Klasifikasi HPLC 122321
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Transcript of Klasifikasi HPLC 122321
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Klasifkasi HPLC
1. Partisi (liquid-liquid chromatography)
2. Adsorpsi (liquid-solidchromatography)
. P!rtukara" io" (io" chromatography)
#. $i%! !&clusio" chromatography'. A"ity chromatography
. Chiral chromatography
$!paratio"m!cha"ism
*yp! o+statio"ary
phas!
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Partisi (liquid-liquid chromatography)
,!"is HPLC ya"g pali"g a"yak digu"aka" asa diam (statio"ary phas!) !rupa caira"
ya"g !rcampur d!"ga" caira" +asa g!rak(moil! phas!)
$ist!m partisi
Liquid-liquidphas!
Liquid-o"d!dphas!
asa diam(adsorpsi fsika)
asa diam(ikata" kimia)
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/ormal phas!
0!!rs!dphas!
asadiam
(polar)
asag!rak
("o"polar)
asa diam("o" polar)
asa g!rak(polar)
*ri!thyl!"!glycolat!r
H!&a"!
Propyl !th!r
Hydrocaro"
at!r3!tha"olAc!to"itril!
*H
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*ypical applicatio" o+ HighP!r+orma"c! Partitio"
chromatographyField Typical mixture separatedPharmac!utical A"tiiotics4 st!roids4 a"alg!sics
5ioch!micals Ami"o acids4 prot!i"s4 carohydrat!s4 lipids
ood products Artifcial s6!!t!"!rs4 a"tio&ida"ts4 a7ato&i"s4
additi!s
8"dustrial ch!micals Co"d!"s!d aromatics4 sur+acta"ts4prop!lla"ts4 dy!s
Polluta"ts P!sticid!s4 h!ricid!s4 ph!"ols4 PC5s
or!"sic ch!mistry 9rugs4 poiso"s4 lood alcohol4 "arcotics
Cli"ical m!dici"! 9rugs m!taolits4 uri"! !&tracts4 !strog!"s4il! acidsL!ih dari :'; aplikasi p!misaha" HPLC
m!"ggu"aka" reversed phase, bonded phase(octyl or octyldecyl-siloxane) packings dan +asa
g!rak"ya a"tara lai" m!ta"ol, acetonitrile, atauTHF
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Adsorpsi (liquid-solid)
chromatography
asa diam !rupa p!rmukaa" padata" polar(silika da" alumi"a)
$ilika l!ih dipilih u"tuk k!a"yaka" aplikasi
kar!"a kapasitas samp!l ya"g l!ih !sar P!"ggu"aa" k!dua +asa diam i"i m!muat
6aktu r!t!"si l!ih lama
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Applicatio" o+ High p!r+orma"c!adsorptio" chromatography
P!misaha" s!"ya6a "o"polar
Water-insoluble organic compounds(3r ='>>>)
Resolve isomeric mixture
(!.g m!ta a"d para sustitut!d !"%!"!sd!riati!s)
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A"ity chromatography
(Kromatograf Af"itas)
8"ol!s coal!"tly o"di"g ar!ag!"t4 call!d a" a"ity liga"d4 to asolid support
*ypical a"ity liga"ds ar! a"tiodi!s4!"%ym! i"hiitors4 or oth!rmol!cul!s that r!!rsily a"d
s!l!cti!ly i"d to a"alyt! mol!cul!si" th! sampl!
*h! r!tai"!d a"alyt!s ca" ! !lut!d
y cha"gi"g th! moil! phas!
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3oil! phas!
*6o disti"ct rol!s?
irst it must support th! stro"g i"di"g o+th! a"alyt! mol!cul!s to th! liga"d
$!co"d4 o"c! th! u"d!sir!d sp!ci!s ar!r!mo!d4 th! moil! phas! must 6!ak!" or!limi"at! th! a"alyt!-liga"d i"t!ractio" sothat th! a"alyt! ca" ! !lut!d
(o+t!" cha"g!s i" pH or io"ic str!"gth ar!us!d to cha"g! th! !lutio" co"ditio" duri"gth! t6o stag!s o+ th! proc!ss)
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$tatio"ary phas!
A solid such as agaros! or a porousglass !ad to 6hich th! a"ityliga"d is immoili%!d
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La"
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