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HistoryofCountercurrentChromatography

TheHistoryofCountercurrentSeparation
TechnologyFromCCDtoEECCCandNotes
onNomenclature
Overview

CountercurrentDistribution(CCD)Development
DCCC(DropletCCC)
CPC(CentrifugalPartitionChromatography)
HSCCC(HighSpeedCCC)
EECCC(ElutionExtrusionCCC),apopularoperationmodeforCPCandHSCCC
AdvantagesofCCC

Introduction
Letsstartatthebeginning
Inordertounderstandpartitionchromatography,onemustfirstunderstandpartitioning(liquid
liquidextraction).Partitioninghasthreestages:
1.MixingThoroughlymixthetwophases
2.SettlingAllowthemixturetosettleintotwophases
3.SeparatingSeparatethephasesfromeachother
Thephysiochemicalpropertythatallowscompoundstobeseparatedthroughpartitioningiscalled
thePartitioncoefficient(K).Thisisdefinedastheconcentration(C)ofacompoundintheupper
phaseofaspecificspecifictwophasesolventsystemdividedbytheconcentrationofthe
compoundinthelowerphaseofthesametwophasesolventsystem(C[upper]/C[lower]).
Advancementsinstrumentationaredrivenbyimprovingpartitioningefficiency!

Nomenclature
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Forhistoricreasonsandduetothelackofharmonization,the"CCC"nomenclatureisrather
fragmentedintheliterature.Duetothehistorichyphenatedspellingof"countercurrent",the
nonhyphenatedspellingof"countercurrent"usedtodaycontinuestobestillabbreviatedas"CC"
ratherthan"C".Accordingly,CCCtypicallystandsfor"counter[]currentchromatography".
Onemajorstreamof"CCC"instrumentdevelopmentledtoawiderangeofcentrifugalpartition
chromatographs("CPC"hydrostaticinstruments)andhasemphasizsepartitioningasits
underlyingmechanism.Atthesametime,thenameofanotherlineofinstruments,generally
termedhighspeedcountercurrentchromatographs("HSCCC"hydrodynamicinstruments)or
variantsthereof,hascontinuedtousetheoriginal"CCC"term.
Notably,bothCPCandHSCCCinstrumentssharethesamebasicprinciplesofoperation
andchromatographictheory,aswellasuserbase.Reflectingthesharedprinciples,wehave
beenusingtheunifyingterms"countercurrentseparation"("CS",syn."CCS")todescribe
bothprincipalmethods.
Seeourreviewarticleformoredetailedinformation:
PauliGF,ProS,FriesenJB
Countercurrentseparationofnaturalproducts
JournalofNaturalProducts71:14891508(2008)
doi:dx.doi.org/10.1021/np800144q
Anupdatedreviewcoveringtheliterature20082014iscurrentlybeingprepared

CountercurrentDistribution
Inthe1940sLymanC.Craiginventedthefirst
apparatus(besidesaseparationfunnel!)toconduct
countercurrentpartitioninghecalledthisCounter
currentDistribution(CCD).Shownhereisan
apparatusbuiltbyHecker(Tuebingen,Germany)that
allowedmanualCCDofsamples.

DropletCCC(DCCC)

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Theconcept:Onephasedripsthroughthesecond
phase,inascending(upperphase=mobile)or
descendingmode(lowerphase=mobile).
Lowflowrate(reliesongravity)makesDCCCa
timeconsumingseparationmethod
Drawback:relativelypoormixingleadsto
reducedseparationefficiency

CentrifugalPartition
Chromatography(CPC)

Basically,CPCishydrostaticlikeDCCC,butwith
improvedmixingandmoretheoreticalplates
Spinningrotor
Centrifugalfieldvs.gravitationalfieldtohold
onephasestationary
Stillhydrostaticseparationconstantgfield
Oneaxisofgyration

MuchfasterseparationsthaninDCCC(flowrates
ca.100foldhigher,FCPC)
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Literature
Murayama,W.etal.,J.Chromatogr.,239,643649
(1982)
Links
http://www.richardscientific.com/fcpc.htm

HighSpeedCCC(HSCCC)

YoichiroIto(NIH,Bethesda,MD)developed

planetary,centrifugalCCC
Increasesgravityimprovesmixingcreated

hydrodynamicequilibrium
Theoreticallythousands(upto70,000perhour)

partitioning/chromatographicsteps
Mixing,Settling,Separating

Tohelpunderstandhowthisworks
HSCCCMovie...comingsoon
noteplanetarymotionofcoils,pinkismixing
zone

ElutionExtrusionCCC(EECCC)
CodevelopedbyAlainBerthod(UnivLyon,
Villeurbanne,France)etal.
TechniqueutilizingmodernHSCCC
Concept:Extrude"column"aftercertainKvalue
Allowshighresolutionofsignificantlylarger
rangeofKvaluesinagiventime

AdvantagesofCCC
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Quick(highthroughputinpreparative
separation)
Inexpensive(onlysolventcosts,whichstillare
significantlylowerthaninHPLC)
Gentleandversatile,forseparationofvaried
compounds,withlesschanceofdecomposition
Abletorangefrommilligramstotensofgrams
onthesameinstrument
Abletoswitchbetweennormalandreversed
phaseatwill
Freeofirreversibleadsorptiontoasolid
support(100%recoveryofsample)

Literature
Armbruster,J.etal.JournalofLiquid
ChromatographyandRelatedTechnology24,1827
1840(2001).
Alvi,K.A.JournalofLiquidChromatography&
RelatedTechnologies24,17651773(2001).

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