Isolation of casein Analysis Of Urine

Analysis of Proteins
usually present in all involves HEATING
Steps Not urine :

Milk K ketones Biuret Test

- " :ñÉÉ•
"

i. weigh -

win &
Water Bath Bicarbonate 10ns
Reagent NaOH 1% US 04
-
:
2. B
↑ Heat destabilizes bond
3 .
Heat t stir between calcium & CASEIN
0
-

Oxalate f) -
Blue
( calcium casemate )

Temp Glucose ( t) Violet

-

4. Monitor G
-

More or less than 40%

micelle
-

will dissociate casein

5- Stop at 40°C & denawriie-ieo.sn
no casein S -

Serum Proteins

6. to drops glacial acetic acid a tests ✗ anthroproleic WÉY

Test
test for
PYtein bonds specific
↳ hydrolyze into _

f gµ , simpler
isaeuire
pephae bonds
precipitations
Test for Protein ( Biuret Test)
Reagent : Cone .
Nitric Acid (HN03)
Proteinuria Ct) -

purple
yellow precipitate
-

7. Note changes c- ) blue ""

(t)
Est;
-
-
¢ nearing

4.65-4.70 Test for Sugar ( Benedict;

8. Measure
pH if exceeds add Clewwse
is a reducing sugar]
positive ,+%✗a% proline
,

Glucosuria -

Test
infyydpin
_

acetic
more glacial -
those wltree
acid Nitz group
net charge is zero

Reagent Copper 11 Sulfate in citrate & carbonate

Reagent Ninhydrin solution
: :
* Isoelectric point neural :
electrically

Filter heterogenous ( liquid ) f) blue ( t) color

deep purple
- -

a.

no .
Squeeze (t) -

red-orange precipitate (t) -

orange ( Proline)fingerprint ↳ detection of

(
reduce [ V11 →
Cv 1)
ii. Discard Filtrate ( liquid)
hydroxylated
Collect & Transfer (Precipitate) Test for ketone Milton's Test
- test for
12 . benzene rings (tyrosine)

"
" " ""

Add 50mL 95% ethanol ' """

acid Milton's reagent
Reagent mixture of glacial acetic
Reagent
-

is in :
.
: I :|
↳ remove unwanted
↳ uses acidified mercuric sulfate
tat
Discard all
and 10% nitroprvsside
*
"' µ .
Stir
iiauialtiitra
Decanter )
te

is
-

Allow to settle (t) -

pinkish ring Enzyme Activity

a- Decant (tt ) -

red ring
residue
Discard the Decantakiia.iq (ttt ) deep purple ring
-

Iodine Test
17 . ' said>

18
. Add 50mL dichloroethane methanol -

"
( t) -
blue
acid
111 ✓
↳ for washing & final drying _
enzyme has already
Bicarbonate Test
Hasler evaporation '
NO blue color the starch → maltose
digested
*
-

19 .
Stir
Reagent
:
2 drops of 3M 1-12504
sulfuric Acid
f) deep orange -

presence of starch
↳
Collect the solids vacuum filtration (t) gas bubbles
-

20 :
-

↳ makes iteasier for

the unwanted solvent
21 . Discard the filtrate to fall
"
""
"

"
"

casein
for chlorine i.mg#iai i.ine:ii i iE
chlorine
✗☒ .
Collect test Naa
Nitric Acid É
' "" '"" ""

6M & 0.1 of Ag NogNitrate

Store
Reagent HNOZ
:
23 .

Acid
Nitric silver

24 .

Weigh (t) :
white bubbles

Test for Ammonia

Reagent 6M NaOH litmus Paper

:
,
↳
dimmer oxide
( t) :
Red → Blue
change in Litmus Paper