Kromatogarfi-Tambahan 2-Eksperimen

8rldglng C0004
kkCMA1CGkAIILksper|men
C0004-KromatograIi-Eksperimen 1
our m|ss|on shou|d you choose to accept |s
to
nalyse coloured markers
Test whether they contain E133 (blue), E122
(pink), E124 (red) or E110 (yellow)
1h|n Layer Chromatography
ample marker
andard food dyes
alonary phase chromaography paper
Moblle phase waer
tructures of L numbers
E110 yellow
E122 pink
E124 red
E133 blue
o what w||| happen?
ach dye wlll ravel up he paper a dlfferen
speeds
1he speed depends on he solubllly of he dye ln
waer and ls lneraclon wlh he paper
1he dyes are all dlfferen molecules wlh dlfferen
characerlslcs
Ca|cu|at|on of resu|ts
Ca|cu|at|on of resu|ts
nalysis
ou must now calculate an Rf value for each spot.
Rf = Distance from the start to the middle of a spot
Distance from start to finish point of the water
Conc|us|ons wr|t|ng up
ne of he key elemens of all sclenlflc
experlmens ls o wrle up your resuls
he end of hls experlmen we would llke
each person o conclude from he 8f values as
o whlch numbers are conalned ln he
markers
Jha ls Chromaography?
Chromofogrophy is o fechnique for
seporofing mixfures info fheir componenfs
in order fo onoIy;e, idenfify, purify,
ond/or quonfify fhe mixfure or
componenfs.
$eparate
Analyze
IdentiIy
PuriIy
QuantiIy
Components Mixture
Dses for Chromaography
Chromofogrophy is used by scienfisfs fo:
AnoIy;e - exomine o mixfure, ifs componenfs,
ond fheir reIofions fo one onofher
Idenfify - defermine fhe idenfify of o mixfure or
componenfs bosed on known componenfs
Purify - seporofe componenfs in order fo isoIofe
one of inferesf for furfher sfudy
Quonfify - defermine fhe omounf of fhe o mixfure
ond/or fhe componenfs presenf in fhe sompIe
Dses for Chromaography
#eoI-Iife exompIes of uses for
chromofogrophy:
PhormoceuficoI Compony - defermine omounf of
eoch chemicoI found in new producf
HospifoI - defecf bIood or oIcohoI IeveIs in o
pofienf's bIood sfreom
Low Enforcemenf - fo compore o sompIe found of
o crime scene fo sompIes from suspecfs
EnvironmenfoI Agency - defermine fhe IeveI of
poIIufonfs in fhe wofer suppIy
Monufocfuring PIonf - fo purify o chemicoI
needed fo moke o producf
eflnllon of Chromaography
efoiIed efinifion:
Chromofogrophy is o Ioborofory fechnique fhof
seporofes componenfs wifhin o mixfure by using fhe
differenfioI offinifies of fhe componenfs for o mobiIe
medium ond for o sfofionory odsorbing medium fhrough
which fhey poss.
TerminoIogy:
ifferenfioI - showing o difference, disfincfive
Affinify - nofuroI offrocfion or force befween fhings
MobiIe Medium - gos or Iiquid fhof corries fhe componenfs
(mobiIe phose)
Sfofionory Medium - fhe porf of fhe opporofus fhof does
nof move wifh fhe sompIe (sfofionory phose)
SimpIified efinifion:
Chromofogrophy seporofes fhe componenfs of o
mixfure by fheir disfincfive offrocfion fo fhe mobiIe
phose ond fhe sfofionory phose.
ExpIonofion:
Compound is pIoced on sfofionory phose
MobiIe phose posses fhrough fhe sfofionory phose
MobiIe phose soIubiIi;es fhe componenfs
MobiIe phose corries fhe individuoI componenfs o
cerfoin disfonce fhrough fhe sfofionory phose,
depending on fheir offrocfion fo bofh of fhe
phoses
eflnllon of Chromaography
llusralon of Chromaography
Components
Affinity to Stutionury
Phuse
Affinity to MobiIe
Phuse
Iue ---------------- InsoIubIe in MobiIe Phose
Iuck
#ed
YeIIow
Mixture Components
$eparation
$tationary Phase
Mobile Phase
Liquid Chromofogrophy - seporofes Iiquid sompIes
wifh o Iiquid soIvenf (mobiIe phose) ond o coIumn
composed of soIid beods (sfofionory phose)
0os Chromofogrophy - seporofes vopori;ed sompIes
wifh o corrier gos (mobiIe phose) ond o coIumn
composed of o Iiquid or of soIid beods (sfofionory
phose)
Poper Chromofogrophy - seporofes dried Iiquid
sompIes wifh o Iiquid soIvenf (mobiIe phose) ond o
poper sfrip (sfofionory phose)
Thin-Loyer Chromofogrophy - seporofes dried Iiquid
sompIes wifh o Iiquid soIvenf (mobiIe phose) ond o gIoss
pIofe covered wifh o fhin Ioyer of oIumino or siIico geI
(sfofionory phose)
%ypes oI Chromatography
(A) uses charge, (B) uses
pores, and (C) uses covalent
bonds to create the differential
affinities among the mixture
components for the stationary
phase.
9rlnclples of 9aper Chromaography
W Caplllary clon he movemen of llquld wlhln he spaces of a
porous maerlal due o he forces of adheslon coheslon and surface
enslon 1he llquld ls able o move up he fller paper because ls
araclon o lself ls sronger han he force of gravly
W olubllly he degree o whlch a maerlal (solue) dlssolves lno a
solven olues dlssolve lno solvens ha have slmllar properles
(Llke dlssolves llke) 1hls allows dlfferen solues o be separaed by
dlfferen comblnalons of solvens
eparalon of componens depends on boh helr solubllly ln he
moblle phase and helr dlfferenlal afflnly o he moblle phase and he
salonary phase
9aper Chromaography xperlmen
hof CoIor is fhof Shorpie7
vervlew of he xperlmen
9urpose
1o lnroduce sudens o he prlnclples and
ermlnology of chromaography and demonsrae
separalon of he dyes ln harple 9ens wlh paper
chromaography
1|me kequ|red
9rep lme 10 mlnues
xperlmen lme 43 mlnues
Costs
Less han $10
Maerlals Lls
W beakers or [ars
W covers or llds
W lsllled P
2
W sopropanol
W Craduaed cyllnder
W srlps of fller paper
W lfferen colors of harple
pens
W 9encll
W 8uler
W clssors
W 1ape
9reparlng he sopropanol olulons
Prepare 15 ml oI the Iollowing isopropanol solutions
in appropriately labeled beakers:
- 0, 5, 10, 20, 50, and 100
9reparlng he Chromaography rlps
W Cu srlps of fller paper
W raw a llne 1 cm above he
boom edge of he srlp
wlh he pencll
W Label each srlp wlh ls
correspondlng solulon
W 9lace a spo from each pen
on your sarlng llne
eveloplng he Chromaograms
W 9lace he srlps ln he beakers
W Make sure he solulon does
no come above your sar llne
W keep he beakers covered
W Le srlps develop unll he
ascendlng solulon fron ls
abou 2 cm from he op of
he srlp
W 8emove he srlps and le
hem dry
eveIoping fhe Chromofogroms
eveIoping fhe Chromofogroms
bservlng he Chromaograms
Concenfrofion of IsoproponoI
0 20 50 70 100
8lack ye
1 yes separaed purple and black
2 no soluble ln low concenralons of
lsopropanol
3 9arlally soluble ln concenralons of
lsopropanol 20
C0004-KromatograIi-Eksperimen 30 Concenfrofion of IsoproponoI
0 20 50 70 100
8lue ye
0 20 50 70 100
I. ye seporofed - bIue
Z. Mof very soIubIe in Iow
concenfrofions of isoproponoI
3. CompIefeIy soIubIe in high
concenfrofions of isoproponoI
Creen ye
0 20 50 70 100
I. ye seporofed - bIue ond yeIIow
Z. 8Iue - SoIubIe in concenfrofions
of isoproponoI Z07
3. YeIIow - SoIubIe in concenfrofions
of isoproponoI 07
8ed ye
1 yes separaed red and yellow
2 ?ellow soluble ln low concenralons of lsopropanol and
less soluble ln hlgh concenralons of lsopropanol
0 20 50 70 100
3. #ed - sIighfIy
soIubIe in Iow
concenfrofions
of isoproponoI,
ond more
soIubIe in
concenfrofions
of isoproponoI
Z07
lernalve xperlmens
W 1est d|fferent samp|es
her markers pens hlghllghers
llower plgmens
lood Colors
W 1est d|fferent so|vents
her alcohols mehanol ehanol
propanol buanol
W 1est d|fferent papers
Coffee fllers
9aper owels
Cardsock
1yplng paper
lernalve xperlmens
lernalve xperlmens
lernalve xperlmens
1k andards
W 1 71 81 uden conducs laboraory lnveslgalons uslng safe
envlronmenally approprlae and ehlcal praclces
W 2 72 82 uden uses sclenlflc lnqulry mehods durlng laboraory
lnveslgalons
W 3 73 83 uden uses crllcal hlnklng and sclenlflc hlnklng and
problem solvlng o make lnformed declslons
W 4 74 84 uden knows how o use a varley of ools and mehods
o conduc sclence lnqulry
W 7 77 89 uden knows ha subsances have physlcal and chemlcal
properles

Kromatogarfi-Tambahan 2-Eksperimen

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