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FormulaforCalculatingtheNumberofTheoreticalPlates
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Introduction
N,thenumberoftheoreticalplates,isoneindexusedtodeterminetheperformanceandeffectivenessofcolumns,andis
calculatedusingequation(1).
1)wheretr:retentiontime,andW:peakwidth

Thispeakwidth,W,isbasedonthebaselineinterceptsoftangentlinestoaGaussianpeak,whichisequivalenttothe
peakwidthat13.4%ofthepeakheight.
However,tosimplifythecalculationandaccommodatenonGaussianpeaks,thefollowingcalculationmethodsareusedin
actualpractice.

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Fig.1PeakWidth

1.TangentLineMethod
Peakwidthisthedistancebetweenpointswherelinestangenttothepeak'sleftandrightinflectionpointsintersectthe
baseline,andiscalculatedusingequation(1).TheUSP(UnitedStatesPharmacopeia)usesthismethod.Thisresultsin
smallNvalueswhenpeakoverlapislarge.
Thisalsopresentsaproblemifthepeakisdistorted,sothatithasmultipleinflectionpoints.
1)

2.HalfPeakHeightMethod
Widthiscalculatedfromthewidthathalfthepeakheight(W0.5).Sincewidthcanbecalculatedeasilybyhand,itisthe
mostwidelyusedmethod.ThisisthemethodusedbytheDAB(GermanPharmacopeia),BP(BritishPharmacopeia),and
EP(EuropeanPharmacopeia).
2)

TheJapanesePharmacopoeia15threvisionissuedinApril2006changedthecoefficientfrom5.55to5.54.
(LCsolutionallowsselectingthecoefficientviathe[ColumnPerformance]setting,wherethecalculation
methodfor5.54is"JP"andfor5.55is"JP2."
Forbroaderpeaks,thehalfpeakheightmethodresultsinlargerNvaluesthanothercalculationmethods.
2)

3.AreaHeightMethod
Widthiscalculatedfromthepeakareaandheightvalues.Thismethodprovidesrelativelyaccurateandreproducible
widths,evenfordistortedpeaks,butresultsinsomewhatlargerNvalueswhenpeakoverlapissignificant.
3)A:Area,H:Height

4.EMGMethod(ExponentiallyModifiedGaussian)
Thismethodintroducesparametersthataccommodatetheasymmetryofpeaks,andusesthepeakwidthat10%ofthe
peakheight(W0.1).Sinceitusesawidthnearthebaseline,itresultsinNvalueslargerthanothermethodsforbroad
peaks.Furthermore,itcannotcalculatethewidthunlessthepeakiscompletelyseparated.
4)a0.1:Widthoffirsthalfofpeakat10%heightb0.1:Widthofsecondhalfofpeakat
10%height

ComparisonofCalculationMethods
GivenaGaussianpeak,eachofthesecalculationmethodsresultsinthesameNvalue.However,normallypeakstendto
havesometailing,whichresultsindifferentNvaluesfordifferentcalculationmethods.
Therefore,thefourcalculationmethodswerecomparedusingchromatograms.ProfileAshowsatypicalchromatogram
(withsometailing),whereasprofileBshowsachromatogramwithsignificanttailing.Thetheoreticalnumberofplates
calculatedusingthefourmethodsareindicatedinthetablebelow.ResultsforNvariedevenforchromatogramA.Also,
peakswithmoresignificantdistortion,suchasatpeak1inprofileB,canresultinNvaluesthatdifferbymanytimes.
Akeyfactorforperformingreliablequantitativeanalysisiswhetherornotseparationispossible,sothereisacommon
opinionthatacalculationmethodthatjudgesbroaderpeaks,suchaswithtailing,moreseverelyismorepractical.
However,unfortunately,thereseemstobenoconsensusonopinionsregardingNandW.
Consequently,ifacertainmethodisalreadybeingusedforevaluation,thentoachievecorrelation,itisprobably
preferabletokeepusingthesamemethod.

Fig.2Chromatograms
ComparisonofTheoreticalNumberofPlates
A(roughlytypicalpeak)

B(significanttailing)

HalfPeakHeightMethod

15649

20444

20389

22245

5972

7917

9957

TangentLineMethod

14061

18516

20309

21447

5773

7692

5795

9707

AreaHeightMethod

13828

19207

17917

21020

4084

7845

6217

8641

EMGMethod

10171

15058

14766

17836

1356

4671

Ahyphenindicatescalculationwasnotpossible.Inthehalfpeakheightmethod,5.54wasusedasthecoefficient.
Shimadzu'sLCworkstationsoftwareisabletooutputperformancereportsusinganyofthemethodsindicatedabove1.tangent
line,2.halfpeakheight(5.54),2'.halfpeakheight(5.55),3.areaheight,orEMG.Werecommendrecordingthecorresponding
columnperformanceresultsalongwithanalyticalresults!

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