It is collective information about Pepsin for my seminar on topic "Preparation, Identification and Analysis of Drug (Pepsin) of natural origin" for completion of my Seminar of the semester seventh of course Batchelor of Pharmacy 4th year.
2. SR. NO. CONTENT PAGE NO.
1 DESCRIPTION 3
2 SCHEMATIC PRESENTATION OF WORKING OF PEPSIN 4
3 Method of Preparation of Pepsin 5
4 Description of pepsin 6
5 Uses 7
6 Case study 8 - 18
7 REFERENCES 19
VIDYAHARTI COLLEGE OF PHARMACY, AMRAVATI 2
INDEX
3. Pepsin
DESCRIPTION:
It is enzyme prepared from mucus membrane of stomach of various animals
like pig, sheep and calf.
The commonly used species of pig is Sus scrofa (Wild boar) belonging to
family Suidae.
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4. VIDYAHARTI COLLEGE OF PHARMACY, AMRAVATI 4
SCHEMATIC PRESENTATION OF WORKING OF PEPSIN
5. VIDYAHARTI COLLEGE OF PHARMACY, AMRAVATI 5
Separation of mucus
membrane from
stomach – by splicing
or crapping off
Placing in acidified
water for autolysis at
370 for 2 hours
Liquid obtained after
hydrolysis consist of
pepsin and peptone
Liquid is filtered and
sodium or ammonium
salts are added till
liquid is half saturated
Only pepsin separates
out
Ppt is collected and
subjected for dialysis
for separation of salts
Remaining amount of
pepsin, if any, in the
solution is precipitated
by adding alcohol into
it
The pepsin is
collected and dried at
room temperature
METHOD OF PREPARATION OF PEPSIN
6. Description of pepsin
Colour: Pale yellow
Odour: Very faint or bland
Taste: Slightly bitter
Solubility: In acids, water, NaCl solution
Storage: Can be stored for 1 – 2 years at 20 – 80 C
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7. VIDYAHARTI COLLEGE OF PHARMACY, AMRAVATI 7
1) It is used in deficiency of gastric secretion.
2) It is used in lab. Analysis of various protein.
3) in the preparation of cheese and protein containing food.
Uses:
8. VIDYAHARTI COLLEGE OF PHARMACY, AMRAVATI 8
The non-boiled liquid is strained red, while boiled liquid remains almost colorless.
Put few shreds of carmine fibers in each and keep it in a thermostat at 380 – 400 for an hour
1st part- boil to deactivate
Divide in two parts
1 gm. Pepsin + 0.002 N HCl
1. Identification Test
9. VIDYAHARTI COLLEGE OF PHARMACY, AMRAVATI 9
0.25 g
sample
+ 1.0 g
of
sodium
chloride
Triturate
and add
slowly
acidified
water
(prepared
by
diluting
65 ml of
1 M
hydroch-
loric acid
to 1000
ml with
water)
Dilute to
1000.0
ml with
the
acidified
water
and
shake for
15
minutes.
Prepare
coagulated
egg
albumin
Coagulated
albumin -
Boil fresh
hen-eggs in
water for 15
minutes,
cooling
rapidly
to room
temperature
by
immersion
in cold
water,
separating
the whites
and rubbing
through a
no. 44
sieve.
Reject
the first
portion
that
passes
through
the sieve
Triturate
15.0 g of
freshly
prepared
coagulat-
ed egg
albumin
with 50
ml of the
acidified
water
Add 50
ml of the
acidified
water
and keep
in a
water-
bath at
51º ± 1º
for 15
minutes
Add
20.0 ml of
the
prepared
solution of
the
substance
under
examina-
tion and
digest at
51º ± 1º for
4 hours,
shaking at
intervals of
15 minutes.
Centrifuge
and decant
off most of
the
clear
supernatant
liquid,
wash the
remainder
into a 10-
ml
graduated
cylinder
and allow
to stand for
30 minutes.
The
volume
of the
undissol-
ved
albumin
is not
more
than 2 ml
2. ASSAY OF PEPSIN
14. VIDYAHARTI COLLEGE OF PHARMACY, AMRAVATI 14
4. Pepsin digestion assay and serum IgE binding analysis
(A) Pepsin digestion assay- Control milk power (left) and BLG-free milk powder (right). Lane 1: molecular weight
markers; lanes 2-6: samples of digests at 0 s, 15 s, 2 min, 30 min and 60 min; lane 7: pepsin control.
(B) IgE-binding capacities of different milk powders to sera from non-allergic volunteers and CMA patients. 1-5:
different non-allergic volunteers; 6-11: different CMA patients; white box: Control milk power; black box: BLG-free
milk powder.
(C) IgE reactivities of different milk powders shown with immunoblotting. Lane 1-2: Control and BLG-free milk
powders. Results are shown as means ± SD. * P < 0.05; ** P < 0.01; *** P < 0.001; NS, not significant.
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5. Investigation of the Binding Between Pepsin and Nucleoside
Analogs by Spectroscopy and Molecular Simulation