::;.

~ect e
;1 th presence of carbohydrate~ faltl a nd protein11 in the following food
Grape11, pafl'ltoell, r ice, butter, biscuit-, m llk, fro undnut;iboiled egg.

fi!EORY
the
'l\e presence of carbohydrates, fats and protein in any food stuff is detected by performing
do not
tt!U for carbohydrates, fats and proteins with the extract of the food stuff. These tests
a 1rferewitheachother.

!,PPARATUS
TC!l lubes, beakers, glass rod, pestle mortar, burner.

PIOCEDURE
hit prepare the extract of the given food stuff by either dry grinding in the morti,,r with a
~lt or by boiling with minimum quantity of water extracting with a small quantity of
an
irpnic: aolvent after grinding the food stuff. The extracts of some
of the food stuffs can be
~ \llredasgivenunder: ••,
For grapes -extractthejuice
Jlorpotatotes -cutinto slicesandboilwithw ater
For nee -boil~thwater
For butter -test directly
For biscuits - boil with water
F<.l!'miJk -test directly
For groundnut -grind in, th~ mortar . . .
For boiled e _ take the white of the egg, gnnd and shake with water.
as
' With the sol:ion or the s~spension perfonn tests for carbohydrates, fats and proteins
l(,d_ 11t Erpts. 10.1, 10.2 and 10.3.
 I. EXPERIMENT 1~.1

To study some simple tests of carbohydrates.

JtEQUIREMENTS
Glucose, sucrose_ (cane-s~ga~), lad~s~ (milk-sugar), starch, Molisch's reagent, Fehling's solution,
Benedict's solut10n and 1od1ne sol~tion. · •

pROCEDURE
1. Molisch's Test
All carbohydrates give this test.
· Take 1-2 ml of aqueous solution of carbohydrate ·(suspension in case of starch) and add
few drops ofMolisch's reagent (10% alcoholic solution of a-naphthol). Put one ml of cone. H 2SO 4
. slowly along the side of the test ~ube. . . ._ . _ _
A red violet ring is produced at the junction of two layers.
Chemistry of the test. Cone: H2Sd; co~verts carbohydrates into furfural or its deriva-
tive which further reacts with a-naphthoLto give a coloured product.
2. Fehling's Test
Take 2 ml of aqueous solution of carbohydrate (nearly 5%) and add 1~2 ml each of Fehling's
solution A and Fehlin,g's solution B. Keep the test tube in boiling water bath.
Reddish ppt. ind~~~tes the presence ~f ii reducing sugar.

PREPARATION OF_FEHLING'S SOLUTION

FEHLING'S SOLUTION A. Dissolve 17 .5 g of CuSO 4 in 250 ml of distilled water containing
few drons
.t"
·· ·4~
of H2.so .
' ·
FEHL!N(}'S Sl>LUTION B. Dissolve 8.65 g of sodium potassium tartarate and 35 g NaOH in
......__150 ml of distilled water . , .

Chemistry of the test

CHO COO-Na+
I -- . - . Tartrate I ·
.- · >
(CHOH) + 2Cu(OH) 2 +NaOH· , rnns (CHOH) + 3H 0 + Cu 20 J,
-I . - 4
. . • I · 4 2
red ppt.
CH 2OH . ,, CH 20H
Glucose ,Sod. salt of
gluconic acid
a,lle
1 nedict's
2
Test
1<.:;- rnl of aqueous solution of carbohydrate in a test-tube add 1_:2 ml of-Benedict's reagent.
p the test-tub~ in a boiling water bath. ·
ll.eddish ppt. indicates the presence of reducing sugar.
:r .,,~F
EDICT'S REAGE NT ,
PREPARATION OF BEN fanhydr ous N 82 C03 in about 80 ml of d· :
Dissolve 17.3 g of sodium citrate and 9 g O
·h te in 100 ml of water. Mix the tw0
18t111
ed ~ate
8 1
Heat if necessary. Dissolve 1. 73 g of copper. ~-ij _a_ :_ • ,_ · , and Il:>.a.Ite tL:
voium,e ~50 ml by adding water. " .... · 1 11
,
· . - · : "that of Fehling's' test. Here citrat •
Note. Chemistry of this test is the same as : . . . • e ions a~
. . . . ,
complexing agent. e Used
. . . ~
4. Tollen' s Test
Take 2-3 ml of aqueou s solution of carbohy drate ~n a teS t -~u}>.e. Add to it 2-3 ml of T
reagent . Keep the test tribe in a boiling water bath for 10 minute s. 0
lle~1
1

A shinin g silver mirror indica~es t~e presen ce of reduc ing carboh ydrat
. -. - - - -. . . e.
PREPA RATIO N OF TOLLE N'S. REAGENT
- ., : ~ • ;: •.{?
. Add NaOH solution to AgN0 ' soluti'on. Then add NH4 0H solution dropwis e till the .
3
dissolve. _The clear solution obtained is' Tolleii's reagent. . PPt. Just
Chemi stry of the test

AgNO 3 + N~/)1:f .~ NH4~0 3 + ~{?H_-

'I
... ' 2AgOH ~ Ag20 + H20
Ag2 O + 2NH4 OH ~ 2 [Ag(NH ) JOH+ 3H O
3 2 2
(Soluble)

. I
.CH2OH7:> . . . ·-.)
:, . COOH .
!
.
I
---~;_ - . - . .
·-" \•, "." -, •' ' -NH:-°OH .
. . (C, HQH).4 ?" ~g2o . -.. 4~·:,➔.·. cct_HOH)_L . -~ ., -·~'.~rg _...
CHO COOH Silver mirror
Glucose ·
\'.:,.· · · .. -,Gluconic ·acid
5; fodine Test.(For s.tarch ·o nly) -- . ..J - - - , . •

To the aqueou s suspens ion of th . . .. .

e sample;
ApIJ}earance ·of blue colour at. add 1-2 d . . ,.
. . . rops of iodine solutio n.
ion indica tes th .
OBSERVATIONS · · e presen ce of starch .

Test
Glucose
1. Taste Lactose
Sw_eet Sucros e Starch
2. Solubil ity ..Sweet
Soluble ·.sweet
3. Molisch 's test Soluble Tasteless
Purple ring .
4. Fehling 's test Soluble Insoluble
Redppt. ·. Purple ring
5. Benedi ct's test Purple ring purple ring
Redppt. ' Red ppt.
6. Iodine test Negativ e Negative
, ,
Negative Red Ppt.
· Negativ e Negativ e Negative
Negativ e Blue colour
 1, ~XPER~NT -~~~i · 1
· To study some simple tes.t s of ~ils and fats.
REQillREMENTS
Chloroform, ethyl alcohol, ~S~ 4 crystal~·, furfural .s olution, Hubie's solution, desi ghee,
vegetable g hee and re~ned 011. _·
; l r _,1 / t ,l•1 ' '

PROCEDURE
1. Solubility Test
This test is based on the fact that oils and fats 'qte sol~ble in organic solvents like chloroform,
alcohol, etc., but are insoluble in wgter. Shake a"s~~l~ ~-~inount of given sample with 5 ml each
of water, alcohol and chloroform in three test-tubes and observe the solubility and draw inferences
as follows: · · "· . ., , · · · ·
.. . ~ z •• -:

Test-tube Solieiit. ~ I t ♦

Inference

1. Water Sample ·is immiscible_. Oil or fat present.

(ii) Sample is•miscible. · Oil or fat absent.
2. Alcohol . (i) . Saihple,fotlns lowe:r,layer,. · Oil or.fat pre~ent..
which dissolves on heating.
.. (ii) Sample·does not dissolve Oil or fat absent.
.~ven on heating.~
3. . Chloroform · (i) Sample .i s miscible. Oil or fat present.
.. (it ) Sample is fmmiscible. :Oil or fat absent: -
2•ri'ran~lusoent Spot Test_ · · ·· · ~ •. · · .
Press a r1ttl f h ·
1
b t ·n the folds of the ,
f ilter paper. On unfoldmg the..fi)ter paper, the
appearan e fo t e slu ~ anct~ r greasy spot on the filter paper indicates the P.resence 9f fat or
1
. ce o trans uscen o . .
011. The t
. 8 po grows 1arger on heating and drying the filter paper. • · . ,
.·
a. Acr-1
ll O . ·
e1n Test .
.
· ent irritating
eat a, 11·ttl f h •th some crystals of J{JI~0 1n ~~est-tube.A pung .. ...,.
Od0 e o t e samp1e w1 , , . · . 4
ur of a cro1e1n . , con 1
- 6irms the presence of fat or 01 .
 T,.,,,,,ffi=,.•P'•~
: :Q: : : : : :r11,on )
gelatin d,aper11mn Millo
n a reagent and Nmh
ydrin re~n t
PRO CED URE
I B,ur et 1e~t
To the dispe nnon of
the subst ance to be
ofNa OH \ulion teste d (s ay 5% solu
80 . Now add 4-6 droJl'I tion of egg albun
nf 1% CuSO , BOlut nn) add about1
minu te•. ion. Warm the mixtu
re for ahooit i.,
Blu ish viold c<>lo
wration indic alltS
t/u, pre,- rnce of
2.Xa ntho prote icTe prote in.
Take abou t 2 ml st
of egg album in dispe
rsion in a test-t ube
and heat. and add a few drops
of rono. ~ .
A y,ello w ppt. lndic
atea the p,...1ern::e
ofpr otei, u.
3.Ni nhyd rinT Ht
T»ke abou t2ml
ofeg galb umir >dill
801u tion.B oilthe ...,nte persi onin ate,, t-tub
eend add3- 4drop
nts. 11ofN inll)'d ru
Inten se blue COW
,,:rotion oonf in,w
l/u, pre,r ence ofpr
oteilJ&.

'fhis l.estisl(ivcnbypro

:::.f:i:::::
tei11. 11oonl.8iningphe nolic
amin oacid s.Ge/ atind
oesno tgiV< lfhiJW
; 111;~: I
prote in,.

MIU. OS'S IU:AG

dispe n<ion ~dd 2 drops
1
gn o-bn ck
ofMi llon's ~age
n,d on boili 4R,
nt.
eonf irmt, tlu pre•"
""'
4 1
HNO,. E..VI' ia
p<ep aredb y.ru.o
Mng5 Keac hofH gNO,
and Hg(NOil,i n 100111
1ofdil_ )