Lab 707 Separating a Mixture of Biphenyl, Benzhydrol, and Benzophenone by ThinLayer Chromatography Intro The purpo e of thi experiment

i to figure out a proper ol!ent that "ill be u ed to eparate a mixture of biphenyl, benzhydrol, and benzophenone by thin-layer #hromatography$ The thin-layer #hromatography "ill be u ed to in!e tigate ol!ent polarity on relati!e #ompound in the mixture, and to identify the pe#ifi# #ompound in an un%no"n mixture #ontaining any #ombination of biphenyl, benzhydrol, and benzophenone$ &a#h eparate #ompound "ill ha!e an 'f !alue that "ill be #al#ulated in order to #ompare the #ombination of the un%no"n to tho e of eparate #ompound in the to#% olution$ The tudent hould already %no" ho" to u e a Bun en burner or mi#roburner$ Thin-layer #hromatography (TLC) i a imple and #heap te#hni*ue that #an effi#iently eparate *uantitie of le than ten mi#rogram of material$ TLC i u ed for the rapid analy i of reagent and produ#t, purity, or to determine the number of #ompound in a mixture$ It+ al o u ed to #ompare un%no"n #ompound beha!ior to tho e of %no"n #ompound o that the un%no"n #ompound #an be bro%en do"n to "hat i in the mixture$ The term #hromatography refer to e!eral related te#hni*ue for identifying, eparating, or analyzing mixture of #ompound $ ,ll #hromatographi# te#hni*ue ha!e a t"o part operation in #ommon$ The fir t i the mobile pha e, "here a ample mixture i pla#ed in a li*uid or ga $ The e#ond i the tationary pha e, "here the mobile pha e #arrie the ample through a olid upport$ The different #ompound in the ample mixture mo!e through the tationary pha e at different rate be#au e of the differen#e in rea#tion bet"een the mobile and tationary pha e $ Thi "ay ea#h #ompound in the mixture "ill be at a different lo#ation "hen the #hromatography i done$ -uring the TLC pro#e , the olid tationary pha e i #alled the ad orbent$ , the mobile pha e (eluent) tra!el up the ad orbent the #ompound "ithin the mixture "ill begin to eparate$ Compound "ith le attra#tion for the ab orbent mo!e rapidly "ith the eluent$ Be#au e TLC ad orbent are typi#ally polar, the more polar a #ompound i in the mixture the tronger it adhere to the ad orbent and the lo"er it mo!e $ The ratio of the di tan#e that a #ompound mo!e to the di tan#e that the eluent front mo!e i #alled the retention fa#tor ('f)$ The e*uation for 'f i . 'f / di tan#e tra!eled by #ompound (mm)0 di tan#e tra!eled by eluent front (mm) Method 1) 2reparing the -e!eloping Chamber

3btain 4 540-ml bea%er and label ea#h bea%er "ith one of the eluet . ethyl a#etate, hexane, methanol, di#hloromethane, and toluene$ 3btain 4 rubber band and pie#e of pla ti# large enough to #o!er the top of the bea%er $ Cut a pie#e of filter paper into a re#tangle "ide enough o extend nearly to the top and long enough o it "rap around 6 of the bea%er$ -o thi for all fi!e bea%er $ 2our

4 ml of the appropriate ol!ent into it bea%er and #o!er ea#h bea%er "ith the pla ti# and et it a ide$ 5) Spotting the TLC 2late

7et fi!e 5$4x7$4 #m TLC ili#a gel plate and be !ery #areful to ma%e ure the #oated urfa#e of the TLC plate are not tou#hed$ 8 ing a pen#il, lightly label the top of ea#h plate "ith the name of one of the fi!e ol!ent $ Mar% the origin on ea#h of the fi!e plate lightly by dra"ing a line a#ro the bottom$ ,l o, mar% t"o #ro hat#h line on the bottom line to indi#ate "here the olution "ill be potted$ Caution: benzhydrol, benzophenone, and biphenyl in a#etone olution are flammable and irritating$ ,!oid u ing the e #ompound near heat our#e and pre!ent any #onta#t$ ,l o, a!oid inhaling !apor or inge ting the e #ompound $ 3btain a !ial of the to#% olution mixture #ontaining all three #ompound $ 2la#e the dra"n out end of a mi#ropipette into the to#% olution and allo" the li*uid to ri e up the #apillary$ Spot the olution onto the TLC plate by *ui#%ly and lightly tou#hing the end of the pipette$ The pot hould be le than 5 mm$ ,llo" the ol!ent to #ompletely dry, and then ma%e a e#ond pot on one of the hat#h mar% $ ,llo" the ol!ent to e!aporate again$ Ma%e ure to do thi for all fi!e plate $ 9) -e!eloping TLC 2late

Che#% to ma%e ure the eluent le!el in the #hamber i belo" the line on the TLC plate $ 2la#e ea#h potted TLC plate into it appropriate #hamber u ing the #hamber "all to upport the plate$ Co!er the #hamber "ith the pla ti# "rap and e#ure it "ith the rubber band$ :hen the eluent ri e to about 1#m under the top of the plate, remo!e the plate from the #hamber and immediately mar% a light line "here the eluent front "a $ Caution: ultra!iolet radiation #an #au e e!ere damage to eye $ :ear goggle and do not loo% dire#tly into the 8; lamp$ :hen all fi!e plate are done, ta%e them out of their #hamber and allo" them to dry off$ Ta%e the plate o!er to the 8; light and ha!e them examined$ Loo% for the eluent that ga!e three eparate pot for the mixture, thi "ill be your ol!ent$ Cir#le the pot on the TLC paper $ <) ,nalyzing an 8n%no"n Mixture

3btain an un%no"n olution and 9 ne" TLC plate $ &a#h plate hould ha!e t"o hat#h mar% for potting$ Label ea#h of the hat#h mar% "ith a eparate olution= to#%, benzhydrol, benzophenone, biphenyl, and an un%no"n$ 2la#e a drop of ea#h of the olution to the #orre ponding hat#h mar%$ 2la#e ea#h of the TLC into the eluent that "a #ho en from the pre!iou tep$ Ma%e ure the TLC plate are upported again t the bea%er "all and if there i more than one TLC plate in the bea%er at a time, that the

plate ne!er tou#h ea#h other$ :hen the eluent of ea#h of plate ri e to about 1 #m under the top of the plate, remo!e it and allo" it to dry$ :hen all three plate are done ta%e them to the 8; lamp and examine them$ Cir#le all pot "ith a pen#il$ The to#% olution hould ha!e three pot , one for biphenyl, benzhydrol, and benzophenone$ Line up the pot from the un%no"n mixture to tho e of biphenyl, benzhydrol, and benzophenone$ Cal#ulate the 'f !alue for all the pot and re#ord the #ompound pre ent in the un%no"n olution$ 'e ult >one of the eluet eparated the to#% olution into three eparate #ompound ex#ept for toluene$ The TLC plate for the un%no"n analy i "ere mar%ed a follo" .

Sto#%

Benzophenone

Benzhydrol

Biphenyl

un%no"n ?

un%no"n <

&a#h eluent front "ent 70 mm$ 'f Benzophenone / 90mm070mm / 0$<9mm 'f Benzhydrol / 14mm070mm / 0$51mm 'f Biphenyl / @0mm070mm / 0$A@mm 'f un%no"n ? / 94mm070mm / 0$40mm 'f un%no"n < / 14mm070mm, 90mm070mm, @0mm070mm / 0$51mm, 0$<9mm, 0$A@mm Con#lu ion The lab at fir t did not go "ell$ >one of the eluet "ere brea%ing the to#% olution into three eparate #ompound $ Binally after 9 trial , toluene "a ho"n to be the proper ol!ent$ The re ult #ame out moothly after that$ The un%no"n ? olution "a made of Benzophenone, and the un%no"n < olution had all three #ompound $