimide [15] 1.1.3.3. Mt s phng php khc Bng 1.5. Mt s phng php khc STTTn phng phpc im phng phpTLTK 1Sc k lp mngnhlng cefoperazon trong ch phm -Pha tnh: silica gel G 60F254 -Pha ng: ethyl acetat - methanol - nc -aceton (3 : 1 : 1 : 2) -Phthin:quanstdinUV220V 50Hz sau khi phun thuc th Dragendorff [13] 2Sckinng mixen nhlngngthicephazolin, cefuroxim,ceftriaxon,cefoperazonv [16] 8 ceftazidim -Ct:maoqunsilicanungchyclp bao polyimid 60cm 75m -Phang:mphosphate-boratpH6,5 ccha10g/lnatridodecylsulfatv17,4 g/l pentanesulfonic acid - Detector UV: = 214 nm 3Quangphhunh quang nh lng cefoperazon trong ch phm -oquangphcbcbatothnhnh phnngviTb3+trongmTrisvipH 8,0 - kch thch = 240 nm; pht x = 485 nm [22] 4Quangphhunh quang nh lng cefoperazon trong ch phm-o quang dn xut coumarin pht quang mu vng c to thnh nh phn ng vi ethyl acetat trong mi trng acid sulfuric - kch thch = 412 nm; pht x = 465 nm [6] 5Somuvphhp th nguyn t nh lng cefoperazon trong ch phm-Somuti525nmhocoquangti 358,6nmdocpiontothnhgiacht phntchvmuiamonireineckat/mi trng acid.- Dung mi: aceton [18] 6in hanhlngcefoperazonetrongchphm v dch sinh hc: Von-ampe sng vung - in cc cng tc: git thy ngn treo- Thi gian tch gp: 60 150 s [17] 9 - Th tch gp: 0 ,3 V- Bc th: 8 mV - Bin xung: 25 mV -Tn s xung: 60 Hz - Cht in ly nn: m acetat pH 4,2 7in hanh lng cefoperazon trong ch phm, t bo vi khun, sa v nc tiu: Von-ampe sng vung - in cc cng tc: git thy ngn treo- Th bt u: 0,0 V - Thi gian tch gp: 1s - Bc th: 5 mV - Bin xung: 25 mV - Tn s xung: 500 Hz -Chtinlynn:mBrittion- Robinson, pH 4,4 [24] 8in hanh lng cefoperazone trong nc tiu:Von-ampeodngtrctipvxungvi phn - in cc cng tc: git thy ngn treo - Th tch gp: 0,4 V -Tcqut:100mV/s(odngtrc tip); 5 mV/s (xung vi phn) - Khong th: 0,4 1,0V - Bin xung: 50 mV - lp ca xung: 0,5 s -Chtinlynn:HClO40,03M;pH= 2,0 [9] 10 9in hanhlngcefoperazonetrongchphm v huyt tng: Von-ampe xung vi phn - in cc cng tc: cacbon than ch- in ly nn: m phosphat 0,2 M pH 2,0 - Tc qut: 20 mV/s - Bin xung: 50 mV - rng xung: 50 ms Von-ampe sng vung - Bin xung: 25 mV - Tn s: 15 Hz - Bc th: 4 mV [10] 1.2.PHNGPHPQUANGPHHPTHTNGOI-KH KIN (UV-Vis) 1.2.1. nh lut Lamber-Beer [1, 3] Khi cho bc x n sc i qua mt mi trng c cha cht hp th quang th hpthcabcxtlvinngcachthpthvchiudycami trnghpth(dungdchhpth).MiquanhnytuntheonhlutLamber-Beer v c biu din bng phng trnh: A (D, E) = lg T1 = lg II0= K.C.l ) 1 . 1 (

A: hp th (hay cn gi l mt quang D, hoc tt E) T:truynqua(haycngilthuquang),ctrngchotrongsut(v mt quang hc) ca dung dch Trong :

T = 0IIx 100 (%) ) 2 . 1 ( 11 I0:cng nh sng n sc ti dung dchI:cng nh sng n sc sau khi truyn qua dung dch K:h s hp th, ph thuc vo bn cht, mt mi trng, bc sng nh sng n sc () v c trng cho bn cht ca cht tan trong dung dch l:b dy ca lp dung dch (cm) C:nng cht tan trong dung dch iu kin p dng nh lut Lamber-Beer: -Chm tia sng phi n sc. -Dung dch phi nm trong khong nng thch hp. -Dung dch phi trong sut. -Cht th phi bn trong dung dch v bn di tc dng ca tia UV-Vis. 1.2.2.nhlngdungdchnhiuthnhphnbngphngphpquangph o hm 1.2.2.1. Phng php quang ph o hm [1, 2] Phng php ph ohm l mt kthut chuyn i ph hp th da trn php ly o hm bc 1, bc 2 hoc bc cao hn ca hp th A i vi bc sng . C s ca phng php ph o hm: Theo nh lut Lamber-Beer ta c: hp thl C K A . . =(1.3) C th ly o hm bc 1, bc 2 hoc bc cao hn ca hp th A i vi bc sng . Ta c: l CddKddA = (1.4) l CdK ddA d =2222

(1.5) 12 V dy l ca lp dung dch lun khng i v ti mt bc sng nht nh o hm caKl mt hng s, nn gi tr o hm ca A ch cn ph thuc tuyn tnh vi nng C ca dung dch. Ph o hm ca hn hp nhiu cht: Ph o hm ca hn hp nhiu cht c tnh cht cng tnh: n hhA A A A + + + = ...2 1

(1.6) ddAddAddAddAn hh+ + + = ...2 1 (1.7) 2222221222... dA ddA ddA ddA dn hh+ + + =

(1.8) Trong : 1, 2,.., n l cht 1, cht 2,, cht n Hnh 1.1. Ph hp th (a), ph o hm bc 2 (b) v bc 4 (c) (di ph nt chm c cng v tr v cng nhng rng gp i); Ph hp th ca trans-stilbene trong cyclohexane (d) v ph o hm bc 2 (e), bc 4 (f) tng ng [21] 13 Hnh 1.1 minh ha cho s chuyn dng t ph hp th UV-Vis sang ph o hm,chothyccphcthtngtnhaukhidngphhpthnhnglirt khc nhau khi dngph o hmdo s cc tr tng khis bc ca pho hm tng. Vic tng phc tp trong ph o hm c th ng dng trong phn tch nh tnh. Phohmbc1phnnhsbinthincahpthAivibc sng . Theo ngha ton hc ca o hm:-Ti cc im cc tr ca ph hp th, o hm bc 1 bng 0. -Ti cc im un ca ph hp th, o hm bc 2 bng 0. Trn ph o hm bc 1 ca cht th nht, ti nhng im c gi tr bng 0 c th tm c gi tr khc 0 trn ph o hm bc 1 ca cht th hai. Vy ti bc sng c th xc nh c cht th hai mt cch c lp, khng b cht th nht lm nh hng. Dng ph o hm bc 2 cng tng t: hai cht c cc i hp th gn nhau nhng cc im un c th xa nhau. Ti bc sng ng vi im un trn ph hp th (im 0 trn ng o hm bc 2) ca cht th nht s tm c gi tr khc 0 trn ng ph o hm bc 2 ca cht th hai. Nh vy, c th xc nh c cht th hai mt cch c lp, khng b cht th nht lm nh hng ti bc sng . 1.2.2.2. Phng php ph o hm t i [4] GiscphhpthUV-ViscamtdungdchchtXnngCxv mt dung dch cht Y nng C0. Ti mi bc sng, ta c mt gi tr ca t s giahpthcachtXvihpthcachtYnngC0.ngbiu din mi lin h gia cc gi tr ny vi bc sng nh sng l ph t i ca cht X so vi cht Y nng C0 v o hm ca ph t i ny c gi l ph o hm t i ca dung dch cht X so vi dung dch cht Y nng C0. Nh vy, ph t i ca cht Y nng Cy so vi cht Y nng C0 theo 14 l thuyt l mt ng thng song song vi trc bc sng. Do vy, o hm ca n lun bng 0. 00 0 ,,=((

=(((

CCddC l kC l kddyyy yii

(1.9) Trong :iyk ,l h s hp th ca cht Y bc sng i

ll b dy cc o, c coi lun bng nhau v thng l 1cm Nudungdchkhostchahaicht:chtXcnngCxvchtYc nng Cy, th o hm ph t i ca n so vi ph ca cht Y nng C0 c tnh nh sau: (((

=(((

+ 0 ,,0 ,,0 ,,C l kC l kddC l kC l kC l kC l kddiiiiiiyx xyx xyy y (1.10) Cng thc ny cho thy gi tr ph o hm t i ti bt k bc sng no cng ch ph thuc vo nng ca X v C0 ca cht Y. iu ny cho php khi C0 bit th c th xc nh c X cn c vo gi tr ph o hm o c ti mt bc sng c chn thch hp (phi c gi tr khc 0). 1.3. PHNG PHP SC K LNG HIU NNG CAO (HPLC) [1] 1.3.1. Nguyn tc chung ca HPLC Scklnghiunngcaolmtphngphptchdatrnilckhc nhau ca cc cht vi hai pha lun tip xc v khng ng tan vi nhau. Pha ng l cht lng chy qua ct vi mt tc nht nh di p sut cao, cn pha tnh l mtchtlngcbaotrnnnmtchtmangrn,hocmtchtmangrn c bin i bng lin kt ha hc vi cc nhm hu c. Pha ng cng vi mu th c bm qua ct di p sut cao, cc cht cn phn tch s di chuyn qua ct vi tc khc nhau. 1.3.2. Cc thng s c trng cho qu trnh sc k 15 a) H s phn b K Tc di chuyn ca cht tan qua pha tnh c xc nh bi h s phn b K: K = MSCC (1.11) Trong :CS: nng mol ca cht tan trong pha tnh CM: nng mol ca cht tan trong pha ng K ty thuc vo bn cht pha tnh, pha ng v cht ha tan. K nh th cht di chuyn nhanh, K ln th cht di chuyn chm. b) Thi gian lu tR -Thi gian lu (tR) l khong thi gian t lc tim mu vo ct n khi xut hin nh ca pic. -Thi gian cht (tM) l thi gian lu ca cht khng b lu gi. -Thi gian lu hiu chnh tR c tnh:tR = tR tM(1.12) Vi iu kin sc k nht nh, tR c trng v hng nh cho mi cht nn c ngha nh tnh. c) H s dung lng k Hsdunglngklthngsmttcdichuyncachtphntch qua ct, n biu th kh nng phn b ca cht tan trong mi pha v cn c gi l h s phn b khi lng gia 2 pha. k = MM RMSMStt tVVKQQ = = (1.13) Trong :QS, QM: lng cht tan trong pha tnh, pha ng VS, VM: th tch pha tnh, pha ng tR, tM: ln lt l thi gian lu, thi gian cht 16 d) H s chn lc H s chn lcoc trng cho tc di chuyn t i ca 2 cht A v B: o = ABKK(1.14) o= M A RM B RABt tt tkk=) () (''(1.15) ocho bit hiu qu tch hai cht ca h thng sc k, vi qui c cht B b lugimnhhnchtA(KB>KA),vvyo >1.Nuo qunh,haichttch khinhaukm.Nuo quln,thigianphntchsdi.Trstiulo = 1,05 2,00. e) H s bt i AF H s bt i AF nh gi tnh bt i xng ca pic:AF = ab(1.16) Trong :b: l chiu rng pha sau ca pic o 1/10 chiu cao ca pic a: l chiu rng pha trc ca pic o 1/10 chiu cao ca pic Gi tr AF cng gn ti 1, pic cng i xng.f) S a l thuyt (N) v chiu cao ca a (H) Hiu lc ca ct thng c o bng hai thng s: S a l thuyt v chiu cao a l thuyt. Ct sc k c coi nh c N tng l thuyt, mi tng s phn b cht tan vo hai pha li t n mt trng thi cn bng mi. Mi tng c gi nh nh l mt lp cht nhi c chiu cao l H. Ta c th tnh s al thuytN theo cng thc sau:N = 16 x 2|.|

\|WtR (1.17) 17 N = 5,54 x 22 / 1||.|

\|WtR(1.18) Trong : W: l chiu rng pic o y pic W1/2: l chiu rng pic o na chiu cao ca nh pic NugiLlchiudicactsck,thchiucao(H)caalthuyt c tnh bng cng thc:NLH = (1.19) N, H: phn nh hiu lc ct. Ct c N ln, H nh th hiu lc ct cao. g) phn gii RS ca ct Hnh 1.2. S tch 2 cht A v B trn 3 ct c phn gii khc nhau phn gii nh gi kh nng tch hai cht trn sc k cho hai pic lin k nhau. 18 RS = ) (] ) ( ) [( 2B AA R B RW Wt t+ (1.20) Phng trnh trn thch hp cho vic tnh RS khi 2 pic tch hn nhau (tc RS 1,5). TrongtrnghpRSnhhn,tc2picbtrnglpmtphnthvicxc nhWA vWB s kh khn hn lW1/2(A) v W1/2(B) lc nyRS sc tnh theo cng thc: RS = ) (] ) ( ) [( 18 , 1) ( 2 / 1 ) ( 2 / 1 B AA R B RW Wt t+ (1.21) RS = ''114BBkk N+oo(1.22) Trong : tRA, tRB : thi gian lu ca 2 pic lin k nhau WA, WB: rng ca pic o cc y pic W1/2A, W1/2B: rng pic o na chiu cao ca nh pic N: s a l thuyt kB: h s dung lng ca cht B Yu cu RS>1, gi tr ti u RS = 1,5: RS = 1,50: hai pic tch gn nh hon ton, ch cn xen ph 0,3%. RS = 1,00: hai pic tch nhau tng i, cn xen ph 0,4%. RS = 0,75: hai pic tch khng tt, cn xen ph nhiu.

19 CHNG 2 I TNG V PHNG PHP NGHIN CU 2.1. I TNG NGHIN CU, NGUYN LIU V THIT B 2.1.1. i tng nghin cu Bng 2.1. i tng nghin cu Tn bit dc c im SulperazonJincetamBacampCefactam Hng sn xutCngty dcphm Pfizer Cngty TNHHdc phm Hankook Korus Cng ty TNHH dcphm Hanlim Cngtydc phm Laboratorios SK20A4052953VN - 8024 - 09VN - 5179 - 10VN - 4852 - 07 S l12902400A017091070104120034 NSX10/02/201221/06/201204/07/201110/2012 HD08/02/201420/06/201503/07/201410/2014

BabitdcSulperazon,JincetamvBacampccngthccho1lthuc bt pha tim gm: -Cefoperazon: 500 mg -Sulbactam: 500 mg 20 Trong , 547 mg sulbactam dng mui tng ng vi 500 mg sulbactam v 517 mg cefoperazon dng mui tng ng vi 500 mg cefoperazon. Cefactam c cng thc cho mt l thuc bt pha tim gm: -Cefoperazon: 1000 mg -Sulbactam: 1000 mg Trong , 1094 mg sulbactam dng mui tng ng vi 1000 mg sulbactam v 1034 mg cefoperazon dng mui tng ng vi 1000 mg cefoperazon. 2.1.2. Nguyn liu v thit b Nguyn liu: -Cht chun (Vin kim nghim thuc Trung ng): +Cefoperazon: 200 mg/l 89,82% C25H26N9 NaO 8S2 SKS: WS.0211230.01(08.07) BQ: 2 - 8C. Trnh nh sng+Sulbactam: 200 mg/l 899,1 g C8H11NO5S/mg (khan) m: 0,45% SKS: 0103159BQ: 2 - 8C. Trnh nh sng -Dungmi:ncct2lnloibion,acetonitriltinhkhitHPLC(Merck- c), nc pH 3,2 c iu chnh bng H3PO4 c. 21 Thit b: -My quang ph hai chm tia UNICAM UV30 (ThermoSpectronic) c kt nivimytnh(chyhiuhnhWindowXP)viphnmmchuyn dng Vision 32. Ch o:Start Lamda: 190 nm - Stop Lamda: 325 nm Data Mode: AbsorbanceBand width: 1,5 nm Scan Speed: Intelliscan nm/min Data Interval: Very High Res -My ct nc 2 ln Hamilton (Hamilton - Anh) -Cuvet thch anh b dy 1 cm -My HPLC: Agilent Technologies 1200 series (Agilent Technologies - M). Ct sc k: Apollo C18 (150mm x 4,6mm; 5 m) -B mng lc mu 0,20 m -Bm tim, l ng mu HPLC -Bm ht chn khng: Vacuubrand GMBH + CO KG (Vacuubrand- c) -B dng c lc chn khng Supelco (Supelco - M) -My o pH: EUTECH INSTRUMENTS pH 510 (EUTECH - c) -My lc nc: Maxima Ultra pure water (Elga - Anh) -My siu m: Ultrasonic LC 60H (ELMA - c) -Cn phn tch Sartorius (Sartorius - c) -Cc dng c khc:22 +Bnh nh mc loi 50 ml +Pipet chnh xc loi 2 ml +Pipet t ng loi 1000 l (NICHIRYO - Nht) +Cc c m loi 100 ml, 500 ml v 1000 ml +Qu bp cao su +a thy tinh 2.2. PHNG PHP NGHIN CU Phngphpquangphohm,phohmtivHPLCcs dng trong ti ny. ti c tin hnh qua cc bc sau: 2.2.1. Xy dng phng php nh lng ng thi cefoperazon v sulbactam bng quang ph o hm ly HPLC lm phng php i chiu Cc phng php quang ph o hm -La chn bc sng nh lng thch hp -Lachnkhongnngcsphthuctuyntnhgianngv cc gi tr PH, PHT -La chn h s lm trn v s ln lm trn -La chn s chia nng vi phng php PHT -Kim tra lp li ca phng php -Kim tra ng ca phng php Phng php HPLC -La chn iu kin sc k: pH v nng m, t l dung mi pha ng, tc dng, bc sng pht hin theo ti liu tham kho [23]. -Thm nh phng php: +Xc nh khong tuyn tnh gia din tch pic v nng cht phn tch 23 +Xc nh lp li ca phng php +Xc nh ng ca phng php 2.2.2.ngdngccphngphpquangphnuvHPLCnhlng ng thi cefoperazon v sulbactam trong ch phm Tinhnhnhlngngthicefoperazonvsulbactamtrongcngmt mu bng cc phng php: - Quang ph UV-Vis: Ph o hm, ph o hm t i - HPLC 2.2.3.X l kt qu thc nghim [5] SphthuctuyntnhcanngviccgitrPHvPHTc thitlpbngphngphpbnhphngtithiuvigitrtuyticahs tng quan ng chun> 0,990. So snh kt qu ca cc phng php quang ph v HPLC v ng v lp bng thng k ton hc (kim nh ANOVA (phn tch phng sai) v Bartlett). Gi tr trung bnh:nXXnii ==1 (2.1) lch chun: SD = ) 1 () (12=nX Xnii (2.2) Phng sai:S2 = 1) (12=nX Xnii (2.3) lch chun tng i:RSD = 100 XSD(2.4) 24 CHNG 3 THC NGHIM, KT QU V BN LUN 3.1. KT QU THC NGHIM V NHN XT 3.1.1. Phng php quang ph o hm 3.1.1.1. Chun b dung mi Nc ct 2 ln loi b ion 3.1.1.2. Chun b mu chun v mu th Chun b mu chun: -Pha dy dung dch CP chun: 10, 15, 20, 25, 30, 35 mg/l +Pha dung dch CP gc 5000 mg/l: cn chnh xc khong 0,2500 g CP, chovobnhnhmc50ml,thmkhong30mlncvlcu, siu m khong 20 pht, b sung nc va . +Pha dy dung dch CP chun trong bnh nh mc 50 ml t dung dch CP gc 5000 mg/l. -Tngt,phadydungdchSBchuntrongbnhnhmc50mlt dung dch SB gc 5000 mg/l. -Pha dung dch hai thnh phn CP (10, 15, 20, 25, 30, 35 mg/l) v SB (20 mg/l) t cc dung dch gc n cht nu. -Pha dung dch hai thnh phn SB (10, 15, 20, 25, 30, 35 mg/l) v CP (20 mg/l). 25 Chun b mu th: Cn chnh xc mt lng bt thuc tim tng ng vi khong 0,2500 g CP v 0,2500 g SB cho vo bnh nh mc 50 ml, thm khong 30 ml nc v lc u, siu m khong 20 pht, b sung nc va . Ly 200 l dung dchva pha cho vo bnh nh mc 50ml, b sung nc va c dung dch th c nng CP 20 mg/l v SB 20 mg/l. 26 3.1.1.3. Xy dng phng php nh lng A. Chn bc sng nh lng a. Xc nh khong cng tnh 0,00,20,40,60,81,01,21,41,6200 220 240 260 280 300 hp thBc sng (nm)CP 20SB 20CP 20 + SB 20Ph cngHnh 3.1. Ph hp th ca SB 20 mg/l, CP 20 mg/l, ph cng v ph hn hp SB 20 mg/l + CP 20 mg/l Hnh 3.1 biu din ph hp th ca cc dung dch chun SB 20 mg/l, CP 20 mg/l, hn hp SB 20 mg/l + CP 20 mg/l. So snh ph cng tng ng ca SB v CP viphhnhpcahaichtnychothySBvCPctnhcngtnhnhsng trong khong bc sng kho st 210 - 300 nm. 27 b. Phng php ph o hm bc 1 XydngPHtphhpthcatngdungdchchunncht(cc nngkhcnhau),lyohmbc1caphhpthcPHbc1(Hnh 3.2).-2,5-2,0-1,5-1,0-0,50,00,51,0210 230 250 270 290Gi tr PH bc 1Bc sng (nm)CP (10 - 35)SB (10 - 35)228,0286,0Hnh 3.2. PH bc 1 ca dy dung dch SB nng thay i (10 - 35 mg/l) v ca dy dung dch CP nng thay i (10 - 35 mg/l) -Trn PHbc 1 ca dydung dch CP c nng thayi t 10mg/l n 35 mg/l, ti = 217,9; 228,0 v 262,7 nm gi tr PH bc 1 ca CP bng 0. Ti cc bc sng ny, gi tr PH bc 1 ca hn hp CP v SB ch cn ph thuc vo gi tr PH bc 1 ca SB, ngha l ch ph thuc vo nng ca SB m khng b nh hng bi CP. Qua kho st, nh lng SB ti = 228,0 nm cho sai s nh hn. 28 -Tng t, trn PH bc 1 ca dy dung dch SB c nng thay i t 10 mg/l n35mg/l,ticc=255,7300,0nmthgitrPHbc1bng0.Ti khongbcsngny,gitrPHbc1cahnhpCPvSBchcnph thuc vo nng ca CP m khng b nhhng bi SB. Qua kho st, nh lng CP ti bc sng 286,0 nm cho sai s nh. c. Phng php ph o hm t i Vi CP -200-1000100200210 230 250 270 290Gi tr PHT bc 1Bc sng (nm)CP (10 - 35) + SB 20 chia SB 20CP (10 - 35) chia SB 20260,9Hnh 3.3. PHT bc 1 ca dy dung dch hn hp CP (10 - 35 mg/l) + SB 20 mg/l vi s chia l SB 20 mg/l v ca dy dung dch CP (10 - 35 mg/l) vi s chia l SB 20 mg/l 29 Ly ph hp th ca dy dung dch hn hp CP (10 - 35 mg/l) v SB 20 mg/l chia cho ph hp th ca dung dich SB 20 mg/l c PT. Ly o hm bc 1 ca ph t i ny c PHT bc 1.Tng t ly PHT bc 1 vi dy dung dch CP (10 - 35 mg/l).Qua kho st, = 260,9 nm c la chn nh lng CP. Vi SB -2,5-2,0-1,5-1,0-0,50,0220 225 230 235 240 245 250Gi tr PHT bc 1Bc sng (nm)CP 20 + SB (10 - 35) chia CP 10SB (10 - 35) chia CP 10236,6 Hnh 3.4. PHT bc 1 ca dy dung dch hn hp SB (10 - 35 mg/l) + CP 20 mg/l vi s chia l CP 10 mg/l v ca dy dung dch SB (10 -35 mg/l) vi s chia l CP 10 mg/l Ly ph hp th ca dy dung dch hn hp SB (10 - 35 mg/l) v CP 20 mg/l chia cho ph hp th ca dung dich CP 10 mg/l c PT. Ly o hm bc 1 ca PT ny c PHT bc 1. Tng t ly PHT bc 1 vi dy dung dch CP 10 - 35 mg/l.30 Qua kho st, = 236,6 nm c la chn nh lng SB. B. Kho st khong tuyn tnh a. Ph o hm bc 1 Vi CP: Tin hnh kho st mi quan h gia gi tr PH bc 1 v nng CP (10 - 35 mg/l) ti = 286,0 nm. Kt qu th hin Bng 3.1 v Hnh 3.5. Bng 3.1. Kt qu kho st khong tuyn tnh ca CP vi gi tr PH bc 1 Nng (mg/l)Gi tr PH bc 1 10 0,543 15 0,836 20 1,089 25 1,371 30 1,640 35 1,899 y = 0,0541x 0,0116r = 0,9998-2,5-2,0-1,5-1,0-0,50,05 10 15 20 25 30 35 40Gi tr PH bc 1Nng (mg/l) Hnh 3.5. th biu din ng chun ca CP (10 - 35 mg/l) vi gi tr PH bc 1 31 Vi SB: Tin hnh kho st mi quan h gia gi tr PH bc 1 v nng SB (10 - 35 mg/l) ti = 228,0 nm. Kt qu th hin Bng 3.2 v Hnh 3.6. Bng 3.2. Kt qu kho st khong tuyn tnh ca SB vi gi tr PH bc 1 Nng (mg/l)Gi tr PH bc 1 10 0,141 15 0,218 20 0,283 25 0,358 30 0,427 35 0,500 y = 0,0143x 0,0001r = 0,9998-0,6-0,5-0,4-0,3-0,2-0,10,00 5 10 15 20 25 30 35 40Gi tr PH bc 1Nng (mg/l) Hnh 3.6. th biu din ng chun ca SB (10 - 35 mg/l) vi gi tr PH bc 1 32 b. Ph o hm t i Vi CP: Tin hnh kho st mi quan h gia gi tr PHT bc 1 v nng CP (10 - 35 mg/l) ti = 260,9 nm. Kt qu th hin Bng 3.3 v Hnh 3.7. Bng 3.3. Kt qu kho st khong tuyn tnh ca CP vi gi tr PHT bc 1 Nng (mg/l)Gi tr PHT bc 1 1023,646 1535,721 2046,128 2558,499 3070,279 3580,954 y = 2,3005x + 0,7769r = 0,99980204060801005 10 15 20 25 30 35 40Gi tr PHT bc 1Nng (mg/l) Hnh 3.7. th biu din ng chun ca CP (10 - 35 mg/l) vi gi tr PHT bc 1 33 Vi SB: Tin hnh kho st mi quan h gia gi tr PHT bc 1 v nng SB (10 - 35 mg/l) ti = 236,6 nm. Kt qu th hin Bng 3.4 v Hnh 3.8. Bng 3.4. Kt qu kho st khong tuyn tnh ca SB vi gi tr PHT bc 1 Nng (mg/l)Gi tr PHT bc 1 10 0,353 15 0,532 20 0,699 25 0,872 30 1,032 35 1,193 y = 0,0336x 0,0251r = 0,9998-1,4-1,2-1,0-0,8-0,6-0,4-0,20,05 10 15 20 25 30 35 40Gi tr PHT bc 1Nng (mg/l) Hnh 3.8. th biu din ng chun ca SB (10 - 35 mg/l) vi gi tr PHT bc 1 34 Tm li, mi quan h gia nng ca CP v SB vi gi tr PH, HT ti cc bc sng nh lng c trnh by trong Bng 3.5. Bng 3.5. Kt qu kho st khong tuyn tnh ca CP v SB Phng php Hot cht Bc sng (nm) Khong tuyn tnh (mg/l) Phng trnh hi quy H s tng quan r PH bc 1 CP286,010 - 35Y = 0,0541x 0,0116 0,9998 SB228,010 - 35Y = 0,0143x 0,0001 0,9998 PHT CP 260,910 - 35 Y =2,3005x + 0,77690,9998 SB236,610 -35Y = 0,0336x 0,0251 0,9998 35 3.1.1.4. lp v ng ca phng php lpvngcaphngphpcxcnhdatrnktqunh lng ca 6hn hp t to CP 20mg/l + SB 20mg/ltrong cngiu kin (Bng 3.6). Bng 3.6. lp v ng ca cc phng php quang ph o hm Hm lng % tm li STT PH bc 1PHT CPSBCPSB 1100,3100,1100,2100,2 299,7101,8101,897,1 3101,3100,7101,9101,9 4101,6102,9101,6101,9 5102,2102,1100,5101,8 6101,3105,099,9101,9 TB101,1102,1101,0100,8 SD0,91,70,91,9 RSD0,91,70,91,9 36 3.1.2. Phng php HPLC 3.1.2.1. Chun b mu nghin cu Dung mi: Nc ct 2 ln loi b ion Dung dch chun v dung dch th: Cc dung dch chun v th sau khi c chun b nh m t trong mc 3.1.1.2 u c lc qua mng c kch thc l lc 0,20 m. 3.1.2.2. Xy dng phng php HPLC -iu kin sc k: Ct: Apollo C18 150 mm x 4,6 mm; 5 m Detector UV: 210 nm Tc dng: 0,8 ml/pht Pha ng: hn hp acetonitril - acid photphoric pH 3,2 (60 : 40, tt/tt) Th tch tim mu: 20 l -Hnh 3.9 biu din sc k ca hn hp chun CP 20 mg/l + SB 20 mg/l.-Cc thng s ca qu trnh sc k c trnh by trong Bng 3.7. Hnh 3.9. Sc k ra gii hn hp chun CP 20 mg/l + SB 20 mg/l 37 Bng 3.7. Thng s qu trnh HPLC ca hn hp CP 20 mg/l + SB 20 mg/l STTThng sCPSB 1 lpRSD < 2% 99 101% R2 > 0,990 AF = 1,2 2 ng 3Khong tuyn tnh 10 - 35 mg/l 4H s bt i 5 phn giiRs = 12 6S a l thuyt/mt44000162000 7Thi gian lu (pht)tR = 4,4tR = 2,7 38 3.1.3. Kt qu nh lng Kt qu nh lng mt s ch phm thuc bt pha tim hin ang lu hnh trn th trngbng quang ph o hmvHPLC c trnh bytrong Bng 3.8. Kt qu cho thy khng c s saikhc c ngha thng k (p > 0,05) v ng (ANOVA)vlp(Bartlett)khisosnhccsliucaquangphohmv HPLC vi cc ch phm em nh lng. Bng 3.8. Kt qu nh lng cc ch phm bng quang ph o hm v HPLC Ch phm Hm lng % so vi nhn HPLCPHPHT Cefactam CP100,1 1,199,4 0,7100,4 0,9 SB99,3 1,298,5 1,799,2 0,8 Sulperazon CP100,4 0,999,9 1,3100,5 0,6 SB100,3 1,299,4 1,899,7 1,3 Jincetam CP99,8 0,7100,6 1,0100,0 0,8 SB99,1 1,098,9 1,699,3 0,9 Bacamp CP100,9 1,1101,6 0,7100,4 0,9 SB100,8 0,8101,2 1,3100,5 1,2 39 3.2. BN LUN Kt qu thc nghim cho thy quang ph o hm c th nh lng c ngthicefoperazonvsulbactamtrongcnghnhpmkhngcnchittch. Ngoi ra, vic s dng nc lm dung mi l mt u im ni bt cacc phng php ny.Viphngphpquangphohm,haiyutchnhnhhngnkt qunhlnglbcsngnhlngvccthngstrongphplyohm, php lm trn ph . Quakhost,bcsngclachnnhlngccchphmth trong ph o hm bc 1 l 286,0 nm vi cefoperazon v 228,0 nm vi sulbactam. V nguyn tc c th tin hnh nh lng cefoperazon ti nhiu bc sng (Hnh 3.2),tuynhinbcsng286,0clachnvsaisinhlngtibcsng nylnhnht.Tngt,bcsng228,0nmclachnnhlng sulbactam. Thng s cho thut ton o hm bc 1 c la chn l: -Php ly o hm: Order: 5; No of coefficient: 9. -Php lm trn: Order: 3; No of coefficient: 125. S khc bit v bc sng nh lng bng ph o hm bc 1 trong nghin cu ny so vi ti liu tham kho [8] c th do s khc nhau v cch x l ph v ly o hm vi tng loi my o quang c s dng. Vi phng php PHT,vic la chn bc sng v nng cht chun lm mu s khi thc hin php chia nh hng nhiu n chnh xc ca kt qu. Tnh thchhp ca thut ton ny c khng nhqua s trng lp v ph ca dy dung dch mt thnh phn v dy dung dch hn hp c nng tng ng (Hnh 3.3 v Hnh 3.4). Qua kho st, bc sng = 260,9 nm v s chia nng SB = 20 mg/l c la chn nh lng cefoperazon, cn bc sng = 236,6 nm v s chia nng CP = 10 mg/l c la chn nh lng sulbactam. 40 Thng s ca thut ton o hm bc 1 ph t i c la chn cng tng t nh thut ton o hm bc 1 nu. Ktquthmnhphngphpquangphohmchothycsph thuc tuyn tnh cht ch gia tnh hiu o hm v nng CP v SB trong khong 10-35mg/l(Bng3.5).Phng php cngcao(tlphn trmtmlitrong khong 100 - 102 %), lp cao (RSD 2%).Trongnghincuny,HPLCcsdunglmphngphpichiuvi quang ph o hm. Cc iu kin sc k c la chn da vo cc cng trnh nghin cu trc y c trnh by trong mc 3.1.2.2. Kt qu thm nh cho thy phng php HPLC ny p ng c yu cu cho php nh lng ng thi CP v SB trong ch phm thuc bt pha tim (Bng 3.7). Ktquthngkvitincy95%chothykhngcskhcbitc ngha (p > 0,05) v ng (ANOVA) v lp (Bartlett) gia quang ph o hm v HPLC. Do vy, phng php quang ph o hm c xy dng hon ton c th thay th HPLC nh lng ng thi cefoperazon v sulbactam trong cc ch phm thuc bt pha tim.

41 KT LUN V KIN NGH Da vo kt qu nghin cu ca ti, chng ti rt ra mt s kt lun nh sau: -Xydngcphpnhlngngthicefoperazonvsulbactam trong cc ch phm thuc bt pha tim mt cch trc tip (khng cn qua giai on chit tch) bng cc phng php: ph o hm bc 1, ph o hm t i vi lp v ng cao. S khc bit v chnh xc( lp v ng) ca cc phng php nu v HPLC khng c ngha thng k vi mc tin cy t 95%. -Cc phng php quang ph o hm nu c k thut n gin, ch s dngdungmivhachtthngthng(ncct2ln)nndthc hinvtitkimkinhph.Do,quangphohmckhnngng dngtrongcngtckimnghimbtthucphatimccha cefoperazon v sulbactam. -Khng c s sai khc ng k gia hm lng thc t ca cc mu thuc c nh lng vi hm lng ghi trn nhn. Trn c s kt qu t c, chng ti kin ngh mt s vn sau: -Tip tc nghin cu ng dng cc phng php quang ph o hm vo kim nghim cc dng bo ch a thnhphn khc ckh nng hp th UV-Vis. -ngdngccphngphpquangphohmnunhlng ngthicefoperazonvsulbactamtrongccchphmthucbtpha tim cha ti cc c s kim nghim c trang b my quang ph hin i. I TI LIU THAM KHO Ti liu ting Vit: 1. 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