klasifikasi hplc 122321

7/26/2019 Klasifikasi HPLC 122321

1/20

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!


2/20

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)


3/20


4/20

/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


5/20


6/20


7/20


8/20

*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


9/20

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


10/20

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)


11/20

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!


12/20

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)


13/20

$tatio"ary phas!

A solid such as agaros! or a porousglass !ad to 6hich th! a"ityliga"d is immoili%!d


14/20


15/20


16/20

La"


17/20


18/20


19/20


20/20

klasifikasi hplc 122321

Documents