Pago: 35
Date: 2022
BASIC IDEA 0F CHROMATO GRAPHY PRINCIPLE,
AND COLUMN CHROMATOGRAPHY.
PAPER_CHROMATORAPRY
Biniple
Chuonatbguashy iu a technique fox sepanating te componeni
0T colutes oj a mmixte oA Hhe basis o the Helative amouna o
lah soute distributed be tueen a
the moble thase ond a contigupu stationauy phase
The Rusian hotanal- Mikkhail Tswett eoined the temm choatogahy
Ie 4acBos joetiue on Hale seyauation pDIOLA jnclude moleular
chauateHNies Helated to ndsomtinn (liquid- soid), panttion
(iauid-colbd) andainihy dieuenca among thein
molecnlax weight.
Beease
longeu in the tatonoy phase and they moe u the
chrowakoqraphy yute,tohile others ja mpidyinto the
mobile phase,and leave the susten aslez
he uelention Jlme u dejuned anas the
the time
time bebueen te sample
inuduubion and the elulion ton the coumn.
Thxee coponank #s Aormthe bau oy ihe crormatogra hy
teehnique -
O Slatiunayphase i- Thi phaseu alway eopbsed o asioa.
akorted. on its susfeee
hase and olayer oiguid ààQdsoree
-PIOnECR
Topic : Page: 3b
Date:
(9 Mobile yhase i
gaseous component
( Sepanated molecules
twheneby ohto watoqraphuy roedues specialiset
Christian hedrieh Schonbein (i65).
-Substames aue disteibuted bebeena stolionay phase and mobile
phase
-(olluloIe layen nfter popr Contain. moahure cwhick ats as
statibaay hase, organd solents/kajen ane sed a mobile phase
he woeiment o a tompound lk dewnibed uing, te etardation
tactor R he Ko a coupound ii descaibed as he distane
the coupound ii eluted (Leonpound), diided by distanee
frou te applicatinn line to tre mobile pha~e tront (Lmakle phass
The laxges Yous qur R the
the oueM
loueN is the interaction
bebueon he compound and Whe statibnay phase.
J he Subclaue moves,ik an mone n the diection
o tne sowent plouo, and cannot move faster han the solvent
-vabutx ange from D(roelutas to i(na ajjinitys.
PIORCCR
Tcfic: Page: 3T
Date
DColunmn Chrowatoqraphey
- Colnan chuoma koqraky ia technique in whieh the substences
to be sepaHated aue ntroduced onto the top o< aacolemn
pocked aoith an adiorbent, tassed huough the coumn at
diguent xates that derend Yjuy o each subslane
Roe the adionbent and or the solvent on sdent mixtune, and
AMe wsuay colected in soation au hey pas romthe column
a4 dijeuent tnes.
the
fkatonay phase ià packed ikb
glau O4 Wetal conn
’ 1he miztue oj analyes is anol the mobile phase,
kioon a the olne paued tugh he column
The thationony phase i tmen coated onto ditcKele smau pautielee
(Hhe mati) ond packed imto tthe colemn
As he eute ous through he column he analyte separala
Hhe bals oy tneir dittributipn Coeliients and ennerge
sin the eluate asit leaves the column
colunnchoMalogyapay
(0 Ton Exchange chromatograhy
2022 to pores moleeules
sthencal slouer
PiCRCER excluston moleukeanelerMedcceuples the retoxde poaea
unhindeNd,
oithout
penetraing the
Date:
l0
6 TOGRAPHy he and ata
fromthe
moleeules. o filspattcle.
Page:38
sie
paxmik is t
volume ager have
form column
CHROMA ha matnix cohich ke
al n caue eoil excluded themobile
that sepoxales
knpeh biologieal Matnx moloculea he
COLDMN_
techwique coith anc
the
bed buerHhe
ilL thwugk
in
e}feen
the
aluo
dieMenCes
raphuy column poroMA
a
beteen
henee entielytebueen
GELFILTRATION Dther equiibratedwithsmaller
o elsluent. pau
ehromatoqraphy,
veutatile"
chromatoq the a peneakion dittributed
and Siejuy
n colunn
phase they
ealed o
volume syaee aHe
ehDIatoge medium tohexeas
oteuu hein mobile moleceelee
the hen appen
and
Hate
the he
ú
a filtration
gel o ted theu in get sieve,
matrix and
filtrahion the iret mopules
OF ehtomatoqaphy
on ta
ted matrix
to
4iltration
DEMONSTRATION based a lngen the
Jepanation
he The teeMed matnx
yached pached olow 0ceDHding
come o}
Ftig
olal1 gel
qel the hus,andSmalottaide
Gel ne
The Jhe The the
is The
Tcpic:
pipete,
chaomatoqaphy paper. od
clannp
pasteus
balanee. to oluw coumn
the an qlas
PiCRCER qraph a
ethanoloilh
ink ehano a
39
Pcge:
Date
d,
padethanol. digital
stamp
glau
glas 96,
size),
omtoh,
slop
q0/.
otfo XAM
at falseougk
hatnd remove
soixled
üntroduned
coik
toith to
90/tekes, clam, pen, on dná gentty
Hue (a0t) Mesh Masher mixed attoched
phae
andin textcoln
and (aDo-4DO wn wese
diadwed were e0a
Red sland toate,glaa coa solient
mobil
appanatuu distiued
ctton beaou
beacs colun
material gel, coton, coith
Silëa eanation glaso inthe
o, Hod. gel
amountamount
silea eotton
Ond coton scaked
qa
toaHA
Requinements
O Byenimental Miseellanepus-
Chemiealu emphy tnetrapped
in the
Smalu
Gau >
olunOAbOut wet
then
)A
to
) () ()
FILTRATION
GEL
OF
CHROMAT0GRAPRY
N COLUMN
DEMONSTRATIO
the
EIOREER° i. bed lt o ntenyal full
thDug wee appue
miYeol Rml action
4O
Page:
raction gel dye toonHhe
uà 907.
ti
to coaU the the colunn,
mn keep
but bee. fesh as
:Date pipette tube
yass wa into jnk o
as elute 5 column, matked
to speed
Hed at coumn coumn
eomn marhed inh mizhune
cate
gteat
adiorbed
the
olect
asteudength auee ae We rom the the
same
ned
qteat to the and
and andsuch pHeLededout to romin to oleeted
coitha and
counan as weeadded MOVed
wee
n conlinedepavate.
saved inh o toith Comes added
seulebubbles, oaA it. but
out
ethanol &m colutions inh
the
ontooduedt wa stanmbed
inh ond
Came
tuene
jeuotically
oy ain gbeeluent qduay
to the ethand eluted
gel ca
hayaDwed blue Siliea anplebue
Jutipn blue coected euens
the hen dyye dye
to 90
Hemone 40%
dy hat and
bxate Qach boh he
such he al
Dnee
the
the duya
ua Such
the
to
coumn and hea be<one
addiional tound
Q0as ml toas
both
cquilio(0
ol
o
together nitaliy l6 1to
ehanol l:
euent tme
10 top Coumn
cas
Qnd ml
appedThen oau Just wil A Total
to R DY
(0
()
4 9 9 )
Topic:
INK
AOR
SCORE
CbLDuk
AD
IAMP
1NK:
PAD
COLOUKB)
DIEEERENT
RIED
OE REDCAMAPABNIKTN
THEScoRING
FRACTION
NMBER
OEELUENTS
SHOWINh
ANDRACTIONS
PH 4
LINE
GRA
BLVE
2
4OLDUR RED BLUESCOREFOR
PIOREER abitretiy
coloui ed
or
Page Date:
ed tblueeluenbs.
weeor by SCORE
FOR
COLOUR
INK
coloue o
either
fration PAP
l2 STAMP +++
to bue PAodueed ++
1 +
fromeither dijenent RED
faction coloux COLOUK
SORE
FOR INK
ol PAD
apeaNanee in
ink STAMP
the o +
Seoxing
pad BluE
au
fo shmp
FinallScoredResut
D NUMBER.
FRACTDN
Tale 1 2 3 12
(5
O
Topic:
BNany Cioufrula!rti! Scoist
moleculea ohereascoLumn
tme
bed
DIRCCR tes ok that
mixte mdoule.
42
Page: cepaLa mate molecule.;
Dato
eight patt towsed moleeeles
that he tiet
sanmle direet ueue oithin
share. eiM Smaler
moleculan
methoc tme
dye
dye
proeet elted
Me eights. more moleuler sied
andn nore beloe Yied Baster
a faster, chOMatographiy
smaler
ink,as
a sìe(onponents luigher beoads laqer and
chronatoqaphy
send
molecularbede coumn duye movd
thei e
to the lagex/ the tso has has pad
wos
Geu to Ahe tendm the takein epauient,
filtration
chOmat
molecul
aceONding
cOxdinghein ikink stamp
the
pOMeA eluedfonm
molocLales ink
euleA
tme filtation
pod
a pad pad
toith
sepaHation and
Conaequently
langeumole slamp
Ted
less stamp
slamp
COelates
he
D
Comment channela
SmaleH lwestisqer
Hhan
ue vo the BlueTedblue
Ihe çn fn
Top:.
