INSTRUMENTAL ANALYSIS AND SEPARATIONS INTRODUCTION TO CHROMATOGRAPHY - LECTURE 10

The Russian botanist Mi hai! Ts"ett #oine$ the te%& #h%o&ato'%a(h) in 1*0+ to $es#%ibe his e,(e%i&ents in se(a%atin' $i--e%ent #o!o%e$ #onstituents o- !ea.es b) (assin' an e,t%a#t o- the !ea.es th%ou'h a #o!u&n o- #a!#iu& #a%bonate/ a!u&ina an$ su#%ose0 In his "o% / the se(a%ate$ #o&(oun$s a((ea%e$ as #o!o%e$ ban$s in the #o!u&n an$ the%e-o%e he #hose the na&e -o% the &etho$ -%o& the G%ee "o%$s chroma &eanin' 1#o!o%1 an$ graphein &eanin' 1to "%ite02 His "o% "ent .i%tua!!) unnoti#e$ -o% $e#a$es/ but to$a) #h%o&ato'%a(h) is (%obab!) the &ost i&(o%tant te#hni3ue e&(!o)e$ in ana!)ti#a! #he&ist%)0 The -i%st ana!)ti#a! use o- #h%o&ato'%a(h) "as $es#%ibe$ b) 4a&es an$ Ma%tin in 1*56/ -o% the use o- 'as #h%o&ato'%a(h) -o% the ana!)sis o- -att) a#i$ &i,tu%es (170 Ch%o&ato'%a(h) #an be $es#%ibe$ as the (%o#ess "he%e ana!)tes a%e se(a%ate$ $ue to thei% .a%)in' $ist%ibution bet"een t"o (hases/ a stationa%) one 8stationa%) (hase7 an$ one that &o.es 8&obi!e (hase70 Co&(oun$s t%a.e!in' in the &obi!e (hase inte%a#t "ith the stationa%) (hase0 Those that a%e st%on'!) %etaine$ b) the stationa%) (hase &o.e s!o"!)/ "hi!e those that inte%a#t on!) "ea !) &o.e %a(i$!)0 Co&(oun$s that &o.e %a(i$!) a%e the%eb) se(a%ate$ -%o& the #o&(oun$s that &o.e s!o"!)0 Most #h%o&ato'%a(h) uses a stationa%) (hase insi$e a #o!u&n but the use o- -!at sheets o&obi!e (hase 8thin !a)e% #h%o&ato'%a(h)7 a%e a!so use$0 Ch%o&ato'%a(h) is a (o"e%-u! se(a%ation too! that is use$ in a!! b%an#hes os#ien#e/ an$ is o-ten the on!) &eans o- se(a%atin' #o&(onents -%o& #o&(!e, &i,tu%es0 Man) t)(es o- #h%o&ato'%a(h) ha.e been $e.e!o(e$0 Hi'h (e%-o%&an#e !i3ui$ #h%o&ato'%a(h) 8HPLC7 is an e,t%e&e!) .e%sati!e te#hni3ue "he%e ana!)tes a%e se(a%ate$ b) (assa'e th%ou'h a #o!u&n (a# e$ "ith &i#%o&ete%-si9e$ (a%ti#!es0 Gas #h%o&ato'%a(h) se(a%ates #o&(oun$s base$ on thei% .o!ati!it) an$ thei% a$so%(tion o% so!ubi!it) in the stationa%) (hase0 Si9e e,#!usion #h%o&ato'%a(h) se(a%ates &o!e#u!es base$ on thei% si9e b) (assin' the sa&(!e th%ou'h a (o%ous st%u#tu%e0 Ions #an be se(a%ate$ base$ on thei% #he&i#a! #ha%'e usin' ion e,#han'e #h%o&ato'%a(h)0 Othe% t)(es o- #h%o&ato'%a(h) in#!u$e e!e#t%o(ho%esis an$ su(e%#%iti#a! -!ui$ #h%o&ato'%a(h)0 An inst%u&ent use$ to (e%-o%& #h%o&ato'%a(h) is #a!!e$ a #h%o&ato'%a(h an$ #onsists o- a in:e#to% 8a (!a#e to a$$ a sa&(!e7/ a #o!u&n 8to se(a%ate the #o&(oun$s7/ an$ a $ete#to% 8to &easu%e the #o&(oun$s70 In a$$ition to these &a:o% #o&(onents/ the%e a%e a!so s)ste&s -o% han$!in' the &obi!e (hase/ s)ste&s -o% #ont%o!!in' te&(e%atu%es/ an$ $e.i#es -o% %e#o%$in' an$ (%o#essin' the $ete#to% out(ut00 Co!u&n #h%o&ato'%a(h)/ o% e!ution #h%o&ato'%a(h) in.o!.es "ashin' sa&(!e #o&(onents th%ou'h the #o!u&n b) #ontinuous a$$ition o- -%esh &obi!e (hase 8e!uent70 Co&(oun$s in the sa&(!e (a%tition bet"een the &obi!e an$ stationa%) (hase/ s(en$in' so&e ti&e in ea#h o- the (hases0 The &o%e ti&e an ana!)te s(en$s in the &obi!e (hase/ the -aste% it "i!! &o.e th%ou'h the #o!u&n0

Lecture 8

I- a $ete#to% is (!a#e$ at the en$ o- the #o!u&n/ the si'na! -%o& the $ete#to% is (!otte$ o.e% ti&e/ the %esu!t "i!! be a se%ies o- (ea s #a!!e$ a #h%o&ato'%a&0 The (osition o- the (ea s "ith ti&e is an i&(o%tant #!ue to the i$enti-i#ation o- #o&(oun$s/ "hi!e the a%ea un$e% the (ea #an be use$ as a 3uantitati.e &easu%e o- the &ass o% #on#ent%ation o- the #o&(oun$0 I- a uni.e%sa! t)(e $ete#to% is use$/ a #h%o&ato'%a& &a) #ontain hun$%e$s o- (ea s/ "hi!e i- a s(e#i-i# $ete#to% is use$/ on!) a Chromatogram -e" (ea Chrom Perfect Report s &a) a((ea%0
Chrom Perfect Chromatogram Report

;?@Othe%@>u%tMa)A0B-%esh-%uitR6Bs000600RAC 1D00

>u%tMa)A0B-%esh-%uitR6Bs

Dete#to% Res(onse

1600

1000

F00

+00

D00

600

60 10 Ti&e Ti&e 0 ;i'u%e 10 Ch%o&ato'%a&s -%o& Gas Ch%o&ato'%a(h) o- St%a"be%%) E,t%a#t -10 to <o!ati!e Co&(oun$s0 I!!ust%atin' Di--e%ent Dete#to% Res(onses -60
0 1 6 A D 5 + E F * 10

Printed on 6/7/02 3:20:01 PM

DISTRIBUTION

Page 1 of 1

-A0 As &entione$ (%e.ious!)/ #o&(oun$s "i!! s(en$ so&e ti&e in the stationa%) -D0ti&e s(ent b) an in$i.i$ua! &o!e#u!e in (hase/ an$ so&e ti&e in the &obi!e (hase0 The ea#h o- the t"o (hases is #a!!e$ the #a(a#it) -a#to% 20 The %atio 0 o% 6 retention D + F factor 10 161 1D 1+ 1F 60 o-66 6D
ti&e s(ent in the t"o (hases is e3ua! to the %atio o- the &ass o- the #o&(oun$ in the t"o (hases0

ti&e s(ent in the stationa%) (hase ti&e s(ent in the &obi!e (hase

Dete#to% Res(onse

Su!-u% Dete#to%

+0 50 D0 A0

;ID Dete#to%

= &ass in the stationa%) (hase &ass in the &obi!e (hase

The a&ount o- ti&e a &o!e#u!e s(en$s in one o- the (hases is $i%e#t!) %e!ate$ to the #on#ent%ation o- the #o&(oun$ in that (hase0 The %atio o- the #on#ent%ation o- a #o&(oun$ in the t"o (hases is #o&&on!) #a!!e$ the (a%tition #oe--i#ient 8>7 "hi#h is a &easu%e o- a #o&(oun$s a--init) -o% the stationa%) (hase0 This .a!ue is so&eti&es #a!!e$ the (a%tition %atio/ $ist%ibution #onstant/ o% e3ui!ib%u& #onstant0

>=

&o!a% #on#ent%ation in stationa%) (hase &o!a% #on#ent%ation in &obi!e (hase

Lecture 8

The -a#to%s %atio0

an$ > a%e %e!ate$ to ea#h othe% base$ on anothe% (a%a&ete% #a!!e$ the (hase

.o!u&e o- &obi!e (hase .o!u&e o- stationa%) (hase an$ > is 'i.en as?

an$ the %e!ationshi(

= >G
This !ast e3uation si&(!) sho"s that the &ass %atio is a -un#tion o- the #on#ent%ation in the t"o (hases an$ the %e!ati.e .o!u&e o- the t"o (hases0 The .a!ue o"hi#h is base$ on &ass is a bette% -a#to% than >8"hi#h is base$ on #on#ent%ation7/ be#ause it is the mass o- the #o&(oun$ that is &o.in' th%ou'h the #o!u&n0 Mo.e&ent Th%ou'h the Co!u&n The %etention -a#to% 8 7 is the %atio o- the &ass o- a #o&(oun$ in the stationa%) an$ &obi!e (hases0 This is %ea!!) the sa&e as the %atio o- the ti&e the #o&(oun$ s(en$s in the t"o (hases0 Put anothe% "a)/ the &o%e &ass o- a #o&(oun$ in the &obi!e (hase/ the -aste% it "i!! &o.e th%ou'h the #o!u&n0 The ti&e it ta es -o% a #o&(oun$ to t%a.e! th%ou'h the #o!u&n 8-%o& "hen an ana!)te is in:e#te$ to "hen it %ea#hes the $ete#to%7 is no"n as the retention time(t%70 I- a #o&(oun$ is not %etaine$ at a!!/ it "i!! sti!! ta e ti&e to t%a.e! th%ou'h the #o!u&n0 The%e-o%e/ in o%$e% to &a e a %e!ationshi( bet"een an$ %etention ti&e/ an a$:ust&ent &ust be &a$e -o% this t%a.e! ti&e0 The ti&e it ta es -o% an un%etaine$ #o&(oun$ to t%a.e! th%ou'h the #o!u&n is o-ten no"n as the dead time8to70 Usin' the $ea$ ti&e/ the %etention -a#to% -o% a #o&(oun$ #an be %e!ate$ to the %etention ti&e0

= 8t% H to7G to
Che%e t% = the %etention ti&e o- the #o&(oun$/ an$ to = the $ea$ ti&e Se!e#ti.it) The %etention ti&e o- a #o&(oun$ is base$ on its .a!ue0 Chen &o%e than one #o&(oun$ is in:e#te$ into the #o!u&n/ the) "i!! t%a.e! th%ou'h the #o!u&n at $i--e%ent %ates base$ on thei% .a!ues0 The %atio bet"een the .a!ues o- t"o #o&(oun$s is no"n as the se!e#ti.it) -a#to% an$ is an in$i#ation o- ho" "e!! the #o&(oun$s "i!! se(a%ate0

Lecture 8

Se!e#ti.it) ;a#to% ?

Che%e kB is the %etention -a#to% -o% the &o%e st%on'!) %etaine$ #o&(oun$ an$ kA is the %etention -a#to% -o% the !ess st%on'!) he!$ #o&(oun$0 (in other words, is always greater than or equal to 1). E--i#ien#) The se!e#ti.it) -a#to% is i&(o%tant to $ete%&inin' ho" #o&(oun$s se(a%ate/ but it is not enou'h b) itse!-0 ;i'u%e 6 sho"s (o%tions o- t"o $i--e%ent #h%o&ato'%a&s "he%e the se!e#ti.it) .a!ue 87 is the sa&e 8the (ea s a%e the sa&e $istan#e a(a%t7/ but in ;i' 6a/ the (ea s a%e se(a%ate$ an$ in ;i'0 6b the (ea s a%e not se(a%ate$0 a7

b7

;i'u%e 60 I!!ust%ation o- the se(a%ation o- 6 (ai%s o- (ea s "ith the sa&e .a!ue0 a7 (ea s a%e -u!!) se(a%ate$ be#ause the) a%e na%%o"0 b7 (ea s a%e not se(a%ate$ be#ause the) a%e "i$e0 The i&(o%tant $i--e%en#e bet"een ;i'0 6a an$ ;i'0 6b is the "i$th o- the (ea s0 I(ea s a%e too "i$e/ the) "onJt se(a%ate "e!!0 The%e-o%e/ "e nee$ a &easu%e o- (ea "i$th a!so no"n as efficiency0 The te%& that is 'ene%a!!) use$ to $es#%ibe #o!u&n e--i#ien#) is 1nu&be% o- theo%eti#a! (!ates2 o% N0 The .a!ue o- 8N7 is %e!ate$ to the #o!u&n !en'th 8L7 an$ the si9e o% hei'ht 8H7 o- an in$i.i$ua! (!ate b) the -o!!o"in' e3uation?

N = LGH
Che%e L =#o!u&n !en'th/ H = (!ate hei'ht 8both in the sa&e units7

Ch%o&ato'%a(h) #o!u&ns "ith hi'h nu&be%s o- theo%eti#a! (!ates (%o$u#e .e%) na%%o" (ea s %esu!tin' in bette% se(a%ation0 The abo.e e3uation sho"s that e--i#ien#) 8N7 #an be in#%ease$ b) usin' !on'e% #o!u&ns/ o% usin' #o!u&ns "ith s&a!! (!ate hei'ht

Lecture 8

The .a!ue -o% N is %e!ate$ to the "i$th o- a (ea b) the -o!!o"in' -o%&u!a?

N = 505D 8t%G"1G676
Che%e t% = the %etention ti&e o- the #o&(oun$/ An$ "1G6 is the (ea "i$th at ha!- hei'ht The .a!ues o- t% an$ "1G6 a%e o-ten both &easu%e$ on the , a,is o- a #h%o&ato'%a&0 The) #ou!$ both t)(i#a!!) ha.e units &easu%e$ in &inutes o% a#tua! $istan#e &easu%e$ "ith a %u!e%/ in "hi#h #ase the units "ou!$ be in #& o% a si&i!a% unit0 It is i&(o%tant that both a%e &easu%e$ in the sa&e unitsK The "i$th o- a #h%o&ato'%a(hi# (ea is t)(i#a!!) &easu%e$ at a (oint ha!- "a) bet"een the base!ine an$ the to( o- the (ea 0 This is be#ause the (ea b%oa$ens %a(i$!) nea% the base!ine an$ it is $i--i#u!t to &easu%e a##u%ate!) 8The fact that the peak width is measured at height instead of at the baseline, is accounted for by the constant 5.5 in the equation70 I- the "i$th at the base!ine is &easu%e$ the e3uation be#o&es?

N = 1+8t%G"76
Che%e t% = the %etention ti&e o- the #o&(oun$/ An$ " is the (ea "i$th at the base!ine0 Reso!ution The (u%(ose o- #h%o&ato'%a(h) is to se(a%ate o% resolve #o&(oun$s0 The se(a%ation o% $istan#e bet"een t"o (ea s is no"n as thei% %eso!ution an$ is a -un#tion othe A -a#to%s $is#usse$ (%e.ious!)? retention 8the ti&e it ta es -o% the ana!)tes to e!ute/ %e!ate$ to 7/ selectivity 8ho" $i--e%ent the ana!)tes a%e -%o& ea#h othe% an$ %e!ate$ to 7/ an$ efficiency 8ho" 'oo$ the #o!u&n is/ %e!ate$ to N70 The%e-o%e/ these th%ee -a#to%s #an be b%ou'ht to'ethe% to $es#%ibe %eso!ution as -o!!o"s?

Rs = L 88-17G7 8 G M17 NN
The%e a%e a nu&be% o- "a) to e,(%ess the .a!ue o- Rs0 Anothe% #o&&on e3uation is so&eti&es use$ to #a!#u!ate Rs -%o& a#tua! &easu%e&ents o- (ea %etention ti&es an$ &easu%e$ (ea "i$ths0 This e3uation is?

Rs = 6 8tR-I H tR-A7G "b-A M "b-I

Che%e? A an$ I a%e the t"o (ea s / tR = %etention ti&e an$ "b = the (ea "i$th at the base o- ea#h (ea

Lecture 8

Cith a %eso!ution .a!ue o- 100/ t"o (ea s o.e%!a( b) about DO0 <a!ues !ess than 100 in$i#ate (ea s that o.e%!a(/ "hi!e at a %eso!ution o- 105/ the (ea s a%e #onsi$e%e$ -u!!) se(a%ate$0 Theoretical plates As (%e.ious!) state$/ #o!u&n e--i#ien#) is %e!ate$ to the 1nu&be% o- theo%eti#a! (!ates2 o% N0 The .a!ue o- 8N7 is %e!ate$ to the #o!u&n !en'th 8L7 an$ the si9e o% hei'ht 8H7 o- an in$i.i$ua! (!ate b) the -o!!o"in' e3uation?

N = LGH
The .a!ue o- L is usua!!) &easu%e$ in #&/ an$ #an .a%) -%o& :ust a -e" #enti&ete%s -o% HPLC #o!u&ns to &an) &ete%s -o% GC #o!u&ns0 The .a!ues o- N a%e usua!!) !a%'e/ in the thousan$s o% e.en &i!!ions/ 'i.in' .a!ues o- H/ t)(i#a!!) in the %an'e o- 1G100 - 1G1000 #enti&ete%0 The si9e o- H a--e#t the e--i#ien#) o- the #o!u&n an$ the%e-o%e it is i&(o%tant to un$e%stan$ "hat a--e#ts the .a!ue o- H0 The .a!ue o- H $e(en$s (%i&a%i!) on -ou% -a#to%s/ 17 the .e!o#it) o- the &obi!e (hase/ 67 &u!ti(ath $i--usion/ A7 the $i--usion o- the #o&(oun$ in the &obi!e (hase/ an$ D7 the t%ans-e% o- the #o&(oun$ bet"een the stationa%) (hase an$ the &obi!e (hase0 ;o% #o!u&ns (a# e$ "ith (a%ti#!es 8HPLC #o!u&ns7/ these -a#to%s #an be e,(%esse$ b) the -o!!o"in' -o%&u!a0

H = A M IGu M 8Cs M C&7 u

Che%e u = the a.e%a'e !inea% &obi!e (hase .e!o#it)/ A is a #onstant e,(%essin' $i--usion $ue to non uni-o%&it) o- the (a# in'0 I is a #onstant e,(%essin' the !on'itu$ina! $i--usion #oe--i#ient in the &obi!e (hase/ Cs is the &ass t%ans-e% te%& in the stationa%) (hase/ C& is the &ass t%ans-e% te%& in the &obi!e (hase The Mu!ti(ath Te%& A0 This -a#to% is a!so #a!!e$ e$$) $i--usion/ but &o%e &o$e%n te,tboo s use the te%& &u!ti(ath0 In (a# e$ #o!u&ns/ (ea -b%oa$enin' is the %esu!t o- a nu&be% o- -a#to%s0 As &o!e#u!es o- the ana!)te &o.e th%ou'h the #o!u&n/ the) ta e &an) $i--e%ent (aths a%oun$ the (a# e$ (a%ti#!es0 So&e o- these (aths a%e un$oubte$!) !on'e% than othe%s so as the &o!e#u!es &o.e th%ou'h the #o!u&n/ the) ten$ to s(%ea$ out0 The a&ount o- s(%ea$in' is a--e#te$ b) the natu%e o- the #o!u&n &ate%ia! an$ ho" "e!! the #o!u&n is (a# e$0 This -a#to% is 'ene%a!!) (%o(o%tiona! to the (a%ti#!e si9e o- the (a# in' &ate%ia!0 This -a#to% &ust be ta en into a##ount -o% (a# e$ #o!u&ns/ but -o% #a(i!!a%) #o!u&ns/ this te%& is not nee$e$ sin#e the%e a%e no (a%ti#!es0

Lecture 8

;!o" Di%e#tion 1 6
Path"a)s o- t"o &o!e#u!es $u%in' e!ution0 Distan#e t%a.e!e$ b) &o!e#u!e 1 is !on'e% than that t%a.e!e$ b) &o!e#u!e 6/ thus &o!e#u!e 1 "i!! ta e !on'e% to e!ute0

;i'u%e A0 T)(i#a! (ath"a)s o- t"o so!ute &o!e#u!es th%ou'h a (a# e$ #o!u&n0 The Lon'itu$ina! Di--usion Te%& B/ Lon'itu$ina! $i--usion a!so #ont%ibutes to (ea b%oa$enin'0 In this (%o#ess/ ana!)tes $i--use -%o& a%eas o- hi'h #on#ent%ation to &o%e $i!ute a%ea in -%ont o- an$ behin$ the &o.in' ban$0 The $e'%ee o- !on'itu$ina! $i--usion is %e$u#e$ to so&e e,tent b) the (a# in' &ate%ia!0 At !o" .e!o#ities !on'itu$ina! $i--usion has a ne'ati.e e--e#t on %eso!ution/ but this e--e#t is ne'!i'ib!e at hi'he% .e!o#ities0 This te%& is .e%) i&(o%tant in 'as #h%o&ato'%a(h) as $i--usion #oe--i#ients in 'asses a%e o%$e%s o- &a'nitu$e hi'he% than in !i3ui$s0 In !i3ui$ #h%o&ato'%a(h)/ this te%& is t)(i#a!!) #!ose to 9e%o %e!ati.e to the othe% te%&s0

Mo!e#u!es $i--use -%o& a%eas o- hi'h #on#ent%ation to a%eas o- !o" #on#ent%ation0

;!o"
O.e% ti&eP0

;!o"

;i'u%e D0 I!!ust%ation o- (ea b%oa$enin' $ue to $i--usion in a #a(i!!a%) #o!u&n0 The Mass T%ans-e% Te%&s ! 0 The &ass t%ans-e% te%& %e!ates to the -a#t that e3ui!ib%iu& bet"een the &obi!e an$ stationa%) (hases is ne.e% %ea!i9e$ in a #h%o&ato'%a(h) #o!u&n0 It ta es ti&e -o% ana!)tes in the &obi!e (hase to &o.e into the Lecture 8 E

stationa%) (hase0 Ie#ause no e3ui!ib%iu& is %ea#he$/ so&e o- the ana!)tes a%e s"e(t ahea$ o- the o- the &ain ban$0 It a!so ta es ti&e -o% &o!e#u!es to &o.e out o- the stationa%) (hase/ an$ so&e o- the ana!)tes &o!e#u!es "i!! be !e-t behin$ b) the %a(i$!) &o.in' &obi!e (hase0 Li e the !on'itu$ina! te%&/ the &ass t%ans-e% te%& is base$ on $i--usion0 Ho"e.e%/ !on'itu$ina! $i--usion ta es (!a#e (a%a!!e! to the $i%e#tion o- -!o"/ an$ the%e-o%e is inversely %e!ate$ to the &obi!e (hase -!o" %ate/ "hi!e &ass t%ans-e% $i--usion ta es (!a#e (e%(en$i#u!a% to the -!o" %ate0 As a #onse3uen#e/ the -aste% the &obi!e (hase &o.es/ the !ess ti&e the%e is -o% e3ui!ib%iu& bet"een the (hases an$ the &ass t%ans-e% e--e#t on (ea b%oa$enin' is directly %e!ate$ to &obi!e (hase .e!o#it)0

P!ate Hei'ht/ H

A M IGu M Cu
Mass T%ans-e% 8both7/ Cu Mu!ti(ath Te%&/ A Lon'itu$ina! $i--usion/ IGu

Linea% <e!o#it)/ u
;i'u%e 50 A .an Dee&te% (!ot -o% a !i3ui$ #h%o&ato'%a(h) #o!u&n0 ;o% hi'h %eso!ution/ these $i--usion -a#to%s shou!$ be &ini&i9e$ 8(!ate hei'ht shou!$ be s&a!!70 The <an Dee&te% (!ot sho"s that the e--e#t o- the .a%ious ban$b%oa$enin' (a%a&ete%s on (!ate hei'ht0 Pea b%oa$enin' $ue to &u!ti(ath $i--usion is %e!ati.e!) #onstant o.e% the no%&a! %an'e o- &obi!e (hase .e!o#ities/ "hi!e (ea b%oa$enin' $ue to &obi!e (hase &ass t%ans-e% in#%eases "ith &obi!e (hase .e!o#it)0 On the #ont%a%)/ (ea b%oa$enin' $ue to !on'itu$ina! $i--usion is hi'h at !o" .e!o#ities an$ has a !esse% e--e#t as &obi!e (hase .e!o#it) in#%eases0 The o.e%a!! e--e#t is that the%e is an inte%&e$iate .e!o#it) that )ie!$s the s&a!!est (!ate hei'ht an$ hen#e the hi'hest %eso!ution0 Ho"e.e%/ in the inte%est o- s(ee$ o- ana!)sis/ %e#o&&en$e$ .e!o#ities a%e o-ten set so&e"hat hi'he% that this0

References"

Lecture 8

Iees!e)/ T0 E0Q Iu'!io/ I0Q S#ott/ R0 P0 C0 !uantitati"e #hromatographic $nalysisQ Ma%#e! De e%/ In#? Ne" Yo% / 6001Q AEF0 G%ant/ D0 C0 #apillary gas chromatographyQ 4ohn Ci!e) R Sons/ Lt$? Cest Susse,/ En'!an$/ 1**+Q 6*50 S oo'/ D0 A0Q Ho!!e%/ ;0 40Q Nie&an/ T0 A0 %rinciples of &nstrumental $nalysis/ 5th e$0Q Ha%#ou%t I%a#e R Co&(an)? Phi!a$e!(hia/ 1**FQ FD*0

Lecture 8