975
KLINICKÁ341804301074588109168189242288347
FARMAKOLOGIE709804301074291138211265312372415474533554598
A13288043010753
FARMACIE1403804301074191138210265316336380
/18298043010732
Klin190680430107446583131
Farmakol2057804301074278105179220258305325
Farm2402804301074178104179
2023;37(3):89-922601804301074386128172186230270290333354368412456485529571
/32178043010732
www.klinickafarmakologie.cz4568804301076513320121425327329234035839843647950254457164568672477179183988990895196610061045
901828026012657116
PŮVODNÍ341207012660132201274348422456
PRÁCE818207012661124196256314
Assay34136501063872104143176
of53236501063970
colistin61736501063372103121153186204245
A,87836501063754
B947365010634
and99636501063879119
colistin113136501063271103120153185203244
methanesulfonate139136501065284117158198239271306346377408449491531559591
in199836501061556
human207036501063980133174215
plasma230136501063263105139193233
by255036501063470
LC-MS/MS26363650106286180134167193245278
and293036501063879119
short-term306436501063172112148180204233265301355
plasma343536501063364106140193234
stability368436501063164100136153184202234273
molecular343621010977129152199242293316361392
weight761621010968115137190242272
is106062101092258
not1145621010951103134
precisely130662101095284130173195232279302347
determined.16806210109529913117720828630834376101095198151170
The54076101094597144
main710761010977121143194
components93076101094191168221273324371423453491
of144776101095077
this155176101093081103139
mixture171676101097698142175226256303
are34390101094373118
colistin47990101094089111132167198219269
A766901010955
and83990101094393145
colistin100190101094189111133168199220270
B,128990101094867
whose1374901010966117167203249
molecular1640901010975125147193235284306349379
weights3431041010967113136188239270307
are677104101094474121
1169,5824104101094786129176192242
and1092104101094496148
1155,4,1267104101094785127172190238257
respectively1551104101093076113166213256289310355401424468
(5,34311810109267088
6).45511810109467087
The566118101094496141
base731118101095196131177
of932118101094976
the1032118101093080125
colistin1181118101094189111132168198219270
molecule14751181010975125147192234284305351
consists185011810109408913934313210109365792123160
of522132101094976
a6171321010943
decapeptide679132101095197138182234280332362383434480
with1178132101096688118168
seven1365132101093681125169220
amino16041321010943118139190240
acid1863132101094384105156
residues343146101092975111132183232278314
in675146101092171
a7631461010944
cyclic824146101094185125147168208
formation1050146101092676106182225255276326376
and1444146101094393145
a16061461010944
fatty1667146101092771101133178
acid1862146101094485106157
tail34316010109307495117
attached479160101094473106150191241286338
to837160101093079
the936160101092979125
tripeptide108116010109305981132179230261282333378
end1479160101094596147
(4,164616010109256988
7).175416010109467189
Due18621601010960111157
to343174101093079
the441174101093080126
toxic587174101092978120141182
side789174101093556108153
effects,962174101094572102147189221258277
colistin1259174101094089111132167198219269
is1548174101092156
used1624174101094985131183
as1826174101094479
an1925174101094393
inactive343188101092170112152183204247290
prodrug,649188101095079128179207257306325
colistin989188101094088109129164194214263
methanesulphonate12671881010974119149198241290334369418438489538587637679708752
(CMS).343202101092575147190215231
CMS5902021010951123167
is772202101092156
hydrolyzed843202101095090140169218239282322366418
spontaneously1276202101093586136185214258307352402452486507550
in1841202101092170
an1927202101094392
aqueous343216101094393142187237286321
solution679216101093584106155184205254303
in997216101092068
vivo1078216101094061101146
and1240216101094392143
in1398216101092068
vitro147921610109406188116162
to1657216101092978
a17502161010943
series1808216101093580110131175211
of343230101095178
partially448230101095399129163186231253275320
methanesulfonated7952301010977124156208253305352389440463491542595640671717771
derivatives1593230101095299130153198244275297342388426
and343244101094393145
colistin507244101094089111132167198219269
(1,79524410109266883
8).89724410109467087
To483258101094590
find599258101092651102154
the779258101093081128
most9332581010975126162194
optimal11532581010950102133155232276298
conditions1478258101094089141193215246268319370407
for1911258101092677108
the343272101093082129
determination500272101095198130176207285308359404435458509561
of1089272101095078
CMS11942721010953128173
and1395272101094496148
colistin,1571272101094191114137174206228280300
various1899272101094389343286101093054107161199
methods5732861010977126159212266321359
have963286101095198143191
been11852861010953102151205
published14212861010953107161186209247300349404
that1856286101093184131163
differ343300101095173100131177208
in571300101092172
sample663300101093681157209232279
preparation,962300101095182128181226256301332354404456476
analytical1458300101094394138161205239261302347369
conditions,1847300101094090142343314101095276109133186240278300
and678314101094598153
measured8663141010977126173212264297345401
concentrations1302314101094193147191239293325358405437461514568607
in1943314101092276
patients’343328101095196127149195247277315335
plasma697328101095174119155231276
samples992328101093681157210233279316
(8,132832810109257192
9).143932810109467290
The1548328101094597144
method17113281010975122153204255308
described343342101095199137179210233287335389
below7603421010952100123175243
is1031342101092259
based1118342101095298135182236
on13823421010950103
already151334210109446798145190243288
published1829342101095210415834335610109214480131178231
and602356101094495147
established777356101094582114160213236258295346393446
methods,12503561010976123155206257310347368
which16453561010967118141182234
we19063561010967113
have343370101094993136180
adapted540370101094395138190220265317
to874370101093079
our970370101094999129
instrumental1116370101092171107138168218293339390419464486
conditions1619370101094089139191212243264314364400
to343384101093082
follow453384101092779103126178247
up7283841010951105
with861384101096891124176
a10653841010945
study1138384101093669121175220
focusing1387384101092779123175212235287341
on17563841010950103
the1888384101093083131
pharmacokinetics3433981010951101145175250294335384427450500546576598639676
of1036398101094975
colistin1129398101093988111132167198220270
used1416398101094985131183
in1616398101092171
critically170439810109407091122143184228250272315
ill34341210109214467
patients.437412101095298129152199251282320341
Colistin8064121010953103126149185218240292
plasma1126412101095175120157234279
concentrations1433412101094191143186232284315346392423445497549586
in343426101092172
critically43842610109407093124146187232255277321
ill78142610109214466
patients869426101095197127149195247277315
generally1206426101095197149195226271294316360
range1588426101092974125177224
from1834426101092758108185
0,6–13343440101094665113163205246
mg/L6104401010975127160204
(3).83544010109257195113
Several483454101094490133177207251273
pitfalls772454101095172103131176197218253
can1041454101094185135
be1192454101095197
encountered1305454101094596136185235285314359389434485
in1807454101092171
the1894454101092979125
determination343468101095196127172202277299349392422443493543
of909468101094976
CMS10074681010952125169
and1198468101094494145
colistin.1365468101094189111132168199220270289
One16764681010962113159
problem1857468101095282131343482101095173119195
is562482101092157
the643482101093080126
adsorption794482101094394130180210262292313363413
of1232482101094976
colistin1333482101094089111132167198219269
to1627482101093079
a17304821010944
range1798482101093074124175221
of343496101095078
materials,4474961010976121152198228251295317354374
including847496101092173115137187240262313365
plastics1239496101095275119156187209251289
and1554496101094496148
glass1729496101095173118154191
in1946496101092173
labware.343510101092165116182226256301320
If685510101092149
colistin756510101094089111132167198219269
adsorbs1048510101094394130180210262297
to1368510101093079
materials14695101010975119148193223245288309345
used1836510101095085131183
in343524101092171
sample439524101093680155207229275
collection,739524101094089111132178219251272322373392
processing,1156524101095181130172216252288309359410429
or1610524101094979
storage,1714524101093667116146190241286305
its34353810109215188
concentrations448538101094089140180225275305335379408429479530565
may10305381010975119160
be1207538101095198
incorrectly1322538101092171112161191221266307339361404
evaluated.1743538101094588132153203247343552101093074126146
Another5105521010955106156185235281311
problem843552101095181131182204250326
is1191552101092156
the1269552101092979125
stability1416552101093666111162184205226256302
of1739552101095076
both18375521010951102132182
substances,343566101093685136172203247297338383419438
CMS8045661010952125169
and996566101094494145
colistin.1164566101094189111132168199220270289
The1476566101094596142
hydrolysis1641566101095091142171221243286322343378
of343580101095076
CMS4455801010952125169
to640580101093079
colistin744580101094190112133168199221271
and1040580101094494146
colistin1211580101094189112133168199220271
stability1507580101093667111163185206227258303
itself183558010109225289135157184
depends343594101095196148195245296332
on6935941010950100
concentration,812594101094089140180225275305335379408429479530549
time,1379594101093051127172191
temperature,158859410109307515120224827832235240234360810109297593
and462608101094393145
the632608101093080126
matrix7836081010975119149179200243
containing1051608101094089140169214235285306357407
the1484608101093080126
substance1635608101093685137173203248298339384
(7,34362210109267181
9,444622101094662
10).525622101094689113130
Experimental3436470014762124195258302333435498568609672703
Chemical3436755014368132191289313367422447
and8196755014354118184
reagents1032675501433592145211269333369415
Colistin4836921010953102124145181212233284
sulfate,793692101093685107134179209254273
Colistinmethate10926921010952101123145180212233284359406436487531561606
Sodium1724692101094495147169219295
and343706101094392143
Polymyxin502706101094795116159234275319341390
B9087061010948
sulfate972706101093584105131175204248
(internal123670610109254595124168197247290311
standard,1563706101093566109159209252281332350
IS)192970610109216589
were3437201010965109137181
purchased539720101095099128166215257291336386
from940720101092655103177
Sigma–Aldrich1132720101094363112185228273327347397426446486534
(St.168172010109256693112
Louis,1808720101094189138158192211
MO,3437341010972133150
USA).5087341010958101154179195
MS7187341010972116
grade849734101095079122172217
water,10827341010965108137181210225
Formic1322734101094492121196216256
acid1594734101094282103153
≥17637341010953
991826734101094692
%,1929734101097090
and343748101094392143
Sulphuric502748101094492114164214263292313354
acid871748101094383104154
961041748101094692
%11447481010971
were12317481010965110139184
purchased14307481010951101130170219262297342393
from1839748101092656105180
VWR3437621010950131180
(Radnor,549762101092574118170220270301316
PA,890762101094899120
USA).10367621010958102157182200
MS12617621010973117
grade1404762101095181125177223
Methanol16527621010973119150200244294345367
was3437761010968115153
purchased5317761010953106138181233280317366420
from987776101092860112192
J.1214776101093555
T.1305776101094662
Baker1403776101095097142189221
(Avantor,1660776101092683127174227258310342359
Gliwice,213862101095881102168190230275293
Poland),245462101094896118162212263289306
and278362101094393144
Sodium295062101094494146167217292
hydroxide326562101095091142171221262284335380
was3668621010966110146
obtained2138761010950102133178199250297349
from251376101092758108184
Lach-Ner272376101094389130181210272319349
(Neratovice,309876101092685132162207237287330353394439457
Czech358176101095394140182233
Republic).213890101094894146196248270291331356373
Chromatographic21381155014368132167229328382418480546581637702767792845
conditions30121155014353115180246270307332396461505
The2278132101094598144
liquid244913210109214395146168220
chromatography2695132101094192122173250295325375428458503556607650
was33711321010967112149
performed354613210109529913016021124221381461010977126180
using2352146101095189112164218
the2604146101093183131
Prominence2769146101095081133212235287336389432479
LC–20A3282146101094497140185233289
HPLC36051461010961112156209
system2138160101093783121154202281
(Shimadzu,2455160101092671123146225271326367420441
Kyoto,2931160101095192144176228248
Japan),3215160101093480134181234259278
and3529160101094597152
an3716160101094598
analytical2138174101094598145170216252276320368392
column2578174101094194118172251305
Arion®2931174101095789114167221260
Polar32391741010949101126173206
C1834931741010953100146
column36871741010941932138188101092272149200
(250×4,6mm;2365188101092671119165232291310358446523541
5mm)29331881010947135212236
purchased31951881010952103134175226270307354407
from3628188101092758108185
Chromservis21382021010954105136187265302350381429452489
(Prague,2654202101092675105151204255303322
Czech3003202101095496143186238
Republic)3268202101094997151203256279301344369
and3664202101094597150
tempered2138216101093074150202248278323375
at2537216101094473
352635216101094690
°C.273721630107252763216101095273
The2861216101094495141
detector3027216101095196127171213245294324
was33752161010967111146
a35462161010943
triple361421610109305981132154200
quadrupole21382301010951102146198228279331383405451
mass26152301010976120157193
spectrometer2834230101093688135177209240290366413444489520
LCMS-80453380230101094394167212243292339388434
(Shimadzu,2138244101092670121142219264317357408427
Kyoto,2592244101094990140171221240
Japan)2858244101093478131176228251
with3136244101096790121172
electrospray3334244101094669115158191221272309361392437480
ionization2138258101092273126148188235266289340393
(ESI).2558258101092671118140167185
The2770258101094699146
mobile29432581010977129183206228276
phases32462581010952105150187235272
consisted35452581010942921451812032402138272101093074126
of2284272101094976
water23792721010966110140185215
containing2613272101094089139169213234284306356406
0,1%3039272101094664105172
formic3230272101092676106182203244
acid3493272101094384105156
and3669272101094393145
methanol21382861010975120150199243292341363
containing2517286101094088138167211232281302352402
0,1%2935286101094563104170
formic3120286101092676105180201242
acid3377286101094384104155
(403548286101092568115
:36742861010919
60,3703286101094693111
v/v).2138300101094574120145164
The2329300101094699146
flow2502300101092753105173
rate2702300101093076107154
was28833001010968114151
0,83061300101094767117
ml/minute.3191300101097799132208231283334366413432
Ions3650300101092275127164
were21383141010966110140184
generated2338314101095095145190220264293337388
using2742314101094984105154205
electrospray2962314101094567112153185214263298349378422463
ionization3441314101092170120141179224253274323373
and2138328101094393145
detected2302328101095196127171213245290342
in2663328101092171
the2753328101092979125
positive28973281010951102138159189210254298
ion3214328101092171122
mode33553281010975125177222
at3596328101094473
the3688328101093080126
following213834210109267698119169235257307358
transitions2514342101092959103153188209240261310361396
of2928342101094976
mass30213421010976119155190
to3229342101093079
charge3325342101094191134164214260
(m/z):3603342101092598130168193211
colistin2138356101094192115137174206228280
A24453561010956
585,552528356101094795142161210256
→28113556011191
101,05;2929356101094791136151200247265
colistin3220356101094292115137174206228280
B35273561010950
578,53604356101094793140160210
→21383696011189
101,15;2250370101094689131146187228244
and2517370101094393144
IS2684370101092166
602,42772370101094693138156203
→29973696011189
101,1;3108370101094689131146187201
120,15;3331370101094686132150191232248
86,15.3602370101094693112153194212
The2138384101094596141
total2295384101093079109153174
analysis2486384101094393136158200236257292
time2795384101092950125171
was29823841010966110146
331443841010946
minutes3206384101097596146196226270306
and3528384101094493145
the3689384101093079125
LabSolutions2138398101094389141185236259309340362412464500
software2664398101093687114148217262292338
(ver.3028398101092669114144160
5,93;3214398101094765110156174
Shimadzu,3414398101094595117193238290330380400
Kyoto,2138412101094886134163211229
Japan)2382412101093375125168217240
was26374121010965108143
used2795412101094983128179
for2989412101092675104
instrument3108412101092170104134163212286330380408
control,3532412101093987136165194242263282
data2138426101095194124168
acquisition,2325426101094484135184205241262292313363413432
and2776426101094494145
processing.2940426101095181131172217253288309359410429
Sample21384515014356111209274300359
preparation25264515014366102159225280315370406431495559
Colistin21384819011053104129153192225250302
The2278496101094698146
volume2450496101094595118169246294
of2770496101095179
1402875496101094790139
µl3027496101095073
of3127496101095078
human32324961010951102179224276
plasma3535496101095275120157234279
was21385101010967113149
treated2313510101093161108152183229282
with2621510101096890121173
202820510101094794
µl2926510101095073
of3025510101095178
a31295101010945
solution3200510101093688110161192214265317
containing35445101010941901421732182402138524101095072122174
0,12338524101094665106
mg/ml24565241010975127159233255
of2737524101095076
IS.283952410109226786
Further2951524101094493123156206252283
preparation3260524101095182127179224254299329350400452
of3737524101095077
all213853810109446789
samples2254538101093682160213236283320
was26015381010968113150
performed27785381010952100131161213244322370423
with3228538101096890122173
Oasis34285381010963109145167204
HLB36595381010960104155
121385521010946
ml2194552101097596
cartridges2305552101094084112145174195245295340375
with2696552101096687116165
302877552101094691
mg29785521010975125
of3118552101094975
sorbent3209552101093584114165210260289
(Waters,35135521010926103144173217246282301
Prague,2138566101094877121172221267285
Czech2448566101095293138179229
Republic).2702566101094894146196247270291331356373
The3099566101094596142
SPE3265566101094592136
extraction3425566101094588121150194235267288338389
consisted2138580101094089140175196231262307359
of2523580101094976
conditioning2624580101094089140191212243264314364386436487
of3136580101095076
cartridges3238580101094084114146176198249300346382
with3645580101096788119169
121385941010946
mL21955941010975119
of2332594101095076
methanol,24275941010975121152201245295345367386
equilibration2832594101094597146168189210261291335365386436487
of3337594101095076
cartridges3432594101094085114147176198249300346382
with2138608101096891123175
123406081010948
ml24016081010977100
of2528608101095179
water26346081010969114145192224
containing2885608101094192145176222244297319371424
0,13336608101094867110
%34596081010973
formic3560608101092779110189211254
acid,2138622101094384105156175
1602332622101094688135
µl2479622101094971
of2569622101094976
sample2664622101093580155206228274
loading,2957622101092171115166188238288308
washing32836221010966110146196217267318
away36196221010944109153195
of2138636101094975
interferences2229636101092171100144174202248277322372412456492
with2737636101096587117166
129196361010946
ml2980636101097596
of3092636101094975
water31836361010966109139183212
containing3411636101094089139168212232282303352403
0,12138650101094665106
%22566501010972
formic2354650101092676107183205246
acid,2625650101094485106158177
and2828650101094494146
finally3000650101092651101145167188232
eluting3257650101094668118149170221272
of3555650101094976
COL36576501010953113157
and2138664101094597150
IS2316664101092269
with2412664101096891123175
0,52615664101094867117
ml27456641010977100
of2873664101095179
methanol29796641010978125157210255307359383
containing3389664101094293145176223245297320372425
0,12138678101094665106
%22566781010971
formic2353678101092777107183205246
acid.2624678101094485106158177
An28276781010955106
amount29596781010943119169220270300
of3285678101094976
103387678101094689
µl3487678101095073
of3585678101095077
the3687678101093081127
sample2138692101093681157209231278
obtained24416921010950103133178200250297349
from2816692101092657107184
the3026692101093080127
elution3178692101094668118149171221272
step3476692101093667112165
was36676921010966111147
injected213870610109217193138180212256308
into2465706101092272101150
HPLC.26347061010959107149199220
CMS21387339011053129177
The22787481010946100148
volume2457748101094496119171250298
of2786748101095179
1402896748101094891141
µl3051748101095174
of3156748101095179
human32677481010951103181227280
blood3578748101095376129181236
plasma2138762101095275119155232277
was24417621010967112148
treated2615762101093161107152182228281
with2922762101096790120171
203120762101094693
µl3225762101095173
of3324762101095078
a34287621010944
solution3498762101093687110161192213264316
containing2138776101094087136165208228277297346396
0,12549776101094563103
mg/ml26627761010974123154226247
of2924776101094874
IS.301377610109216483
Acid3111776101095492113163
hydrolysis3289776101094989138167215236278313333367
was36717761010965108143
performed393462101095197126155204234309354406
by435562101095193
adding446462101094393144165214265
15474462101094686
µl484162101094971
of492862101094975
15019621010945
M5080621010972
sulfuric516862101093584105131181211232272
acid545662101094383104154
to393476101093079
a4032761010943
plasma409476101095173117152227271
sample438476101093680155207229274
containing467776101094189140169214235285306356407
CMS.5103761010952125169188
After531076101095581114159189
30551876101094692
minutes,393490101097697148198229274311331
30429190101094693
µl439590101095173
of449490101095077
1M4597901010946115
sodium473890101093686139161211287
hydroxide505190101095092143173223265287339385
was5462901010967112148
added3934104101094598152199253
to4215104101093182
stop4325104101093769120174
hydrolysis.4527104101095295147178230253298336358395416
The4971104101094699147
subsequent5146104101093788142179227281333380433464
sample3934118101093783160214237284
treatment4245118101093162109154185263310363393
was46651181010968114151
the4843118101093082130
same5000118101093683160208
as5235118101094481
for5343118101092779110
the5480118101093183130
colistin3934132101094089111132167198219269
samples.4222132101093680156207229275311330
Validation39341575014361113138162228282318342406471
Calibration4074174101095194114134184213256284304353402
curves4490174101094088117162205239
for4744174101092674103
colistin4862174101093987108128162192212260
and5137174101094391141
CMS52931741010951121164
were547217410109651083934188101092975
constructed4031188101094189140175206236286327359404456
in4509188101092171
the4603188101093080126
range4751188101093074124175221
of4994188101095076
0,15–305093188101094665106146193238284
mg/L,53891881010975126158201221
where39342021010966116161191235
each4185202101094588128178
calibration4379202101094083104125176206249279299349399
point47932021010951101123172201
of5010202101094975
the5101202101093079124
curve5241202101094089119165209
was54652021010966110145
measured39342161010975121165200249279324376
at4334216101094373
least4430216101092267111147177
six463121610109365799
times.4754216101092950126172208227
All500521610109557697
points51262161010951102123173203240
in5389216101092272
the5484216101093080126
calibration3934230101094084105126178207251281302351402
series4352230101093581111132178213
were45812301010966111141186
used4783230101094985130182
to4981230101093079
determine5076230101095096126170200276297347392
the5484230101093080126
precision3934244101095283129171193229250301353
and4313244101094495147
accuracy4486244101094486126177207252294340
of4852244101095077
the4955244101093181128
inter-day5109244101092273103149179205260304346
(intraday)5482244101092647983934258101093059104155198240264
measurements.42182581010976121165201250280325400446497526563583
Samples4821258101094488163215237283319
at5160258101094373
2,5253258101094665
10,5338258101094689107
and5465258101094394145
303934272101094590
mg/L40342721010974124155197
were42462721010965108137181
prepared4442272101095180124174217246290340
for4797272101092674103
intraday491527210109217098127170220262303
(inter-day)523327210109254493122165194219271313353377
precision3934286101095079123163183217237285334
and4282286101094391141
accuracy4438286101094281120168196239278321
measurements.47742861010973118160194242270314388432481509545563
Plasma5352286101094766108142215258
samples3934300101093579153204225270305
with4255300101096586116165
colistin4435300101094088109130165195215265
and4715300101094392142
CMS48733001010951123166
concentrations5055300101094088137177221271300329372401421470520555
of3934314101094975
0,07;4025314101094563109153167
0,1;4208314101094664104117
0,15;4341314101094563104144160
0,5;4516314101094664111127
1;4659314101094659
and4733314101094393143
248923141010946
mg/L49483141010975125156199
were51623141010966110140184
prepared53623141010950801251762193934328101092975127
for4080328101092676106
limit4206328101092142117138168
of4394328101094976
quantification44893281010951101144194224245272296336380410431481531
(LOQ)5040328101092567128191217
and5276328101094494145
limit5441328101092142117138169
of3934342101094976
detection4029342101095196127172213245266316367
(LOD)4415342101092567127188212
determination.4646342101095197127172202278299349393422443493544563
Stability3934367501435690145211237261286323380
of4343367501436395
COL44673675014368145199
and46953675014354118184
CMS49083675014368162217
samples39343815014345100198264290348393
testing4356381501433793138177201265331
Studies4074398101094676128182204252290
for4394398101092779111
short-term,4534398101093789142173208235267314345424445
long-term,5009398101092174127180210242289321400420
and5459398101094597151
freeze-thaw3934412101092759106154196243275307359404472
stability4434412101093769115169192214237268315
were47774121010968115147194
also4998412101094568105157
performed51824121010953102133164216247326374428
on3934426101094999
the4048426101092978123
samples.4187426101093579153203225269305324
The4526426101094494139
validation4681426101094386107127177220249270319368
procedure5065426101095080128169213264312341386
was54664261010965109143
derived3934440101095095124145188232282
from4232440101092655104178
the4425440101093079124
European4564440101094491120169219265308357
Medicines49364401010971117167188228248297342377
Agency53284401010955104148198238282
(EMA)3934454101092567138192216
recommendations.4164454101092972112159233306350399449491519539588637670689
Control4867454101095198147176204252273
plasma5154454101095071113147220263
samples54314541010935781513934468101095172117152
from4102468101092655104179
three4296468101092978107152197
female4508468101092671146188209254
and4778468101094292142
three4935468101093079108152198
male51484681010974117138183
donors5346468101095099149198228264
obtained39344821010949100129173193243288339
from4288482101092656104179
the4482482101093079124
Transfusion4621482101094468111161195222272307327376426
Department50624821010959105155199228260334379429457
of5535482101094975
the3934496101093080126
Olomouc4086496101096285135211261312353
University44644961010959109130174219249286307337383
Hospital48734961010958109145196218249293315
were52134961010967112142188
used5426496101095086132184
for3934510101092676106
calibration4059510101094185106127178209253282303353404
and4482510101094393145
validation.4646510101094387108129181224254275325375395
Results39345350014768131181249281323375
and43365350014761130200
discussion45645350014769100151205274324374405473542
No40745521010959110
pure42095521010952102132177
colistin4412552101094089111132168199220270
A47085521010955
and4789552101094393145
B49595521010949
reference5034552101092975102148178223274315359
standards54195521010936661111613934566101095194124175210
were41625661010966111141186
available.4366566101094486130151172216267289335354
Therefore,4738566101094495141171216243293323368387
the5143566101092979125
pharmaceutical528656610109511021461752512953934580101094084133163184224268289
secondary4238580101093681122170220270313343388
standard4641580101093666110159210253282332
of4989580101094975
colistin5080580101094088109130165195216265
sulfate5361580101093584105132176205249
was39345941010966110146
used4098594101094985131182
for4298594101092777107
the4423594101092979125
method45665941010975121152201251303
determination.4887594101095197127172202278299349393422443493544563
The5468594101094596142
purity39346081010951101130152181226
of4176608101094975
the4267608101093080125
standard4408608101093566110159210254283333
was47576081010966110145
determined4918608101095095126170200275296346391443
by5376608101095193
the5485608101093079125
manufacturer39346221010975118167216243287327358408437482511
(11)4461622101092567104123
to4600622101093078
be4694622101095196
914806622101094689
%49056221010971
by4992622101095092
HPLC-UV51006221010958106147197220279330
analysis,54466221010943921353934636101092164100121156176
which41326361010966117138178228
contained4382636101094189140169214235285331383
30,464787636101094692110157205
%50036361010971
colistin5096636101094189111133168199220270
A53886361010955
and5465636101094393145
53,83934650101094690108157
%41026501010971
colistin4192650101094089111132167198219269
B.4481650101094868
As4074664101095592
shown4182664101093585135200251
in4449664101092171
Figure4536664101094464115164194239
1.a,4791664101094660105125
the4932664101092979125
retention5073664101092974104149199228249299349
time5438664101093051126172
of3934678101095077
colistin4037678101094090112134169201222273
A43366781010955
and4417678101094495147
B45906781010949
and4665678101094394146
IS4837678101092267
are4930678101094373119
2,11;5075678101094766107145159
2,09,5260678101094766113161177
and5463678101094495147
2,13934692101094665106
minutes,4066692101097597147197227272308328
respectively.4419692101093075112163210251284305349394416459474
Calibration4919692101095297118139191221266295317367417
curves5362692101094190120167212248
were39347061010966111141186
constructed4141706101094089139175205235285326358403455
in4616706101092272
the4708706101093080126
0,15–304854706101094665106146193238284
mg/L51497061010976126158202
range.5371706101093074124175220239
Plotting3934720101094868118148181202252303
the4259720101093080126
peak4408720101095197141183
area4614720101094473118162
versus4799720101094388118154204239
concentration,5061720101094089140180225275305335379408429479530549
linear393473410109214292138182211
calibration4168734101094185106127179209253283304353404
curves4595734101094090120166211247
were48657341010966111141186
obtained50747341010950101132176197247293345
with5442734101096688118168
a39347481010943
confidence3999748101094189140166190241287338378423
value4444748101094387109158204
of4670748101094976
R2476874810109484818748706427
=48677481010954
0,99974943748101094664109156203247
for5212748101092777107
colistin5341748101094089111132167198219269
A39347621010955
and4009762101094393144
R2417376210109484223762706427
=42697621010954
0,99894342762101094665109156203249
for4611762101092676106
colistin4736762101094189111132168198219269
B.5025762101094968
In5113762101092171
the5204762101093080125
case5349762101094084120166
of5534762101095076
hydrolysed3934776101095091141170219241283319364416
CMS43667761010951123167
to4549776101093078
COL,46437761010952111154174
the4833776101092979124
confidence4973776101094088138164188239284334375419
value5407776101094387108157203