Xây dựng quy trình kỹ thuật để định lượng tobramycin bằng phương pháp HPLC

8132019 Xacircy dựng quy trigravenh kỹ thuật để định lượng tobramycin bằng phương phaacutep HPLC

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AacuteN TT NGHIP

tagravei ldquoXacircy dng quy trigravenh k thut

nh lư ng tobramycin bng

phươ ng phaacutep HPLC rdquo

wwwdaykemquynhonucozcom

wwwfacebookcomdaykemquynhon



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MC LC

PHN 1 TNG QUAN - 3 -11 TNG QUAN V TOBRAMYCIN - 7 -111 Cocircng th c cu to - 7 -112 Tiacutenh cht lyacute hoacutea - 7 -113 Ngun gc - 7 -114 Dư c ng hc - 7 -115 Taacutec dng vagrave cơ ch taacutec dng - 8 -116 Ch nh - 8 -117 Chng ch nh - 9 -118 Dng bagraveo ch vagrave liu lư ng - 9 -12 MT S PHƯƠ NG PHAacuteP NH LƯ NG TOBRAMYCIN - 9 -121 nh lư ng tobramycin bng phươ ng phaacutep vi sinh - 9 -1211 Phươ ng phaacutep 1 [15] - 9 -1212 Phươ ng phaacutep 2 [3] - 10 -1213 Phươ ng phaacutep 3[14] [20] - 10 -122 nh lư ng tobramycin bng phươ ng phaacutep o quang ph hp th UV-VIS- 11 -1221 Phươ ng phaacutep 1 [4] - 11 -1222 Phươ ng phaacutep 2 [19] - 11 -123 nh lư ng tobramycin bng phươ ng phaacutep HPLC - 12 -1221 Phươ ng phaacutep 1 [14] - 12 -1222 Phươ ng phaacutep 2 [20] [23] - 12 -1223 Phươ ng phaacutep 3 [15] - 13 -1224 Phươ ng phaacutep 4 [16] - 13 -1225 Phươ ng phaacutep 5 [15] - 14 -1226 Phươ ng phaacutep 6 [5] - 14 -1227 Phươ ng phaacutep 7 [9] - 15 -13 TNG QUAN V PHƯƠ NG PHAacuteP SC KIacute LNG HIU NĂNGCAO (HPLC) - 16 -131 Nguyecircn tc - 16 -132 Cơ s lyacute thuyt - 17 -133 Nguyecircn tc cu to h thng HPLC - 17 -

1331 H th ng bơ m - 17 -1332 Bigravenh ch a dung mocirci vagrave h th ng x lyacute dung mocirci - 17 -1333 H tiecircm m u - 17 -1334 C t s c kyacute lng HPLC - 18 -1335 Detector trong HPLC - 18 -1336 Thi t b hi n th k t qu - 18 -





- 5 -

134 Caacutec thocircng s c trư ng ca quaacute trigravenh sc kyacute - 19 -135 Cơ s lyacute thuyt ca vic l a chn iu kin sc kyacute - 21 -1351 L a chn ct (pha t ĩ nh) [12] [18] - 21 -1352 L a chn pha ng cho HPLC [12] [18] - 22 -

1353 Chn m pH - 23 -1354 T c dograveng - 23 -136 Caacutech aacutenh giaacute pic - 23 -137 ng dng ca HPLC - 24 -1371 nh tiacutenh vagrave th tinh khi t - 24 -1372 S c kyacute i u ch - 24 -1371 nh lư ng - 24 -PHN 2 TH C NGHIM VAgrave KT QU - 26 -21 NGUYEcircN VT LIU VAgrave PHƯƠ NG PHAacuteP TH C NGHIM - 26 -211 Nguyecircn vt liu - 26 -

2111 Nguyecircn liu vagrave hoaacute cht - 26 -2112 Dng c - 26 -212 Ni dung vagrave phươ ng phaacutep nghiecircn c u - 26 -2121 Phươ ng phaacutep nghiecircn cu - 26 -2122 Ni dung nghiecircn cu - 27 -22 TH C NGHIM VAgrave KT QU - 28 -221 Xacircy d ng quy trigravenh k thut nh lư ng tobramycin bngphươ ng phaacutep HPLC - 28 -2211 Nguyecircn tc l a chn iu kin sc kyacute - 28 -2212 Kho saacutet l a chn iu kin sc kiacute - 28 -a Kho saacutet chn bư c soacuteng thiacutech h p - 28 -Higravenh 1 Ph hp th ca dung dch tobramycin 002 - 29 -b La chn ct- 29 -c La chn m - 29 -d La chn pha ng - 30 -e La chn tc dograveng - 30 - iu kin sc kyacute la chn - 31 -222 Qui trigravenh nh lư ng - 31 -2221 Kho saacutet tiacutenh thiacutech h p ca h thng - 32 -2222 Xacircy dng phươ ng phaacutep nh lư ng - 32 -2223 iu kin sc kyacute - 33 -2224 Tiacutenh kt qu - 33 -223 nh lư ng tobramycin nguyecircn liu bng phươ ng phaacutep m i xacircyd ng - 34 -224 aacutenh giaacute phươ ng phaacutep nh lư ng - 35 -2241 Tiacutenh tuyn tiacutenh - 35 -





- 6 -

2242 Tiacutenh chiacutenh xaacutec - 37 -2243 Tiacutenh uacuteng - 37 - Bng 6 K t qu aacutenh giaacute tiacutenh uacuteng ca phươ ng phaacutep - 38 -2244 Tiacutenh c hiu - 39 -

23 BAgraveN LUN VAgrave KT QU - 40 -KT LUN VAgrave XUT - 43 -31 KT LUN - 43 -32 XUT - 43 -





- 7 -

PHN 1 TNG QUAN11 TNG QUAN V TOBRAMYCIN

111 Cocircng th c cu to

C18H37N5O9 PTL 4675

Tecircn khoa hc 4-O-(3-Amino-deoxy-α-D-glucopyranosyl)-2-deoxy-6-O-(26-

diamino-236-trideoxy-α-D-ribo-hexopyranosyl)-L-streptamine [21]

112 Tiacutenh cht lyacute hoacutea

Bt magraveu trng hoc trng ngagrave D tan trong nư c rt khoacute tan trong

ethanol thc t khocircng tan trong cloroform vagrave ether

Goacutec quay cc riecircng [α]D20 +1380 n +1480[7]

113 Ngun gc

Chit xut t mocirci trư ng nuocirci cy Streptomyces tenebrarius coacute th

baacuten tng h p t kanamycin B [7]

114 Dư c ng hc

Tobramycin hu như khocircng hp thu qua ư ng tiecircu hoaacute nhưng hp thu

tt qua ư ng tiecircm bp tiecircm t ĩ nh mch Thuc iacutet liecircn kt v i protein huyt





- 8 -

tươ ng Do phacircn t phacircn cc mnh necircn khoacute i vagraveo caacutec mocirc k c natildeo Thuc

t nng cao trong v thn Khi s dng cho ph n mang thai thuc tiacutech

lũy trong thai gacircy c cho c m vagrave con Tobramycin agraveo thi ch yu qua

thn necircn phi gim liu khi suy thn thư ng da vagraveo creatinin huyt thanh traacutenh c tiacutenh Hin nay tobramycin ư c dugraveng v i ch mt liu cao vagrave

duy nht trong ngagravey Caacutech dugraveng nagravey cho thy hiu qu iu tr cao hơ n vagrave iacutet

c tiacutenh hơ n so v i dugraveng liu nh vagrave nhiu ln trong ngagravey như trư c acircy i

v i caacutec ca nng (nhim Pseu aeruginosa ngư i gim neutrophile vagrave caacutec ca

suy thn) cn khong caacutech liu dagravei hơ n 24 gi Th i gian baacuten thi ca

tobramycin 2 - 5 gi [6]

115 Taacutec dng vagrave cơ ch taacutec dng

Tobramycin lagrave mt khaacuteng sinh nhoacutem aminoglycosid coacute taacutec dng trecircn

nhiu vi khun Gr (-) hiu khiacute vagrave mt s vi khun Gr (+) hiu khiacute Thuc

khocircng coacute taacutec dng v i Chlamydia nm virus vagrave a s caacutec vi khun ym khiacute

Tobramycin rt ging gentamycin v tiacutenh cht sinh hc vagrave c tiacutenh

chuacuteng coacute cugraveng na i thi tr nng nh trong huyt thanh iacutet liecircn kt v i

protein th tiacutech phacircn b vagrave s bagravei tit ch yu qua lc cu thn

im quan trng nht ca tobramycin lagrave coacute hot tiacutenh i v i phn l n

caacutec chng Pseudomonas aeruginose mnh hơ n c gentamycin

Cơ ch taacutec dng Tobramycin c ch s tng h p protein caacutec vi

khun nhy cm bng caacutech gn khocircng thun nghch v i caacutec tiu phacircn 30S ca

ribosom [2] [11]

116 Ch nh

ư c ch nh trong caacutec bnh nhim khun nng e da t i tiacutenh mngc bit v i caacutec bnh magrave nguyecircn nhacircn chưa rotilde ragraveng hoc b nhim khun

huyt do vi khun Gr (-)





- 9 -

Trong iu tr caacutec bnh nhim khun nng hoc trong caacutec bnh nhim

khun nng toagraven thacircn do Pseudomonas sp gacircy ra tobramycin coacute th dugraveng

phi h p v i mt khaacuteng sinh nhoacutem beta-lactam

Trong bnh viecircm ni tacircm mc do Streptococcus faecalis hoc alpha -

Streptococcus gacircy ra coacute th dugraveng tobramycin phi h p v i ampicilin hoc

benzylpenicilin nhưng phi tiecircm riecircng r [2]

117 Chng ch nh

Ngư i coacute tin s d ng v i caacutec khaacuteng sinh loi aminoglycosid ngư i

nghe keacutem vagrave ngư i coacute bnh thn [2]

118 Dng bagraveo ch vagrave liu lư ng

Tobramycin sulphat Dung dch tiecircm bp tiecircm t ĩ nh mch 40 mgml

(ngư i l n) 10 mgml (tr em) Bt pha tiecircm 30 - 40 mgl

L 5 ml 03 nh mt Tuyacutep 35 g m tra mt 03

Dng thuc hiacutet qua ư ng ming bng maacutey khi dugraveng iu tr nhim

P aeruginosa ư ng hocirc hp bnh nhacircn b xơ nang hoacutea [11]

12 MT S PHƯƠ NG PHAacuteP NH LƯ NG TOBRAMYCIN

121 nh lư ng tobramycin bng phươ ng phaacutep vi sinh

1211 Phươ ng phaacutep 1 [15]

- Chng vi khun Bacillus subtilis CMCC(B)63501

- Dung dch m phosphat pH 78 plusmn 01

Dikali hydrophosphat 559 g

Kali dihydrophosphat 041 g

- Mocirci trư ng nh lư ng

Pepton 50 gCao tht 30 g


Thch 150 - 200 g





- 10 -

Nư c ct 1000 ml

pH sau khi tit trugraveng 78 plusmn 02

- Chun b dung dch chun vagrave th sau oacute tin hagravenh th vagrave tiacutenh kt qu

theo [15]1212 Phươ ng phaacutep 2 [3]

- Chng vi khun Bacillus pumilus NCTC 8241



Kali dihydrophosphat 0523g


Cao men bia 30 g

Pepton 60 g

Casein pancreatic 40 g

Glucose 10 g

Cao tht 15 g

Thch 150 g

Nư c ct 100 mlpH sau khi tit trugraveng 82 plusmn 02

- Chun b dung dch chun vagrave th

Cacircn chiacutenh xaacutec khong 250 mg tobramycin hogravea tan trong dung dch

m phosphat pH 80 coacute nng tobramycin chiacutenh xaacutec khong 10 IUml

20 IUml vagrave 40 IUml

- Tin hagravenh th vagrave tiacutenh toaacuten kt qu theo phươ ng phaacutep hot lc khaacuteng

sinh - DVN III1213 Phươ ng phaacutep 3[14] [20]

- Xaacutec nh hot lc khaacuteng sinh ca tobramycin bng phươ ng phaacutep vi

sinh vt theo BP 2000 hay JP14





- 11 -

- Chng vi khun Bacillus subtilis ATCC 6633

- Mocirci trư ng nuocirci cy Mocirci trư ng c

- Dung dch chun

Cacircn chiacutenh xaacutec mt lư ng tobramycin chun tươ ng ươ ng khong 25mg (hiu lc) hoagrave tan trong dung dch m phosphat 01 moll pH 8 thagravenh

chiacutenh xaacutec 25 ml vagrave oacute chiacutenh lagrave dung dch chun gc Gi dung dch chun

gc nhit t 5οC n 15οC vagrave s dng trong vograveng 30 ngagravey Pha loatildeng

dung dch bng dung dch chun m phosphat 01M pH 8 ư c caacutec dung

dch coacute nng 8microgml vagrave 2microgml (theo hiu lc)

- Dung dich th

Cacircn chiacutenh xaacutec mt lư ng tobramycin tươ ng ươ ng khong 25 mg(hiu lc) hoagrave tan trong dung dch m phosphat 01 moll pH 8 thagravenh chiacutenh

xaacutec 25ml Pha loatildeng dung dch bng dung dch chun m phosphat 01M

pH 8 ư c caacutec dung dch coacute nng 8microgml vagrave 2microgml (theo hiu lc)

122 nh lư ng tobramycin bng phươ ng phaacutep o quang ph hp th

UV-VIS


Cacircn mt lư ng tobramycin sulfat tươ ng ng v i khong 03 gtobramycin nguyecircn liu cho vagraveo bigravenh nh mc coacute dung tiacutech 100 ml b sung

nư c ct 2 ln va n vch trn u Ly 1 ml dung dch nagravey cho vagraveo

bigravenh nh mc coacute dung tiacutech 20 ml thecircm 5 ml dung dch NaOH 001N 2 ml

dung dch KMnO4 Dung dch va pha em un caacutech thu 400C trong 60

phuacutet

Song song tin hagravenh iu ch dung dch chun bng caacutech dugraveng cht i

chiu tobramycin

Mu trng tin hagravenh như mu th nhưng khocircng coacute tobramycin

Sau khi un caacutech thu 400C th i gian 60 phuacutet em o mt quang

ca caacutec dung dch nagravey bư c soacuteng 425 nm cuvet dagravey 1cm






- 12 -

Da trecircn cơ s to ra sn phm mu xanh v i 3-methyl-2

benzothiazolinon hydrazon hydrochlorid vagrave FeCl3 Sn phm coacute hp th

l n nht 645 nm nh lut Lambert-Beer ư c aacutep dng trong khong nng

t 50-500 mUIml123 nh lư ng tobramycin bng phươ ng phaacutep HPLC


- Ct styren - divinylbenzen copolymer (46 x 250 mm 8microm)

- Nhit ct 55 0C

- Pha ng Hn h p pha trong 1l nư c atilde loi CO2 gm 52 g natri

sulfat khan 15 g natri octansulfonat 3 ml tetrahydrofuran vagrave 50 ml dung

dch kali dihydrophosphat 02 M atilde ư c chnh pH v 30 bng acid

phosphoric

- Tc dograveng 1 mlphuacutet

- Dung dch thecircm vagraveo sau ct Dung dch natri hydroxyd 2 pha trong

nư c atilde loi CO2 Tc 03 mlphuacutet

- Detector Detector ampe k hoc caacutec thit b tươ ng t

- Th tiacutech tiecircm 20 microl

- Nng dung dch chun vagrave th 01 mgml

1222 Phươ ng phaacutep 2 [20] [23]

- Ct RP 18 (39 mm x 30 cm)

- Pha ng Hogravea tan 20 g tris (hydroxymethyl) aminomethan trong

khong 800 ml nư c thecircm vagraveo dung dch nagravey 20 ml dung dch acid sulfuric

1N sau oacute pha loatildeng bng acetonitril t i 2000 ml lc u

- Thuc th 24-dinitrofluorobenzen Dung dch 24dinitrofluorobenzen1 trong ethanol 96


- Detector UV 365 nm





- 13 -


- Nng dung dch chun vagrave th Khong 022 mgml trong dung

dch acid sulfuric 0004 N

- Caacutec dung dch chun vagrave th ư c to dn xut v i caacutec thuc th 24-

dinitrofluorobenzen vagrave tris (hydroxymethyl) aminomethan trư c khi tiecircm sc

kyacute


- Ct Purospherreg STARRP-18e (4 x 55 mm 3 microm Merck)

- Pha ng Acetonitril - nư c (5050)



- Nng dung dch chun vagrave th Khong 5 microgml trong nư c

- Dung dch chun vagrave dung dch th ư c to dn xut v i dung dch

thuc th 1-naphthyl isothiocyanat trong pyridin 700C


- Ct Xterrareg RP - 18 (21 x 250 mm 5 microm)

- Nhit ct 300

C- Pha ng A Thecircm 07 ml dung dch amoni hydroxyd 28 - 30 vagraveo 1

liacutet nư c Milli - Qreg S dng dung dch natri hydroxyd 25 N chnh pH pha

ng A n 110

- Pha ng B Acetonitril

- Tin hagravenh sc kyacute theo chươ ng trigravenh gradient dung mocirci như sau

Th i gian

(phuacutet)

Pha ng A

()

Pha ng B

()

0 100 0

10 0 100





- 14 -


- Detector Khi ph

- Th tiacutech tiecircm 4microl


dch NaCl 09


- Ct Ultrasphera RP 8 (46 x 250 mm 5microm)

- Pha ng Acetonitril - m phosphat 005 M pH 35 [6238]


- Thuc th to dn cht Acid 246-trinitrobenzen sulfomic



- Nng dung dch chun vagrave th Khong 002 mgml trong m

phosphat pH 74

- Dung dch chun vagrave th ư c to dn xut v i thuc th acid

246-trinitrobenzen sulfomic 70 0C

1226 Phươ ng phaacutep 6 [5]- Ct RP -18 brava ODS (150 x 46 mm5microm)

- Detector huỳnh quang ExEm= 338 nm 455 nm

- Tc dograveng 1mlphuacutet

- Tc thuc th 07mlphuacutet

- Th tiacutech tiecircm 20 microm

- Pha ng Hoagrave tan 1775 g natri sulfat khan 305g natri

pentansulfonat vagrave 1ml acid acetic băng trong 1000 ml nư c lc u lc qua

magraveng lc 045microm

- Thuc th to dn xut sau ct Hoagrave tan 05g o-phthalaldehyd trong

100 ml methanol thecircm 10 ml 2-mercaptoethanol lc 3 phuacutet Thecircm 15 ml





- 15 -

dung dch polyoxyethylen lauryl ether 12 vagrave 350 ml dung dch m borat

pH 104 lc u

- Dung dch m borat pH 104 Hoagrave tan 2464 g acid boric trong 900

ml nư c iu chnh pH n 104 bng dung dch NaOH 40 thecircm nư c t i

va 1000 ml lc u lc qua magraveng lc 045 microm (pha theo BP 2005)


Dung d ch th

Cacircn chiacutenh xaacutec khong 200 mg tobramycin nguyecircn liu cho vagraveo bigravenh

nh mc 200 ml Hogravea tan vagrave pha loatildeng bng nư c va n vch Ly

chiacutenh xaacutec 50 ml dung dch trecircn cho vagraveo bigravenh nh mc 200 ml Thecircm 05 ml

thuc th X em un caacutech thy 80 0C trong 60 phuacutet Thecircm dung mocirci va

n vch lc u Lc qua magraveng lc 045 microm

Dung d ch chu n ( i chi u)

Tin hagravenh tươ ng t như dung dch th nhưng thay tobramycin nguyecircn

liu bng tobramycin chun (i chiu)

- Ct sc kyacute Nucleosil C18 (250 x 4 mm 5microm)

- Detector UV = 215 nm- Pha ng Methanol m phosphat 001 M pH 3 = 20 80



- Nhit phacircn tiacutech Nhit phograveng thiacute nghim

Nhn xeacutet Qua caacutec phươ ng phaacutep nh lư ng necircu trecircn chuacuteng tocirci nhn thy

- nh lư ng bng phươ ng phaacutep vi sinh R ơ n gin khocircng cn trang

thit b phc tp nhưng coacute như c im l n lagrave mt nhiu th i gian vagrave keacutem dc

hiu do s coacute mt ca caacutec cht khaacuteng vi sinh vt khaacutec tiacutenh chiacutenh xaacutec khocircng

cao





- 16 -

- nh lư ng bng phươ ng phaacutep o hp th UV-VIS Coacute chiacutenh xaacutec

cao tin hagravenh ơ n gin khocircng yecircu cu maacutey moacutec quaacute phc tp hoaacute cht

thocircng dng vagrave coacute th aacutep dng nh lư ng caacutec cơ s kim nghim trong c

nư c Tuy nhiecircn như c im ca phươ ng phaacutep lagrave Tiacutenh c hiu chưa cao (s coacute mt ca cht khaacutec coacute kh năng hp th t ngoi kh kin như

benzalkonium clorid mt cht bo qun hay dugraveng s nh hư ng kt qu nh

lư ng) hoc phi dugraveng thuc th nhp ngoi to dn cht

- nh lư ng bng phươ ng phaacutep HPLC

Ư u im Noacutei chung coacute tiacutenh c hiu cao taacutech riecircng ư c hot cht

Như c im Tin hagravenh phc tp ogravei hi coacute trang thit b vagrave thuc th t

tin vigrave detector UV thư ng khocircng dugraveng ư c magrave thư ng phi to dn cht

Phươ ng phaacutep HPLC pha o v i detector UV rt hay s dng phacircn

tiacutech caacutec h p cht phacircn cc vagrave hp th UV Nhưng v i caacutec h p cht hp th

UV yu như tobramycin thigrave thư ng phi to dn cht hoc phi s dng

detector loi c bit t tin

Vigrave vy v i caacutec trang thit b sn coacute phograveng thiacute nghim chuacuteng tocirci t

vn nghiecircn cu xacircy dng mt quy trigravenh nh lư ng tobramycin bng

phươ ng phaacutep HPLC ơ n gin d thc hin vagrave kinh phiacute thp hơ n khocircng s

dng thuc th to dn xut như caacutec phươ ng phaacutep hin nay v i hi vng coacute th

ưa vagraveo s dng trong cocircng taacutec kim nghim - goacutep phn qun lyacute cht lư ng

caacutec sn phm cha tobramycin

13 TNG QUAN V PHƯƠ NG PHAacuteP SC KIacute LNG HIU NĂNG

CAO (HPLC)

131 Nguyecircn tcPhươ ng phaacutep HPLC lagrave 1 phươ ng phaacutep phacircn tiacutech hoacutea lyacute dugraveng taacutech

vagrave nh lư ng caacutec thagravenh phn trong hn h p da trecircn aacutei lc khaacutec nhau gia

caacutec cht v i 2 pha luocircn tip xuacutec nhưng khocircng hogravea ln vagraveo nhau Pha t ĩ nh





- 17 -

(trong ct hiu năng cao) vagrave pha ng (dung mocirci ra gii) Khi dung dch ca

hn h p caacutec cht cn phacircn tiacutech ưa vagraveo ct chuacuteng s ư c hp ph hoc phacircn

b vagraveo pha t ĩ nh tugravey thuc vagraveo bn cht ca ct vagrave ca cht cn phacircn tiacutech Khi

ta bơ m dung mocirci pha ng vagraveo ct thigrave tugravey thuc vagraveo aacutei lc ca caacutec cht v ihai pha chuacuteng s di chuyn qua ct v i vn tc khaacutec nhau dn n s phacircn

taacutech

Caacutec cht sau khi ra khi ct s ư c phaacutet hin b i b phn phaacutet hin

gi lagrave detector vagrave ư c chuyn qua b x lyacute kt qu Kt qu cui cugraveng ư c

hin th trecircn magraven higravenh hoc ưa ra maacutey in [8]

132 Cơ s lyacute thuyt

Quaacute trigravenh phacircn taacutech trong k thut HPLC lagrave do quaacute trigravenh vn chuyn vagravephacircn b ca caacutec cht tan gia 2 pha khaacutec nhau Khi pha ng di chuyn v i

mt tc nht nh qua ct sc kyacute s y caacutec cht tan b pha t ĩ nh lưu gi ra

khi ct Tugravey theo bn cht pha t ĩ nh cht tan vagrave pha ng magrave quaacute trigravenh ra

gii taacutech ư c caacutec cht khi ra khi ct sc kyacute Nu ghi quaacute trigravenh taacutech sc kyacute

chuacuteng ta coacute sc [8] [11]

133 Nguyecircn tc cu to h thng HPLC

H thng HPLC ơ n gin vagrave lagravem vic theo k thut HPLC baogm 6 b phn chiacutenh sau

1331 H th ng bơ m

bơ m pha ng vagraveo ct taacutech Bơ m nagravey phi iu chnh ư c aacutep sut

(0 - 400 bar)

1332 Bigravenh ch a dung mocirci vagrave h th ng x lyacute dung mocirci

Bigravenh cha dung mocirci thư ng bng thy tinh hoc theacutep khocircng g Dung

mocirci chy sc kyacute ư c lc qua magraveng lc (thư ng magraveng lc c l 045 microm ) vagraveui khiacute hogravea tan

1333 H tiecircm m u

ưa mu phacircn tiacutech vagraveo ct





- 18 -

Coacute nhiu phươ ng phaacutep tiecircm mu khaacutec nhau ơ n gin nht lagrave s dng

mt van tiecircm Trong caacutec h thng sc kyacute hin i lagrave h tiecircm mu t ng

Mu th (dng lng hay dng rn) u phi hoagrave tan trong dung mocirci thiacutech h p

ri lc qua magraveng lc 045microm trư c khi tiecircm

1334 C t s c kyacute lng HPLC

Ct ư c ch to bng theacutep c bit trơ v i hoacutea cht chu ư c v i aacutep

sut cao n vagravei trăm bar

- Chiu dagravei ct 10 15 hoc 25 cm thiacutech h p v i caacutec tiu phacircn pha t ĩ nh coacute

ư ng kiacutenh rt nh (3 5 10 microm)

- ư ng kiacutenh ct 4 hoc 46 mm

Noacutei chung ct HPLC coacute tui th khaacute dagravei nu ta s dng uacuteng caacutech

Nu s dng khocircng uacuteng caacutech tui th ca ct s gim Viacute d dung mocirci coacute

tiacutenh cht acid mnh hoc base mnh hoc tiecircm liecircn tip caacutec mu sinh hc hay

nguyecircn liu bn thigrave tui th ca ct s b gim

1335 Detector trong HPLC

Lagrave b phn phaacutet hin cht phacircn tiacutech tugravey theo bn cht ca cht cn

phacircn tiacutech magrave s dng detector thiacutech h p Detector hay s dng lagrave detector hp

th UV - VIS

Nguyecircn lyacute hot ng ca detector UV - VIS da trecircn nguyecircn lyacute hot

ng ca phươ ng phaacutep o quang ph hp th Ngoagravei ra cograven coacute detector khuacutec

x huỳnh quang in hoacutea

1336 Thi t b hi n th k t qu

Coacute nhiu loi nhưng ơ n gin vagrave ph bin nht lagrave maacutey t ghi caacutec tiacuten

hiu dư i dng pic [8] [14]

Sơ khi h thng HPLC ư c tng quaacutet higravenh 11

983107ộ983156

983106983145983150983144

983139983144ứ983137

983152983144983137

độ983150983143

983106ơ983149

983108983141983156983141983139983156983151983154 983106ộ 983152983144ậ983150 983156ự 983143983144983145

983126983137983150 983156983145983274983149





- 19 -

Higravenh 11 S ơ kh i t ng quaacutet h th ng HPLC

134 Caacutec thocircng s c trư ng ca quaacute trigravenh sc kyacute

Higravenh 12 S c kyacute ca 2 ch t vagrave caacutec thocircng s c tr ư ng

Kt qu ca quaacute trigravenh taacutech caacutec cht ư c detector phaacutet hin phoacuteng i

vagrave ghi thagravenh sc kyacute v i caacutec thocircng s

tR (Th i gian lưu) th i gian k t khi cht phacircn tiacutech ư c bơ m vagraveo ct cho

n khi ư c phaacutet hin nng cu i ca noacute

t0 (Th i gian cht) th i gian cn thit pha ng chy qua h thng sc

kyacute

tRrsquo (Th i gian lưu thc) tRrsquo = tR - t0 W05 rng ca pic na chiu cao

Wb rng aacutey pic

t

t

Wb2

05-2W

05-1W

b1W

R2

R2t

tR1

R1t

o





- 20 -

Th i gian lưu lagrave thocircng tin v mt nh tiacutenh ca sc noacute lagrave mt hng

s i v i mt cht nht nh khi tin hagravenh sc kyacute trong iu kin khocircng i

1341 H s dung lư ng k rsquo

[8][14]

Nu k nh thigrave tR cũng nh vagrave s taacutech keacutem Trong thc t k nm trong

khong 2 - 5 lagrave tt nht Hai cht ch ư c taacutech ra khi nhau nu chuacuteng coacute giaacute

tr khaacutec nhau Trong phacircn tiacutech thư ng chn ct pha ng vagrave caacutec iu kin

phacircn tiacutech sao cho 1 lt k lt 8

1342 chn lc α (selectivity - factor)

α=tt ttkk

01R

02R

1

2

minusminus= (k2

gt k1 ) [8] [14]

α khaacutec 1 cagraveng nhiu thigrave kh năng taacutech cagraveng rotilde ragraveng ti ưu t 15 n 2

1343 phacircn gii (resolution)

c trưng cho mc taacutech hai cht ra khi nhau trecircn mt iu kin sc

kyacute phacircn gii ca 2 pic cnh nhau ư c tiacutenh theo cocircng thc

=R

α

minusα=

+

minus=

+

minus

+ k1

k

ww

tt

ww

ttB

B

A2 1B2 1

ARBR

AB

ARBR 1

4

N1812 [8] [14]

phacircn gii cơ bn t ư c khi R = 15

1344 H s b t i AF

Cho bit mc cacircn i ca pic sc kyacute noacute ư c tiacutenh theo cocircng thc

a2AF w 20 1= [8] [14]

Trong oacute

W120 lagrave chiu rng ca pic ư c o 120 chiu cao ca pica khong caacutech t ư ng vuocircng goacutec h t nh pic n meacutep

ư ng cong phiacutea trư c ti v triacute 120 chiu cao ca pic

Trong pheacutep nh lư ng yecircu cu 09 le AF le 2 [1] [18] [22]

1tt

ttt

ttk

0

R

0

0R

0

R minus=minus

==





- 21 -

1345 S ĩ a lyacute thuy t N

c trưng cho hiu lc ct

N = 16

w

t

B

R

2

hay N=554

w

t

B2 1

R

2

[8] [12] [22]

Nu gi L lagrave chiu cao ct sc kyacute thigrave chiu cao ca ĩ a lyacute thuyt H

ư c tiacutenh bng cocircng thc =HN

L

135 Cơ s lyacute thuyt ca vic l a chn iu kin sc kyacute

1351 L a chn ct (pha t ĩ nh) [12] [18]

Pha t ĩ nh trong HPLC lagrave cht nhi ct coacute nhim v taacutech mt hn h p coacute

nhiu thagravenh phn Noacute lagrave mt cht rn xp coacute kiacutech thư c ht rt nh ư ng

kiacutenh c ht t 3 - 10 microm din tiacutech b mt riecircng t 50 - 500 m2 g

Yecircu cu pha t ĩ nh trong HPLC

Phi trơ vagrave bn vng trong mocirci trư ng HPLC

Coacute kh năng taacutech mt hn h p cht tan nht nh

Tiacutenh cht b mt n nh

Cacircn bng ng hc ca s taacutech din ra nhanh vagrave lp li tt C ht phi ng nht

Pha t ĩ nh thư ng ư c ch to trecircn nn silica (SiO2) nn oxyd nhocircm

(Al2O3) nn h p cht cao phacircn t (cellulose) hay trecircn nn mch carbon

Trong sc kyacute hp ph pha t ĩ nh trecircn nn silica coacute nhiu ưu im vagrave s dng

nhiu nht Coacute th phacircn loi cht nhi ct theo gc siloxan

|

0 CH3

| |

Si 0 Si R

| |





- 22 -

0 CH3

| R lagrave nhoacutem phacircn cc (ưa nư c) nhoacutem hydroxyl (-0H) hoc caacutec

alkylamin (-CH2NH2) alkylnitril (-CH2CN) Loi nagravey s dng lagravem pha t ĩ nhtrong sc kyacute pha thun phacircn tiacutech caacutec cht iacutet hoc khocircng phacircn cc

R lagrave caacutec nhoacutem iacutet phacircn cc caacutec nhoacutem octyl octadecyl phenyl ư c

ch bng caacutech alkyl hoacutea caacutec nhoacutem OH trecircn b mt silica trung tiacutenh bng caacutec

gc alkyl - R ca mch carbon (C2 C8 C18) hay caacutec gc carbon vograveng

(phenyl) Do caacutec nhoacutem OH thacircn nư c ư c thay bng caacutec gc R k nư c necircn

b mt tr necircn iacutet phacircn cc Silica atilde alkyl hoacutea ư c s dng trong sc kyacute pha

o phacircn tiacutech caacutec cht phacircn cc iacutet phacircn cc hay sc kyacute cp ion1352 L a chn pha ng cho HPLC [12] [18]

Pha ng lagrave dung mocirci ra gii cht phacircn tiacutech ra khi ct sc kyacute lagrave

yu t th hai quyt nh hiu sut taacutech ca mt hn h p Pha ng coacute th lagrave

nư c dung mocirci hu cơ hay hn h p dung mocirci theo t l nht nh

Pha ng trong HPLC nh hư ng t i rt nhiu thocircng s ca quaacute trigravenh

sc kyacute như chn lc α th i gian lưu tR hiu lc taacutech ca ct phacircn gii

RS rng ca pic sc kyacute Chiacutenh vigrave vy vic la chn pha ng thiacutech h p

lagrave rt quan trng

Yecircu cu ca pha ng

Phi trơ v i pha t ĩ nh

Hogravea tan ư c cht cn phacircn tiacutech

Bn vng theo th i gian

Coacute tinh khit cao

Nhanh t cacircn bng trong quaacute trigravenh sc kyacute

Phugrave h p v i detector Coacute tiacutenh kinh t vagrave m bo mocirci trư ng





- 23 -

Trong sc kyacute pha thun pha ng thư ng lagrave caacutec dung mocirci hu cơ iacutet

phacircn cc (k nư c) như n-hexan n-heptan benzen cloroform Caacutec pha

ng nagravey thư ng ư c batildeo hoagrave

Trong sc kyacute pha o pha ng lagrave h dung mocirci hu cơ phacircn cc nhưnư c methanol acetonitril hay hn h p ca chuacuteng Caacutec dung mocirci nagravey coacute

th hogravea tan thecircm mt lư ng nh acid hay base hu cơ

Bn yu t chiacutenh cn chuacute yacute khi la chn pha ng

Bn cht ca dung mocirci chy pha ng

Thagravenh phn ca caacutec cht trong pha ng

pH ca pha ng

Tc dograveng ca pha ng

1353 Chn m pH

Trong sc kyacute to cp ion sc kyacute trao i ion sc kyacute hp ph magrave cht

tan coacute tiacutenh acid hay base thư ng phi thecircm m vagraveo pha ng n nh pH

cho quaacute trigravenh sc kyacute

Giaacute tr pH thiacutech h p s lagravem tăng hiu lc taacutech ca sc kyacute [12] [18]

1354 T c dograveng

Sau khi coacute pha t ĩ nh pha ng pH thiacutech h p thigrave mt yu t cn lachn quaacute trigravenh taacutech tt hơ n lagrave tc dograveng [12] [18]

136 Caacutech aacutenh giaacute pic

aacutenh giaacute din tiacutech pic Din tiacutech ca mt cht tươ ng ng v i tng

lư ng cht oacute tiacutenh din tiacutech pic hin nay ngư i ta thư ng dugraveng maacutey tiacutech

phacircn in t gn v i maacutey vi tiacutenh (sai s khong 05 ) hoc maacutey phacircn tiacutech cơ

hc (sai s 13 ) Phươ ng phaacutep nagravey coacute th dugraveng cho caacutec pic khocircng b trocirci

ư ng nn vagrave c pic coacute ư ng nn b trocirci Phươ ng phaacutep nagravey ch cn im uim cui ca pic ư c nhn ra chiacutenh xaacutec vagrave cho vagrave cho kt qu tt v i nng

va trung bigravenh vagrave cao





- 24 -

aacutenh giaacute chi u cao pic Khi pic coacute dng khocircng i thigrave chiu cao pic

(khong caacutech gia ư ng nn vagrave nh pic) lagrave mt i lư ng t l v i din tiacutech

pic vagrave noacute cũng coacute th dugraveng aacutenh giaacute ph Phươ ng phaacutep ch aacutep dng khi caacutec

ch s k lagrave hng nh

137 ng dng ca HPLC

Sc kyacute noacutei chung vagrave HPLC noacutei riecircng coacute ba ng dng chiacutenh

1371 nh tiacutenh vagrave th tinh khi t

Th i gian lưu ca cht th trecircn sc phi tươ ng ng v i th i gian lưu

ca cht chun i chiu trecircn sc kyacute

1372 S c kyacute i u ch

Qua quacutea trigravenh sc kyacute caacutec cht ư c taacutech ra vagrave dch ra gii ư c hng

riecircng ri cho bc hơ i dung mocirci thu ly cht

1371 nh lư ng

Sc kyacute lng hiu năng cao coacute ng rt l n trong nh lư ng cht trong

hn h p vagrave xaacutec nh gi i hn tp cht

Caacutec phươ ng phaacutep nh lư ng bng sc kyacute u da trecircn nguyecircn tc

nng ca mt cht t l v i chiu cao hoc din tiacutech pic ca noacute Mt s

phươ ng phaacutep nh lư ng hay aacutep dng trong HPLC

Phươ ng phaacutep chun ngoi (external standard method)

acircy lagrave phươ ng phaacutep nh lư ng cơ bn trong oacute c hai mu

chun vagrave mu th u ư c tin hagravenh sc kyacute trong cugraveng iu kin Sau

oacute em so saacutenh din tiacutech (hoc chiu cao) pic thu ư c t mu th v i

pic ca mu chun atilde bit nng

Phươ ng phaacutep chun ni (internal standard method)Lagrave phươ ng phaacutep cho thecircm nhng lư ng ging nhau ca cht chun th

hai coacute th i gian lưu vagrave aacutep ng gn ging mu th vagraveo c mu chun

vagrave





- 25 -

mu th ri tin hagravenh sc kyacute Cht chun th hai gi lagrave cht chun ni

Phươ ng phaacutep chun hoacutea din tiacutech pic (area normalisation)

+ Nguyecircn tc Hagravem lư ng phn trăm ca mt cht trong hn h p

nhiu thagravenh phn ư c tiacutenh bng t l phn trăm din tiacutech pic ca noacute sov i tng din tiacutech ca tt c caacutec pic thagravenh phn trecircn sc

Phươ ng phaacutep nagravey yecircu cu tt c caacutec thagravenh phn u ư c ra

gii vagrave ư c phaacutet hin vagrave tt c caacutec thagravenh phn u coacute aacutep ng

detector như nhau

+ Aacutep dng Iacutet dugraveng nh lư ng ch dugraveng trong xaacutec nh hagravem

lư ng tp cht khi coi aacutep ng ca caacutec cht lagrave như nhau [12] [18]





- 26 -

PHN 2 TH C NGHIM VAgrave KT QU

21 NGUYEcircN VT LIU VAgrave PHƯƠ NG PHAacuteP TH C NGHIM

211 Nguyecircn vt liu

2111 Nguyecircn liu vagrave hoaacute cht

- Nguyecircn liu Tobramycin ca cocircng ty CPDP Hagrave Tacircy

- Cht i chiu Tobramycin ca vin kim nghim trung ươ ng hagravem

lư ng 9496

- KH2PO4 (PA)

- H3PO4 (PA)

- Methanol dugraveng cho HPLC (Meck)

- Nư c ct tinh khit 2 ln

- Mt vagravei hoaacute cht khaacutec

2112 Dng c

- Maacutey sc kyacute lng hiu năng cao DIONEX - detector UV Diode array -

PDA 100

- Maacutey quang ph UV- VIS Lambda EZ210

- Maacutey siecircu acircm SONOREX- Cacircn phacircn tiacutech Mettler AB 204 coacute chiacutenh xaacutec 01mg

- Maacutey lc chacircn khocircng Satorius

- Bigravenh nh mc caacutec loi

- Cc coacute m

- Pipet chiacutenh xaacutec pipet thư ng

- L ng mu magraveng lc 045 microm

212 Ni dung vagrave phươ ng phaacutep nghiecircn c u

2121 Phươ ng phaacutep nghiecircn cu

- Tham kho caacutec phươ ng phaacutep nh lư ng tobramycin atilde cocircng b

- Tham kho caacutec phươ ng phaacutep nh lư ng tobramycin bng HPLC





- 27 -

- Da vagraveo cu to phacircn t tiacutenh cht hoaacute lyacute ca tobramycin cũng như

iu kin thc nghim ca nư c ta hin nay xacircy dng quy trigravenh k

thut nh lư ng tobramycin bng HPLC

2122 Ni dung nghiecircn cu Nghiecircn cu caacutec iu kin tin hagravenh xacircy dng phươ ng phaacutep

Chn ct pha ng bư c soacuteng phaacutet hin vagrave tc dograveng th tiacutech tiecircm

thiacutech h p nh lư ng

Xacircy dng aacutenh giaacute phươ ng phaacutep nh lư ng tobramycin nguyecircn

liu

- Kho saacutet tiacutenh thiacutech h p ca h thng sc kyacute

- aacutenh giaacute s ph thuc tuyn tiacutenh gia nng tobramycin vagrave din

tiacutech pic trecircn sc kyacute

- aacutenh giaacute tiacutenh chiacutenh xaacutec ca phươ ng phaacutep

- aacutenh giaacute tiacutenh uacuteng ca phươ ng phaacutep

- aacutenh giaacute tiacutenh c hiu ca phươ ng phaacutep

Mt s cocircng thc tiacutenh toaacuten trong x lyacute thng kecirc kt qu

- Giaacute tr trung bigravenh sum==

n

1iixn

1

x (21)

- lch chun( )

1n

xS

n

1i

2

ixminus

minus

=sum

= (22)

- lch chun tươ ng i (RSD) 100x

Ss x

times= (23)

- Sai s chunn

S

sx

= (24)

- Sai s tươ ng i ( ) 100x

st x)1n(

timestimes=εminusα

(25)

- Khong tin cy x plusmn tαSx (26)





- 28 -

22 TH C NGHIM VAgrave KT QU

221 Xacircy d ng quy trigravenh k thut nh lư ng tobramycin bng

phươ ng phaacutep HPLC

2211 Nguyecircn tc l a chn iu kin sc kyacuteChn phươ ng phaacutep sc kyacute Tobramycin sulfat lagrave cht hu cơ phacircn cc

tan nhiu trong nư c Vigrave vy phacircn tiacutech vagrave nh lư ng tt chuacuteng tocirci chn

phươ ng phaacutep sc kyacute lng hiu năng cao pha o

Chn ct sc kyacute Trong phươ ng phaacutep nh lư ng HPLC pha o ct

sc kyacute thocircng dng lagrave Nucleosil C8 C18 V i iu kin sn coacute ca phograveng thiacute

nghim chuacuteng tocirci s dng ct Nucleosil C18 (250 x 4 mm 5 microm)

Chn pha ng Trong phươ ng phaacutep HPLC pha o pha ng thocircng

dng nht lagrave acetonitril methanol nư c hay hn h p ca methanol acetonitril

v i nư c Do tobramycin sulfat tan nhiu trong nư c ruacutet ngn th i gian

lưu thiacutech h p ca cht cn phacircn tiacutech pha ng cn phi coacute nư c chuyn

tobramycin sang ht dng ion cn s dng m pH acid

Chn bư c soacuteng cu truacutec tobramycin coacute 5 nhoacutem amin

(-NH2) tuy nhiecircn kh năng hp th UV ca tobramycin khocircng cao vigrave vybư c soacuteng nghiecircn cu d kin lagrave bư c soacuteng thp

2212 Kho saacutet l a chn iu kin sc kiacute

a Kho saacutet chn bư c soacuteng thiacutech h p

xaacutec nh bư c soacuteng thiacutech h p phaacutet hin ra tobramycin chuacuteng tocirci

tin hagravenh queacutet ph UV dung dch tobramycin 02 mgml trecircn maacutey quang ph

UV- VIS Lambda EZ210

Tin hagravenh Cacircn chiacutenh xaacutec khong 20 mg tobramycin nguyecircn liu chovagraveo bigravenh nh mc 1000 ml thecircm nư c va n vch lc u

Queacutet ph hp th t ngoi ca dung dch nagravey Kt qu ghi higravenh 1





- 29 -

200 250 300

nm

To bra 002

00

05

Abs

Higravenh 1 Ph hp th ca dung dch tobramycin 002

Nhn xeacutet

T ph hp th ca dung dch tobramycin chuacuteng tocirci thy Cc i hp

th ca dung dch cht nagravey khong 194 nm gim nh hư ng ca dung mocirci

vagrave tp cht nhưng vn m bo nh lư ng ư c tobramycin chuacuteng tocirci chn

bư c soacuteng o lagrave 195 nm

b La chn ct

S dng ct Nucleosil C18 (250x4 mm) c ht 5 microm sn coacute b mocircn

Hoaacute dư c trư ng i hc Dư c Hagrave Ni

c La chn m

Chuacuteng tocirci tin hagravenh kho saacutet pha ng lagrave dung dch m phosphat 001M

caacutec pH khaacutec nhau kt qu cho thy

V i pH=35 thigrave h s bt i ca pic tobramycin lagrave 295



Nhn xeacutet









- 32 -

2221 Kho saacutet tiacutenh thiacutech h p ca h thng

Cacircn chiacutenh xaacutec khong 300 mg tobramycin i chiu cho vagraveo bigravenh nh

mc 100ml Hoagrave tan vagrave pha loatildeng bng nư c va n vch lc u Lc

qua magraveng lc 045 microm dugraveng dch lc tiecircm sc kyacute

Tiecircm 5 ln mu i chiu vagraveo h thng HPLC tin hagravenh sc kyacute theo

iu kin atilde chn trecircn Ghi kt qu th i gian lưu din tiacutech vagrave h s bt i

ca pic tobramycin

Kt qu kho saacutet tiacutenh thiacutech h p ca h thng sc kyacute ư c trigravenh bagravey như

bng 2

Bng 2 K t qu kho saacutet tiacutenh thiacutech h p ca h th ng

STTPic ca tobramycin

Th i gian lưu (phuacutet) Din tiacutech pic (mAU) H s bt i (AF)

1 294 122985 152

2 295 123001 153

3 296 122805 151

4 296 122230 152

5 296 121916 152RSD() 030 040

Nhn xeacutet

Kt qu kho saacutet tiacutenh thiacutech h p ca h thng HPLC cho thy

lch chun tươ ng i (RSD ) ca th i gian lưu vagrave din tiacutech pic

trong caacutec pheacutep th lagrave 030 vagrave 040 u nh hơ n 2

Caacutec giaacute tr ca h s bt i giao ng t 151 n 153

Kt lun

H thng HPLC magrave chuacuteng tocirci s dng lagrave thiacutech h p nh lư ng

tobramycin trong nguyecircn liu

2222 Xacircy dng phươ ng phaacutep nh lư ng





- 33 -

a Dung dch th


nh mc 100 ml Hoagrave tan vagrave pha loatildeng bng nư c va n vch lc u

Lc qua magraveng lc 045 microm

b Dung dch chun (i chiu)

Cacircn chiacutenh xaacutec khong 300 mg tobramycin i chiu cho vagraveo bigravenh



c Mu trng

Nư c ct 2 ln lc qua magraveng lc 045 microm siecircu acircm ui khiacute 15 phuacutet

d Pha ng

Dung dch m phosphat 001 M pH 30

Caacutech pha Hoagrave tan 068g KH2PO4 trong nư c vagrave pha loatildeng bng nư c

va 500 ml iu chnh v pH 30 bng H3PO4 c Lc qua magraveng lc

045microm siecircu acircm ui khiacute 15 phuacutet

2223 iu kin sc kyacute

- Ct Nucleosil C18 (250 x 46 mm 5microm)- Pha ng m phosphat 001 M pH 30




- Nhit phacircn tiacutech Nhit phograveng

2224 Tiacutenh kt qu

Ghi din tiacutech pic thu ư c trecircn sc kyacute ca mu chun vagrave mu th

tươ ng ng Tiacutenh hagravem lư ng tobramycin coacute trong mu th theo cocircng thc

sau





- 34 -

X = PmAmA

tc

ct timestimes

times

Trong oacute At Ac lagrave din tiacutech pic tươ ng ng ca mu th vagrave mu chun

mt mc lagrave lư ng cacircn tươ ng ng ca mu th vagrave mu chun (mg)P lagrave hagravem lư ng ghi trecircn nhatilden ca tobramycin chun

223 nh lư ng tobramycin nguyecircn liu bng phươ ng phaacutep m i xacircy

d ng

Chun b vagrave cho h thng HPLC chy n nh v i pha ng trong 30

phuacutet Tiecircm ln lư t 30 microl mi dung dch atilde chun b vagraveo h thng sc kyacute

Tin hagravenh nh lư ng 5 mu th vagrave 1 mu chun tobramycin theo caacutec iu

kin atilde chn Ghi caacutec giaacute tr din tiacutech pic thu ư c trecircn sc kyacute ca mu

chun vagrave mu th tươ ng ng Kt qu nh lư ng ư c trigravenh bagravey bng 3

Bng 3 K t qu nh lư ng tobramycin nguyecircn liu

Khi lư ng cacircn(mg)

Din tiacutechpic(mAUmin)

Hagravem lư ng

296 119207 9097297 119585 9095299 119165 9002

301 119975 9003303 120954 9017

m( chun) =290 121916 9496

S liu thng kecircGiaacute tr trung bigravenh 9043 X =

lch chun S = 046

lch chun tươ ng i RSD = 051

Sai s chun Sx= 021( 1) 05962n x

X t S α minus

∆ = times =

Khong tin cy = 9043 plusmn 059

Kt lun





- 35 -

Kt qu trigravenh bagravey bng trecircn cho thy nguyecircn liu tobramycin em

nh lư ng coacute hagravem lư ng lagrave 9043 plusmn 059

224 aacutenh giaacute phươ ng phaacutep nh lư ng

Chuacuteng tocirci tin hagravenh aacutenh giaacute phươ ng phaacutep nh lư ng tobramycinnguyecircn liu bng HPLC m i xacircy dng da vagraveo caacutec ch tiecircu sau

- aacutenh giaacute tiacutenh tuyn tiacutenh

- aacutenh giaacute tiacutenh chiacutenh xaacutec

- aacutenh giaacute tiacutenh uacuteng

- aacutenh giaacute tiacutenh c hiu

2241 Tiacutenh tuyn tiacutenh

Tin hagravenh kho saacutet s ph thuc tuyn tiacutenh gia nng v i din tiacutech

pic ca dn cht tobramycin (tiacutenh theo lư ng tobramycin ưa vagraveo) trecircn cht

chun tobramycin

Caacutech pha Cacircn chiacutenh xaacutec khong 375 mg tobramycin chun cho vagraveo

bigravenh nh mc 500 ml Hoagrave tan vagrave pha loatildeng bng nư c va n vch

Dung dch thu ư c coacute nng 75 mgml Tin hagravenh pha loatildeng theo bng 4

thu ư c caacutec dung dch coacute nng ln lư t lagrave 24 27 30 33 36 mgml

Lc qua magraveng lc 045microm

Bng 4 Caacutech pha caacutec dung d ch kho saacutet khong tuy n tiacutenh

Tiecircm ln lư t 30 microl caacutec dung dch atilde chun b vagraveo h thng sc kyacute

Tin hagravenh sc kyacute theo iu kin atilde chn Ghi caacutec giacutea tr din tiacutech pic o ư c

Bigravenh 1 2 3 4 5

Tobramycin (mg) 80 90 100 110 120

Nư c ct va (ml) 25 25 25 25 25

Nng (mgml) 24 27 30 33 36





- 36 -

ng v i caacutec nng ca caacutec dung dch trecircn Kt qu kho saacutet khong tuyn

tiacutenh ư c trigravenh bagravey bng 5

Bng 5 K t qu kho saacutet khong tuy n tiacutenh

T caacutec kt qu thu ư c bng trecircn chuacuteng tocirci thit lp mi tươ ng quan

gia din tiacutech pic thu ư c trecircn sc kyacute vagrave nng ca dung dch (tiacutenh theo

lư ng tobramycin ưa vagraveo)

Phươ ng trigravenh hi quy biu th mi tươ ng quan nagravey lagrave y= 39878 x + 0039 v i

h s tươ ng quan r= 09999 lch chun y_intercept Sb= 0064

Khong tin cy 95 ca y_intercept 0039 plusmn 0204

hay -0165 le y_intercept le 0243 cha gc to

th biu din s ph thuc tuyn tiacutenh gia nng dung dch vagrave din tiacutech

pic ca tobramycin ư c ghi higravenh 2

Nng (mgml) 24 27 30 33 36

Din tiacutech pic (mAUmin) 96023 107959 120322 131989 143825

Tiacutenh tuyến tiacutenh

y = 39878x + 0039

R2 = 09999

0

4

8

12

16

0 1 2 3 4

Nồng độ (mgml)

D i ệ n t iacute c h p i c ( m A U m i n )





- 37 -

Higravenh 2 th biu din s ph thuc tuyn tiacutenh gia nng dung dch vagrave

din tiacutech pic ca tobramycin

T kt qu thu ư c higravenh 2 cho thy

Tt c caacutec giaacute tr thu ư c nm trecircn ư ng hi quy hoc phacircn b ngu c 2 phiacutea ca ư ng hi quy

H s tươ ng quan r= 09999

Khong tin cy ca y_intercept cha gc to

Nhn xeacutet

Phươ ng phaacutep tuyn tiacutenh trong khong nng kho saacutet t 24 n 36

mgml (tiacutenh theo lư ng tobramycin ưa vagraveo) hay tuyn tiacutenh trong khong

nng t 80 n 120 nng lagravem vic2242 Tiacutenh chiacutenh xaacutec

Nguyecircn tc Tiacutenh chiacutenh xaacutec ca phươ ng phaacutep ư c aacutenh giaacute bng

lch chun tươ ng i ca 5 pheacutep th song song

Caacutech tin hagravenh vagrave kt qu ư c trigravenh bagravey như phn 223

Kt qu th chiacutenh xaacutec ca phươ ng phaacutep cho thy trong caacutec iu

kin sc kyacute atilde chn lch chun tươ ng i ca caacutec kt qu rt nh (RSD =

051 ltlt 2) Như vy phươ ng phaacutep lagrave chiacutenh xaacutec

2243 Tiacutenh uacuteng

Nguyecircn tc aacutenh giaacute tiacutenh uacuteng ca phươ ng phaacutep ư c tin hagravenh trecircn

vic kho saacutet kh năng tigravem li ca phươ ng phaacutep

Chun b caacutec dung dch chun coacute nng t 80 n 120 nng

nh lư ng Khi lư ng cacircn như bng 6

Tin hagravenh nh lư ng caacutec mu nagravey theo iu kin atilde chn mi munh lư ng 3 ln ly kt qu trung bigravenh Tiacutenh hagravem lư ng ca tobramycin

coacute trong mu th da trecircn chun atilde bit hagravem lư ng t oacute tiacutenh tigravem li ca

mu th







- 39 -

lch chun tươ ng i RSD = 052 (ltlt 2)

- Phươ ng trigravenh hi quy tuyn tiacutenh v mi tươ ng quan gia lư ng cacircn vagrave

lư ng hot cht tigravem li

Phươ ng trigravenh y = 09686x + 06252

lch chun ca a Sa = 001

lch chun ca y_intercept Sb = 0298

- V i tin cy 95 ta coacute

Khong tin cy ca dc a plusmn 0127 hay 0840 lt a lt 1095

(a cha 1=gt khocircng coacute sai s h thng t l)

Khong tin cy ca y_intercept bplusmn 3786

hay -316 le y_intercept le 4393 (y cha gc ta =gt khocircng coacute sai

s hng nh)

Kt lun

Qua caacutec kt qu trecircn ta thy phươ ng phaacutep uacuteng trong khong kho saacutet

2244 Tiacutenh c hiu

kho saacutet tiacutenh c hiu ca phươ ng phaacutep chuacuteng tocirci tin hagravenh như

sauChun b mu trng (nư c ct hai ln) vagrave mu th

Tin hagravenh sc kyacute mu trng vagrave mu th theo quy trigravenh atilde chn

Sc kyacute mu trng vagrave mu th ư c trigravenh bagravey như higravenh 4 vagrave 5





- 40 -

Higravenh 4Sc kiacute mu trng

Higravenh 5 Sc kiacute mu th Nhn xeacutet

Sc kyacute ca mu i chiu cho mt pic chiacutenh v i tR=296 phuacutet cacircn

i

Trecircn sc kyacute mu trng trong khong th i gian lưu ca pic to b i

tobramycin khocircng coacute pic nagraveo xut hin

Vy phươ ng phaacutep nh lư ng tobramycin trong nguyecircn liu ư c xacircy

dng coacute tiacutenh c hiu

23 BAgraveN LUN VAgrave KT QU







- 42 -


nh mc 100ml Hoagrave tan vagrave pha loatildeng bng nư c va n vch lc

u Lc qua magraveng lc 045 microm

Mu trngNư c ct 2 ln lc qua magraveng lc 045 microm siecircu acircm ui khiacute 15 phuacutet

Tiacutenh kt qu Hagravem lư ng tobramycin trong nguyecircn liu ư c tiacutenh

theo cocircng thc

X = PmAmA

tc

ct timestimes

times

At Ac Din tiacutech pic ca mu th vagrave mu chun

mt mc Khi lư ng cacircn ca mu th vagrave mu chun (mg)

P Lagrave hagravem lư ng ghi trecircn nhatilden ca tobramycin chun

T kt qu thc nghim vagrave x lyacute s liu thng kecirc cho thy phươ ng

phaacutep m i xacircy dng lagrave chiacutenh xaacutec (RSD = 051) uacuteng tuyn tiacutenh trong

khong nng kho saacutet vagrave c hiu Vigrave vy coacute th aacutep dng phươ ng phaacutep

nagravey nh lư ng tobramycin nguyecircn liu Do th i gian coacute hn chuacuteng tocirci

chưa tip tc nghiecircn cu aacutep dng phươ ng phaacutep nagravey nh lư ng tobramycin

trong caacutec ch phm





- 43 -

KT LUN VAgrave XUT

31 KT LUN

Trong quaacute trigravenh lagravem khoacutea lun tt nghip chuacuteng tocirci atilde thu ư c mt s

kt qu

1 Xacircy dng ư c mt phươ ng phaacutep nh lư ng tobramycin nguyecircn

liu bng phươ ng phaacutep sc kiacute lng hiu năng cao (HPLC) ơ n gin d thc

hin vagrave kinh phiacute thp Kt qu nghiecircn cu cho thy phươ ng phaacutep aacutep ng y

caacutec ch tiecircu ca mt phươ ng phaacutep nh lư ng (tiacutenh c hiu tiacutenh chiacutenh

xaacutec tiacutenh uacuteng vagrave tiacutenh tuyn tiacutenh trong khong nng kho saacutet)

2 Xaacutec nh ư c hagravem lư ng ca tobramycin trong nguyecircn liu

3 Hc ư c caacutech tigravem tagravei liu lagravem thc nghim vagrave trigravenh bagravey văn bn

mt caacutech khoa hc v mt vn nghiecircn cu Tigravem hiu phươ ng phaacutep HPLC

lagravem quen vagrave s dng ư c maacutey HPLC nh lư ng mt sn phm bit caacutech

aacutenh giaacute mt phươ ng phaacutep nh lư ng

32 XUT

1 Coacute th aacutep dng phươ ng phaacutep nagravey ti caacutec cơ s kim tra cht lư ng

trong c nư c xaacutec nh v mt hagravem lư ng tobramycin trong nguyecircn liu2 Tip tc nghiecircn cu kho saacutet aacutep dng nh lư ng tobramycin

trong caacutec thagravenh phm như thuc nh mt thuc tiecircm








- 4 -

MC LC

PHN 1 TNG QUAN - 3 -11 TNG QUAN V TOBRAMYCIN - 7 -111 Cocircng th c cu to - 7 -112 Tiacutenh cht lyacute hoacutea - 7 -113 Ngun gc - 7 -114 Dư c ng hc - 7 -115 Taacutec dng vagrave cơ ch taacutec dng - 8 -116 Ch nh - 8 -117 Chng ch nh - 9 -118 Dng bagraveo ch vagrave liu lư ng - 9 -12 MT S PHƯƠ NG PHAacuteP NH LƯ NG TOBRAMYCIN - 9 -121 nh lư ng tobramycin bng phươ ng phaacutep vi sinh - 9 -1211 Phươ ng phaacutep 1 [15] - 9 -1212 Phươ ng phaacutep 2 [3] - 10 -1213 Phươ ng phaacutep 3[14] [20] - 10 -122 nh lư ng tobramycin bng phươ ng phaacutep o quang ph hp th UV-VIS- 11 -1221 Phươ ng phaacutep 1 [4] - 11 -1222 Phươ ng phaacutep 2 [19] - 11 -123 nh lư ng tobramycin bng phươ ng phaacutep HPLC - 12 -1221 Phươ ng phaacutep 1 [14] - 12 -1222 Phươ ng phaacutep 2 [20] [23] - 12 -1223 Phươ ng phaacutep 3 [15] - 13 -1224 Phươ ng phaacutep 4 [16] - 13 -1225 Phươ ng phaacutep 5 [15] - 14 -1226 Phươ ng phaacutep 6 [5] - 14 -1227 Phươ ng phaacutep 7 [9] - 15 -13 TNG QUAN V PHƯƠ NG PHAacuteP SC KIacute LNG HIU NĂNGCAO (HPLC) - 16 -131 Nguyecircn tc - 16 -132 Cơ s lyacute thuyt - 17 -133 Nguyecircn tc cu to h thng HPLC - 17 -

1331 H th ng bơ m - 17 -1332 Bigravenh ch a dung mocirci vagrave h th ng x lyacute dung mocirci - 17 -1333 H tiecircm m u - 17 -1334 C t s c kyacute lng HPLC - 18 -1335 Detector trong HPLC - 18 -1336 Thi t b hi n th k t qu - 18 -





- 5 -








- 6 -







- 7 -



C18H37N5O9 PTL 4675







113 Ngun gc



114 Dư c ng hc







- 8 -





















ribosom [2] [11]

116 Ch nh







- 9 -







117 Chng ch nh



















Thch 150 - 200 g





- 10 -

Nư c ct 1000 ml









Cao men bia 30 g

Pepton 60 g


Glucose 10 g

Cao tht 15 g

Thch 150 g














- 11 -



- Dung dch chun






- Dung dich th





UV-VIS






phuacutet


chiu tobramycin









- 12 -







- Nhit ct 55 0C




phosphoric








- Ct RP 18 (39 mm x 30 cm)











- 13 -






kyacute











- Nhit ct 300



ng A n 110



Th i gian

(phuacutet)

Pha ng A

()

Pha ng B

()

0 100 0

10 0 100





- 14 -


- Detector Khi ph



dch NaCl 09









phosphat pH 74

















- 15 -


pH 104 lc u





Dung d ch th


















cao





- 16 -






















CAO (HPLC)








- 17 -





taacutech












1331 H th ng bơ m


(0 - 400 bar)




1333 H tiecircm m u






- 18 -


















th UV - VIS








983107ộ983156

983106983145983150983144

983139983144ứ983137

983152983144983137

độ983150983143

983106ơ983149

983108983141983156983141983139983156983151983154 983106ộ 983152983144ậ983150 983156ự 983143983144983145

983126983137983150 983156983145983274983149





- 19 -









kyacute


Wb rng aacutey pic

t

t

Wb2

05-2W

05-1W

b1W

R2

R2t

tR1

R1t

o





- 20 -




[8][14]






α=tt ttkk

01R

02R

1

2

minusminus= (k2

gt k1 ) [8] [14]





=R

α

minusα=

+

minus=

+

minus

+ k1

k

ww

tt

ww

ttB

B

A2 1B2 1

ARBR

AB

ARBR 1

4

N1812 [8] [14]


1344 H s b t i AF


a2AF w 20 1= [8] [14]

Trong oacute




1tt

ttt

ttk

0

R

0

0R

0

R minus=minus

==





- 21 -



N = 16

w

t

B

R

2

hay N=554

w

t

B2 1

R

2

[8] [12] [22]



L


1351 L a chn ct (pha t ĩ nh) [12] [18]













|

0 CH3

| |

Si 0 Si R

| |





- 22 -

0 CH3



























- 23 -









pH ca pha ng


1353 Chn m pH





1354 T c dograveng













- 24 -





137 ng dng ca HPLC








1371 nh lư ng













vagrave





- 25 -







detector như nhau







- 26 -







lư ng 9496

- KH2PO4 (PA)

- H3PO4 (PA)




2112 Dng c


PDA 100





- Cc coacute m











- 27 -








liu









n

1iixn

1

x (21)

- lch chun( )

1n

xS

n

1i

2

ixminus

minus

=sum

= (22)


Ss x

times= (23)

- Sai s chunn

S

sx

= (24)


st x)1n(


(25)






- 28 -




























- 29 -

200 250 300

nm

To bra 002

00

05

Abs


Nhn xeacutet





b La chn ct



c La chn m






Nhn xeacutet









- 32 -







ca pic tobramycin


bng 2




1 294 122985 152

2 295 123001 153

3 296 122805 151

4 296 122230 152

5 296 121916 152RSD() 030 040

Nhn xeacutet





Kt lun








- 33 -

a Dung dch th








c Mu trng


d Pha ng














sau





- 34 -

X = PmAmA

tc

ct timestimes

times




d ng









Hagravem lư ng

296 119207 9097297 119585 9095299 119165 9002

301 119975 9003303 120954 9017

m( chun) =290 121916 9496


lch chun S = 046


Sai s chun Sx= 021( 1) 05962n x

X t S α minus

∆ = times =


Kt lun





- 35 -












chun tobramycin









Bigravenh 1 2 3 4 5

Tobramycin (mg) 80 90 100 110 120

Nư c ct va (ml) 25 25 25 25 25

Nng (mgml) 24 27 30 33 36





- 36 -













Nng (mgml) 24 27 30 33 36



y = 39878x + 0039

R2 = 09999

0

4

8

12

16

0 1 2 3 4

Nồng độ (mgml)






- 37 -







Nhn xeacutet

















mu th







- 39 -












s hng nh)

Kt lun











- 40 -




i












- 42 -






theo cocircng thc

X = PmAmA

tc

ct timestimes

times














- 43 -

KT LUN VAgrave XUT

31 KT LUN


kt qu











32 XUT











- 5 -








- 6 -







- 7 -



C18H37N5O9 PTL 4675







113 Ngun gc



114 Dư c ng hc







- 8 -





















ribosom [2] [11]

116 Ch nh







- 9 -







117 Chng ch nh



















Thch 150 - 200 g





- 10 -

Nư c ct 1000 ml









Cao men bia 30 g

Pepton 60 g


Glucose 10 g

Cao tht 15 g

Thch 150 g














- 11 -



- Dung dch chun






- Dung dich th





UV-VIS






phuacutet


chiu tobramycin









- 12 -







- Nhit ct 55 0C




phosphoric








- Ct RP 18 (39 mm x 30 cm)











- 13 -






kyacute











- Nhit ct 300



ng A n 110



Th i gian

(phuacutet)

Pha ng A

()

Pha ng B

()

0 100 0

10 0 100





- 14 -


- Detector Khi ph



dch NaCl 09









phosphat pH 74

















- 15 -


pH 104 lc u





Dung d ch th


















cao





- 16 -






















CAO (HPLC)








- 17 -





taacutech












1331 H th ng bơ m


(0 - 400 bar)




1333 H tiecircm m u






- 18 -


















th UV - VIS








983107ộ983156

983106983145983150983144

983139983144ứ983137

983152983144983137

độ983150983143

983106ơ983149

983108983141983156983141983139983156983151983154 983106ộ 983152983144ậ983150 983156ự 983143983144983145

983126983137983150 983156983145983274983149





- 19 -









kyacute


Wb rng aacutey pic

t

t

Wb2

05-2W

05-1W

b1W

R2

R2t

tR1

R1t

o





- 20 -




[8][14]






α=tt ttkk

01R

02R

1

2

minusminus= (k2

gt k1 ) [8] [14]





=R

α

minusα=

+

minus=

+

minus

+ k1

k

ww

tt

ww

ttB

B

A2 1B2 1

ARBR

AB

ARBR 1

4

N1812 [8] [14]


1344 H s b t i AF


a2AF w 20 1= [8] [14]

Trong oacute




1tt

ttt

ttk

0

R

0

0R

0

R minus=minus

==





- 21 -



N = 16

w

t

B

R

2

hay N=554

w

t

B2 1

R

2

[8] [12] [22]



L


1351 L a chn ct (pha t ĩ nh) [12] [18]













|

0 CH3

| |

Si 0 Si R

| |





- 22 -

0 CH3



























- 23 -









pH ca pha ng


1353 Chn m pH





1354 T c dograveng













- 24 -





137 ng dng ca HPLC








1371 nh lư ng













vagrave





- 25 -







detector như nhau







- 26 -







lư ng 9496

- KH2PO4 (PA)

- H3PO4 (PA)




2112 Dng c


PDA 100





- Cc coacute m











- 27 -








liu









n

1iixn

1

x (21)

- lch chun( )

1n

xS

n

1i

2

ixminus

minus

=sum

= (22)


Ss x

times= (23)

- Sai s chunn

S

sx

= (24)


st x)1n(


(25)






- 28 -




























- 29 -

200 250 300

nm

To bra 002

00

05

Abs


Nhn xeacutet





b La chn ct



c La chn m






Nhn xeacutet









- 32 -







ca pic tobramycin


bng 2




1 294 122985 152

2 295 123001 153

3 296 122805 151

4 296 122230 152

5 296 121916 152RSD() 030 040

Nhn xeacutet





Kt lun








- 33 -

a Dung dch th








c Mu trng


d Pha ng














sau





- 34 -

X = PmAmA

tc

ct timestimes

times




d ng









Hagravem lư ng

296 119207 9097297 119585 9095299 119165 9002

301 119975 9003303 120954 9017

m( chun) =290 121916 9496


lch chun S = 046


Sai s chun Sx= 021( 1) 05962n x

X t S α minus

∆ = times =


Kt lun





- 35 -












chun tobramycin









Bigravenh 1 2 3 4 5

Tobramycin (mg) 80 90 100 110 120

Nư c ct va (ml) 25 25 25 25 25

Nng (mgml) 24 27 30 33 36





- 36 -













Nng (mgml) 24 27 30 33 36



y = 39878x + 0039

R2 = 09999

0

4

8

12

16

0 1 2 3 4

Nồng độ (mgml)






- 37 -







Nhn xeacutet

















mu th







- 39 -












s hng nh)

Kt lun











- 40 -




i












- 42 -






theo cocircng thc

X = PmAmA

tc

ct timestimes

times














- 43 -

KT LUN VAgrave XUT

31 KT LUN


kt qu











32 XUT











- 6 -







- 7 -



C18H37N5O9 PTL 4675







113 Ngun gc



114 Dư c ng hc







- 8 -





















ribosom [2] [11]

116 Ch nh







- 9 -







117 Chng ch nh



















Thch 150 - 200 g





- 10 -

Nư c ct 1000 ml









Cao men bia 30 g

Pepton 60 g


Glucose 10 g

Cao tht 15 g

Thch 150 g














- 11 -



- Dung dch chun






- Dung dich th





UV-VIS






phuacutet


chiu tobramycin









- 12 -







- Nhit ct 55 0C




phosphoric








- Ct RP 18 (39 mm x 30 cm)











- 13 -






kyacute











- Nhit ct 300



ng A n 110



Th i gian

(phuacutet)

Pha ng A

()

Pha ng B

()

0 100 0

10 0 100





- 14 -


- Detector Khi ph



dch NaCl 09









phosphat pH 74

















- 15 -


pH 104 lc u





Dung d ch th


















cao





- 16 -






















CAO (HPLC)








- 17 -





taacutech












1331 H th ng bơ m


(0 - 400 bar)




1333 H tiecircm m u






- 18 -


















th UV - VIS








983107ộ983156

983106983145983150983144

983139983144ứ983137

983152983144983137

độ983150983143

983106ơ983149

983108983141983156983141983139983156983151983154 983106ộ 983152983144ậ983150 983156ự 983143983144983145

983126983137983150 983156983145983274983149





- 19 -









kyacute


Wb rng aacutey pic

t

t

Wb2

05-2W

05-1W

b1W

R2

R2t

tR1

R1t

o





- 20 -




[8][14]






α=tt ttkk

01R

02R

1

2

minusminus= (k2

gt k1 ) [8] [14]





=R

α

minusα=

+

minus=

+

minus

+ k1

k

ww

tt

ww

ttB

B

A2 1B2 1

ARBR

AB

ARBR 1

4

N1812 [8] [14]


1344 H s b t i AF


a2AF w 20 1= [8] [14]

Trong oacute




1tt

ttt

ttk

0

R

0

0R

0

R minus=minus

==





- 21 -



N = 16

w

t

B

R

2

hay N=554

w

t

B2 1

R

2

[8] [12] [22]



L


1351 L a chn ct (pha t ĩ nh) [12] [18]













|

0 CH3

| |

Si 0 Si R

| |





- 22 -

0 CH3



























- 23 -









pH ca pha ng


1353 Chn m pH





1354 T c dograveng













- 24 -





137 ng dng ca HPLC








1371 nh lư ng













vagrave





- 25 -







detector như nhau







- 26 -







lư ng 9496

- KH2PO4 (PA)

- H3PO4 (PA)




2112 Dng c


PDA 100





- Cc coacute m











- 27 -








liu









n

1iixn

1

x (21)

- lch chun( )

1n

xS

n

1i

2

ixminus

minus

=sum

= (22)


Ss x

times= (23)

- Sai s chunn

S

sx

= (24)


st x)1n(


(25)






- 28 -




























- 29 -

200 250 300

nm

To bra 002

00

05

Abs


Nhn xeacutet





b La chn ct



c La chn m






Nhn xeacutet









- 32 -







ca pic tobramycin


bng 2




1 294 122985 152

2 295 123001 153

3 296 122805 151

4 296 122230 152

5 296 121916 152RSD() 030 040

Nhn xeacutet





Kt lun








- 33 -

a Dung dch th








c Mu trng


d Pha ng














sau





- 34 -

X = PmAmA

tc

ct timestimes

times




d ng









Hagravem lư ng

296 119207 9097297 119585 9095299 119165 9002

301 119975 9003303 120954 9017

m( chun) =290 121916 9496


lch chun S = 046


Sai s chun Sx= 021( 1) 05962n x

X t S α minus

∆ = times =


Kt lun





- 35 -












chun tobramycin









Bigravenh 1 2 3 4 5

Tobramycin (mg) 80 90 100 110 120

Nư c ct va (ml) 25 25 25 25 25

Nng (mgml) 24 27 30 33 36





- 36 -













Nng (mgml) 24 27 30 33 36



y = 39878x + 0039

R2 = 09999

0

4

8

12

16

0 1 2 3 4

Nồng độ (mgml)






- 37 -







Nhn xeacutet

















mu th







- 39 -












s hng nh)

Kt lun











- 40 -




i












- 42 -






theo cocircng thc

X = PmAmA

tc

ct timestimes

times














- 43 -

KT LUN VAgrave XUT

31 KT LUN


kt qu











32 XUT











- 7 -



C18H37N5O9 PTL 4675







113 Ngun gc



114 Dư c ng hc







- 8 -





















ribosom [2] [11]

116 Ch nh







- 9 -







117 Chng ch nh



















Thch 150 - 200 g





- 10 -

Nư c ct 1000 ml









Cao men bia 30 g

Pepton 60 g


Glucose 10 g

Cao tht 15 g

Thch 150 g














- 11 -



- Dung dch chun






- Dung dich th





UV-VIS






phuacutet


chiu tobramycin









- 12 -







- Nhit ct 55 0C




phosphoric








- Ct RP 18 (39 mm x 30 cm)











- 13 -






kyacute











- Nhit ct 300



ng A n 110



Th i gian

(phuacutet)

Pha ng A

()

Pha ng B

()

0 100 0

10 0 100





- 14 -


- Detector Khi ph



dch NaCl 09









phosphat pH 74

















- 15 -


pH 104 lc u





Dung d ch th


















cao





- 16 -






















CAO (HPLC)








- 17 -





taacutech












1331 H th ng bơ m


(0 - 400 bar)




1333 H tiecircm m u






- 18 -


















th UV - VIS








983107ộ983156

983106983145983150983144

983139983144ứ983137

983152983144983137

độ983150983143

983106ơ983149

983108983141983156983141983139983156983151983154 983106ộ 983152983144ậ983150 983156ự 983143983144983145

983126983137983150 983156983145983274983149





- 19 -









kyacute


Wb rng aacutey pic

t

t

Wb2

05-2W

05-1W

b1W

R2

R2t

tR1

R1t

o





- 20 -




[8][14]






α=tt ttkk

01R

02R

1

2

minusminus= (k2

gt k1 ) [8] [14]





=R

α

minusα=

+

minus=

+

minus

+ k1

k

ww

tt

ww

ttB

B

A2 1B2 1

ARBR

AB

ARBR 1

4

N1812 [8] [14]


1344 H s b t i AF


a2AF w 20 1= [8] [14]

Trong oacute




1tt

ttt

ttk

0

R

0

0R

0

R minus=minus

==





- 21 -



N = 16

w

t

B

R

2

hay N=554

w

t

B2 1

R

2

[8] [12] [22]



L


1351 L a chn ct (pha t ĩ nh) [12] [18]













|

0 CH3

| |

Si 0 Si R

| |





- 22 -

0 CH3



























- 23 -









pH ca pha ng


1353 Chn m pH





1354 T c dograveng













- 24 -





137 ng dng ca HPLC








1371 nh lư ng













vagrave





- 25 -







detector như nhau







- 26 -







lư ng 9496

- KH2PO4 (PA)

- H3PO4 (PA)




2112 Dng c


PDA 100





- Cc coacute m











- 27 -








liu









n

1iixn

1

x (21)

- lch chun( )

1n

xS

n

1i

2

ixminus

minus

=sum

= (22)


Ss x

times= (23)

- Sai s chunn

S

sx

= (24)


st x)1n(


(25)






- 28 -




























- 29 -

200 250 300

nm

To bra 002

00

05

Abs


Nhn xeacutet





b La chn ct



c La chn m






Nhn xeacutet









- 32 -







ca pic tobramycin


bng 2




1 294 122985 152

2 295 123001 153

3 296 122805 151

4 296 122230 152

5 296 121916 152RSD() 030 040

Nhn xeacutet





Kt lun








- 33 -

a Dung dch th








c Mu trng


d Pha ng














sau





- 34 -

X = PmAmA

tc

ct timestimes

times




d ng









Hagravem lư ng

296 119207 9097297 119585 9095299 119165 9002

301 119975 9003303 120954 9017

m( chun) =290 121916 9496


lch chun S = 046


Sai s chun Sx= 021( 1) 05962n x

X t S α minus

∆ = times =


Kt lun





- 35 -












chun tobramycin









Bigravenh 1 2 3 4 5

Tobramycin (mg) 80 90 100 110 120

Nư c ct va (ml) 25 25 25 25 25

Nng (mgml) 24 27 30 33 36





- 36 -













Nng (mgml) 24 27 30 33 36



y = 39878x + 0039

R2 = 09999

0

4

8

12

16

0 1 2 3 4

Nồng độ (mgml)






- 37 -







Nhn xeacutet

















mu th







- 39 -












s hng nh)

Kt lun











- 40 -




i












- 42 -






theo cocircng thc

X = PmAmA

tc

ct timestimes

times














- 43 -

KT LUN VAgrave XUT

31 KT LUN


kt qu











32 XUT











- 8 -





















ribosom [2] [11]

116 Ch nh







- 9 -







117 Chng ch nh



















Thch 150 - 200 g





- 10 -

Nư c ct 1000 ml









Cao men bia 30 g

Pepton 60 g


Glucose 10 g

Cao tht 15 g

Thch 150 g














- 11 -



- Dung dch chun






- Dung dich th





UV-VIS






phuacutet


chiu tobramycin









- 12 -







- Nhit ct 55 0C




phosphoric








- Ct RP 18 (39 mm x 30 cm)











- 13 -






kyacute











- Nhit ct 300



ng A n 110



Th i gian

(phuacutet)

Pha ng A

()

Pha ng B

()

0 100 0

10 0 100





- 14 -


- Detector Khi ph



dch NaCl 09









phosphat pH 74

















- 15 -


pH 104 lc u





Dung d ch th


















cao





- 16 -






















CAO (HPLC)








- 17 -





taacutech












1331 H th ng bơ m


(0 - 400 bar)




1333 H tiecircm m u






- 18 -


















th UV - VIS








983107ộ983156

983106983145983150983144

983139983144ứ983137

983152983144983137

độ983150983143

983106ơ983149

983108983141983156983141983139983156983151983154 983106ộ 983152983144ậ983150 983156ự 983143983144983145

983126983137983150 983156983145983274983149





- 19 -









kyacute


Wb rng aacutey pic

t

t

Wb2

05-2W

05-1W

b1W

R2

R2t

tR1

R1t

o





- 20 -




[8][14]






α=tt ttkk

01R

02R

1

2

minusminus= (k2

gt k1 ) [8] [14]





=R

α

minusα=

+

minus=

+

minus

+ k1

k

ww

tt

ww

ttB

B

A2 1B2 1

ARBR

AB

ARBR 1

4

N1812 [8] [14]


1344 H s b t i AF


a2AF w 20 1= [8] [14]

Trong oacute




1tt

ttt

ttk

0

R

0

0R

0

R minus=minus

==





- 21 -



N = 16

w

t

B

R

2

hay N=554

w

t

B2 1

R

2

[8] [12] [22]



L


1351 L a chn ct (pha t ĩ nh) [12] [18]













|

0 CH3

| |

Si 0 Si R

| |





- 22 -

0 CH3



























- 23 -









pH ca pha ng


1353 Chn m pH





1354 T c dograveng













- 24 -





137 ng dng ca HPLC








1371 nh lư ng













vagrave





- 25 -







detector như nhau







- 26 -







lư ng 9496

- KH2PO4 (PA)

- H3PO4 (PA)




2112 Dng c


PDA 100





- Cc coacute m











- 27 -








liu









n

1iixn

1

x (21)

- lch chun( )

1n

xS

n

1i

2

ixminus

minus

=sum

= (22)


Ss x

times= (23)

- Sai s chunn

S

sx

= (24)


st x)1n(


(25)






- 28 -




























- 29 -

200 250 300

nm

To bra 002

00

05

Abs


Nhn xeacutet





b La chn ct



c La chn m






Nhn xeacutet









- 32 -







ca pic tobramycin


bng 2




1 294 122985 152

2 295 123001 153

3 296 122805 151

4 296 122230 152

5 296 121916 152RSD() 030 040

Nhn xeacutet





Kt lun








- 33 -

a Dung dch th








c Mu trng


d Pha ng














sau





- 34 -

X = PmAmA

tc

ct timestimes

times




d ng









Hagravem lư ng

296 119207 9097297 119585 9095299 119165 9002

301 119975 9003303 120954 9017

m( chun) =290 121916 9496


lch chun S = 046


Sai s chun Sx= 021( 1) 05962n x

X t S α minus

∆ = times =


Kt lun





- 35 -












chun tobramycin









Bigravenh 1 2 3 4 5

Tobramycin (mg) 80 90 100 110 120

Nư c ct va (ml) 25 25 25 25 25

Nng (mgml) 24 27 30 33 36





- 36 -













Nng (mgml) 24 27 30 33 36



y = 39878x + 0039

R2 = 09999

0

4

8

12

16

0 1 2 3 4

Nồng độ (mgml)






- 37 -







Nhn xeacutet

















mu th







- 39 -












s hng nh)

Kt lun











- 40 -




i












- 42 -






theo cocircng thc

X = PmAmA

tc

ct timestimes

times














- 43 -

KT LUN VAgrave XUT

31 KT LUN


kt qu











32 XUT











- 9 -







117 Chng ch nh



















Thch 150 - 200 g





- 10 -

Nư c ct 1000 ml









Cao men bia 30 g

Pepton 60 g


Glucose 10 g

Cao tht 15 g

Thch 150 g














- 11 -



- Dung dch chun






- Dung dich th





UV-VIS






phuacutet


chiu tobramycin









- 12 -







- Nhit ct 55 0C




phosphoric








- Ct RP 18 (39 mm x 30 cm)











- 13 -






kyacute











- Nhit ct 300



ng A n 110



Th i gian

(phuacutet)

Pha ng A

()

Pha ng B

()

0 100 0

10 0 100





- 14 -


- Detector Khi ph



dch NaCl 09









phosphat pH 74

















- 15 -


pH 104 lc u





Dung d ch th


















cao





- 16 -






















CAO (HPLC)








- 17 -





taacutech












1331 H th ng bơ m


(0 - 400 bar)




1333 H tiecircm m u






- 18 -


















th UV - VIS








983107ộ983156

983106983145983150983144

983139983144ứ983137

983152983144983137

độ983150983143

983106ơ983149

983108983141983156983141983139983156983151983154 983106ộ 983152983144ậ983150 983156ự 983143983144983145

983126983137983150 983156983145983274983149





- 19 -









kyacute


Wb rng aacutey pic

t

t

Wb2

05-2W

05-1W

b1W

R2

R2t

tR1

R1t

o





- 20 -




[8][14]






α=tt ttkk

01R

02R

1

2

minusminus= (k2

gt k1 ) [8] [14]





=R

α

minusα=

+

minus=

+

minus

+ k1

k

ww

tt

ww

ttB

B

A2 1B2 1

ARBR

AB

ARBR 1

4

N1812 [8] [14]


1344 H s b t i AF


a2AF w 20 1= [8] [14]

Trong oacute




1tt

ttt

ttk

0

R

0

0R

0

R minus=minus

==





- 21 -



N = 16

w

t

B

R

2

hay N=554

w

t

B2 1

R

2

[8] [12] [22]



L


1351 L a chn ct (pha t ĩ nh) [12] [18]













|

0 CH3

| |

Si 0 Si R

| |





- 22 -

0 CH3



























- 23 -









pH ca pha ng


1353 Chn m pH





1354 T c dograveng













- 24 -





137 ng dng ca HPLC








1371 nh lư ng













vagrave





- 25 -







detector như nhau







- 26 -







lư ng 9496

- KH2PO4 (PA)

- H3PO4 (PA)




2112 Dng c


PDA 100





- Cc coacute m











- 27 -








liu









n

1iixn

1

x (21)

- lch chun( )

1n

xS

n

1i

2

ixminus

minus

=sum

= (22)


Ss x

times= (23)

- Sai s chunn

S

sx

= (24)


st x)1n(


(25)






- 28 -




























- 29 -

200 250 300

nm

To bra 002

00

05

Abs


Nhn xeacutet





b La chn ct



c La chn m






Nhn xeacutet









- 32 -







ca pic tobramycin


bng 2




1 294 122985 152

2 295 123001 153

3 296 122805 151

4 296 122230 152

5 296 121916 152RSD() 030 040

Nhn xeacutet





Kt lun








- 33 -

a Dung dch th








c Mu trng


d Pha ng














sau





- 34 -

X = PmAmA

tc

ct timestimes

times




d ng









Hagravem lư ng

296 119207 9097297 119585 9095299 119165 9002

301 119975 9003303 120954 9017

m( chun) =290 121916 9496


lch chun S = 046


Sai s chun Sx= 021( 1) 05962n x

X t S α minus

∆ = times =


Kt lun





- 35 -












chun tobramycin









Bigravenh 1 2 3 4 5

Tobramycin (mg) 80 90 100 110 120

Nư c ct va (ml) 25 25 25 25 25

Nng (mgml) 24 27 30 33 36





- 36 -













Nng (mgml) 24 27 30 33 36



y = 39878x + 0039

R2 = 09999

0

4

8

12

16

0 1 2 3 4

Nồng độ (mgml)






- 37 -







Nhn xeacutet

















mu th







- 39 -












s hng nh)

Kt lun











- 40 -




i












- 42 -






theo cocircng thc

X = PmAmA

tc

ct timestimes

times














- 43 -

KT LUN VAgrave XUT

31 KT LUN


kt qu











32 XUT











- 10 -

Nư c ct 1000 ml









Cao men bia 30 g

Pepton 60 g


Glucose 10 g

Cao tht 15 g

Thch 150 g














- 11 -



- Dung dch chun






- Dung dich th





UV-VIS






phuacutet


chiu tobramycin









- 12 -







- Nhit ct 55 0C




phosphoric








- Ct RP 18 (39 mm x 30 cm)











- 13 -






kyacute











- Nhit ct 300



ng A n 110



Th i gian

(phuacutet)

Pha ng A

()

Pha ng B

()

0 100 0

10 0 100





- 14 -


- Detector Khi ph



dch NaCl 09









phosphat pH 74

















- 15 -


pH 104 lc u





Dung d ch th


















cao





- 16 -






















CAO (HPLC)








- 17 -





taacutech












1331 H th ng bơ m


(0 - 400 bar)




1333 H tiecircm m u






- 18 -


















th UV - VIS








983107ộ983156

983106983145983150983144

983139983144ứ983137

983152983144983137

độ983150983143

983106ơ983149

983108983141983156983141983139983156983151983154 983106ộ 983152983144ậ983150 983156ự 983143983144983145

983126983137983150 983156983145983274983149





- 19 -









kyacute


Wb rng aacutey pic

t

t

Wb2

05-2W

05-1W

b1W

R2

R2t

tR1

R1t

o





- 20 -




[8][14]






α=tt ttkk

01R

02R

1

2

minusminus= (k2

gt k1 ) [8] [14]





=R

α

minusα=

+

minus=

+

minus

+ k1

k

ww

tt

ww

ttB

B

A2 1B2 1

ARBR

AB

ARBR 1

4

N1812 [8] [14]


1344 H s b t i AF


a2AF w 20 1= [8] [14]

Trong oacute




1tt

ttt

ttk

0

R

0

0R

0

R minus=minus

==





- 21 -



N = 16

w

t

B

R

2

hay N=554

w

t

B2 1

R

2

[8] [12] [22]



L


1351 L a chn ct (pha t ĩ nh) [12] [18]













|

0 CH3

| |

Si 0 Si R

| |





- 22 -

0 CH3



























- 23 -









pH ca pha ng


1353 Chn m pH





1354 T c dograveng













- 24 -





137 ng dng ca HPLC








1371 nh lư ng













vagrave





- 25 -







detector như nhau







- 26 -







lư ng 9496

- KH2PO4 (PA)

- H3PO4 (PA)




2112 Dng c


PDA 100





- Cc coacute m











- 27 -








liu









n

1iixn

1

x (21)

- lch chun( )

1n

xS

n

1i

2

ixminus

minus

=sum

= (22)


Ss x

times= (23)

- Sai s chunn

S

sx

= (24)


st x)1n(


(25)






- 28 -




























- 29 -

200 250 300

nm

To bra 002

00

05

Abs


Nhn xeacutet





b La chn ct



c La chn m






Nhn xeacutet









- 32 -







ca pic tobramycin


bng 2




1 294 122985 152

2 295 123001 153

3 296 122805 151

4 296 122230 152

5 296 121916 152RSD() 030 040

Nhn xeacutet





Kt lun








- 33 -

a Dung dch th








c Mu trng


d Pha ng














sau





- 34 -

X = PmAmA

tc

ct timestimes

times




d ng









Hagravem lư ng

296 119207 9097297 119585 9095299 119165 9002

301 119975 9003303 120954 9017

m( chun) =290 121916 9496


lch chun S = 046


Sai s chun Sx= 021( 1) 05962n x

X t S α minus

∆ = times =


Kt lun





- 35 -












chun tobramycin









Bigravenh 1 2 3 4 5

Tobramycin (mg) 80 90 100 110 120

Nư c ct va (ml) 25 25 25 25 25

Nng (mgml) 24 27 30 33 36





- 36 -













Nng (mgml) 24 27 30 33 36



y = 39878x + 0039

R2 = 09999

0

4

8

12

16

0 1 2 3 4

Nồng độ (mgml)






- 37 -







Nhn xeacutet

















mu th







- 39 -












s hng nh)

Kt lun











- 40 -




i












- 42 -






theo cocircng thc

X = PmAmA

tc

ct timestimes

times














- 43 -

KT LUN VAgrave XUT

31 KT LUN


kt qu











32 XUT











- 11 -



- Dung dch chun






- Dung dich th





UV-VIS






phuacutet


chiu tobramycin









- 12 -







- Nhit ct 55 0C




phosphoric








- Ct RP 18 (39 mm x 30 cm)











- 13 -






kyacute











- Nhit ct 300



ng A n 110



Th i gian

(phuacutet)

Pha ng A

()

Pha ng B

()

0 100 0

10 0 100





- 14 -


- Detector Khi ph



dch NaCl 09









phosphat pH 74

















- 15 -


pH 104 lc u





Dung d ch th


















cao





- 16 -






















CAO (HPLC)








- 17 -





taacutech












1331 H th ng bơ m


(0 - 400 bar)




1333 H tiecircm m u






- 18 -


















th UV - VIS








983107ộ983156

983106983145983150983144

983139983144ứ983137

983152983144983137

độ983150983143

983106ơ983149

983108983141983156983141983139983156983151983154 983106ộ 983152983144ậ983150 983156ự 983143983144983145

983126983137983150 983156983145983274983149





- 19 -









kyacute


Wb rng aacutey pic

t

t

Wb2

05-2W

05-1W

b1W

R2

R2t

tR1

R1t

o





- 20 -




[8][14]






α=tt ttkk

01R

02R

1

2

minusminus= (k2

gt k1 ) [8] [14]





=R

α

minusα=

+

minus=

+

minus

+ k1

k

ww

tt

ww

ttB

B

A2 1B2 1

ARBR

AB

ARBR 1

4

N1812 [8] [14]


1344 H s b t i AF


a2AF w 20 1= [8] [14]

Trong oacute




1tt

ttt

ttk

0

R

0

0R

0

R minus=minus

==





- 21 -



N = 16

w

t

B

R

2

hay N=554

w

t

B2 1

R

2

[8] [12] [22]



L


1351 L a chn ct (pha t ĩ nh) [12] [18]













|

0 CH3

| |

Si 0 Si R

| |





- 22 -

0 CH3



























- 23 -









pH ca pha ng


1353 Chn m pH





1354 T c dograveng













- 24 -





137 ng dng ca HPLC








1371 nh lư ng













vagrave





- 25 -







detector như nhau







- 26 -







lư ng 9496

- KH2PO4 (PA)

- H3PO4 (PA)




2112 Dng c


PDA 100





- Cc coacute m











- 27 -








liu









n

1iixn

1

x (21)

- lch chun( )

1n

xS

n

1i

2

ixminus

minus

=sum

= (22)


Ss x

times= (23)

- Sai s chunn

S

sx

= (24)


st x)1n(


(25)






- 28 -




























- 29 -

200 250 300

nm

To bra 002

00

05

Abs


Nhn xeacutet





b La chn ct



c La chn m






Nhn xeacutet









- 32 -







ca pic tobramycin


bng 2




1 294 122985 152

2 295 123001 153

3 296 122805 151

4 296 122230 152

5 296 121916 152RSD() 030 040

Nhn xeacutet





Kt lun








- 33 -

a Dung dch th








c Mu trng


d Pha ng














sau





- 34 -

X = PmAmA

tc

ct timestimes

times




d ng









Hagravem lư ng

296 119207 9097297 119585 9095299 119165 9002

301 119975 9003303 120954 9017

m( chun) =290 121916 9496


lch chun S = 046


Sai s chun Sx= 021( 1) 05962n x

X t S α minus

∆ = times =


Kt lun





- 35 -












chun tobramycin









Bigravenh 1 2 3 4 5

Tobramycin (mg) 80 90 100 110 120

Nư c ct va (ml) 25 25 25 25 25

Nng (mgml) 24 27 30 33 36





- 36 -













Nng (mgml) 24 27 30 33 36



y = 39878x + 0039

R2 = 09999

0

4

8

12

16

0 1 2 3 4

Nồng độ (mgml)






- 37 -







Nhn xeacutet

















mu th







- 39 -












s hng nh)

Kt lun











- 40 -




i












- 42 -






theo cocircng thc

X = PmAmA

tc

ct timestimes

times














- 43 -

KT LUN VAgrave XUT

31 KT LUN


kt qu











32 XUT











- 12 -







- Nhit ct 55 0C




phosphoric








- Ct RP 18 (39 mm x 30 cm)











- 13 -






kyacute











- Nhit ct 300



ng A n 110



Th i gian

(phuacutet)

Pha ng A

()

Pha ng B

()

0 100 0

10 0 100





- 14 -


- Detector Khi ph



dch NaCl 09









phosphat pH 74

















- 15 -


pH 104 lc u





Dung d ch th


















cao





- 16 -






















CAO (HPLC)








- 17 -





taacutech












1331 H th ng bơ m


(0 - 400 bar)




1333 H tiecircm m u






- 18 -


















th UV - VIS








983107ộ983156

983106983145983150983144

983139983144ứ983137

983152983144983137

độ983150983143

983106ơ983149

983108983141983156983141983139983156983151983154 983106ộ 983152983144ậ983150 983156ự 983143983144983145

983126983137983150 983156983145983274983149





- 19 -









kyacute


Wb rng aacutey pic

t

t

Wb2

05-2W

05-1W

b1W

R2

R2t

tR1

R1t

o





- 20 -




[8][14]






α=tt ttkk

01R

02R

1

2

minusminus= (k2

gt k1 ) [8] [14]





=R

α

minusα=

+

minus=

+

minus

+ k1

k

ww

tt

ww

ttB

B

A2 1B2 1

ARBR

AB

ARBR 1

4

N1812 [8] [14]


1344 H s b t i AF


a2AF w 20 1= [8] [14]

Trong oacute




1tt

ttt

ttk

0

R

0

0R

0

R minus=minus

==





- 21 -



N = 16

w

t

B

R

2

hay N=554

w

t

B2 1

R

2

[8] [12] [22]



L


1351 L a chn ct (pha t ĩ nh) [12] [18]













|

0 CH3

| |

Si 0 Si R

| |





- 22 -

0 CH3



























- 23 -









pH ca pha ng


1353 Chn m pH





1354 T c dograveng













- 24 -





137 ng dng ca HPLC








1371 nh lư ng













vagrave





- 25 -







detector như nhau







- 26 -







lư ng 9496

- KH2PO4 (PA)

- H3PO4 (PA)




2112 Dng c


PDA 100





- Cc coacute m











- 27 -








liu









n

1iixn

1

x (21)

- lch chun( )

1n

xS

n

1i

2

ixminus

minus

=sum

= (22)


Ss x

times= (23)

- Sai s chunn

S

sx

= (24)


st x)1n(


(25)






- 28 -




























- 29 -

200 250 300

nm

To bra 002

00

05

Abs


Nhn xeacutet





b La chn ct



c La chn m






Nhn xeacutet









- 32 -







ca pic tobramycin


bng 2




1 294 122985 152

2 295 123001 153

3 296 122805 151

4 296 122230 152

5 296 121916 152RSD() 030 040

Nhn xeacutet





Kt lun








- 33 -

a Dung dch th








c Mu trng


d Pha ng














sau





- 34 -

X = PmAmA

tc

ct timestimes

times




d ng









Hagravem lư ng

296 119207 9097297 119585 9095299 119165 9002

301 119975 9003303 120954 9017

m( chun) =290 121916 9496


lch chun S = 046


Sai s chun Sx= 021( 1) 05962n x

X t S α minus

∆ = times =


Kt lun





- 35 -












chun tobramycin









Bigravenh 1 2 3 4 5

Tobramycin (mg) 80 90 100 110 120

Nư c ct va (ml) 25 25 25 25 25

Nng (mgml) 24 27 30 33 36





- 36 -













Nng (mgml) 24 27 30 33 36



y = 39878x + 0039

R2 = 09999

0

4

8

12

16

0 1 2 3 4

Nồng độ (mgml)






- 37 -







Nhn xeacutet

















mu th







- 39 -












s hng nh)

Kt lun











- 40 -




i












- 42 -






theo cocircng thc

X = PmAmA

tc

ct timestimes

times














- 43 -

KT LUN VAgrave XUT

31 KT LUN


kt qu











32 XUT











- 13 -






kyacute











- Nhit ct 300



ng A n 110



Th i gian

(phuacutet)

Pha ng A

()

Pha ng B

()

0 100 0

10 0 100





- 14 -


- Detector Khi ph



dch NaCl 09









phosphat pH 74

















- 15 -


pH 104 lc u





Dung d ch th


















cao





- 16 -






















CAO (HPLC)








- 17 -





taacutech












1331 H th ng bơ m


(0 - 400 bar)




1333 H tiecircm m u






- 18 -


















th UV - VIS








983107ộ983156

983106983145983150983144

983139983144ứ983137

983152983144983137

độ983150983143

983106ơ983149

983108983141983156983141983139983156983151983154 983106ộ 983152983144ậ983150 983156ự 983143983144983145

983126983137983150 983156983145983274983149





- 19 -









kyacute


Wb rng aacutey pic

t

t

Wb2

05-2W

05-1W

b1W

R2

R2t

tR1

R1t

o





- 20 -




[8][14]






α=tt ttkk

01R

02R

1

2

minusminus= (k2

gt k1 ) [8] [14]





=R

α

minusα=

+

minus=

+

minus

+ k1

k

ww

tt

ww

ttB

B

A2 1B2 1

ARBR

AB

ARBR 1

4

N1812 [8] [14]


1344 H s b t i AF


a2AF w 20 1= [8] [14]

Trong oacute




1tt

ttt

ttk

0

R

0

0R

0

R minus=minus

==





- 21 -



N = 16

w

t

B

R

2

hay N=554

w

t

B2 1

R

2

[8] [12] [22]



L


1351 L a chn ct (pha t ĩ nh) [12] [18]













|

0 CH3

| |

Si 0 Si R

| |





- 22 -

0 CH3



























- 23 -









pH ca pha ng


1353 Chn m pH





1354 T c dograveng













- 24 -





137 ng dng ca HPLC








1371 nh lư ng













vagrave





- 25 -







detector như nhau







- 26 -







lư ng 9496

- KH2PO4 (PA)

- H3PO4 (PA)




2112 Dng c


PDA 100





- Cc coacute m











- 27 -








liu









n

1iixn

1

x (21)

- lch chun( )

1n

xS

n

1i

2

ixminus

minus

=sum

= (22)


Ss x

times= (23)

- Sai s chunn

S

sx

= (24)


st x)1n(


(25)






- 28 -




























- 29 -

200 250 300

nm

To bra 002

00

05

Abs


Nhn xeacutet





b La chn ct



c La chn m






Nhn xeacutet









- 32 -







ca pic tobramycin


bng 2




1 294 122985 152

2 295 123001 153

3 296 122805 151

4 296 122230 152

5 296 121916 152RSD() 030 040

Nhn xeacutet





Kt lun








- 33 -

a Dung dch th








c Mu trng


d Pha ng














sau





- 34 -

X = PmAmA

tc

ct timestimes

times




d ng









Hagravem lư ng

296 119207 9097297 119585 9095299 119165 9002

301 119975 9003303 120954 9017

m( chun) =290 121916 9496


lch chun S = 046


Sai s chun Sx= 021( 1) 05962n x

X t S α minus

∆ = times =


Kt lun





- 35 -












chun tobramycin









Bigravenh 1 2 3 4 5

Tobramycin (mg) 80 90 100 110 120

Nư c ct va (ml) 25 25 25 25 25

Nng (mgml) 24 27 30 33 36





- 36 -













Nng (mgml) 24 27 30 33 36



y = 39878x + 0039

R2 = 09999

0

4

8

12

16

0 1 2 3 4

Nồng độ (mgml)






- 37 -







Nhn xeacutet

















mu th







- 39 -












s hng nh)

Kt lun











- 40 -




i












- 42 -






theo cocircng thc

X = PmAmA

tc

ct timestimes

times














- 43 -

KT LUN VAgrave XUT

31 KT LUN


kt qu











32 XUT











- 14 -


- Detector Khi ph



dch NaCl 09









phosphat pH 74

















- 15 -


pH 104 lc u





Dung d ch th


















cao





- 16 -






















CAO (HPLC)








- 17 -





taacutech












1331 H th ng bơ m


(0 - 400 bar)




1333 H tiecircm m u






- 18 -


















th UV - VIS








983107ộ983156

983106983145983150983144

983139983144ứ983137

983152983144983137

độ983150983143

983106ơ983149

983108983141983156983141983139983156983151983154 983106ộ 983152983144ậ983150 983156ự 983143983144983145

983126983137983150 983156983145983274983149





- 19 -









kyacute


Wb rng aacutey pic

t

t

Wb2

05-2W

05-1W

b1W

R2

R2t

tR1

R1t

o





- 20 -




[8][14]






α=tt ttkk

01R

02R

1

2

minusminus= (k2

gt k1 ) [8] [14]





=R

α

minusα=

+

minus=

+

minus

+ k1

k

ww

tt

ww

ttB

B

A2 1B2 1

ARBR

AB

ARBR 1

4

N1812 [8] [14]


1344 H s b t i AF


a2AF w 20 1= [8] [14]

Trong oacute




1tt

ttt

ttk

0

R

0

0R

0

R minus=minus

==





- 21 -



N = 16

w

t

B

R

2

hay N=554

w

t

B2 1

R

2

[8] [12] [22]



L


1351 L a chn ct (pha t ĩ nh) [12] [18]













|

0 CH3

| |

Si 0 Si R

| |





- 22 -

0 CH3



























- 23 -









pH ca pha ng


1353 Chn m pH





1354 T c dograveng













- 24 -





137 ng dng ca HPLC








1371 nh lư ng













vagrave





- 25 -







detector như nhau







- 26 -







lư ng 9496

- KH2PO4 (PA)

- H3PO4 (PA)




2112 Dng c


PDA 100





- Cc coacute m











- 27 -








liu









n

1iixn

1

x (21)

- lch chun( )

1n

xS

n

1i

2

ixminus

minus

=sum

= (22)


Ss x

times= (23)

- Sai s chunn

S

sx

= (24)


st x)1n(


(25)






- 28 -




























- 29 -

200 250 300

nm

To bra 002

00

05

Abs


Nhn xeacutet





b La chn ct



c La chn m






Nhn xeacutet









- 32 -







ca pic tobramycin


bng 2




1 294 122985 152

2 295 123001 153

3 296 122805 151

4 296 122230 152

5 296 121916 152RSD() 030 040

Nhn xeacutet





Kt lun








- 33 -

a Dung dch th








c Mu trng


d Pha ng














sau





- 34 -

X = PmAmA

tc

ct timestimes

times




d ng









Hagravem lư ng

296 119207 9097297 119585 9095299 119165 9002

301 119975 9003303 120954 9017

m( chun) =290 121916 9496


lch chun S = 046


Sai s chun Sx= 021( 1) 05962n x

X t S α minus

∆ = times =


Kt lun





- 35 -












chun tobramycin









Bigravenh 1 2 3 4 5

Tobramycin (mg) 80 90 100 110 120

Nư c ct va (ml) 25 25 25 25 25

Nng (mgml) 24 27 30 33 36





- 36 -













Nng (mgml) 24 27 30 33 36



y = 39878x + 0039

R2 = 09999

0

4

8

12

16

0 1 2 3 4

Nồng độ (mgml)






- 37 -







Nhn xeacutet

















mu th







- 39 -












s hng nh)

Kt lun











- 40 -




i












- 42 -






theo cocircng thc

X = PmAmA

tc

ct timestimes

times














- 43 -

KT LUN VAgrave XUT

31 KT LUN


kt qu











32 XUT











- 15 -


pH 104 lc u





Dung d ch th


















cao





- 16 -






















CAO (HPLC)








- 17 -





taacutech












1331 H th ng bơ m


(0 - 400 bar)




1333 H tiecircm m u






- 18 -


















th UV - VIS








983107ộ983156

983106983145983150983144

983139983144ứ983137

983152983144983137

độ983150983143

983106ơ983149

983108983141983156983141983139983156983151983154 983106ộ 983152983144ậ983150 983156ự 983143983144983145

983126983137983150 983156983145983274983149





- 19 -









kyacute


Wb rng aacutey pic

t

t

Wb2

05-2W

05-1W

b1W

R2

R2t

tR1

R1t

o





- 20 -




[8][14]






α=tt ttkk

01R

02R

1

2

minusminus= (k2

gt k1 ) [8] [14]





=R

α

minusα=

+

minus=

+

minus

+ k1

k

ww

tt

ww

ttB

B

A2 1B2 1

ARBR

AB

ARBR 1

4

N1812 [8] [14]


1344 H s b t i AF


a2AF w 20 1= [8] [14]

Trong oacute




1tt

ttt

ttk

0

R

0

0R

0

R minus=minus

==





- 21 -



N = 16

w

t

B

R

2

hay N=554

w

t

B2 1

R

2

[8] [12] [22]



L


1351 L a chn ct (pha t ĩ nh) [12] [18]













|

0 CH3

| |

Si 0 Si R

| |





- 22 -

0 CH3



























- 23 -









pH ca pha ng


1353 Chn m pH





1354 T c dograveng













- 24 -





137 ng dng ca HPLC








1371 nh lư ng













vagrave





- 25 -







detector như nhau







- 26 -







lư ng 9496

- KH2PO4 (PA)

- H3PO4 (PA)




2112 Dng c


PDA 100





- Cc coacute m











- 27 -








liu









n

1iixn

1

x (21)

- lch chun( )

1n

xS

n

1i

2

ixminus

minus

=sum

= (22)


Ss x

times= (23)

- Sai s chunn

S

sx

= (24)


st x)1n(


(25)






- 28 -




























- 29 -

200 250 300

nm

To bra 002

00

05

Abs


Nhn xeacutet





b La chn ct



c La chn m






Nhn xeacutet









- 32 -







ca pic tobramycin


bng 2




1 294 122985 152

2 295 123001 153

3 296 122805 151

4 296 122230 152

5 296 121916 152RSD() 030 040

Nhn xeacutet





Kt lun








- 33 -

a Dung dch th








c Mu trng


d Pha ng














sau





- 34 -

X = PmAmA

tc

ct timestimes

times




d ng









Hagravem lư ng

296 119207 9097297 119585 9095299 119165 9002

301 119975 9003303 120954 9017

m( chun) =290 121916 9496


lch chun S = 046


Sai s chun Sx= 021( 1) 05962n x

X t S α minus

∆ = times =


Kt lun





- 35 -












chun tobramycin









Bigravenh 1 2 3 4 5

Tobramycin (mg) 80 90 100 110 120

Nư c ct va (ml) 25 25 25 25 25

Nng (mgml) 24 27 30 33 36





- 36 -













Nng (mgml) 24 27 30 33 36



y = 39878x + 0039

R2 = 09999

0

4

8

12

16

0 1 2 3 4

Nồng độ (mgml)






- 37 -







Nhn xeacutet

















mu th







- 39 -












s hng nh)

Kt lun











- 40 -




i












- 42 -






theo cocircng thc

X = PmAmA

tc

ct timestimes

times














- 43 -

KT LUN VAgrave XUT

31 KT LUN


kt qu











32 XUT











- 16 -






















CAO (HPLC)








- 17 -





taacutech












1331 H th ng bơ m


(0 - 400 bar)




1333 H tiecircm m u






- 18 -


















th UV - VIS








983107ộ983156

983106983145983150983144

983139983144ứ983137

983152983144983137

độ983150983143

983106ơ983149

983108983141983156983141983139983156983151983154 983106ộ 983152983144ậ983150 983156ự 983143983144983145

983126983137983150 983156983145983274983149





- 19 -









kyacute


Wb rng aacutey pic

t

t

Wb2

05-2W

05-1W

b1W

R2

R2t

tR1

R1t

o





- 20 -




[8][14]






α=tt ttkk

01R

02R

1

2

minusminus= (k2

gt k1 ) [8] [14]





=R

α

minusα=

+

minus=

+

minus

+ k1

k

ww

tt

ww

ttB

B

A2 1B2 1

ARBR

AB

ARBR 1

4

N1812 [8] [14]


1344 H s b t i AF


a2AF w 20 1= [8] [14]

Trong oacute




1tt

ttt

ttk

0

R

0

0R

0

R minus=minus

==





- 21 -



N = 16

w

t

B

R

2

hay N=554

w

t

B2 1

R

2

[8] [12] [22]



L


1351 L a chn ct (pha t ĩ nh) [12] [18]













|

0 CH3

| |

Si 0 Si R

| |





- 22 -

0 CH3



























- 23 -









pH ca pha ng


1353 Chn m pH





1354 T c dograveng













- 24 -





137 ng dng ca HPLC








1371 nh lư ng













vagrave





- 25 -







detector như nhau







- 26 -







lư ng 9496

- KH2PO4 (PA)

- H3PO4 (PA)




2112 Dng c


PDA 100





- Cc coacute m











- 27 -








liu









n

1iixn

1

x (21)

- lch chun( )

1n

xS

n

1i

2

ixminus

minus

=sum

= (22)


Ss x

times= (23)

- Sai s chunn

S

sx

= (24)


st x)1n(


(25)






- 28 -




























- 29 -

200 250 300

nm

To bra 002

00

05

Abs


Nhn xeacutet





b La chn ct



c La chn m






Nhn xeacutet









- 32 -







ca pic tobramycin


bng 2




1 294 122985 152

2 295 123001 153

3 296 122805 151

4 296 122230 152

5 296 121916 152RSD() 030 040

Nhn xeacutet





Kt lun








- 33 -

a Dung dch th








c Mu trng


d Pha ng














sau





- 34 -

X = PmAmA

tc

ct timestimes

times




d ng









Hagravem lư ng

296 119207 9097297 119585 9095299 119165 9002

301 119975 9003303 120954 9017

m( chun) =290 121916 9496


lch chun S = 046


Sai s chun Sx= 021( 1) 05962n x

X t S α minus

∆ = times =


Kt lun





- 35 -












chun tobramycin









Bigravenh 1 2 3 4 5

Tobramycin (mg) 80 90 100 110 120

Nư c ct va (ml) 25 25 25 25 25

Nng (mgml) 24 27 30 33 36





- 36 -













Nng (mgml) 24 27 30 33 36



y = 39878x + 0039

R2 = 09999

0

4

8

12

16

0 1 2 3 4

Nồng độ (mgml)






- 37 -







Nhn xeacutet

















mu th







- 39 -












s hng nh)

Kt lun











- 40 -




i












- 42 -






theo cocircng thc

X = PmAmA

tc

ct timestimes

times














- 43 -

KT LUN VAgrave XUT

31 KT LUN


kt qu











32 XUT











- 17 -





taacutech












1331 H th ng bơ m


(0 - 400 bar)




1333 H tiecircm m u






- 18 -


















th UV - VIS








983107ộ983156

983106983145983150983144

983139983144ứ983137

983152983144983137

độ983150983143

983106ơ983149

983108983141983156983141983139983156983151983154 983106ộ 983152983144ậ983150 983156ự 983143983144983145

983126983137983150 983156983145983274983149





- 19 -









kyacute


Wb rng aacutey pic

t

t

Wb2

05-2W

05-1W

b1W

R2

R2t

tR1

R1t

o





- 20 -




[8][14]






α=tt ttkk

01R

02R

1

2

minusminus= (k2

gt k1 ) [8] [14]





=R

α

minusα=

+

minus=

+

minus

+ k1

k

ww

tt

ww

ttB

B

A2 1B2 1

ARBR

AB

ARBR 1

4

N1812 [8] [14]


1344 H s b t i AF


a2AF w 20 1= [8] [14]

Trong oacute




1tt

ttt

ttk

0

R

0

0R

0

R minus=minus

==





- 21 -



N = 16

w

t

B

R

2

hay N=554

w

t

B2 1

R

2

[8] [12] [22]



L


1351 L a chn ct (pha t ĩ nh) [12] [18]













|

0 CH3

| |

Si 0 Si R

| |





- 22 -

0 CH3



























- 23 -









pH ca pha ng


1353 Chn m pH





1354 T c dograveng













- 24 -





137 ng dng ca HPLC








1371 nh lư ng













vagrave





- 25 -







detector như nhau







- 26 -







lư ng 9496

- KH2PO4 (PA)

- H3PO4 (PA)




2112 Dng c


PDA 100





- Cc coacute m











- 27 -








liu









n

1iixn

1

x (21)

- lch chun( )

1n

xS

n

1i

2

ixminus

minus

=sum

= (22)


Ss x

times= (23)

- Sai s chunn

S

sx

= (24)


st x)1n(


(25)






- 28 -




























- 29 -

200 250 300

nm

To bra 002

00

05

Abs


Nhn xeacutet





b La chn ct



c La chn m






Nhn xeacutet









- 32 -







ca pic tobramycin


bng 2




1 294 122985 152

2 295 123001 153

3 296 122805 151

4 296 122230 152

5 296 121916 152RSD() 030 040

Nhn xeacutet





Kt lun








- 33 -

a Dung dch th








c Mu trng


d Pha ng














sau





- 34 -

X = PmAmA

tc

ct timestimes

times




d ng









Hagravem lư ng

296 119207 9097297 119585 9095299 119165 9002

301 119975 9003303 120954 9017

m( chun) =290 121916 9496


lch chun S = 046


Sai s chun Sx= 021( 1) 05962n x

X t S α minus

∆ = times =


Kt lun





- 35 -












chun tobramycin









Bigravenh 1 2 3 4 5

Tobramycin (mg) 80 90 100 110 120

Nư c ct va (ml) 25 25 25 25 25

Nng (mgml) 24 27 30 33 36





- 36 -













Nng (mgml) 24 27 30 33 36



y = 39878x + 0039

R2 = 09999

0

4

8

12

16

0 1 2 3 4

Nồng độ (mgml)






- 37 -







Nhn xeacutet

















mu th







- 39 -












s hng nh)

Kt lun











- 40 -




i












- 42 -






theo cocircng thc

X = PmAmA

tc

ct timestimes

times














- 43 -

KT LUN VAgrave XUT

31 KT LUN


kt qu











32 XUT











- 18 -


















th UV - VIS








983107ộ983156

983106983145983150983144

983139983144ứ983137

983152983144983137

độ983150983143

983106ơ983149

983108983141983156983141983139983156983151983154 983106ộ 983152983144ậ983150 983156ự 983143983144983145

983126983137983150 983156983145983274983149





- 19 -









kyacute


Wb rng aacutey pic

t

t

Wb2

05-2W

05-1W

b1W

R2

R2t

tR1

R1t

o





- 20 -




[8][14]






α=tt ttkk

01R

02R

1

2

minusminus= (k2

gt k1 ) [8] [14]





=R

α

minusα=

+

minus=

+

minus

+ k1

k

ww

tt

ww

ttB

B

A2 1B2 1

ARBR

AB

ARBR 1

4

N1812 [8] [14]


1344 H s b t i AF


a2AF w 20 1= [8] [14]

Trong oacute




1tt

ttt

ttk

0

R

0

0R

0

R minus=minus

==





- 21 -



N = 16

w

t

B

R

2

hay N=554

w

t

B2 1

R

2

[8] [12] [22]



L


1351 L a chn ct (pha t ĩ nh) [12] [18]













|

0 CH3

| |

Si 0 Si R

| |





- 22 -

0 CH3



























- 23 -









pH ca pha ng


1353 Chn m pH





1354 T c dograveng













- 24 -





137 ng dng ca HPLC








1371 nh lư ng













vagrave





- 25 -







detector như nhau







- 26 -







lư ng 9496

- KH2PO4 (PA)

- H3PO4 (PA)




2112 Dng c


PDA 100





- Cc coacute m











- 27 -








liu









n

1iixn

1

x (21)

- lch chun( )

1n

xS

n

1i

2

ixminus

minus

=sum

= (22)


Ss x

times= (23)

- Sai s chunn

S

sx

= (24)


st x)1n(


(25)






- 28 -




























- 29 -

200 250 300

nm

To bra 002

00

05

Abs


Nhn xeacutet





b La chn ct



c La chn m






Nhn xeacutet









- 32 -







ca pic tobramycin


bng 2




1 294 122985 152

2 295 123001 153

3 296 122805 151

4 296 122230 152

5 296 121916 152RSD() 030 040

Nhn xeacutet





Kt lun








- 33 -

a Dung dch th








c Mu trng


d Pha ng














sau





- 34 -

X = PmAmA

tc

ct timestimes

times




d ng









Hagravem lư ng

296 119207 9097297 119585 9095299 119165 9002

301 119975 9003303 120954 9017

m( chun) =290 121916 9496


lch chun S = 046


Sai s chun Sx= 021( 1) 05962n x

X t S α minus

∆ = times =


Kt lun





- 35 -












chun tobramycin









Bigravenh 1 2 3 4 5

Tobramycin (mg) 80 90 100 110 120

Nư c ct va (ml) 25 25 25 25 25

Nng (mgml) 24 27 30 33 36





- 36 -













Nng (mgml) 24 27 30 33 36



y = 39878x + 0039

R2 = 09999

0

4

8

12

16

0 1 2 3 4

Nồng độ (mgml)






- 37 -







Nhn xeacutet

















mu th







- 39 -












s hng nh)

Kt lun











- 40 -




i












- 42 -






theo cocircng thc

X = PmAmA

tc

ct timestimes

times














- 43 -

KT LUN VAgrave XUT

31 KT LUN


kt qu











32 XUT











- 19 -









kyacute


Wb rng aacutey pic

t

t

Wb2

05-2W

05-1W

b1W

R2

R2t

tR1

R1t

o





- 20 -




[8][14]






α=tt ttkk

01R

02R

1

2

minusminus= (k2

gt k1 ) [8] [14]





=R

α

minusα=

+

minus=

+

minus

+ k1

k

ww

tt

ww

ttB

B

A2 1B2 1

ARBR

AB

ARBR 1

4

N1812 [8] [14]


1344 H s b t i AF


a2AF w 20 1= [8] [14]

Trong oacute




1tt

ttt

ttk

0

R

0

0R

0

R minus=minus

==





- 21 -



N = 16

w

t

B

R

2

hay N=554

w

t

B2 1

R

2

[8] [12] [22]



L


1351 L a chn ct (pha t ĩ nh) [12] [18]













|

0 CH3

| |

Si 0 Si R

| |





- 22 -

0 CH3



























- 23 -









pH ca pha ng


1353 Chn m pH





1354 T c dograveng













- 24 -





137 ng dng ca HPLC








1371 nh lư ng













vagrave





- 25 -







detector như nhau







- 26 -







lư ng 9496

- KH2PO4 (PA)

- H3PO4 (PA)




2112 Dng c


PDA 100





- Cc coacute m











- 27 -








liu









n

1iixn

1

x (21)

- lch chun( )

1n

xS

n

1i

2

ixminus

minus

=sum

= (22)


Ss x

times= (23)

- Sai s chunn

S

sx

= (24)


st x)1n(


(25)






- 28 -




























- 29 -

200 250 300

nm

To bra 002

00

05

Abs


Nhn xeacutet





b La chn ct



c La chn m






Nhn xeacutet









- 32 -







ca pic tobramycin


bng 2




1 294 122985 152

2 295 123001 153

3 296 122805 151

4 296 122230 152

5 296 121916 152RSD() 030 040

Nhn xeacutet





Kt lun








- 33 -

a Dung dch th








c Mu trng


d Pha ng














sau





- 34 -

X = PmAmA

tc

ct timestimes

times




d ng









Hagravem lư ng

296 119207 9097297 119585 9095299 119165 9002

301 119975 9003303 120954 9017

m( chun) =290 121916 9496


lch chun S = 046


Sai s chun Sx= 021( 1) 05962n x

X t S α minus

∆ = times =


Kt lun





- 35 -












chun tobramycin









Bigravenh 1 2 3 4 5

Tobramycin (mg) 80 90 100 110 120

Nư c ct va (ml) 25 25 25 25 25

Nng (mgml) 24 27 30 33 36





- 36 -













Nng (mgml) 24 27 30 33 36



y = 39878x + 0039

R2 = 09999

0

4

8

12

16

0 1 2 3 4

Nồng độ (mgml)






- 37 -







Nhn xeacutet

















mu th







- 39 -












s hng nh)

Kt lun











- 40 -




i












- 42 -






theo cocircng thc

X = PmAmA

tc

ct timestimes

times














- 43 -

KT LUN VAgrave XUT

31 KT LUN


kt qu











32 XUT











- 20 -




[8][14]






α=tt ttkk

01R

02R

1

2

minusminus= (k2

gt k1 ) [8] [14]





=R

α

minusα=

+

minus=

+

minus

+ k1

k

ww

tt

ww

ttB

B

A2 1B2 1

ARBR

AB

ARBR 1

4

N1812 [8] [14]


1344 H s b t i AF


a2AF w 20 1= [8] [14]

Trong oacute




1tt

ttt

ttk

0

R

0

0R

0

R minus=minus

==





- 21 -



N = 16

w

t

B

R

2

hay N=554

w

t

B2 1

R

2

[8] [12] [22]



L


1351 L a chn ct (pha t ĩ nh) [12] [18]













|

0 CH3

| |

Si 0 Si R

| |





- 22 -

0 CH3



























- 23 -









pH ca pha ng


1353 Chn m pH





1354 T c dograveng













- 24 -





137 ng dng ca HPLC








1371 nh lư ng













vagrave





- 25 -







detector như nhau







- 26 -







lư ng 9496

- KH2PO4 (PA)

- H3PO4 (PA)




2112 Dng c


PDA 100





- Cc coacute m











- 27 -








liu









n

1iixn

1

x (21)

- lch chun( )

1n

xS

n

1i

2

ixminus

minus

=sum

= (22)


Ss x

times= (23)

- Sai s chunn

S

sx

= (24)


st x)1n(


(25)






- 28 -




























- 29 -

200 250 300

nm

To bra 002

00

05

Abs


Nhn xeacutet





b La chn ct



c La chn m






Nhn xeacutet









- 32 -







ca pic tobramycin


bng 2




1 294 122985 152

2 295 123001 153

3 296 122805 151

4 296 122230 152

5 296 121916 152RSD() 030 040

Nhn xeacutet





Kt lun








- 33 -

a Dung dch th








c Mu trng


d Pha ng














sau





- 34 -

X = PmAmA

tc

ct timestimes

times




d ng









Hagravem lư ng

296 119207 9097297 119585 9095299 119165 9002

301 119975 9003303 120954 9017

m( chun) =290 121916 9496


lch chun S = 046


Sai s chun Sx= 021( 1) 05962n x

X t S α minus

∆ = times =


Kt lun





- 35 -












chun tobramycin









Bigravenh 1 2 3 4 5

Tobramycin (mg) 80 90 100 110 120

Nư c ct va (ml) 25 25 25 25 25

Nng (mgml) 24 27 30 33 36





- 36 -













Nng (mgml) 24 27 30 33 36



y = 39878x + 0039

R2 = 09999

0

4

8

12

16

0 1 2 3 4

Nồng độ (mgml)






- 37 -







Nhn xeacutet

















mu th







- 39 -












s hng nh)

Kt lun











- 40 -




i












- 42 -






theo cocircng thc

X = PmAmA

tc

ct timestimes

times














- 43 -

KT LUN VAgrave XUT

31 KT LUN


kt qu











32 XUT











- 21 -



N = 16

w

t

B

R

2

hay N=554

w

t

B2 1

R

2

[8] [12] [22]



L


1351 L a chn ct (pha t ĩ nh) [12] [18]













|

0 CH3

| |

Si 0 Si R

| |





- 22 -

0 CH3



























- 23 -









pH ca pha ng


1353 Chn m pH





1354 T c dograveng













- 24 -





137 ng dng ca HPLC








1371 nh lư ng













vagrave





- 25 -







detector như nhau







- 26 -







lư ng 9496

- KH2PO4 (PA)

- H3PO4 (PA)




2112 Dng c


PDA 100





- Cc coacute m











- 27 -








liu









n

1iixn

1

x (21)

- lch chun( )

1n

xS

n

1i

2

ixminus

minus

=sum

= (22)


Ss x

times= (23)

- Sai s chunn

S

sx

= (24)


st x)1n(


(25)






- 28 -




























- 29 -

200 250 300

nm

To bra 002

00

05

Abs


Nhn xeacutet





b La chn ct



c La chn m






Nhn xeacutet









- 32 -







ca pic tobramycin


bng 2




1 294 122985 152

2 295 123001 153

3 296 122805 151

4 296 122230 152

5 296 121916 152RSD() 030 040

Nhn xeacutet





Kt lun








- 33 -

a Dung dch th








c Mu trng


d Pha ng














sau





- 34 -

X = PmAmA

tc

ct timestimes

times




d ng









Hagravem lư ng

296 119207 9097297 119585 9095299 119165 9002

301 119975 9003303 120954 9017

m( chun) =290 121916 9496


lch chun S = 046


Sai s chun Sx= 021( 1) 05962n x

X t S α minus

∆ = times =


Kt lun





- 35 -












chun tobramycin









Bigravenh 1 2 3 4 5

Tobramycin (mg) 80 90 100 110 120

Nư c ct va (ml) 25 25 25 25 25

Nng (mgml) 24 27 30 33 36





- 36 -













Nng (mgml) 24 27 30 33 36



y = 39878x + 0039

R2 = 09999

0

4

8

12

16

0 1 2 3 4

Nồng độ (mgml)






- 37 -







Nhn xeacutet

















mu th







- 39 -












s hng nh)

Kt lun











- 40 -




i












- 42 -






theo cocircng thc

X = PmAmA

tc

ct timestimes

times














- 43 -

KT LUN VAgrave XUT

31 KT LUN


kt qu











32 XUT











- 22 -

0 CH3



























- 23 -









pH ca pha ng


1353 Chn m pH





1354 T c dograveng













- 24 -





137 ng dng ca HPLC








1371 nh lư ng













vagrave





- 25 -







detector như nhau







- 26 -







lư ng 9496

- KH2PO4 (PA)

- H3PO4 (PA)




2112 Dng c


PDA 100





- Cc coacute m











- 27 -








liu









n

1iixn

1

x (21)

- lch chun( )

1n

xS

n

1i

2

ixminus

minus

=sum

= (22)


Ss x

times= (23)

- Sai s chunn

S

sx

= (24)


st x)1n(


(25)






- 28 -




























- 29 -

200 250 300

nm

To bra 002

00

05

Abs


Nhn xeacutet





b La chn ct



c La chn m






Nhn xeacutet









- 32 -







ca pic tobramycin


bng 2




1 294 122985 152

2 295 123001 153

3 296 122805 151

4 296 122230 152

5 296 121916 152RSD() 030 040

Nhn xeacutet





Kt lun








- 33 -

a Dung dch th








c Mu trng


d Pha ng














sau





- 34 -

X = PmAmA

tc

ct timestimes

times




d ng









Hagravem lư ng

296 119207 9097297 119585 9095299 119165 9002

301 119975 9003303 120954 9017

m( chun) =290 121916 9496


lch chun S = 046


Sai s chun Sx= 021( 1) 05962n x

X t S α minus

∆ = times =


Kt lun





- 35 -












chun tobramycin









Bigravenh 1 2 3 4 5

Tobramycin (mg) 80 90 100 110 120

Nư c ct va (ml) 25 25 25 25 25

Nng (mgml) 24 27 30 33 36





- 36 -













Nng (mgml) 24 27 30 33 36



y = 39878x + 0039

R2 = 09999

0

4

8

12

16

0 1 2 3 4

Nồng độ (mgml)






- 37 -







Nhn xeacutet

















mu th







- 39 -












s hng nh)

Kt lun











- 40 -




i












- 42 -






theo cocircng thc

X = PmAmA

tc

ct timestimes

times














- 43 -

KT LUN VAgrave XUT

31 KT LUN


kt qu











32 XUT











- 23 -









pH ca pha ng


1353 Chn m pH





1354 T c dograveng













- 24 -





137 ng dng ca HPLC








1371 nh lư ng













vagrave





- 25 -







detector như nhau







- 26 -







lư ng 9496

- KH2PO4 (PA)

- H3PO4 (PA)




2112 Dng c


PDA 100





- Cc coacute m











- 27 -








liu









n

1iixn

1

x (21)

- lch chun( )

1n

xS

n

1i

2

ixminus

minus

=sum

= (22)


Ss x

times= (23)

- Sai s chunn

S

sx

= (24)


st x)1n(


(25)






- 28 -




























- 29 -

200 250 300

nm

To bra 002

00

05

Abs


Nhn xeacutet





b La chn ct



c La chn m






Nhn xeacutet









- 32 -







ca pic tobramycin


bng 2




1 294 122985 152

2 295 123001 153

3 296 122805 151

4 296 122230 152

5 296 121916 152RSD() 030 040

Nhn xeacutet





Kt lun








- 33 -

a Dung dch th








c Mu trng


d Pha ng














sau





- 34 -

X = PmAmA

tc

ct timestimes

times




d ng









Hagravem lư ng

296 119207 9097297 119585 9095299 119165 9002

301 119975 9003303 120954 9017

m( chun) =290 121916 9496


lch chun S = 046


Sai s chun Sx= 021( 1) 05962n x

X t S α minus

∆ = times =


Kt lun





- 35 -












chun tobramycin









Bigravenh 1 2 3 4 5

Tobramycin (mg) 80 90 100 110 120

Nư c ct va (ml) 25 25 25 25 25

Nng (mgml) 24 27 30 33 36





- 36 -













Nng (mgml) 24 27 30 33 36



y = 39878x + 0039

R2 = 09999

0

4

8

12

16

0 1 2 3 4

Nồng độ (mgml)






- 37 -







Nhn xeacutet

















mu th







- 39 -












s hng nh)

Kt lun











- 40 -




i












- 42 -






theo cocircng thc

X = PmAmA

tc

ct timestimes

times














- 43 -

KT LUN VAgrave XUT

31 KT LUN


kt qu











32 XUT











- 24 -





137 ng dng ca HPLC








1371 nh lư ng













vagrave





- 25 -







detector như nhau







- 26 -







lư ng 9496

- KH2PO4 (PA)

- H3PO4 (PA)




2112 Dng c


PDA 100





- Cc coacute m











- 27 -








liu









n

1iixn

1

x (21)

- lch chun( )

1n

xS

n

1i

2

ixminus

minus

=sum

= (22)


Ss x

times= (23)

- Sai s chunn

S

sx

= (24)


st x)1n(


(25)






- 28 -




























- 29 -

200 250 300

nm

To bra 002

00

05

Abs


Nhn xeacutet





b La chn ct



c La chn m






Nhn xeacutet









- 32 -







ca pic tobramycin


bng 2




1 294 122985 152

2 295 123001 153

3 296 122805 151

4 296 122230 152

5 296 121916 152RSD() 030 040

Nhn xeacutet





Kt lun








- 33 -

a Dung dch th








c Mu trng


d Pha ng














sau





- 34 -

X = PmAmA

tc

ct timestimes

times




d ng









Hagravem lư ng

296 119207 9097297 119585 9095299 119165 9002

301 119975 9003303 120954 9017

m( chun) =290 121916 9496


lch chun S = 046


Sai s chun Sx= 021( 1) 05962n x

X t S α minus

∆ = times =


Kt lun





- 35 -












chun tobramycin









Bigravenh 1 2 3 4 5

Tobramycin (mg) 80 90 100 110 120

Nư c ct va (ml) 25 25 25 25 25

Nng (mgml) 24 27 30 33 36





- 36 -













Nng (mgml) 24 27 30 33 36



y = 39878x + 0039

R2 = 09999

0

4

8

12

16

0 1 2 3 4

Nồng độ (mgml)






- 37 -







Nhn xeacutet

















mu th







- 39 -












s hng nh)

Kt lun











- 40 -




i












- 42 -






theo cocircng thc

X = PmAmA

tc

ct timestimes

times














- 43 -

KT LUN VAgrave XUT

31 KT LUN


kt qu











32 XUT











- 25 -







detector như nhau







- 26 -







lư ng 9496

- KH2PO4 (PA)

- H3PO4 (PA)




2112 Dng c


PDA 100





- Cc coacute m











- 27 -








liu









n

1iixn

1

x (21)

- lch chun( )

1n

xS

n

1i

2

ixminus

minus

=sum

= (22)


Ss x

times= (23)

- Sai s chunn

S

sx

= (24)


st x)1n(


(25)






- 28 -




























- 29 -

200 250 300

nm

To bra 002

00

05

Abs


Nhn xeacutet





b La chn ct



c La chn m






Nhn xeacutet









- 32 -







ca pic tobramycin


bng 2




1 294 122985 152

2 295 123001 153

3 296 122805 151

4 296 122230 152

5 296 121916 152RSD() 030 040

Nhn xeacutet





Kt lun








- 33 -

a Dung dch th








c Mu trng


d Pha ng














sau





- 34 -

X = PmAmA

tc

ct timestimes

times




d ng









Hagravem lư ng

296 119207 9097297 119585 9095299 119165 9002

301 119975 9003303 120954 9017

m( chun) =290 121916 9496


lch chun S = 046


Sai s chun Sx= 021( 1) 05962n x

X t S α minus

∆ = times =


Kt lun





- 35 -












chun tobramycin









Bigravenh 1 2 3 4 5

Tobramycin (mg) 80 90 100 110 120

Nư c ct va (ml) 25 25 25 25 25

Nng (mgml) 24 27 30 33 36





- 36 -













Nng (mgml) 24 27 30 33 36



y = 39878x + 0039

R2 = 09999

0

4

8

12

16

0 1 2 3 4

Nồng độ (mgml)






- 37 -







Nhn xeacutet

















mu th







- 39 -












s hng nh)

Kt lun











- 40 -




i












- 42 -






theo cocircng thc

X = PmAmA

tc

ct timestimes

times














- 43 -

KT LUN VAgrave XUT

31 KT LUN


kt qu











32 XUT











- 26 -







lư ng 9496

- KH2PO4 (PA)

- H3PO4 (PA)




2112 Dng c


PDA 100





- Cc coacute m











- 27 -








liu









n

1iixn

1

x (21)

- lch chun( )

1n

xS

n

1i

2

ixminus

minus

=sum

= (22)


Ss x

times= (23)

- Sai s chunn

S

sx

= (24)


st x)1n(


(25)






- 28 -




























- 29 -

200 250 300

nm

To bra 002

00

05

Abs


Nhn xeacutet





b La chn ct



c La chn m






Nhn xeacutet









- 32 -







ca pic tobramycin


bng 2




1 294 122985 152

2 295 123001 153

3 296 122805 151

4 296 122230 152

5 296 121916 152RSD() 030 040

Nhn xeacutet





Kt lun








- 33 -

a Dung dch th








c Mu trng


d Pha ng














sau





- 34 -

X = PmAmA

tc

ct timestimes

times




d ng









Hagravem lư ng

296 119207 9097297 119585 9095299 119165 9002

301 119975 9003303 120954 9017

m( chun) =290 121916 9496


lch chun S = 046


Sai s chun Sx= 021( 1) 05962n x

X t S α minus

∆ = times =


Kt lun





- 35 -












chun tobramycin









Bigravenh 1 2 3 4 5

Tobramycin (mg) 80 90 100 110 120

Nư c ct va (ml) 25 25 25 25 25

Nng (mgml) 24 27 30 33 36





- 36 -













Nng (mgml) 24 27 30 33 36



y = 39878x + 0039

R2 = 09999

0

4

8

12

16

0 1 2 3 4

Nồng độ (mgml)






- 37 -







Nhn xeacutet

















mu th







- 39 -












s hng nh)

Kt lun











- 40 -




i












- 42 -






theo cocircng thc

X = PmAmA

tc

ct timestimes

times














- 43 -

KT LUN VAgrave XUT

31 KT LUN


kt qu











32 XUT











- 27 -








liu









n

1iixn

1

x (21)

- lch chun( )

1n

xS

n

1i

2

ixminus

minus

=sum

= (22)


Ss x

times= (23)

- Sai s chunn

S

sx

= (24)


st x)1n(


(25)






- 28 -




























- 29 -

200 250 300

nm

To bra 002

00

05

Abs


Nhn xeacutet





b La chn ct



c La chn m






Nhn xeacutet









- 32 -







ca pic tobramycin


bng 2




1 294 122985 152

2 295 123001 153

3 296 122805 151

4 296 122230 152

5 296 121916 152RSD() 030 040

Nhn xeacutet





Kt lun








- 33 -

a Dung dch th








c Mu trng


d Pha ng














sau





- 34 -

X = PmAmA

tc

ct timestimes

times




d ng









Hagravem lư ng

296 119207 9097297 119585 9095299 119165 9002

301 119975 9003303 120954 9017

m( chun) =290 121916 9496


lch chun S = 046


Sai s chun Sx= 021( 1) 05962n x

X t S α minus

∆ = times =


Kt lun





- 35 -












chun tobramycin









Bigravenh 1 2 3 4 5

Tobramycin (mg) 80 90 100 110 120

Nư c ct va (ml) 25 25 25 25 25

Nng (mgml) 24 27 30 33 36





- 36 -













Nng (mgml) 24 27 30 33 36



y = 39878x + 0039

R2 = 09999

0

4

8

12

16

0 1 2 3 4

Nồng độ (mgml)






- 37 -







Nhn xeacutet

















mu th







- 39 -












s hng nh)

Kt lun











- 40 -




i












- 42 -






theo cocircng thc

X = PmAmA

tc

ct timestimes

times














- 43 -

KT LUN VAgrave XUT

31 KT LUN


kt qu











32 XUT











- 28 -




























- 29 -

200 250 300

nm

To bra 002

00

05

Abs


Nhn xeacutet





b La chn ct



c La chn m






Nhn xeacutet









- 32 -







ca pic tobramycin


bng 2




1 294 122985 152

2 295 123001 153

3 296 122805 151

4 296 122230 152

5 296 121916 152RSD() 030 040

Nhn xeacutet





Kt lun








- 33 -

a Dung dch th








c Mu trng


d Pha ng














sau





- 34 -

X = PmAmA

tc

ct timestimes

times




d ng









Hagravem lư ng

296 119207 9097297 119585 9095299 119165 9002

301 119975 9003303 120954 9017

m( chun) =290 121916 9496


lch chun S = 046


Sai s chun Sx= 021( 1) 05962n x

X t S α minus

∆ = times =


Kt lun





- 35 -












chun tobramycin









Bigravenh 1 2 3 4 5

Tobramycin (mg) 80 90 100 110 120

Nư c ct va (ml) 25 25 25 25 25

Nng (mgml) 24 27 30 33 36





- 36 -













Nng (mgml) 24 27 30 33 36



y = 39878x + 0039

R2 = 09999

0

4

8

12

16

0 1 2 3 4

Nồng độ (mgml)






- 37 -







Nhn xeacutet

















mu th







- 39 -












s hng nh)

Kt lun











- 40 -




i












- 42 -






theo cocircng thc

X = PmAmA

tc

ct timestimes

times














- 43 -

KT LUN VAgrave XUT

31 KT LUN


kt qu











32 XUT











- 29 -

200 250 300

nm

To bra 002

00

05

Abs


Nhn xeacutet





b La chn ct



c La chn m






Nhn xeacutet









- 32 -







ca pic tobramycin


bng 2




1 294 122985 152

2 295 123001 153

3 296 122805 151

4 296 122230 152

5 296 121916 152RSD() 030 040

Nhn xeacutet





Kt lun








- 33 -

a Dung dch th








c Mu trng


d Pha ng














sau





- 34 -

X = PmAmA

tc

ct timestimes

times




d ng









Hagravem lư ng

296 119207 9097297 119585 9095299 119165 9002

301 119975 9003303 120954 9017

m( chun) =290 121916 9496


lch chun S = 046


Sai s chun Sx= 021( 1) 05962n x

X t S α minus

∆ = times =


Kt lun





- 35 -












chun tobramycin









Bigravenh 1 2 3 4 5

Tobramycin (mg) 80 90 100 110 120

Nư c ct va (ml) 25 25 25 25 25

Nng (mgml) 24 27 30 33 36





- 36 -













Nng (mgml) 24 27 30 33 36



y = 39878x + 0039

R2 = 09999

0

4

8

12

16

0 1 2 3 4

Nồng độ (mgml)






- 37 -







Nhn xeacutet

















mu th







- 39 -












s hng nh)

Kt lun











- 40 -




i












- 42 -






theo cocircng thc

X = PmAmA

tc

ct timestimes

times














- 43 -

KT LUN VAgrave XUT

31 KT LUN


kt qu











32 XUT















- 32 -







ca pic tobramycin


bng 2




1 294 122985 152

2 295 123001 153

3 296 122805 151

4 296 122230 152

5 296 121916 152RSD() 030 040

Nhn xeacutet





Kt lun








- 33 -

a Dung dch th








c Mu trng


d Pha ng














sau





- 34 -

X = PmAmA

tc

ct timestimes

times




d ng









Hagravem lư ng

296 119207 9097297 119585 9095299 119165 9002

301 119975 9003303 120954 9017

m( chun) =290 121916 9496


lch chun S = 046


Sai s chun Sx= 021( 1) 05962n x

X t S α minus

∆ = times =


Kt lun





- 35 -












chun tobramycin









Bigravenh 1 2 3 4 5

Tobramycin (mg) 80 90 100 110 120

Nư c ct va (ml) 25 25 25 25 25

Nng (mgml) 24 27 30 33 36





- 36 -













Nng (mgml) 24 27 30 33 36



y = 39878x + 0039

R2 = 09999

0

4

8

12

16

0 1 2 3 4

Nồng độ (mgml)






- 37 -







Nhn xeacutet

















mu th







- 39 -












s hng nh)

Kt lun











- 40 -




i












- 42 -






theo cocircng thc

X = PmAmA

tc

ct timestimes

times














- 43 -

KT LUN VAgrave XUT

31 KT LUN


kt qu











32 XUT











- 33 -

a Dung dch th








c Mu trng


d Pha ng














sau





- 34 -

X = PmAmA

tc

ct timestimes

times




d ng









Hagravem lư ng

296 119207 9097297 119585 9095299 119165 9002

301 119975 9003303 120954 9017

m( chun) =290 121916 9496


lch chun S = 046


Sai s chun Sx= 021( 1) 05962n x

X t S α minus

∆ = times =


Kt lun





- 35 -












chun tobramycin









Bigravenh 1 2 3 4 5

Tobramycin (mg) 80 90 100 110 120

Nư c ct va (ml) 25 25 25 25 25

Nng (mgml) 24 27 30 33 36





- 36 -













Nng (mgml) 24 27 30 33 36



y = 39878x + 0039

R2 = 09999

0

4

8

12

16

0 1 2 3 4

Nồng độ (mgml)






- 37 -







Nhn xeacutet

















mu th







- 39 -












s hng nh)

Kt lun











- 40 -




i












- 42 -






theo cocircng thc

X = PmAmA

tc

ct timestimes

times














- 43 -

KT LUN VAgrave XUT

31 KT LUN


kt qu











32 XUT











- 34 -

X = PmAmA

tc

ct timestimes

times




d ng









Hagravem lư ng

296 119207 9097297 119585 9095299 119165 9002

301 119975 9003303 120954 9017

m( chun) =290 121916 9496


lch chun S = 046


Sai s chun Sx= 021( 1) 05962n x

X t S α minus

∆ = times =


Kt lun





- 35 -












chun tobramycin









Bigravenh 1 2 3 4 5

Tobramycin (mg) 80 90 100 110 120

Nư c ct va (ml) 25 25 25 25 25

Nng (mgml) 24 27 30 33 36





- 36 -













Nng (mgml) 24 27 30 33 36



y = 39878x + 0039

R2 = 09999

0

4

8

12

16

0 1 2 3 4

Nồng độ (mgml)






- 37 -







Nhn xeacutet

















mu th







- 39 -












s hng nh)

Kt lun











- 40 -




i












- 42 -






theo cocircng thc

X = PmAmA

tc

ct timestimes

times














- 43 -

KT LUN VAgrave XUT

31 KT LUN


kt qu











32 XUT











- 35 -












chun tobramycin









Bigravenh 1 2 3 4 5

Tobramycin (mg) 80 90 100 110 120

Nư c ct va (ml) 25 25 25 25 25

Nng (mgml) 24 27 30 33 36





- 36 -













Nng (mgml) 24 27 30 33 36



y = 39878x + 0039

R2 = 09999

0

4

8

12

16

0 1 2 3 4

Nồng độ (mgml)






- 37 -







Nhn xeacutet

















mu th







- 39 -












s hng nh)

Kt lun











- 40 -




i












- 42 -






theo cocircng thc

X = PmAmA

tc

ct timestimes

times














- 43 -

KT LUN VAgrave XUT

31 KT LUN


kt qu











32 XUT











- 36 -













Nng (mgml) 24 27 30 33 36



y = 39878x + 0039

R2 = 09999

0

4

8

12

16

0 1 2 3 4

Nồng độ (mgml)






- 37 -







Nhn xeacutet

















mu th







- 39 -












s hng nh)

Kt lun











- 40 -




i












- 42 -






theo cocircng thc

X = PmAmA

tc

ct timestimes

times














- 43 -

KT LUN VAgrave XUT

31 KT LUN


kt qu











32 XUT











- 37 -







Nhn xeacutet

















mu th







- 39 -












s hng nh)

Kt lun











- 40 -




i












- 42 -






theo cocircng thc

X = PmAmA

tc

ct timestimes

times














- 43 -

KT LUN VAgrave XUT

31 KT LUN


kt qu











32 XUT













- 39 -












s hng nh)

Kt lun











- 40 -




i












- 42 -






theo cocircng thc

X = PmAmA

tc

ct timestimes

times














- 43 -

KT LUN VAgrave XUT

31 KT LUN


kt qu











32 XUT











- 40 -




i












- 42 -






theo cocircng thc

X = PmAmA

tc

ct timestimes

times














- 43 -

KT LUN VAgrave XUT

31 KT LUN


kt qu











32 XUT













- 42 -






theo cocircng thc

X = PmAmA

tc

ct timestimes

times














- 43 -

KT LUN VAgrave XUT

31 KT LUN


kt qu











32 XUT











- 43 -

KT LUN VAgrave XUT

31 KT LUN


kt qu











32 XUT









Xây dựng quy trình kỹ thuật để định lượng tobramycin bằng phương pháp HPLC

Documents

Transcript of Xây dựng quy trình kỹ thuật để định lượng tobramycin bằng phương pháp HPLC