Couise ,Modein Anulyticul Metlods in Clemicul Industiy

Cliomutogiuplic Teclniques
Piinciple
Cos cbromoloqroby [CC}.
Hiqb rossoro
ligoio cbromoloqroby [HPLC}.
Tbin loyor cbromoloqroby [TLC}.
nob||o
hoso
qosos
[H
2
, N
2
,
/r, Ho}
H
2
C,
orqonic
solvonls
voor o
orqonic
solvonls
in|oclion
orl
slolionory boso
[soorolion o lbo mixloro}
nob||o hoso
nob||o hoso
somlo somlo somlo somlo
[mixloro} [mixloro} [mixloro} [mixloro}
ooloclor
oo|oc|os
(oxon|os;
TCD, FlD,
MSD
\-\lS, Rl,
MSD
olicol
ooloclion
[\-\lS,
loorosconco}
s|o||ooo,
hoso
coillory or
oc|oo
colomns
wilb
soosliloloo
siloxonos
imroqnoloo
lolos
o||cob||||,
vololilo orqonic
comoonos,
ormononl qosos
orqonic
comoonos ono
inorqonic solls
orqonic
comoonos
|oqo oonbo o| s|ooooo|zoo ocoooos (oons; |o |ooos||o| ooo oo|oonoo|o| ooo|,s|s
Fundumentuls ol Cliomutogiuply
l
C
- ,ooolb limo ~ rolonlion o o comoono
wilb no inloroclion wilb lbo slolionory boso
l
R[1}
- lolol rolonlion limo o comononl 1
l
R[2}
- lolol rolonlion limo o comononl 2
l'
R[1}
- nol rolonlion limo o comononl 1 [~ l
R[1}
- l
C
}
l'
R[2}
- nol rolonlion limo o comononl 2 [~ l
R[2}
- l
C
}

[1}
- ooso oo| wilb o comononl 1

[2}
- ooso oo| wilb o comononl 2
s|qoo|
||no |
Comononl 1
Comononl 2
|
0
|
(|;
|
(2;
|
(|;
|
(2;

[!;

[2;
somlo in|oclion
[l ~ C}
Equutions in Cliomutogiuply (1)
Cooc||, |oc|o |
(o||o bo|&ooo |ho os|oooco ||nos o| |ho son|o |o |ho s|o||ooo, ooo |ho nob||o hoso;
1
t
t
t
t t
t
t
k
0
R
0
0 R
0
R
=

o|o||o o|oo||oo/Sooo||oo |oc|o/So|oc||||,

) 1 (
) 2 (
k
k

=
Soorolion o lwo comononls is only ossiolo i > 1
L - lonqlb o lbo colomn
l
C
- oooo limo
F - Fooo rolio o lbo mooilo boso |ml/s|
g - roo cross-soclionol oroo o lbo colomn
L|ooo o|oc||, o
q
F
t
L
u
0
= =
Equutions in Cliomutogiuply (2)
fonoob||||, K
0
2
t p
L
p
L u
K

=

=
Soc|||c onoob||||, K
0
1000
d
K K
p
T
0
= =
- rossoro oioronco
oolwoon inlol ono oollol o o colomn
lorqo K - wronq orliclo oc|inq, low K - cloqqinq
- viscosily o lbo mooilo boso
o

- orliclo oiomolor o lbo oc|inq

foos||, o| o co|ono
T
r - rooios o lbo colomn
\
R
- volomo o lbo omly colomn
R
0
2
0
2
T
V
t F
L i
t F
i
q
=

=

=
Equutions in Cliomutogiuply (3)
E||oc||o oonbo o| |o|os N
o||.
(|oooooooo| oo conoooo|s o| |ho son|o;
2
R
2
. ell
' t
16
1 ' k
' k
N N

+
=
foo| booooo|oq
N - nomoor o lolos
- ooso oo| wilb

C.5
- oo| wilb ol bol oo| boiqbl
H - boiqbl o lolos
2
) i ( , S . 0 ) i ( R
2
) i ( ) i ( R
) t (
S4 . S
) t (
16
N

=

=
N
L
H =
Equutions in Cliomutogiuply (4)
oso|o||oo bo|&ooo 2 oo|s

=
' t
' t ' t
2 R
) 2 ( ) 1 (
) 1 ( R ) 2 ( R
[or oo|s closo ono lo lbo onolbor}
R ~ 18 oo| soorolion, R ~ 1.5 olly sooroloo oo|s
N
1 ' k
' k
) 1 (
4
1
R
) 2 (
) 2 (

=
[ - 1}/ - soloclivily lorm
[oolorminoo oy
slolionory ono mooilo bosos}
|'
[2}
/[|'
[2}
1} - coocily lorm
[only siqniiconl
or C |'
[2}
5,
or lorqo |'
[2}
1}
N
1/2
- ooclivily lorm
[oolorminoo oy o

, L ono o}
Equutions in Cliomutogiuply (S)
\oo Doon|o ooo||oo
u C
u
B
A H + + =
Eooy oiosion lorm
[oo| orooooninq oy
oioronl woys o somlo
molocolos in o colomn}
oxiol oiosion lorm
[slolislic oxiol oislrioolion
o somlo molocolos}
lorm or moss oxcbonqo
oolwoon slolionory ono mooilo bosos
[oosorlion/oosorlion, oiosion/ooc| oiosion}
|
o
H ~ / 8/o C o
C o
8/o
/
Son|os. - qosooos ono ligoio somlos
- limilolions. comoono wbicb sboolo oo onolyzoo,
sboolo oo sloolo onoor CC oorolion conoilions ono
sboolo bovo o voor rossoro siqniiconlly biqbor lbon zoro
- somlo roorolion. illrolion, oxlroclion, i nocossory oorivolisolion
[~ convorsion o ,crilicol soosloncos lo socb wilb biqbor
slooilily ono voor rossoro, o.q. coroon ocios lo oslors}
/ob||o hosos.
- Ho, H
2
, /r, N
2
[orily . or oollor}
Doo||oo o| oo ooo|,s|s.
- 5 ... C min [slonooro CC}, 2 min [Micro CC}
/||co||oo. - orily conlrol, goolily monoqomonl ono corliicolion [wioo olicolion}
- onvironmonlol ono bormocoolicol onolysis
- onolysis o moin ono lroco comononls [ lo m}
Gus Cliomutogiuply
in|oclion
orl
colomn wilb slolionory boso
[insorloo in o CC ovon}
nob||o hoso
somlo somlo somlo somlo
[mixloro} [mixloro} [mixloro} [mixloro}
ooloclor
low
conlrollors
GC Setup
CC ooo
&||h
co|ono(s;
/o|oson|o
|o ||o|o
son|os
Ko,booo
|o coo|o|||oq
||o& o|os ooo
|onoo|oos
Cono|o |o
CC ooo||oo ooo
oo|o hooo||oq
S|o|os o|s|o,
|o ooo||oo
oono|os
F|D ooo TCD
|ooc||oo |o|s
o| oo|oson|o
/SD
|ooc|o
Sumple Injection
L|o|o son|os.
- monool [osinq o syrinqo, C.1 - 1C 0l, ioonliicolion o comoonos}
- oolomoloo oy oolosomlor [nooo o 1 - 2 ml somlo sololion, biqb rorooocioilily,
oso or goonlilolivo onolysis}
- on colomn" in|oclion or somlos wilb low lbormol slooilily
[in|oclion oiroclly on lo o lbo colo colomn, lbon slow boolinq}
biqb rocision, ool oonqor o colomn ovorloooinq ono ollolion
- lomoroloro roqrommoo voorizolion - PT\
[in|oclion inlo lbo colo in|oclor, lbon lomoroloro roqrommoo boolinq ono voorizolion}
biqb sonsilivily ono rorooocioilily, roloclion o lbo colomn
- booo soco" in|oclion - somlinq o lbo voor boso ovor lbo somlo
[osool i lbo somlo conloins solio orliclos}
Cosooos son|os
- monool [osinq o qos-oonso syrinqo, 5 - 5C 0l}
- oolomoloo oy qos somlinq volvos [C.25 - 5 ml}
|ooc|o |,os.
- slil-slilloss in|oclor
- vololilo inloroco
Rolonlion oobovior ooonos on.
- olorily o somlo ono slolionory boso
- vololilily o lbo somlo comoono
- volocily o lbo corrior qos [mooilo boso}
- lomoroloro
lomoroloro roqroms
S|o||ooo, hosos
- silicon olymors [olysiloxonos, Si-C-R}
R ~ molbyl - non-olor,
R ~ bonyl or cyonoroyl -
inlormooiolo olorily,
R ~ olbylono qlycol or loorinoloo
byorocoroon - olor,
voriolion o olorily oy co-olymorizolion}
- PLCT bosos [oroos loyor oon looolor}
smoll orliclos immooilizoo in lbo woll,
or soorolion o biqb vololilo comoonos,
lyicol slolionory bosos.
/l
2
C
3
, molsiovo 5/,
olyslyrono-oivinyloonzono [D\8}
- Solocloo colomn monoocloros.
1 & W [/qilonl},
Cbromoc|, Roslo|, Soolco
GC Columns
Co|||o,
co|onos
lbin loyor o
slolionory
boso
foc|oo
co|onos
o ~ C.1 - C.53 mm,
l ~1C - 5C m,
ilm lbic|noss.
C.3 - 5C 0m
o ~ C.53 mm,
l ~C.5 - 1C cm,
orliclo oiomolor.
45 - 12C mosb
smoll orliclos
o
slolionory
boso
GC Detectois
1C
4
C.1 - 2C q/s
[ooonoinq on
lbo olomonl}
lonoolo or ony olomonl /ED
/lom omission ooloclor
1C
3
>1 nq
[ooonoinq on
oosorlion}
molocolor viorolions [o.q.
orqonic comoonos}
FTlRD
Foorior lronsormoo inroroo
ooloclor
1C
4
C.4 q N/s
C.2 q P/s
N, P, bolorooloms NPD
Nilroqon bosboros ooloclor
[Tbormoionic ooloclor}
1C
4
C.1 q Cl/s boloqons ECD
Eloclron coloro ooloclor
1C
5
1C nq [SC/N}
1C q [SlM}
lonoolo or ony socios MSD
Moss soloclivo ooloclor
1C
7
5 q C/s moloriols lbol oro ionizoo in
oir/H
2
lomos
[o.q. byorocoroons}
FlD
Flomo ionizolion ooloclor
1C

4CC q/ml
corrior
non-soloclivo
[onylbinq wbol oiors rom
lbo corrior qos}
TCD
Tbormol conooclivily ooloclor
L|ooo o,oon|c
ooqo
T,|co| n|o|non
oo|oc|ob|o |oo|
So|oc||||, Nono
Tleimul Conductivity Detectoi (TCD)
Flume Ionizution Detectoi (FID)
Muss Selective Detectoi (MSD)
E|oc|oo
sooco
Son|o boon |on /SD |o|o|oco
(ossoo oooc||oo |o ocoon;
Cooooo| noss ||||o
Do|oc|o
(SC/N o S|/ nooo;
|oo|zo||oo ooo
Foqnoo|o||oo o| |ho
son|o no|oco|os b,
|ho o|oc|oo boon
|oo|zo||oo ooo |oqnoo|o||oo.
[1} M o
-
M

2 o
-
[yioloinq molor oo| M

}
[2} M

[M - Z}
Z

[Z - }
..
[yioloinq cboroclorislic moss oo|s
ono moss oioroncos}
[3} Roorronqomonl rooclions
cboroclorislic moss soclro,
ioonliicolion o soosloncos ossiolo
GC Applicution Exumples
fo|ochon|co| ooo|,s|s.
o||oo, qos
fhonocoo||co| ooo|,s|s.
/||o|o|o s|oo| ooqs
Eo|oonoo|o| ooo|,s|s.
Ch|o|oo|oo os||c|oos
Higl Piessuie Liquid Cliomutogiuply (HPLC)
orl
in|oclion volvo
colomn wilb slolionory boso
[normolly ol room lomoroloro}
nob||o hoso
somlo somlo somlo somlo
[mixloro} [mixloro} [mixloro} [mixloro}
ooloclor
biqb rossoro
oms
Son|os. - ligoio somlos
- limilolions. solooilily in lbo mooilo boso, no lbormol roslriclions
- somlo roorolion. illrolion, oxlroclion
Doo||oo o| oo ooo|,s|s.
- 5 ... C min
/||co||oo. - orily conlrol, goolily monoqomonl ono corliicolion [wioo olicolion}
- onvironmonlol ono bormocoolicol onolysis
- onolysis o moin ono lroco comononls [ lo m}
S|o||ooo, fhoso /ob||o fhoso
non-olor olor [H
2
C, ooor sololions, olcobols,
ocolonilrilo ono mixloros o lbom}
Rovorsoo Pboso HPLC
olor non-olor [o.q. byorocoroons} Normol Pboso HPLC
0.0! - !0 n|/n|o
o| ossoos
o |o 500 bos
HPLC Setup
S|ooqo |o nob||o hoso
(o|ooo|;
F|||o||oo ooo ooqoss|oq
fooo||oo o|
o|ooo| n|x|oos
fon
/o|oson|o
Co|ono box
|ooc||oo o|o
Coo|o| oo||
Do|oc|o oo|| &||h J\-\|S |on
Coo||oo nooos.
- |soco||c. coos|oo| o|ooo| conos|||oo
- qoo|oo|. coo||oooos o|o||oo o|
o|ooo| n|x|oo oo|oq ooo|,s|s
HPLC Injectois
o| |ooc||oo o|o
(s|n||o |o qos son||oq o|os |o CC;
Comoinolion wilb oolosomlors or biqb rorooociolo somlo in|oclion
HPLC Columns
Normol boso HPLC
Rovorsoo boso HPLC
S|o||ooo, hoso /ob||o hoso /||co||oo
/l
2
C
3
, SiC
2
omino boso
[SiC
2
-[CH
2
}
n
-NH
2
}
oiol boso
[SiC
2
-[CH
2
}
n
-CH[CH}-CH
2
CH}
cyono boso
[SiC
2
-[CH
2
}
n
-CN}
SiC
2
-[CH
2
}
n
-CH
3
[n ~ 8 - RP 8 or 18 - RP 18}
onocooo colomns"
~ goonlilolivo solorolion
o oll CH qroos oy -CH
3
}
o|o||,
byorocoroons,
iso-roonol
wolor,
molbonol,
ocolonilrilo
non-olor
comoonos
[o.q. byorocoroons,
bolobyorocoroons,
olbors}
olor comoonos
[o.q. olcobols,
coroon ocios}
Cnly woo|
inloroclions
oolwoon lbo somlo
ono lbo slolionory
boso is rogoiroo.
HPLC Detectois
1C
5
no oolo ovoiloolo - mosl soloclivo ooloclor or HPLC
- slronq rogoiromonls or lbo inloroco
[lronsilion rom lbo biqb colomn rossoro
lo vocoom insioo lbo MSD}
- biqb cosls
/oss so|oc||o oo|oc|o
!
1C
3
1C
-8
- sociic low-cosl ooloclor or comoonos
oissocioloo inlo ions [o.q. inorqonic ono
orqonic solls, lonsioos, omino ocios}
E|oc||c cooooc||||,
oo|oc|o
1C
4
5 1C
-1C
- non-sociic low-cosl ooloclor o|oc||oo |ooox
oo|oc|o
1C
3
1C
-1C
. 1C
-
- oolocls loorosconco rooiolion omilloo oy
lbo somlo comoonos
- sociic or biqbly conoonsoo orqonic
molocolos li|o P/H
F|oooscooco oo|oc|o
!
5 1C
4
5 1C
-1C
- or lorqor orqonic molocolos ono
lronsilion molol comoono wbicb oosoro
\-\lS liqbl
- limo rosolvoo rocoroinq o \-\lS
soclro, ossioilily o ooconvoloolion o
non-sooroloo oo|s
J\-\|S oo|oc|o/
D|ooo oo, oo|oc|o
!
L|ooo o,oon|c
ooqo
T,|co| n|o|non
oo|oc|ob|o |oo| jq/n|j
So|oc||||, Nono
1 - soiloolo or qrooionl locbnigoos
Speciul HPLC Teclniques
|oo Chono|oqoh,/|oo Exchooqo Chono|oqoh, (|C/|EC;.
- slolionory boso. ion oxcbonqo rosins
[R-SC
3
-
or R-CCC
-
or colion onolysis, R-NH
3

or R-N[ol|yl}
3

or onion onolysis}
- mooilo boso. ogoooos sololions
[oiloloo minorol ocios or colion onolysis,
byoroqon coroonolo ooor or onion onolysis}
- ooloclor. oloclric conooclivily ooloclor wilb ro-inslolloo sorossion colomn
- olicolion. osl onolysis o inorqonic ono orqonic solls in wolor
[moinly ol|oli ono ol|olino oorlb molol solls}
- ooloclion limil. C.5 m
Co|||o, E|oc|ohoos|s (CE;.
- oorolion o lbo colomn in on oloclricol iolo
- Movomonl o lbo ions is orivon oy oloclricol ollroclion.
- coolinq wilb lyicol HPLC ooloclors [incl. MSD}
HPLC Applicution Exumples
8|o|ochoo|oq|co| ooo|,s|s.
/n|oo oc|os
fhonocoo||co| ooo|,s|s.
8os|c ooqs
Eo|oonoo|o| ooo|,s|s.
fo|,c,c||c
oono||c h,oocoboos
Quuntitutive Anulysis
Ex|ooo| s|ooooo
[1} Coliorolion or o |nown sooslonco oy in|oclion o oioronl |nown sooslonco
conconlrolions, colcololion o o roqrossion corvo
[2} ln|oclion o lbo somlo wilb lbo on|nown conconlrolion,
ooc| colcololion o lbo conconlrolion oy osinq lbo coliorolion onclion
|o|ooo| s|ooooo (o||n|oo||oo o| soos|||||, o|o||oos;
[1} /ooinq o o |nown ogool omoonl o o sooslonco wbicb is nol o orl o lbo somlo
lo oocb coliorolion somlo ono lo lbo somlo wilb lbo on|nown conconlrolion
[2} Normolizolion o lbo siqnol rosonso [oo| oroo or boiqbl} o oll comononls
on ooso o o conslonl siqnol o lbo oxlornol slonooro comoono [ ~
norm
/
rool
}
S|ooooo ooo|||oo
[1} ln|oclion o lbo somlo wilb lbo on|nown conconlrol|oo
[2} /ooinq o |nown omoonls o lbo sooslonco wbicb sboolo oo onolyzoo ono
ororminq o now onolysis olor oocb oooilion
[3} Colcololion o lbo on|nown conconlrolion
oy sollinq ~ C or lbo conconlrolion-rosonso-onclion