713
www.klinickafarmakologie.cz340804301076513320121425327329234035839843647950254457164568672477179183988990895196610061045
/27048043010732
Klin278180430107446583131
Farmakol2932804301074279105179220258305325
Farm3277804301074178105179
2023;37(3):89-923476804301074487128172186230270290333354368412456485529571
/40928043010732
KLINICKÁ4170804301074488109168189242288346
FARMAKOLOGIE4538804301074191138210265312372414474533554598
A51578043010753
FARMACIE5231804301074291138211266317337381
9156558026012657112
PŮVODNÍ4821207012660132201274348422456
PRÁCE5298207012661124196256314
Assay199536501063872104143176
of218636501063970
colistin227136501063372103121153186204245
A,253236501063754
B2601365010634
and265036501063879119
colistin278536501063271103120153185203244
methanesulfonate304536501065284117158198239271306346377408449491531559591
in365236501061556
human372436501063980133174215
plasma395536501063263105139193233
by420436501063470
LC-MS/MS42893650106296281135168194246279
and458436501063879119
short-term471836501063172112148180204233265301355
plasma508936501063364106140193234
stability533836501063164100136153184202234273
of343244101094976
R24432441010948492244706427
=5432441010954
0,9995621244101094664109156203248
for893244101092676106
colistin1023244101094189111132168198219270
A13162441010956
and1395244101094494145
R2156424410109491614244706427
=16652402016268
0,99821770244101094664109156203248
for343258101092676106
colistin469258101094089111132167198219269
B.758258101094868
The845258101094596142
results1007258101092974110159181211248
of1274258101095076
the1370258101093080125
inter-day1515258101092171101145175201254298339
and1874258101094393145
intraday34327210109217098127170220262303
measurements6612721010973118161195243272316390434484512548
for1224272101092674103
method13422721010974119148197246296
validation1653272101094386106127177219248268317366
are343286101094475121
accurate491286101094486127178209254285331
and848286101094596149
precise,1024286101095283129172194230278297
with1347286101096890122173
an1546286101094596
error1669286101094677108159190
not18852861010951103134
exceeding343300101094587126170216267288337388
15746300101094686
%8433001010970
with929300101096687117166
the1111300101092979124
LOQ12503001010943102165
set1430300101093681111
at1556300101094373
0,151644300101094664104145
mg/L.17993001010975125157199220
The343314101094595141
validation500314101094387108129180223253274323373
confirmed889314101094089139165189218294339391
the1296314101093079125
reliability143731410109297496117160211233254275305349
of1802314101095076
the1894314101093079125
LC-MS343328101094293116190234
method5943281010975121151201251303
for913328101092777107
measuring10363281010975121165200249279301351402
concentrations1454328101094089139180225275305334379408429479530565
of343342101094976
colistin441342101094189111132168199220270
in733342101092171
human827342101095099174218268
plasma.1118342101095173116152227271290
Summary1431342101094493169244288317363
information1816342101092171981481783433561010976121151173224275
on6453561010950101
the773356101093081128
analysis927356101094495139161205242264300
parameters1253356101095297128173249296327373404441
is1720356101092258
given1804356101095173118163215
in343370101092171
Table433370101094583134156202
1.654370101094661
The483384101094595141
CMS6403841010952124168
concentration824384101094189139180224274304333377407427477527
was13673841010966110145
measured15283841010975121164200249278323375
indirectly191938410109217134339810109507199143184215236278
by636398101095092
acid743398101094282102153
hydrolysis,911398101094989139167216237279314334369388
for1314398101092674104
which14333981010965114135174223
we16713981010966110
found1796398101092674123172223
it343412101092151
most4124121010975125161192
useful622412101094985130157208230
to869412101093079
use966412101095085131
151115412101094686
µl1213412101094971
of1302412101094976
113964121010946
M14604121010972
sulfuric1550412101093685106133184213235276
acid.1843412101094485106157176
After343426101095581114159189
30549426101094692
minutes,658426101097596146196226271307326
the1001426101093080126
hydrolysis1144426101095091142171221243286322343378
was15394261010966110146
stopped1702426101093566115167219265317
by343440101095193
adding461440101094495146168218269
30µl754440101094793154175
of954440101095076
1M10554401010946114
sodium1194440101093686138159209284
hydroxide.1503440101095091142171221262283335380399
As1927440101095592
this343454101093080102137
is506454101092157
an588454101094494
indirect708454101092172123145175220262294
method,10284541010976122153203253306325
it1378454101092252
is1456454101092156
necessary1538454101095096138183219255300330376
to1940454101092979
back-calculate343468101095195136178204247291313354403425468498543
the908468101093080126
CMS10574681010952125169
concentrations1249468101094089139180225275305334379408429479530565
from1837468101092657106182
the343482101093080126
difference:48848210109507298128174203249299340384403
CMS3735023010950121162
=5515023010954
COLtotal6215023010950108148768508906416446188101
–8865023010945
COLbefore94750230109501081481095508906428526896114138
hydrolysis1243508906428517996124137161182194214
where3435241010966117163193238
COLtotal6015241010952111155755530706417476591104
is879524101092156
the955524101093080126
concentration1101524101094089139180225275304334378408429479529
after1650524101094470103148178
hydrolysis1848524101094991141343538101092979101144180201237
of605538101094976
CMS7075381010952124169
to901538101093079
colistin1006538101094089111132167198219269
and1301538101094394145
COLbefore1471538101095311215616275447064305874104122149
hydrolysis17915447064295484102132145171193206227
is343552101092156
the415552101093080125
circulating55655210109406191131180202245275295345396
concentration968552101094089139180224274304333377407428477528
of1512552101094975
colistin1603552101094189111132167198219269
formed18885521010926751053435661010976123176
by545566101095295
endogenous666566101094698150201253300352403454490
transformation1182566101093061106157193221272303380425455477527579
of1787566101095078
the1891566101093081128
prodrug343580101095181131183212262313
to675580101093079
its77358010109215188
active880580101094484117138181226
form.1125580101092676106182202
Long-term483594101094496149203234266313345425
stability943594101093669116170194217240272320
studies1298594101093770122176199247286
of1619594101095179
colistin1733594101094293117140177210233286
showed343608101093789141209256310
no6806081010951103
degradation811608101095299153184230283329360382434487
in1325608101092174
stock1426608101093669120163207
solutions1660608101093789112163195217269322359
and343622101094393145
patient510622101095195125146192242272
plasma804622101095173117152227271
samples1097622101093680155207229274310
stored1429622101093667116146191243
at1694622101094473
−701789622101095496140
°C1940622301072519666221010952
for343636101092676106
at467636101094373
least558636101092167110146177
90752636101094694
days.864636101095094135171191
Also,1073636101095576111162179
no12706361010950100
degradation1388636101095196148177221273316346367417467
was18736361010966110146
observed3436501010950104141188219266312366
in735650101092273
three835650101093082112159206
freeze-thaw1068650101092758105152193240271302354398466
cycles1560650101094288129151198235
(data1822650101092676121151197
not343664101094998128
shown).486664101093584133198248270287
The788664101094495139
short-term943664101093584133162194219249293322396
stability1355664101093565108159180201221251295
of1665664101094975
colistin1755664101094088109130164194215264
was3436781010966110145
measured5056781010975120164199249278323375
for896678101092776106
three1018678101093080110155201
concentration1235678101094089139180224275304334378407428478529
points17806781010951101123173202239
at343692101094373
room435692101092979129205
temperature659692101093074150201248278322351401431476
(RT)1154692101092674120148
and1321692101094393145
371485692101094689
°C1585692301072516116921010952
(Table1682692101092673111163185230
2).193169210109467087
After343706101095581114159189
24557706101094791
hours6737061010950100150180216
at915706101094373
RT,1013706101094995109
the1147706101093080126
degradation1298706101095197148178222273317347368418468
of1792706101094976
the1893706101093080126
samples343720101093680156207230275311
reached680720101092975119159209255307
almost1013720101094365140191226257
101296720101094689
%.1396720101097292
However,15147201010958109175220264308338353
the1893720101093080126
degradation343734101095196148178222273317346367417468
of835734101095076
colistin936734101094089111132167198219269
was12307341010966110146
more14007341010975125155201
significant1625734101093657108158179206229270314364394
at343748101094474
37443748101094690
°C.5447483010725571748101095374
Already671748101095577107152197248292
after989748101094471104149180
301194748101094793
minutes,1313748101097698148198229274311330
degradation1669748101095197150180224276320343762101093051102153
reaching521762101093076120161211232283334
up8817621010950102
to1008762101093080
251114762101094693
%12187621010972
was13167621010967111147
observed,14887621010950102139185215262307359378
the1892762101093081127
average343776101094385129158202252297
degradation655776101095196147176219270313342363412462
for1132776101092676105
all125377610109436484
samples1353776101093579153204226271306
was16757761010966109144
111834776101094683
%.1928776101097191
CMS21387481010952125169
stability2324748101093566111162184205226256301
in2642748101092171
plasma2730748101095173117152227271
samples3018748101093580155207229274310
at3345748101094373
a34357481010943
selected3495748101093581103149190222267319
concentration2138762101094088138178222271300329373402422471521
of2675762101094975
10mg/L2765762101094688163213245287
was30687621010965109143
studied3227762101093565114165186231282
at3524762101094372
room3612762101092978127202
temperature2138776101093074150201247277321351400430475
and2629776101094393145
372790776101094689
°C.289177630107252917776101095273
As3006776101095592
expected,3114776101094588140186227259304356375
the3505776101093079125
conversion36467761010940891393934748101094386115151171220269
of4218748101094975
CMS43087481010951122166
to4489748101092977
colistin4581748101094087108129163193213262
is4858748101092155
more49287481010974123152197
significant5140748101093555105154174200223263306355383
at5538748101094372
373934762101094689
°C4034762301072540597621010952
than4126762101093079122172
at4313762101094372
RT.4401762101094893107
At4524762101095482
elevated4622762101094566111154197226271322
temperatures,4959762101093074149199245275318347396426470506525
we54997621010966110
observe39347761010949100135180209255299
the4248776101092978123
conversion4387776101094088137179222252288308357407
of4809776101094975
CMS48997761010952123167
into508177610109217099147
its524477610109205086
colistin5345776101094088110130165195215265
Tab.2140244201124993148172
1.2332244201125071
Summary2450244701073985157229275302342
information281024470107196589134161233279306325371418
on3246244701074593
the3357244701072673112
analysis3487244701074593138158196227245276
parameters3781244701074592118162234273300338365397
Chromatographic217125840984793122168237278307352399429470517564585622
conditions281025840983681127175196225246292338372
HPLC:217127070955091126170184
Shimadzu,35382707095378097162199243276320334
Prominence388927070954164106172189232272315351389
LC–20A429627070953679111147185231
Column:217128270954687105148214257270
Arion®35382827095477188131175205
Polar37612827095408199136159
C18393828270954682117
(25040722827095226099138
×4220282709548
4,642782827095395293
mm;4381282709565131144
54543282709539
mm)4592282709565131150
Mobile2171294709562106151168185224
phase241329470954488124154194
A:262429470954762
Mobile2171304709562106151168185224
phase241330470954488124154194
B:262430470954154
0,1%35382947095395388142
formic36982947095216388154171207
acid39232947095367289133
in407429470951659
water415129470955895119157181
0,1%35383047095395388142
formic36983047095216388154171207
acid39233047095367289133
in407430470951659
methanol4151304709565105130173210253296314
(40:60,51112997095215899114154194208
v/v)53362997095366399120
Flow21713167095375397155
rate:23433167095246185124138
0,835383167095395394
ml/minute364231670956583112176193235278303342
Column217132870954687105148214257
temperature:244532870952563129173214237275299342366405418
35353832870953976
°C362432870952470
The217134070953782121
volume23103407095367996139205244
of257234070954264
injection:26543407095165976115152180197240283297
10353834070953975
µl362334070954260
Analysis217135270954790127144180210227256
time:244535270952542107147161
33538352709539
minutes359535270956582125167193231261
MS/MS2171364409866108135201244
detection243136440984789118160197227248294341
Mass217137670956299129159
spectrometer:234837670952973114151178202244311350375414437453
Shimadzu,35383767095378097162199243276320334
LCMS-8045388937670953680143180205246286327364
Ionization217138870951760104121154191216233276320
mode:2508388709566109154194207
ESI35383887095377692
positive364738870954589119135161178215253
Ion217140070951760104
transition22924007095254987130159176201218261305
monitored:2614400709566109153170195237261300345358
Colistin353840070954687105122151178195238
A3793400709547
585,55385740070953978117129169206
→4080400109879
101,05417740070953975110120160199
Colistin353841270954687105122151178195238
B3793412709540
578,5385141270953977115129169
→4037412109879
101,15413441270953975110120155189
IS353842470951753
602,4360942470953979116130170
→3796424109879
101,1;389242470953975110120155164
120,15;407442470953972111125159193205
86,1542974247095397993128162
Validation217143640984887109130177218247268314360
parameters254843640984788118159227270299340370405
Calibration217144870954683100117161185223247264307351
curve253944870953679103142181
range273744870952461104149188
(mg/l)294344870952185129158177198
0,15–3035384487095395388121163199238
Limit217146070953653119136161
of234946070954365
quantification243146070954386123166190207229247283320344362405448
–2897460709540
LOQ295446070953688143
(mg/l)311446070952285130159177199
0,235384607095395393
Limit217147270953653119136161
of234947270954365
detection243147270954483108146183211228271314
–2763472709540
LOD282047270953688141
(µg/l)297847270952164109138156178
4,735384727095395291
Recovery217148470954080117159195234258296
(%)24854847095217998
81353848470953975
Tab.2140504501124992146170
2.2328504501125075
Stability2448505001073964107152171189207233272
of2736505001074568
colistin282150500107378099117147173191237
A30745050010748
and3139505001074591136
B32925050010743
(%)3351505001072486107
at3474505001074672
three356250500107267297134173
concentration3751505001073780127164201247272298340366384429476
levels424350500107195795132151180
(2;443950500107246478
10;453450500107417894
204644505001074282
mg/L)47375050010772117148185209
in4962505001071864
human5042505001074792163208255
plasma5313505001074564109139209254
Colistin342851870984792114135169198219265
A3710518709851
Colistin477551870984792114135169198219265
B5057518709846
Time261053070984264132175
23028530709843
mg/L3081530709868115142183
10345653070984383
mg/L3549530709868115143183
20390353070984386
mg/L3999530709868115143183
24373530709843
mg/L4426530709868114142183
10480153070984383
mg/L4894530709868115143183
20524853070984386
mg/L5344530709868115142183
RT217154270984689
0,526255430095395392
h2727543009543
99,2830615430095397990131169
101,28349454300953975110120161199
100,18394354300953975116129164199
107,01439454300953975111117157193
105,14484154300953975113126161196
103,41528854300953975112125165199
12651555009539
h2700555009543
98,9030605550095397993131171
97,8635125550095397682123163
99,0539585550095397990130169
92,9344085550095397790128165
97,2848585550095397682123161
104,80528255500953975116129170210
22651567009539
h2700567009543
96,9230625670095397993131168
97,6635135670095397682123163
96,0839565670095397993133173
92,1544115670095397790125159
95,3148565670095397890130165
99,8953025670095397990132170
52651579009539
h2700579009543
93,2830625790095397689130168
92,1635145790095397790125161
95,6439565790095397890131172
92,2244055790095397790131170
94,1548585790095398093128162
96,5753025790095397993133171
24263359100953975
h2718591009543
90,5730605910095398093133171
92,7135115910095397790129166
91,3739625910095397585125161
90,1544105910095398093128162
93,9348575910095397689127164
96,4653005910095397993133174
37217160270984384
°C226560270982876
30258760300953978
min268360300956582125
91,6530646030095397585125164
92,7635116030095397790129165
91,2139626030095397585126161
74,9644086030095397387124164
92,7648566030095397790129165
92,1753086030095397790125158
RT2171614901074483
–22726149010745
room2335614901072671118190
temperature2543614901072764136183222249293319366392430
Tab.2140635501124992146170
3.2328635501125073
Stability2447636001073964107152171189207233272
of2735636001074568
CMS28206360010748116154
(%)2990636001072486107
at3113636001074672
103201636001074179
mg/L32916360010771117147184
concentration3492636001073780127164200247272297340366384429476
level398463600107185795132150
in4151636001071864
human4231636001074792163208255
plasma4502636001074564109139209254
37344564970984384
°C353964970982876
RT482764970984689
°C493364970982775
Time217166170984264132175
COL308366200954697134
A3234662009547
COL378166200954697134
B3932662009541
COL447166200954697134
A4622662009547
COL516966200954697134
B5320662009541
0,521716740095395392
h2273674009543
103,7308867370984383126145188
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h2220686009543
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22171698009539
h2220698009543
114,0309169770984379119138182
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h2220710009543
117,6309670970984379117130174
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111,7448370970984379116132175
113,8517370970984379118138182
RT2171721801074482
–22707218010745
room2331721801072670116187
temperature2534721801072763135180219244287313359384421
Fig.34360601124771125149
1.51060601125070
Representative62561101074482128154190221259305330374400418456493
chromatograms113461101073783109152224269295339385410453524554
of170461101074568
calibration178861101073882100119164190232258276321368
point217261101074591110156181
10236961101074279
mg/L2459611010772117147185
(1.a)26606110107246173118140
and281661101074592137
patient296961101074591117135173219245
sample323061101073176147192211249
(1.b)34956110107246173118140
1.a34378201125070120
1.b298278201125070126
Time136922530974060127169
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intensity3391515471297186390130175207226253293
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colistin26051582095367795112141168185228
B2850158209540
colistin26051707095367795112141168185228
A2850170709547
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colistin52641707095367795112141168185228
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Time402022530974060127169
(min)420622530972287107151173
Absolute297118364712974995127172192236264304
intensity29711515471297186390130175207226253293