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Experiment #6 Chromatography
Introduction :
This experiment is designed to separate components of a mixture through a process caed
chromatography! This is done "y the distri"ution of ions and different organic compound "y
appying t#o phases: stationary and mo$ing! Chromatography can "e compared to so$ent
extraction #hich their distri"ution reso$es around t#o so$ent and their sou"iity #hie
chromatography focuses on ho# strong a phase a"sor" the ions %depends on the mixture
poarities& to mo$e through! Thin ayered is cheap and easy #hie high performance i'uid
chromatography is compex and expensi$e! Thin ayered and coumn chromatography #i "e
used in this experiment to sho# the principes of separation of organic compounds! The coumn
chromatography is perform #ith acti$ated aumina using ()*+)) si,e partices! This compound
shoud "e dried and stored o$ernight in an o$er at --) C in a seaed container! .n thin ayered
chromatography/ fexi"e siica ge pate from 0hatman #ith a fuorescent indicator #i "e
needed in this experiment!
Procedure 3A :
-! 1"tain a -)x2!3cm TLC pate and "e cautious a"out "ending the pate!
+! Dra# a ine across the pate a"out -cm from the "ottom #ith a ead penci and #ith a
ruer mo$e the index a"out )!6cm in from of the edge pate and separate them into fi$e
-cm interas!
3! 4repare fi$e micropipettes and a TLC de$eopment cham"er #ith methyene choride
and a "ea5er co$ered #ith auminum foi #ith a scre#*cap "otte!
6! 7d8ust fiter paper iner so it does not go competey around the inside of container %+!2
inches&!
2! Loo5ing from eft to right/ identify fuorene/ fuoreno/ fuorenone and the un5no#n
mixture!
6! 7ppy each sampe "y touching the pipette ighty to the thin*ayered pate %9+cm&!
:! 4ace TLC pate in de$eopment center #ithout touching the fiter paper!
(! ;emo$e the so$ent #hen the front is -*+cm from the top of the pate and mar5 the ine
#ith pen!
<! Dry the pate and pace it in a 8ar containing iodine and cap it unti spots appear!
-)! ;emo$e pate and outine a the spots that appeared and measure the distance
tra$eed!
--! Cacuate ;f and each compound and identify the compound!
Procedure B :
-! 1"tain )!2mL of each t#o and one containing "oth compounds!
+! 4repare three thin ayered pate i5e in procedure 7/ each pate shoud "e -)x3!3cm and
three -cm apart!
3! =pot t#o of the indi$idua compound and the mixture of "oth/ use different pipette for
each!
6! 4ace each pate in de$eoping cham"ers and remo$e #hen so$ent is -*+cm from the
top!
2! Dry the pate and use a ead penci to ma5e the position of the so$ent!
6! 1"ser$e resut under short #a$e >? ight in a dar5 hood and outine any spots !
:! 4ace pate into a 8ar of iodine and ta5e it off #hen spots start appearing!
(! @ar5 in mm the distance tra$eed and and cacuate ;f!
Procedure C:
-! 0or5 #ith a partner and prepapre + thin ayers pate/ one #ith -)x2!3cm and -)x6!3cm!
+! @ar5 fi$e -cm apart for the first pate and 6cm inter$a for the second!
3! Ta5e fi$e sampes from reaction mixtures %)/ -2/ 3)/ 6)/ -+)&! Three shoud appear on
the arge and t#o on the sma pate!
6! =pot each pate #ith fuorenone and fuoreno and mar5 the reference point of the
compounds!
2! >sing the same steps as procedure 7/ prepare : micropipettes to spot pates!
6! 7dd sodium "orohydride to the reaction mixture/ ta5e sampe at time indicated!
:! =pot each sampe once and use different pipette for each trias!
(! 7dd a magnetic spin $ane into the 2*mL conica $ia!
<! 7dd )!+)g of fuorenone and 6mL of methano to the $ia and add a magnetic stirrer
using an auminum "oc5 or a camp to hod the $ia
-)! =tir mixture unti a soid disso$es then ta5e first sampe at )s to spot!
--! 0eigh )!+)g of sodium "orohydride and add it the mixture then "egin timing!
-+! >se micropipette to remo$e at: -2/ 3)/ 6)/ -+)s!
-3! =pot t#o reference soution of fuorenone and fuoreno in acetone!
-6! Dry and $isuai,e spots #ith iodine and s5etch resut!
-2! Transfer reaction #ith a 4asteur pipette to a -)mL Erenmeyer fas5!
-6! 7dd -mL of #ater and heat mixture amost to "oiing point for +min then coo it to
temperature!
-:! 4ace it in a #ater "ath to crystai,e the reaction and coect crysta through $accum
fitration!
-(! 0ash crysta #ith three -!)mL portion of an ice*cod mixture of ()A methano/ +)A
#ater!
-<! 0eigh and determine meting point!
Observations, Data & Results:
Conlusion: