Page No.
: 36
Prautfcal No- 1O
FreN cise involving isolation and dtection
acttue pridciples. Caf feine rom tea dus8]
BLyuine menis :
Calassuare Beakex, masuring cyinde, glass Yod
ptri-plate qlass stidete.
Chanmicals- Chloxa fotm, methanol etayt acetote,
artic acid, silita ql maak s*um oxicle
cthanot, Hasoy To dine
Crude dugs Caffeine From tea dust
Stan dond- Caffeine
Tpstrument- Heating metal, hot air oven.
Principle
+Lsolaion 0f CafPeine
Caffeine 3,7tdmethylxanthin is a
punine base Present 1elated
bases like
Coffee
fhuaphyline Yaeabnomine
tea, cocoa quatana, kola & mate.
Alhough Caffún is aey p10d uced
it s usually iolatec fiom
Seeol
tea leaves from coffee
duing decaffeinaton Mas ftrs discoverec
by Kabiquet in coffee t A +in 321
Page No. : 34
mid Loter in. 1827
teo leave s..
Procedunei
) Pont A- Extracton OF caffene
Weigh 10 of tc leaves and trOns fr to
250 distille d water
Boil ta 0atuy foY 30nsnutes with
DLCasion
stining. decoction is efFected
(ompleke allouo
to cool Ailtey the solution
Ailbrate a
Supanattng funnel e to
It add 100m) chloxofom
that to bal caffeine will ba trasferged
to chlogoform
Seposate chlonoform layen Evapo1ate ch lono form
oatn bath white emys tals will collect
the bottom.
Method 2
" Poyo der the
tea leaves to cOarse sized
portcles andextTat oith wwatey for
/ halP an hour
Filter ta entract treat oith lead acetate
to preupftates tanninsftltun the solution
yemove
and add dit. Ho SO4. to soluion to
Page No. : 39
eSLcess Lead actate in the form of Leac
Swphate
Ciltuy; add charcool to YeMOve the colouring
mattuy iltex this Soluion.
Take Hhis filtate a seporatng funnel
and extrat with 3 portfons 0f chloro for
witth mild shaking. Sepodate and com bine
th chloxoform laers and eveporate
preupfate olite ezystal of cafFeine
TeUrystallize with atohol
2) Port B- Thin Lan ChYomatog1aphy of Caffine
"Statfo nary phase- s+ca
Mobile phase- ttayl acetatemthano l-aceic actol
(80 :1o :16)
Dís solue 1lmg 0f cafune in Lm o£ chlorofo
tx mothanol spot tae Sample on E TLC plte
3na Lnposwm to iodine vapour coffein
spot at Rand
clwlqps a spot R voue 0-4)
#Tests Alkalbids
) DaaqindaoFF's Test
2-3 flExateadd feuo cops
T2agent brown PPt
is formd. onang
Page No.: 39
DMauen's Test
2- filbzate oith feo drops Myu's.
Tgurt qives ppb.
Cnt Wagner 's Test
2-rd filtyate uit fuo daops of
TRagent qivy yeddish broen ppt 00gner's
(iMu erÍ de Tet for Puxine Alkadoids
3-4m tut
Of coDc. HNO solutions add 3-4 dnops
Evap onate to
[Link] an d
2daaps OF NH40H Purple colour is olbserved
() Ammonu Tuna clete Teut.
Add o-2 hydnooylamine to a
Solution of ammoium
saturateol
run atat cnd
aidity with dil HCL alkaloido qlue
pink ppt, The ppt is Soluble in 5by
Qctone oich Is ed to ewys tallize it.
It
(vi) Tannins aid Tst
Test Solutio n tyeated_with tannic aid
solution4ive buff coloun ed_ppt.
vi) Picxolonic acd Test
Tist solution on treatant with pi oYo lonic
Page No. : u0
Reswt :
Re valu of caffein was found
be
Refernce
Bireo Shah A-k TRrt
o0k oF Phorma tognosy
and Phyto Chenmästry 2nc ediiony
